U.S. patent application number 17/205508 was filed with the patent office on 2021-08-05 for biological fluid filtration system.
This patent application is currently assigned to Astrin Biosciences, Inc.. The applicant listed for this patent is Astrin Biosciences, Inc.. Invention is credited to Jayant Parthasarathy.
Application Number | 20210237068 17/205508 |
Document ID | / |
Family ID | 1000005464416 |
Filed Date | 2021-08-05 |
United States Patent
Application |
20210237068 |
Kind Code |
A1 |
Parthasarathy; Jayant |
August 5, 2021 |
Biological Fluid Filtration System
Abstract
A biological fluid filtration system for scanning a biological
fluid so as to filter potentially undesirable constituents from the
biological fluid for therapeutic or diagnostic purposes. The
biological fluid filtration system generally includes a fluid
receiving device adapted to receive a biological fluid. A valve
including an inlet, a first outlet, and a second outlet is fluidly
connected to the fluid receiving device. The biological fluid
within the fluid receiving device is scanned by a scanner to
produce scanned data relating to the biological fluid. A control
unit in communication with the scanner and the valve receives the
scanned data and controls the valve based on the scanned data. The
valve is controlled to direct the biological fluid through either
the first or second outlet of the valve depending upon the
constituents of the biological fluid identified by the control
unit.
Inventors: |
Parthasarathy; Jayant;
(Excelsior, MN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Astrin Biosciences, Inc. |
Excelsior |
MN |
US |
|
|
Assignee: |
Astrin Biosciences, Inc.
|
Family ID: |
1000005464416 |
Appl. No.: |
17/205508 |
Filed: |
March 18, 2021 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
17162450 |
Jan 29, 2021 |
|
|
|
17205508 |
|
|
|
|
62968476 |
Jan 31, 2020 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
B01D 46/0067 20130101;
B01D 46/522 20130101; B01D 53/78 20130101; B01L 3/502753 20130101;
B01D 46/0013 20130101; B01L 3/502715 20130101; B01L 2300/0681
20130101; B01D 46/2414 20130101; B01D 46/0028 20130101; B01L
2300/0654 20130101; B01D 46/0024 20130101 |
International
Class: |
B01L 3/00 20060101
B01L003/00; B01D 46/24 20060101 B01D046/24; B01D 46/00 20060101
B01D046/00; B01D 46/52 20060101 B01D046/52; B01D 53/78 20060101
B01D053/78 |
Claims
1. A biological fluid filtration system, comprising: a fluid
receiving device adapted to receive a biological fluid from a
patient; a valve comprising an inlet, a first outlet, and a second
outlet, wherein the inlet of the valve is fluidly connected to the
fluid receiving device; a scanner configured to scan the biological
fluid within the fluid receiving device to produce a scanned data
relating to the biological fluid within the fluid receiving device;
and a control unit in communication with the scanner and the valve,
wherein the control unit is configured to receive the scanned data
from the scanner, and wherein the control unit is configured to
control the valve based on the scanned data from the scanner;
wherein the control unit is configured to compare the scanned data
to a reference data, wherein the reference data includes a
characteristic of at least one desirable constituent from an
individual other than the patient; wherein the control unit is
configured to control the valve to (a) direct the biological fluid
through the first outlet if the control unit determines that the
scanned data indicates a presence of only the at least one
desirable constituent within the biological fluid or (b) direct the
biological fluid through the second outlet if the control unit
determines that the scanned data does not indicate a presence of
only the at least one desirable constituent within the biological
fluid.
2. A method of filtering a biological fluid using the biological
fluid filtration system of claim 1, comprising the steps of:
identifying the characteristic of the at least one desirable
constituent from the individual other than the patient; storing the
characteristic of the at least one desirable constituent in the
reference data; receiving the biological fluid by the fluid
receiving device; scanning the biological fluid within the fluid
receiving device by the scanner to produce the scanned data;
comparing the scanned data to the reference data by the control
unit; directing the biological fluid through the first outlet if
the control unit determines that the scanned data indicates a
presence of only the at least one desirable constituent within the
biological fluid; and directing the biological fluid through the
second outlet if the control unit determines that the scanned data
does not indicate a presence of only the at least one desirable
constituent within the biological fluid.
3. The biological fluid filtration system of claim 1, wherein the
at least one desirable constituent is comprised of a healthy
cell.
4. The biological fluid filtration system of claim 1, wherein the
at least one desirable constituent is comprised of a benign
bacterium.
5. A biological fluid filtration system, comprising: a fluid
receiving device adapted to receive a biological fluid from a
patient; a valve comprising an inlet, a first outlet, and a second
outlet, wherein the inlet of the valve is fluidly connected to the
fluid receiving device; a scanner configured to scan the biological
fluid within the fluid receiving device to produce a scanned data
relating to the biological fluid within the fluid receiving device;
and a control unit in communication with the scanner and the valve,
wherein the control unit is configured to receive the scanned data
from the scanner, and wherein the control unit is configured to
control the valve based on the scanned data from the scanner;
wherein the control unit is configured to compare the scanned data
to a reference data, wherein the reference data includes an image
of at least one desirable constituent from an individual other than
the patient; wherein the control unit is configured to control the
valve to (a) direct the biological fluid through the first outlet
if the control unit determines that the scanned data indicates a
presence of only the at least one desirable constituent within the
biological fluid or (b) direct the biological fluid through the
second outlet if the control unit determines that the scanned data
does not indicate a presence of only the at least one desirable
constituent within the biological fluid.
6. A method of filtering a biological fluid using the biological
fluid filtration system of claim 5, comprising the steps of:
identifying the image of the at least one desirable constituent
from the individual other than the patient; storing the image of
the at least one desirable constituent in the reference data;
receiving the biological fluid from the patient by the fluid
receiving device; scanning the biological fluid within the fluid
receiving device by the scanner to produce the scanned data;
comparing the scanned data to the reference data by the control
unit; directing the biological fluid through the first outlet if
the control unit determines that the scanned data indicates a
presence of only the at least one desirable constituent within the
biological fluid; and directing the biological fluid through the
second outlet if the control unit determines that the scanned data
does not indicate a presence of only the at least one desirable
constituent within the biological fluid.
7. The biological fluid filtration system of claim 5, wherein the
at least one desirable constituent is comprised of a healthy
cell.
8. The biological fluid filtration system of claim 5, wherein the
at least one desirable constituent is comprised of a benign
bacterium.
9. A biological fluid filtration system, comprising: a fluid
receiving device adapted to receive a biological fluid from a
patient; a valve comprising an inlet, a first outlet, and a second
outlet, wherein the inlet of the valve is fluidly connected to the
fluid receiving device; a scanner configured to scan the biological
fluid within the fluid receiving device to produce a scanned data
relating to the biological fluid within the fluid receiving device;
and a control unit in communication with the scanner and the valve,
wherein the control unit is configured to receive the scanned data
from the scanner, and wherein the control unit is configured to
control the valve based on the scanned data from the scanner;
wherein the control unit is configured to compare the scanned data
to a reference data, wherein the reference data includes a
characteristic of at least one desirable constituent from the
patient; wherein the control unit is configured to control the
valve to (a) direct the biological fluid through the first outlet
if the control unit determines that the scanned data indicates a
presence of only the at least one desirable constituent within the
biological fluid or (b) direct the biological fluid through the
second outlet if the control unit determines that the scanned data
does not indicate a presence of only the at least one desirable
constituent within the biological fluid.
10. A method of filtering a biological fluid using the biological
fluid filtration system of claim 9, comprising the steps of:
identifying the characteristic of the at least one desirable
constituent from the patient; storing the characteristic of the at
least one desirable constituent in the reference data; directing
the biological fluid through the first outlet if the control unit
determines that the scanned data indicates a presence of only the
at least one desirable constituent within the biological fluid; and
directing the biological fluid through the second outlet if the
control unit determines that the scanned data does not indicate a
presence of only the at least one desirable constituent within the
biological fluid.
11. The method of claim 10, wherein the characteristic of the at
least one desirable constituent is identified during a
pre-filtration session.
12. The method of claim 11, further comprising the steps of:
receiving the biological fluid from the patient by the fluid
receiving device during a filtration session, wherein the
filtration session occurs after the pre-filtration session;
scanning the biological fluid within the fluid receiving device by
the scanner to produce the scanned data; and comparing the scanned
data to the reference data by the control unit.
13. The biological fluid filtration system of claim 9, wherein the
at least one desirable constituent is comprised of a healthy
cell.
14. The biological fluid filtration system of claim 9, wherein the
at least one desirable constituent is comprised of a benign
bacterium.
15. A biological fluid filtration system, comprising: a fluid
receiving device adapted to receive a biological fluid from a
patient; a valve comprising an inlet, a first outlet, and a second
outlet, wherein the inlet of the valve is fluidly connected to the
fluid receiving device; a scanner configured to scan the biological
fluid within the fluid receiving device to produce a scanned data
relating to the biological fluid within the fluid receiving device;
and a control unit in communication with the scanner and the valve,
wherein the control unit is configured to receive the scanned data
from the scanner, and wherein the control unit is configured to
control the valve based on the scanned data from the scanner;
wherein the control unit is configured to compare the scanned data
to a reference data, wherein the reference data includes an image
of at least one desirable constituent from the patient; wherein the
control unit is configured to control the valve to (a) direct the
biological fluid through the first outlet if the control unit
determines that the scanned data indicates a presence of only the
at least one desirable constituent within the biological fluid or
(b) direct the biological fluid through the second outlet if the
control unit determines that the scanned data does not indicate a
presence of only the at least one desirable constituent within the
biological fluid.
16. A method of filtering a biological fluid using the biological
fluid filtration system of claim 15, comprising the steps of:
identifying the image of the at least one desirable constituent
from the patient; storing the image of the at least one desirable
constituent in the reference data; directing the biological fluid
through the first outlet if the control unit determines that the
scanned data indicates a presence of only the at least one
desirable constituent within the biological fluid; and directing
the biological fluid through the second outlet if the control unit
determines that the scanned data does not indicate a presence of
only the at least one desirable constituent within the biological
fluid.
17. The method of claim 16, wherein the image of the at least one
desirable constituent is identified during a pre-filtration
session.
18. The method of claim 17, further comprising the steps of:
receiving the biological fluid from the patient by the fluid
receiving device during a filtration session, wherein the
filtration session occurs after the pre-filtration session;
scanning the biological fluid within the fluid receiving device by
the scanner to produce the scanned data; and comparing the scanned
data to the reference data by the control unit.
19. The biological fluid filtration system of claim 15, wherein the
at least one desirable constituent is comprised of a healthy
cell.
20. The biological fluid filtration system of claim 15, wherein the
at least one desirable constituent is comprised of a benign
bacterium.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation of U.S.
application Ser. No. 17/162,450 filed on Jan. 29, 2021 (Docket No.
PART-004), which claims priority to U.S. Provisional Application
No. 62/968,476 filed Jan. 31, 2020 (Docket No. PART-002). Each of
the aforementioned patent applications, and any applications
related thereto, is herein incorporated by reference in their
entirety.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] Not applicable to this application.
BACKGROUND
Field
[0003] Example embodiments in general relate to a biological fluid
filtration system for optically scanning a biological fluid so as
to filter potentially harmful constituents from the biological
fluid for therapeutic or diagnostic purposes.
Related Art
[0004] Any discussion of the related art throughout the
specification should in no way be considered as an admission that
such related art is widely known or forms part of common general
knowledge in the field.
[0005] Eight million people die from cancer each year and it is
predicted that 13.2 million patients will do so in 2030. 90% of
deaths are caused by metastases. Cancer metastasis spreads when
Circulating Tumor Cells (CTCs) dislodge from the primary tumor site
and travel via the circulatory system (or lymphatic system) and
lodge at another site. Certain cancer types disseminate as single
cells, while others--such as oral squamous cell carcinoma,
colorectal carcinoma, melanoma, breast cancer, endometrial
carcinoma, and pancreatic cancer, do so by collective cell
(CTC-cluster) migration. Significant variability has also been
found from transcriptional profiling and surface marker analysis,
leading to an understanding that CTCs are highly heterogeneous. In
2012, it was shown that CTC-variability is highly consistent with
tumor tissue variability, meaning that phenotyping of CTCs will
bring to light many of the characteristics present in the tumor
itself.
[0006] It is estimated that every day, tumors release thousands of
cells into the circulation where CTCs survive for about 1-2.5
hours. Most CTCs undergo apoptosis upon release or remain dormant,
and only a few (0.1%) are able to survive the effect of stressors
and ultimately then form distant metastases. Using the only
FDA-approved system for CTC separation, CellSearch (Menarini
Silicon Biosystems), researchers detected a single CTC (median 1,
range from 0 to 4) in 7.5 mL of blood in 13% of analyzed samples,
while analyzing 30-mL blood volume samples allowed a detection rate
of 2 CTCs (range from 0 to 9) in 47% of the analyzed samples. Thus,
rarity is a significant challenge for the identification and
separation of CTCs and CTC-clusters. 10 mL of a peripheral blood
sample from a metastatic cancer patient typically contains 0-100
single CTCs and roughly 0-5 CTC-clusters (5-20% of all CTCs) among
approximately 50.times.10.sup.9 red blood cells, 80.times.10.sup.6
white blood cells and 3.times.10.sup.9 platelets. Furthermore, some
CTCs overlap in size with white blood cells, making size based
separation challenging.
[0007] Yet, due to the clinical significance of CTCs in cancer
prognosis and treatment, research in techniques to separate CTCs is
very active. Separation strategies are categorized as positive
enrichment, negative enrichment, and label-free techniques.
Positive enrichment typically refers to a process that selects for
the CTCs while leaving the other particles behind. This typically
has a very high accuracy, and one of the most common methods
involves antibody tagging surface antigens (e.g. EpCAM) on the
CTC.
[0008] Negative enrichment involves targeting and removing other
types of cells, in this case, white and red blood cells. This
generally leads to lower purity but does not have the challenge of
removal of binding probes from the surface of the CTC, and is often
able to bypass the challenge of CTC heterogeneity. A label-free
technique is one that avoids biochemically tagging the desired
molecule. This means that rather than using immune affinity for
capture or sample cleaning, another method that does not involve
labeling cells is used such as size-based, mechanical property
based, acoustic, or optical. Some researchers have proposed
combining several techniques, such as optical detection of
fluorescently tagged CTCs followed by size based microfluidic
separation, to achieve better results. Recently, building upon
advances in AI and machine learning, methods have been published to
automatically detect CTCs by training machine learning algorithms
on microscopic images of CTCs in sample blood.
SUMMARY
[0009] An example embodiment is directed to a biological fluid
filtration system. The biological fluid filtration system generally
includes a fluid receiving device adapted to receive a biological
fluid. A valve including an inlet, a first outlet, and a second
outlet is fluidly connected to the fluid receiving device. The
biological fluid within the fluid receiving device is scanned by a
scanner to produce scanned data relating to the biological fluid. A
control unit in communication with the scanner and the valve
receives the scanned data and controls the valve based on the
scanned data. The valve is controlled to direct the biological
fluid through either the first or second outlet of the valve
depending upon the constituents of the biological fluid identified
by the control unit.
[0010] There has thus been outlined, rather broadly, some of the
embodiments of the biological fluid filtration system in order that
the detailed description thereof may be better understood, and in
order that the present contribution to the art may be better
appreciated. There are additional embodiments of the biological
fluid filtration system that will be described hereinafter and that
will form the subject matter of the claims appended hereto. In this
respect, before explaining at least one embodiment of the
biological fluid filtration system in detail, it is to be
understood that the biological fluid filtration system is not
limited in its application to the details of construction or to the
arrangements of the components set forth in the following
description or illustrated in the drawings. The biological fluid
filtration system is capable of other embodiments and of being
practiced and carried out in various ways. Also, it is to be
understood that the phraseology and terminology employed herein are
for the purpose of the description and should not be regarded as
limiting.
BRIEF DESCRIPTION OF THE DRAWINGS
[0011] Example embodiments will become more fully understood from
the detailed description given herein below and the accompanying
drawings, wherein like elements are represented by like reference
characters, which are given by way of illustration only and thus
are not limitative of the example embodiments herein.
[0012] FIG. 1 is a perspective view of a biological fluid
filtration system in accordance with an example embodiment.
[0013] FIG. 2 is a perspective view of a biological fluid
filtration system in accordance with an example embodiment.
[0014] FIG. 3 is a perspective view of a biological fluid
filtration system in accordance with an example embodiment.
[0015] FIG. 4 is a perspective view of a biological fluid
filtration system in accordance with an example embodiment.
[0016] FIG. 5 is a perspective drawing of a fluid receiving device
with microfluidic channels in accordance with an example
embodiment.
[0017] FIG. 6 is a schematic drawing of a fluid receiving device
with microfluidic channels in accordance with an example
embodiment.
[0018] FIG. 7 is a schematic drawing of a fluid receiving device
with microfluidic channels in accordance with an example
embodiment.
[0019] FIG. 8 is a perspective drawing of a fluid receiving device
with a microwell array in accordance with an example
embodiment.
[0020] FIG. 9 is a schematic drawing of a fluid receiving device
with a microwell array in accordance with an example
embodiment.
[0021] FIG. 10 is a schematic drawing of a fluid receiving device
with a microwell array in accordance with an example
embodiment.
[0022] FIG. 11 is a histogram of typical cell count data in
blood.
[0023] FIG. 12 is a block diagram of an aphaeretic system and
method of removing undesirable constituents using microfluidic
channels in accordance with an example embodiment.
[0024] FIG. 13 is a block diagram of an aphaeretic system and
method of removing undesirable constituents using microwell arrays
in accordance with an example embodiment.
[0025] FIG. 14 is a block diagram of a diagnostic system in
accordance with an example embodiment.
[0026] FIG. 15 is a block diagram of an aphaeretic system and
method of removing undesirable pathogens from component blood fluid
in accordance with an example embodiment.
[0027] FIG. 16 is a block diagram of an aphaeretic system and
method incorporating image data of known pathogens and optical
artifacts into a reference data in accordance with an example
embodiment.
[0028] FIG. 17 is a block diagram of an aphaeretic system and
method of removing undesirable CTCs from blood in accordance with
an example embodiment.
[0029] FIG. 18 is a block diagram of an aphaeretic system and
method incorporating image data of CTCs and CTC-clusters into a
reference data in accordance with an example embodiment.
[0030] FIG. 19 is a block diagram of an aphaeretic system and
method of removing undesirable CTCs from whole blood fluid in
accordance with an example embodiment.
[0031] FIG. 20 is a block diagram of an aphaeretic system and
method of incorporating image data of CTCs and CTC-clusters into a
reference data in accordance with an example embodiment.
[0032] FIG. 21 is a block diagram of an aphaeretic system and
method for removal of undesirable CTCs and CTC-clusters using blood
samples from representative individuals other than the patient to
obtain a reference data in accordance with an example
embodiment.
[0033] FIG. 22 is a block diagram of an aphaeretic system and
method incorporating image data of pathogens into a reference data
in accordance with an example embodiment.
[0034] FIG. 23 is a block diagram of filtration of cytokines from
blood component fluid in accordance with an example embodiment.
[0035] FIG. 24 is a block diagram of immunomagnetic filtration of
CTCs from blood component fluid in accordance with an example
embodiment.
[0036] FIG. 25 is a block diagram of molecular and genetic
profiling of undesirable CTCs removed from blood in accordance with
an example embodiment.
[0037] FIG. 26 is a graphical display of cell characteristics based
on image data of cells.
[0038] FIG. 27 is a block diagram of an aphaeretic system and
method of computer processing of reference image data obtained from
optic scans of biological fluid components from fluid samples of
representative individuals other than patient.
[0039] FIG. 28 is a block diagram of an aphaeretic system and
method of computer processing of reference image data obtained from
optic scans of biological fluid components from fluid sample of
patient.
[0040] FIG. 29 is a block diagram of an aphaeretic system and
method which utilizes a certifying/verification algorithm to remove
undesirable pathogens from component blood fluid in accordance with
an example embodiment.
[0041] FIG. 30 is a flowchart illustrating a system and method of
filtering a biological fluid utilizing both a pre-filtration
session and a filtration session in accordance with an example
embodiment.
[0042] FIG. 31 is a block chart illustrating a scanner utilizing
digital holographic microscopy of a biological fluid filtration
system in accordance with an example embodiment.
[0043] FIG. 32 is a flowchart illustrating a system and method of
filtering a biological fluid for use in connection with
leukapheresis in accordance with an example embodiment.
[0044] FIG. 33 is a flowchart illustrating a system and method of
filtering a biological fluid for use in connection with the removal
of stem cells in accordance with an example embodiment.
[0045] FIG. 34 is a block diagram illustrating a system and method
of filtering a biological fluid for use in connection with testing
of drugs and treatments in accordance with an example
embodiment.
[0046] FIG. 35 is a flowchart illustrating a multi-stage system and
method of filtering a biological fluid in accordance with an
example embodiment.
[0047] FIG. 36A is a block diagram illustrating a system and method
of filtering a biological fluid including a fluid receiving device
utilizing droplet sorting in accordance with an example
embodiment.
[0048] FIG. 36B is a block diagram illustrating additional channels
added after an isolation path of a biological fluid filtration
system in accordance with an example embodiment.
[0049] FIG. 37 is a flowchart illustrating a system and method of
filtering a biological fluid for use in connection with filtration
of subsets of healthy cells in accordance with an example
embodiment.
[0050] FIG. 38 is a flowchart illustrating a system and method of
filtering a biological fluid utilizing a presorting stage in
accordance with an example embodiment.
[0051] FIG. 39 is a flowchart illustrating a system and method of
filtering a biological fluid utilizing a presorting stage and
having multiple outlet ports in accordance with an example
embodiment.
[0052] FIG. 40 is a flowchart illustrating a system and method of
filtering a biological fluid utilizing a presorting stage for
diagnostics in accordance with an example embodiment.
[0053] FIG. 41 is a flowchart illustrating a system and method of
filtering a biological fluid utilizing a presorting stage for
diagnostics with healthy cells being returned in accordance with an
example embodiment.
[0054] FIG. 42 is a flowchart illustrating a system and method of
filtering a biological fluid utilizing a presorting stage for
diagnostics with healthy cells being returned in accordance with an
example embodiment.
[0055] FIG. 43 is a block diagram illustrating a system and method
of filtering a biological fluid utilizing multiple pre-sorting
stages in accordance with an example embodiment.
[0056] FIG. 44A is a block diagram illustrating a body-worn device
of a biological fluid filtration system worn on a patient in
accordance with an example embodiment.
[0057] FIG. 44B is a block diagram illustrating a body-worn device
of a biological fluid filtration system including anti-coagulant
and buffer fluid worn on a patient in accordance with an example
embodiment.
[0058] FIG. 44C is a perspective view of a portable filtration
device of a biological fluid filtration system in accordance with
an example embodiment.
[0059] FIG. 45 is a block diagram illustrating a body-worn device
of a biological fluid filtration system in accordance with an
example embodiment.
[0060] FIG. 46 is a flowchart illustrating the usage of a body-worn
device of a biological fluid filtration system in accordance with
an example embodiment.
[0061] FIG. 47 is a flowchart illustrating the usage of a body-worn
device with a pre-sorting stage of a biological fluid filtration
system in accordance with an example embodiment.
[0062] FIG. 48 is a flowchart illustrating the usage of a body-worn
device with drug infuser in the return path of a biological fluid
filtration system in accordance with an example embodiment.
[0063] FIG. 49 is a block diagram illustrating a closed-loop
filtering, monitoring, and testing system of a biological fluid
filtration system in accordance with an example embodiment.
[0064] FIG. 50 is a block diagram illustrating a closed-loop
filtering, monitoring, and testing system of a biological fluid
filtration system utilizing drug choice optimization in accordance
with an example embodiment.
[0065] FIG. 51 is a block diagram illustrating a biological fluid
filtration system being connected to a dialysis unit in accordance
with an example embodiment.
[0066] FIG. 52 is a block diagram illustrating a biological fluid
filtration system being connected to a blood bag in accordance with
an example embodiment.
[0067] FIG. 53 is a block diagram illustrating the usage of a
body-worn device to filter and sequester small samples of blood
containing CTCs and CTC-clusters of a biological fluid filtration
system in accordance with an example embodiment.
DETAILED DESCRIPTION
A. Overview
[0068] An example biological fluid filtration system 10 generally
comprises a receiver path 20 adapted to receive a biological fluid
16 from a biological fluid source 17. The receiver path 20 may
comprise various types of conduits, channels, or the like known in
the art for transferring a fluid between two locations such as, for
example, an apheresis catheter, an indwelling line, or a venous
catheter. Other examples could include two intravenous (IV) lines:
one for access to an artery and the other for access to a vein. The
systems and methods described herein may be utilized for filtering
a wide range of biological fluids 16, such as but not limited to
blood, lymphatic fluid, cerebrospinal fluid, sweat, urine,
pericardial fluid, stools, saliva, and the like. The biological
fluid source 17 may comprise the patient 12 herself or, in some
embodiments, a separate reservoir in which the biological fluid 16
is temporarily stored after being drawn from the patient 12.
[0069] As shown throughout the figures, a fluid receiving device 30
may be fluidly connected to the receiver path 20 so as to receive
the biological fluid 16 from the receiver path 20. The fluid
receiving device 30 generally comprises a structure in which the
biological fluid 16 is temporarily stored or channeled to be
optically scanned by a scanner 70. As discussed in more detail
below, the fluid receiving device 30 may vary in different
embodiments. In a first exemplary embodiment, the fluid receiving
device 30 may comprise one or more microfluidic channels 31. In a
second exemplary embodiment, the fluid receiving device 30 may
comprise a microwell array 32 comprised of a plurality of
microwells. In yet another exemplary embodiment, the fluid
receiving device 30 may comprise a microfluidic droplet generator
34 to scan cell encapsulating droplets 39.
[0070] A valve 40 may be positioned downstream of the fluid
receiving device 30, with the valve 40 being configured to direct
flow of the biological fluid 16 after being scanned within the
fluid receiving device 30. The valve 40 may comprise an inlet 41
which is fluidly connected to the fluid receiving device 30. The
valve 40 may comprise a first outlet 42 which is fluidly connected
to an isolation path 50 and, in some embodiments, a second outlet
43 which is fluidly connected to a return path 60. In some
embodiments, the return path 60 may be omitted, with the biological
fluid 16 being returned to the biological fluid source 17 through
the receiver path 20 rather than a separate return path 60. In some
embodiments, instead of or in addition to a return path 60 back to
the biological fluid source 17, a reprocessing path 62 may be
utilized to return back to the fluid receiving device 30 for
further processing.
[0071] It should be appreciated that the valve 40 may have a
default state in which flow is directed towards one of the two
outlets 42, 43 in the absence of control of the valve 40. For
example, the valve 40 may be a spring-based valve 40 that, by
default, directs flow to the first outlet 42. Such a valve 40 may
be adjusted to direct flow to the second outlet 43, such as by
activation of a spring. Upon release of the spring, such a valve 40
may revert back to its original, default state in which the valve
40 directs flow to the first outlet 42. Thus, the valve 40 may not
need to be controlled if the default state of the valve 40 already
directs flow to the desired outlet 42, 43.
[0072] The isolation path 50 is fluidly connected to the first
outlet 42 of the valve 40 such that the biological fluid 16
contents of the fluid receiving device 30 may be isolated or
sequestered from the biological fluid source 17 if undesirable
constituents 14 are identified in the biological fluid 16 during
scanning by the scanner 70 as discussed in more detail below. The
isolation path 50 may comprise one or more channels which are
fluidly connected to the first outlet 42 of the valve 40 such that
any scanned biological fluid 16 containing undesirable constituents
14 may be sequestered or isolated. The isolation path 50 may be
fluidly connected to a reservoir, cartridge, container, vessel, or
any device capable of holding a fluid. Such sequestered or isolated
biological fluids 16 may be utilized for diagnostics or may be
disposed of.
[0073] The scanner 70 is generally oriented toward the fluid
receiving device 30. Although the singular term is used throughout,
it should be appreciated that, in some embodiments, multiple
scanners 70 may be utilized. For example, in an embodiment in which
the fluid receiving device 30 is comprised of a microwell array 32,
a first scanner 70 could be oriented toward a first portion of the
microwell array 32 and a second scanner 70 could be oriented toward
a second portion of the microwell array 32. In any case, the
scanner 70 is adapted to optically scan the biological fluid 16
within the fluid receiving device 30 so as to derive a scanned data
90 of the biological fluid 16.
[0074] In an exemplary embodiment, the scanner 70 may utilize
digital holographic microscopy (DHM) to scan the contents of the
fluid receiving device 30. In such an embodiment, the scanner 70
may comprise a light source 72 which is adapted to illuminate the
biological fluid 16 to be scanned. More specifically, the scanner
70 may comprise a light source 72 such as a laser or a
light-emitting diode. In addition to the light source 72, the
scanner 70 in such an embodiment will generally include a
microscope objective 73 adapted to gather light from the biological
fluid 16 and an image sensor 76 so as to produce a holographic
image. Thus, the microscope objective 73 will generally comprise a
lens which functions to collect an object wave 77 front created by
the light source 72 as discussed below.
[0075] The control unit 80 in such an embodiment may then function
as a digital lens to calculate a viewable image of the object wave
77 front. Since the microscope objective 73 in such a digital
holographic microscopy embodiment is only used to collect light,
rather than to form an image, the microscope objective 73 may be
omitted entirely in some embodiments. Other example embodiments
could rely on other arrangements to generate holograms of the
object being scanned.
[0076] A control unit 80 may be communicatively connected to the
scanner 70 so as to receive the scanned data 90 of the biological
fluid 16 from the scanner 70. The control unit 80 may be
operatively connected to the valve 40 such that the control unit 80
may direct the opening or closing of the inlet 41 and outlets 42,
43 of the valve 40.
[0077] The control unit 80 is adapted to compare the scanned data
90 of the biological fluid 16 with a reference data 91. As
discussed below, the reference data 91 may comprise patterns,
criteria, images, and/or other characteristics of desirable
constituents 15 such as red blood cells, white blood cells, and the
like. The desirable constituents 15 may comprise biological fluid
constituents 13 that are desirable for a particular patient 12 or
for a particular application. It should be appreciated that a
biological fluid constituent 13 which is desirable for a first
patient 12 may be undesirable for a second patient 12. Further, a
biological fluid constituent 13 which is desirable for a particular
application may be undesirable for other applications.
[0078] Thus, the control unit 80 may be configured to determine if
a particular sample of biological fluid 16 consists only of such
desirable constituents, with the sample of biological fluid 16
being directed through the first outlet 42 of the valve 40 if only
desirable constituents are identified. If the sample of biological
fluid 16 does not consist exclusively of such desirable
constituents, the sample of the biological fluid 16 may be directed
instead through the second outlet 43 of the valve 40.
[0079] The control unit 80 is adapted to direct flow of the
biological fluid 16 from the fluid receiving device 30 by operation
of the valve 40. If the scanned data 90 of the biological fluid 16
contains only desirable constituents 15 exhibiting criteria that
match with any of the healthy or desirable constituents of the
reference data 91, the control unit 80 will switch the valve 40 so
as to direct the biological fluid 16 to the biological fluid source
17 by a return path 60, such as by the receiver path 20 in reverse.
If the scanned data 90 of the biological fluid 16 includes one or
more undesirable constituents 14 not exhibiting criteria that match
with any known desirable constituents 15 of the reference data 91,
the control unit 80 will switch the valve 40 so as to direct the
biological fluid 16 to the isolation path 50 to be isolated or
sequestered from the biological fluid source 17 for further
diagnostic or therapeutic processing. Alternatively, if the scanned
data 90 includes one or more undesirable constituents 14, the valve
40 may be switched so as to direct the biological fluid 16 back to
the fluid receiving device 30 by a reprocessing path 62 for further
processing. The scanned and reference data 90, 91 could include
images and/or morphological data of the cells in the images.
[0080] In an exemplary embodiment, the biological fluid filtration
system 10 may comprise a fluid receiving device 30 adapted to
receive a biological fluid 16 from a biological fluid source 17. A
valve 40 may be fluidly connected to the fluid receiving device 30,
with the valve 40 including an inlet 41, a first outlet 42, and a
second outlet 43. The inlet 41 of the valve 40 is fluidly connected
to the fluid receiving device 30. A scanner 70 is configured to
scan the biological fluid 16 within the fluid receiving device 30
to produce a scanned data 90 relating to the biological fluid 16
within the fluid receiving device 30. A control unit 80 is in
communication with the scanner 70 and the valve 40, with the
control unit 80 being configured to receive the scanned data 90
from the scanner 70 and to control the valve 40 based on the
scanned data 90 from the scanner 70. The control unit 80 is
configured to control the valve 40 to (a) direct the biological
fluid 16 through the first outlet 42 if the control unit 80
determines that the scanned data 90 indicates a presence of an
undesirable constituent 14 within the biological fluid 16 or (b)
direct the biological fluid 16 through the second outlet 43 if the
control unit 80 determines that the scanned data 90 does not
indicate a presence of an undesirable constituent 14 within the
biological fluid 16.
[0081] The fluid receiving device 30 may comprise a microfluidic
channel 31. An inlet valve 33 may be fluidly connected to an inlet
of the microfluidic channel 31 for pausing flow of the biological
fluid 16 into the microfluidic channel 31. The undesirable
constituent 14 may be comprised of an unknown constituent of the
biological fluid 16 that is not recognized by the control unit 80.
The undesired constituent 14 may be comprised of a tumor cell. The
control unit 80 may be configured to compare the scanned data 90 to
a reference data 91. The reference data 91 may be comprised of a
characteristic or an image of a healthy cell. The scanned data 90
may be comprised of an image.
[0082] The fluid receiving device 30 may comprise a plurality of
microfluidic channels 31. Each of the microfluidic channels 31 may
be arranged in parallel. The scanner 70 may be configured to scan
each of the plurality of microfluidic channels 31. The biological
fluid 16 may be comprised of blood, cerebrospinal fluid, or
lymphatic fluid. The first outlet 42 of the valve 40 may be fluidly
connected to a reservoir. The second outlet 43 of the valve 40 may
be fluidly connected to a biological fluid source 17. The scanner
70 may be comprised of an optical scanner such as a digital
holographic microscope. The scanner 70 may include a monochromatic
laser.
[0083] In an exemplary embodiment of the biological fluid
filtration system 10, the control unit 80 is configured to control
the valve 40 to (a) direct the biological fluid 16 through the
first outlet 42 if the control unit determines that the scanned
data 90 indicates that the biological fluid 16 consists of only
desirable constituents 15 or (b) direct the biological fluid 16
through the second outlet 43 if the control unit 80 determines that
the scanned data 90 does not indicate that the biological fluid 16
consists of only desirable constituents 15. A microfluidic
separation module 100 may be fluidly connected to an inlet of the
fluid receiving device 30 for removing one or more biological fluid
constituents 13 from the biological fluid 16 prior to scanning.
[0084] In an exemplary embodiment, the control unit 80 is
configured to determine that the scanned data 90 indicates the
presence of the undesirable constituent 14 when the control unit 80
detects the undesirable constituent 14 within the scanned data 90.
In another exemplary embodiment, the control unit 80 is configured
to determine that the scanned data 90 indicates the presence of the
undesirable constituent 14 when the control unit 80 detects an
unknown constituent within the scanned data 90. In another
exemplary embodiment, the control unit 80 is configured to
determine that the scanned data 90 indicates the presence of the
undesirable constituent 14 when the control unit 80 detects only
desirable constituents 15 within the scanned data 90.
[0085] An exemplary method of filtering a biological fluid 16 using
the biological fluid filtration system 10 may comprise the steps of
receiving the biological fluid 16 by the fluid receiving device 30;
scanning the biological fluid 16 within the fluid receiving device
30 by the scanner 70; directing the biological fluid 16 through the
first outlet 42 if the control unit 80 determines that the scanned
data 90 indicates a presence of an undesirable constituent 14
within the biological fluid 16; and directing the biological fluid
16 through the second outlet 43 if the control unit 80 determines
that the scanned data 90 indicates a presence of an undesirable
constituent 14 within the biological fluid 16.
B. Biological Fluid Filtration System
[0086] As shown in FIG. 30, an exemplary embodiment of a biological
fluid filtration system 10 may comprise an automated optofluidic
system for removal of undesirable constituents 14 from a biological
fluid 16 by optically inspecting the biological fluid constituents
13 and sequestering any undesirable constituents 14 that do not
meet recognized criterial of known desirable constituents 15 of the
biological fluid 16 such as biological fluid constituents 13 that
are desirable for a particular patient 12 or application. It should
be appreciated that the biological fluid constituents 13 may
comprise biological constituents or may comprise non-biological
constituents such as dyes and the like.
[0087] The systems and methods described herein may be utilized for
the filtration of a wide range of biological fluids 16, such as but
not limited to blood, lymphatic fluid, cerebrospinal fluid, sweat,
urine, pericardial fluid, stools, saliva, and any other
organism-derived extracellular fluid that retains or transports
nutrients, cells, waste products, or foreign bodies, or that is
susceptible to pathogenic infection. It should be appreciated that
the systems and methods described herein are further not intended
to be limited to humans, and could be utilized in connection with
veterinary treatment of a wide range of animals in some
embodiments.
[0088] The systems and methods described herein may be utilized for
both diagnostic and therapeutic applications. In some embodiments,
multiple biological fluids 16 may be processed from the same
individual (animal or human). As a non-limiting example, there are
cases in which cancer has spread from the original tumor site
(e.g., breast, lung, etc.) to the meninges surrounding the brain
and/or spinal cord. In such cases, the systems and methods
described herein may be utilized to process and filter CTCs and
CTC-clusters from multiple biological fluids 16, such as from blood
as well as from cerebrospinal fluid. In some embodiments, these
multiple biological fluids 16 may be aphaeretically processed
simultaneously, particularly in a diagnostic setting.
[0089] After sequestration of the output, the sequestered output
may be reprocessed additional times using an optical filtration
system (without the aphaeretic components) to further isolate CTCs,
CTC-clusters, WBC's, and cell-free plasma. By utilizing multiple
valves and output ports, the isolated CTCs, CTC-clusters, WBC's,
and cell-free plasma could be held separately for further
processing.
[0090] In some embodiments, rather than the biological fluid 16
being directly sourced from a person or animal, the biological
fluid 16 could be a sample with or without a return path 60 back to
its source. Such embodiments could include the use in diagnostic
systems to separate pathogens, CTCs, or CTC-clusters from
biological samples for downstream diagnostics (e.g., genomic,
transcriptomic, metabolomics, drug sensitivity, drug resistance,
etc.) and characterization.
[0091] In another exemplary embodiment, the biological fluid source
17 could be an aphaeretic extract from a therapeutic aphaeresis or
leukapheresis machine. Such leukapheresis machines are routinely
utilized in labs, such as where WBC's are separated from a patient.
In the case of cancer patients, such leukapheresis extracts may
contain CTCs or CTC-clusters and some platelets in addition to
WBC's. FIG. 32 illustrates an exemplary method of optical
filtration being utilized to process leukapheresis extracts and
filter CTCs and CTC-clusters.
[0092] In yet another exemplary embodiment, tumor cell contaminants
may be identified and removed from autologous stem-cell transplant
products. Autologous stem cell transplants are typically used in
patients who need to undergo high doses of chemotherapy and
radiation to cure their diseases. These treatments could be toxic
and damage the bone marrow. An autologous stem cell transplant aids
to replace the damaged bone marrow, but it is often reported that
the process to collect stem cells from the patient could lead to
contamination of such products with tumor cells. FIG. 33
illustrates an exemplary method of optical filtration being
utilized to process stem cell extracts.
[0093] Biological fluid constituents 13 of the biological fluid 16
may comprise desirable constituents 15 and undesirable constituents
14. The desirable constituents 15 may comprise entities within the
biological fluid 16 that are recognized as being normally found in
the subject biological fluid 16 or that are desirable for a
particular patient 12 or application. Such desirable constituents
15 will typically play a role in the proper functioning of tissues,
organs, and body systems. In some embodiments, desirable
constituents 15 may comprise biological fluid constituents 13 which
are known to be benign or otherwise non-harmful.
[0094] The undesirable constituents 14 may comprise any biological
fluid constituents 13 which are not recognized as being benign,
healthy, or otherwise non-harmful. Undesirable constituents 14 may
comprise any entity that is not a healthy constituent for a
particular patient 12. It should be appreciated that biological
fluid constituents 13 may be healthy or desirable for a first
patient 12 and unhealthy or undesirable for a second patient 12.
Thus, the identification of a particular biological fluid
constituent 13 as either healthy/desirable or unhealthy/undesirable
will typically be on a patient-by-patient basis. By way of example
and without limitation, undesirable constituents 14 may comprise
circulating tumor cells (CTCs), CTC-clusters, and pathogens,
including bacterial and fungal organisms, protozoa, extracellular
vesicles, lipids, cholesterol, dyes, drugs, and infectious viral
agents. In some embodiments, any biological fluid constituent 13
which is unrecognized or unknown may be assumed to be an
undesirable constituent 14.
[0095] FIG. 1 is a schematic diagram illustrating a configuration
of one embodiment of biological fluid filtration system 10. The
illustrated exemplary embodiment includes a receiver path 20
adapted to receive a biological fluid 16 from a biological fluid
source 17; a fluid receiving device 30 fluidly connected to the
receiver path 20 so as to receive the biological fluid 16 from
receiver path 20; a valve 40 comprising an inlet 41, a first outlet
42, and a second outlet 43, wherein the inlet 41 is fluidly
connected to the fluid receiving device 30; an isolation path 50
fluidly connected to first outlet 42; a return path 60 in fluid
communication with second outlet 43; a scanner 70 oriented toward
the fluid receiving device 30, and a control unit 80
communicatively connected to the scanner 70 and operatively
connected to the valve 40.
[0096] The scanner 70 is adapted to optically scan the biological
fluid 16 within the fluid receiving device 30 so as to derive a
scanned data 90 of the biological fluid 16 and relay the scanned
data 90 to the control unit 80. The control unit 80 is adapted to
compare the scanned data 90 of the biological fluid 16 with a
reference data 91, the reference data 91 comprising recognized data
patterns characteristic of desirable constituents 15 of the
biological fluid 16. In some embodiments, both the scanned and
reference data 90, 91 may comprise images which are compared to
each other. In other embodiments, the scanned and reference data
90, 91 may comprise characteristics such as cellular
characteristics including but not limited to cell solidity, cell
surface characteristics, nucleus-cytoplasmic ratio, convexity,
luminescence, circularity, elongation, size, inner diameter and
roundness. In other embodiments, the scanned and reference data 90,
91 may comprise both images as well as characteristics of the cells
in those images, such as cellular characteristics including but not
limited to cell solidity, cell surface characteristics,
nucleus-cytoplasmic ratio, convexity, luminescence, circularity,
elongation, size, inner diameter and roundness.
[0097] The control unit 80 is adapted to switch the valve 40 so as
to direct the biological fluid 16 to the biological fluid source 17
by the return path 60 if the scanned data 90 of the biological
fluid 16 includes only desirable constituents 15 having one or more
recognized characteristics of any of the desirable constituents 15
of the reference data 91. The control unit 80 is further adapted to
switch the valve 40 so as to direct the biological fluid 16 to the
isolation path 50 if the scanned data 90 of the biological fluid 16
includes one or more undesirable constituents 14 lacking one or
more recognized characteristics of any of the desirable
constituents 15 of the reference data 91. In some embodiments, a
reprocessing path 62 may be included to route the biological fluid
16 back to the fluid receiving device 30 for further processing.
The return path 60 and reprocessing path 62 may comprise any device
or conduit suitable for transferring a fluid.
[0098] The return path 60 may comprise a channel or a plurality of
channels through which the biological fluid 16 may be returned to
the biological fluid source 17. For example, the return path 60 may
comprise a catheter, one or more intravenous lines, an indwelling
line, or a venous catheter. Similarly, the reprocessing path 60 may
comprise a channel or a plurality of channels through which the
biological fluid 16 may be returned to the fluid receiving device
30 for further processing.
[0099] The receiver path 20 includes any suitable vessel for
sterile delivery or transfer of a biological fluid 16 for various
purposes such as clinical analysis or processing. For example, in
the aphaeretic system of the preferred embodiment, the receiver
path 20 may be an apheresis catheter, two intravenous lines (one
for the artery and the other for a vein), an indwelling line, or a
venous catheter.
[0100] The fluid receiving device 30 includes any microfluidic
device or system suitable for optic imaging or microscopy. The
fluid receiving device 30 will generally include an inlet through
which biological fluid 16 is received into the fluid receiving
device 30 and an outlet through which biological fluid 16 may exit
the fluid receiving device 30. In some embodiments, the fluid
receiving device 30 may include multiple inlets and/or multiple
outlets. The inlet of the fluid receiving device 30 may be fluidly
connected to the biological fluid source 17, and the outlet of the
fluid receiving device 30 may be fluidly connected to an isolation
path 50, return path 60, reprocessing path 62, or cartridge 126,
146.
[0101] In one embodiment, which is shown schematically in FIGS.
5-7, the fluid receiving device 30 comprises one or more
microfluidic channels 31. In the illustrated embodiment, the
microfluidic channels 31 are configured for illustration purposes
as a batch of planar, parallel channels with a linear geometry.
However, microfluidic channels suitable for use with systems and
methods described herein may have any number of topographies,
geometries and patterns. For example, the microfluidic channels 31
may be etched or molded in a microfluidic chip.
[0102] In an alternative embodiment, which is shown in FIGS. 8 and
9, the fluid receiving device 30 comprises a microwell array 32.
Other suitable microfluidic devices include, e.g., glass capillary
systems. In a further embodiment, best shown in FIG. 36A, the fluid
receiving device 30 may comprise a microfluidic droplet generator
34.
[0103] The valve 40 is preferably a router-type micro-valve, but
could include other types as well such as diaphragm-type
micro-valve with electromagnetic actuation, piezo electric,
thermoplastic, etc. The valve 40 is actuated by a control signal 81
sent by the control unit 80. The valve 40 can be actuated by
various methods such as, for example, mechanically, electrically,
piezo, electro-thermally, pneumatically, electromagnetically, by
phase changes, or by introduction of external force. The valve 40
is preferably a diaphragm-type micro-valve with electromagnetic
actuation. For fluid receiving devices 30 comprising multiple
microfluidic channels 31 or a microwell array 32, a corresponding
number of valves 40, in which each valve 40 exclusively directs the
flow of contents out of a single corresponding channel or well, is
preferred. However, in some example embodiments, multiple wells
could share a valve 40. In other example embodiments, rather than
valves 40, pipettes could be utilized to extract contents from
microwells.
[0104] The isolation path 50 includes any suitable vessel or
microwells for sterile delivery or transfer of a biological fluid
16 for various purposes such as in connection with clinical
analysis or processing. In a preferred embodiment, biological fluid
16 directed to the isolation path is transported and stored in
accordance with appropriate hematology procedures for diagnostic or
conformational analysis, as described in more detail below.
[0105] The scanner 70 may comprise any device capable of scanning a
biological fluid 16. The scanner 70 may comprise an optical
scanner, including but not limited to a digital holographic
microscope. The scanner 70 generally comprises a light source and
optical detector. The scanner 70 is preferably configured to
develop a multi-modal reference data 91 from which patterns
characteristic of desirable constituents 15 of biological fluid 16
may be can be recognized. In an example embodiment, the scanner 70
comprises a Differential Interference Contrast Microscopy System
(DIC Microscopy System). Generally, DIC Microscopy generates
contrast by translating refractive index gradients of different
areas of a specimen into amplitude variations that are visualized
as differences in brightness. In that regard, the DIC Microscopy
System is adapted to create enhanced contrast images, and is
particularly suited for use in imaging unstained cell specimens
exhibiting little natural visible contrast. DIC Microscopy's
contrast-enhanced imaging yields information concerning cell
characteristics highly relevant for many of the embodiments
presented herein, including but not limited to cell solidity, cell
surface characteristics, nucleus-cytoplasmic ratio, convexity,
luminescence, circularity, elongation, size, inner diameter and
roundness.
[0106] Other imaging devices and techniques suitable for use
individually or in combination as a scanner 70, for example
include, Phase contrast microscopy (PCM), Hoffman modulation,
polarized light microscopy, holographic microscopy, confocal
scanning optic microscopy (CSOM), or laser scanning optic
microscopy (SOM) to measure voxel fluorescence, bright-field
microscopy, dark-field illumination, Raman spectrometry to measure
Raman Scattering, Optical interferometry to measure optical
interference, total internal reflection fluorescence microscopy to
measure evanescent effect, planar waveguides for refractive index
detection, photonic crystal biosensors for measure of biomolecules
on cell surfaces, and light property modulation detections such as
surface plasmon resonance (SPR) detection.
[0107] The optical detector of the scanner 70 may comprise a single
lens, or multiple linear lenses or lens arrays, and may include
lenses of different shapes, for example, PCX lenses or Fresnel
lenses.
[0108] In alternative embodiment, the scanner 70 and fluid
receiving device 30 are combined in individual modules in a modular
optofluidic system (MOPS), which permits modification or
reconfiguration of modules to suit a given application.
[0109] The control unit 80 may be local, remote, or cloud-hosted
such as through distributed networking. As shown in FIG. 29, the
control unit 80 follows an image processing software program
comprising an algorithm 82 (or series of algorithms) adapted to
identify patterns in reference data 91 of images taken of desirable
constituents 15 of a biological fluid 16 and to recognize those
patterns in processing the scanned data 90 obtained by the scanner
70.
[0110] If all biological fluid constituents 13 of the scanned
biological fluid 16 present in the fluid receiving device 30 at
that moment sufficiently match one or more characteristics of
desirable constituents 15 recognized by the algorithm 82, then the
image processing software program generates a first control signal
instruction 83 to the control unit 80 to relay a control signal 81
to the valve 40 to route the scanned biological fluid 16 to the
return path 60.
[0111] If, on the other hand, one or more of the constituents 13 of
the scanned biological fluid 16 present in the fluid receiving
device 30 at that moment fails to sufficiently match patterns of
desirable constituents 15 recognized by algorithm 82, then the
image processing software program generates a second control signal
instruction 84 to the control unit 80 to relay a control signal 81
to the valve 40 to route the scanned biological fluid 16 to the
isolation path 50.
[0112] The control unit 80 may optionally also follow an additional
image processing software program adapted to recognize and search
for patterns in the scanned data 90 indicating the presence of an
undesirable constituent 14 such as a disease-related constituent in
the scanned biological fluid 16. The additional processing software
program preferably comprises a certifying algorithm 85 adapted to
provide validation for each control signal instruction based on a
verification condition. For example, a control signal instruction
to route the scanned biological fluid 16 to the return path 60 is
validated only on condition that no patterns were detected in the
scanned data 90 to indicate the presence of an undesirable
constituent 14 such as a targeted disease-related constituent in
the scanned biological fluid 16. If the condition is not satisfied,
a corrected instruction is issued to route the scanned biological
fluid 16 to the isolation path 50.
[0113] In one embodiment, the reference data 91 is derived based on
normative data for recognized desirable constituents 15 of the
biological fluid 16 within a relevant reference population. In
another embodiment, the reference data 91 is derived by imaging
biological fluid 16 samples obtained from representative
individuals in the relevant reference population. In other
embodiments, the reference data 91 may at least partially be
derived from a pre-filtration session with a particular patient
12.
[0114] In such an embodiment, as illustrated in FIGS. 12-17 and 19,
the reference data 91 is obtained based on imaging of a fluid
sample of the subject patient's biological fluid system during a
pre-filtration session. Because the reference data 91 of the
embodiment is patient specific, data ambiguity based on constituent
heterogeneity is avoided. Accordingly, the algorithm 82, which may
rely upon machine learning, is configured to more precisely
identify patterns of desirable constituents 15 in the
subject-specific reference data 91 and thus more precisely
recognize desirable constituents 15 of the scanned data 90 taken of
the subject biological fluid source 17 during filtration.
[0115] In some embodiments, machine learning and/or artificial
intelligence models may be utilized to develop and optimize the
reference data 91. By way of example, samples of biological fluids
16 from the patient 12 or from others may be analyzed to identify
desirable constituents 15 of the biological fluid 16 through
machine learning. Such a system may be more easily accomplished
with relation to healthy cells which are common to all patients 12,
such as red blood cells, white blood cells, and the like. This is a
result of consistency of the characteristics or images of such
healthy cells and the availability of a large sample size. With
respect to undesirable constituents 14, machine learning and/or
artificial intelligence models may also be applied, though with a
smaller sample size and with the caveat that certain unhealthy
cells such as CTCs may have different characteristics in different
patients 12. Additionally, the scanned data 90 may also be analyzed
using machine learning and/or artificial intelligence models.
Further, the process of comparing the scanned data 90 with the
reference data 91, and the resulting determination by the control
unit 80, may also utilize machine learning and/or artificial
intelligence models.
[0116] In another embodiment, as illustrated in FIG. 16, the
reference data 91 includes imaging data allowing the control unit
80 to discern optical artifacts, e.g., halos, air bubbles, or
blurring, to avoid errors where characteristics of such artifacts
in the imaging data might otherwise trigger an erroneous second
control signal instruction 84 to direct fluid containing only
desirable constituents 15 to the isolation path 50. For the same
reason, the reference data 91 may include imaging data allowing the
control unit 80 to discern non-targeted biological entities, e.g.,
benign bacteria.
[0117] In another embodiment, the biological fluid filtration
system 10 conducts a continuous-flow microfluidic operation. In
such an embodiment, the biological fluid 16 may continuously flow
through the fluid receiving device 30 as the biological fluid 16 is
scanned. In an alternative embodiment, as illustrated in FIGS.
12-16, the microfluidic channel 31 includes an inlet valve 33 such
as a channel lock adapted to suspend fluid flow while biological
fluid 16 in the microfluidic channel 31 is scanned.
[0118] In yet another embodiment, which is illustrated in FIGS. 2,
15, 17, 18, and 20-22, the biological fluid filtration system 10
performs pre-processing on the biological fluid 16 using a
microfluidic separation module 100 to separate particular cellular
constituents of the biological fluid 16 for promotion to the fluid
receiving device 30. Suitable pre-processing devices and techniques
suitable may rely on a variety of cell-characteristics to perform
separation including, size, density, inertial hydrodynamic, antigen
binding affinity, acoustics, motility, electric charge, electric
dipole moment, centrifugation, or magnetism. Some techniques might
also involve the use of buffer solutions.
[0119] In one embodiment, in which blood is filtered for removal of
CTCs, whole blood is pre-processed to separate leukocytes from
smaller-sized blood constituents (i.e., erythrocytes, thrombocytes,
plasma) and large CTCs or CTC-clusters. The separated,
leukocyte-enriched blood, which contains small CTCs, is promoted to
the fluid receiving device 30 for scanning and filtering. CTC-free
leukocytes along with the other separated healthy blood
constituents are returned to the subject's circulatory system, and
the separated large and small CTCs or CTC-clusters are
sequestered.
[0120] The other separated blood constituents can optionally be
subjected to additional filtering before being returned to the
subject's circulatory system. For example, as illustrated in FIGS.
23-26, separated small blood constituents can be subject to
antigen-based filtration to remove proteins that promote
metastasis, including aiding in CTC extravasation and evasion of
immune response, e.g., cytokines, chemokines, and growth factors
released by tumor-associated macrophages. Antigen-based filtration
may also be optionally used to compliment both pre-processing and
optical scanning and filtering processes to remove any remaining
CTCs in pre-processed or optically scanned and filtered biological
fluids 16 before being returned to the subject.
[0121] In an exemplary embodiment, which is shown in FIGS. 1-4, the
biological fluid 16 is blood, the biological fluid source 17 is the
patient 12, and the biological fluid filtration system 10 is an
aphaeretic system that circulates blood from the circulatory system
of a patient 12, removes undesirable constituents 14 from the
blood, and then returns the filtered, healthy blood back to the
circulatory system. Such a biological fluid filtration system 10
has equal application to other biological fluids 16, including, for
example, cerebrospinal fluid, lymphatic fluid, and the various
other fluids described herein.
[0122] The biological fluid filtration system 10 can be applied as
a therapeutic, a diagnostic system, or both. In one embodiment, a
patient's pathology is determined and the particular
cancer-associated CTCs or disease causative agent identified, and
the system is adapted to filter out the known CTCs or agents from
patients the biological fluid system. In such an embodiment as
illustrated in FIGS. 3 and 4, the receiver path 20 is adapted to
receive and facilitate flow of biological fluid 16 from the patient
12 via, e.g., a cannula and venous catheter assembly, to the fluid
receiving device 30.
[0123] As shown in FIGS. 1 and 2, the scanner 70 is adapted to
optically scan the biological fluid 16 on or within the fluid
receiving device 30 so as to derive a scanned data 90 of the
biological fluid 16 and relay the scanned data 90 to the control
unit 80. The control unit 80 is adapted to compare the scanned data
90 of the biological fluid 16 with a reference data 91 to relay a
control signal 81 to the valve 40 to route healthy biological fluid
16 to the return path 60 and sequester biological fluid 16
containing CTCs or undesirable disease causative agents to the
isolation path 50, as described herein. The return path 60 is
adapted to receive and facilitate flow of healthy biological fluid
16 from the fluid receiving device 30 via the valve 40, and
returning the fluid 16 to the patient 12 via, e.g., a cannula and
venous catheter assembly. In another embodiment, the sequestered
biological fluid 16 is diagnostically processed for conformational
analysis.
[0124] In another embodiment, which is shown in FIG. 14, a
subject's pathology is undetermined, and the system is adapted to
filter out undesirable constituents 14 not recognized by the system
as meeting pre-determined criteria characteristic of desirable
constituents 15 of the subject biological fluid 16, as described
herein. As with the therapeutic system described above, the scanner
70 of the diagnostic system is adapted to optically scan the
biological fluid 16 within the fluid receiving device 30 so as to
derive the scanned data 90 of the biological fluid and relay the
scanned data 90 to the control unit 80, and, in turn, the control
unit 80 is adapted to compare the scanned data 90 of the biological
fluid 16 with the reference data 91 and relay a control signal 81
to the valve 40 to route healthy biological fluid 16 to the return
path 60 and sequester biological fluid 16 containing undesirable
constituents 14, such as undesirable, indeterminate biological
agents, as described herein. If used for diagnostic purposes, in
some embodiments, such a system could be used for early detection
of the signs of cancer in the patient. In such diagnostic systems,
even the filtered fluid containing healthy components may not be
returned back to the subject.
[0125] The sequestered biological fluid 16 is then subject to
diagnostic testing to identify the undesirable constituents 14
present in the sequestered biological fluid 16. In the diagnostic
embodiment, the receiver path 20 is optionally adapted to receive
and facilitate flow of biological fluid 16 from the patient's
biological fluid system and the return path 60 is optionally
adapted to return healthy biological fluid 16 to the patient's
biological fluid system. In one embodiment, the receiver path 20
and return path 60 are respectively adapted to receive and return
biological fluid 16 from the subject's biological fluid system, the
diagnostic system is adapted to both diagnose and filter out
undesirable constituents 14 in the subject's biological fluid
system. In an alternative embodiment, the receiver path 20 is
adapted to receive and facilitate flow of a biological fluid 16
sample taken of the subject's biological fluid system, and the
healthy biological fluid 16 routed to the return path 60 is
processed for storage or disposal.
[0126] In some embodiments such as shown in FIG. 36B, the isolation
path 50 may further bifurcate into additional channels and valves
40, with each such bifurcated additional channel including its own
fluid receiving device 30 and scanner 70. The geometry of such
additional channels could be of various shapes, including without
limitation rectangular, serpentine, spiral, circular, square, or
combinations thereof. The geometry of the exemplary embodiment
shown in FIG. 36B should thus not be construed as limiting.
[0127] Continuing to reference FIG. 36B, additional channels may be
added after the isolation path 50 of any of the embodiments
described and shown herein. Contents from such an isolation path 50
are scanned in multiple additional stages for further enrichment of
the biological fluid 16. As shown in FIG. 36B, contents from an
isolation path 50 enter an additional fluid receiving device 30 and
are scanned again by a scanner 70. The resulting scanned data 90 is
processed by the control unit 80. Valves 40 are provided to direct
the contents into additional stages, with the valves 40 being
operated by the control unit 80 based on the results of its
processing of the contents. In exemplary embodiments utilized in an
aphaeretic setting, any contents including only healthy cells
(e.g., desirable constituents 15) are carried forward, such as back
to the biological fluid source 17 via a return path 60.
[0128] Contents which include a mixture of healthy and non-healthy
cells (e.g., any contents including undesirable constituents 14)
are directed along an additional channel to an additional fluid
receiving device 30 and are scanned again by a scanner 70. After
the additional fluid receiving device 30, valves 40 are provided to
bifurcate the additional channel such that contents including only
healthy cells may be diverted towards a return path 60. Contents
including non-healthy cells such as undesirable constituents 14 are
directed along an additional channel to an additional fluid
receiving device 30 and are scanned again by a scanner 70. This
process may be repeated any number of times, with healthy cells
being carried forward to be isolated or returned, and any unhealthy
cells being scanned additional times.
[0129] As shown in FIG. 36B, pumps 46 may be utilized to regulate
flow rates of the biological fluid 16 through the various channels.
The pumps 46 may comprise pressure pumps, syringes, peristaltic,
and the like. The scanning techniques could vary across successive
stages or sections (e.g., DHM, digital inline holography, etc.) and
the algorithms utilized to identify cells may also vary (e.g.,
neural network type classifiers to decision trees, etc.). In other
example embodiments, such an arrangement could involve droplets 39
as opposed to cells in media.
C. Fluid Receiving Device
[0130] As described and shown herein, a wide range of fluid
receiving devices 30 may be utilized for the biological fluid
filtration system 10 in combination with various types of
biological fluids 16, including but not limited to blood and/or
leukapheresis extracts. The fluid receiving device 30 may comprise
a microfluidic platform. In a first exemplary embodiment as shown
in FIG. 5, the fluid receiving device 30 may comprise one or more
microfluidic channels 31. In a second exemplary embodiment as shown
in FIG. 8, the fluid receiving device 30 may comprise a microwell
array 32. In a third exemplary embodiment as shown in FIG. 36A, the
fluid receiving device 30 may comprise a microfluidic droplet
generator 34. It should also be appreciated that various other
types of fluid receiving devices 30 may be utilized.
[0131] It should be appreciated that, in some embodiments, multiple
types of fluid receiving devices 30 may be grouped together. For
example, an exemplary fluid receiving device 30 may comprise both
microfluidic channels 31 and a microwell array 32, arranged
together in parallel or in series. As a further example, a fluid
receiving device 30 may comprise both microfluidic channels 31 and
a droplet generator 34. As yet another example, a fluid receiving
device 30 may comprise microfluidic channels 31, a microwell array
32, and a droplet generator 34 all configured to work in concert.
Additionally, in some embodiments, there could be multiple
successive valves 40 and fluid receiving devices 30 attached to a
single outlet to enrich the contents of a microchannel.
[0132] As shown in FIG. 5, an exemplary embodiment of the fluid
receiving device 30 may comprise one or more microfluidic channels
31. The microfluidic channels 31 will generally comprise networks
of one or multiple channels through which biological fluids 16 may
be routed to be processed using the systems and methods described
herein. In some embodiments, a single microfluidic channel 31 may
be utilized, such as with an embodiment effectuating single-stage
processing of a single biological fluid 16. In other embodiments,
multiple microfluidic channels 31 may be utilized, such as with an
embodiment effectuating multi-stage processing of a single
biological fluid 16, or single-stage processing of multiple
biological fluids 16 simultaneously.
[0133] By way of example, multiple fluid receiving devices 30 may
be arranged in parallel so as to allow for multiple biological
fluids 16, either from the same or different biological fluid
sources 17, to be simultaneously scanned and then routed
accordingly. In this manner, multiple types of biological fluids 16
from the same or multiple biological fluid sources 17 may be
processed simultaneously. By way of example, a first biological
fluid 16 may be processed in a first fluid receiving device 30 and
a second biological fluid may be processed in a second fluid
receiving device 30. Additional fluid receiving devices 30 may
similarly be operated in parallel to suit any number of biological
fluids 16.
[0134] As a further example, multiple fluid receiving devices 30
may be arranged in series such that a single biological fluid 16,
or multiple biological fluids 16, may be scanned multiple times in
multiple stages. For example, a first fluid receiving device 30 may
process a biological fluid 16 a first time, and then the biological
fluid 16 may be transferred to a second fluid receiving device 30
in series with the first fluid receiving device 30 so as to process
the biological fluid 16 a second time. Subsequent fluid receiving
devices 30 may also be added, allowing for multi-stage separation
of the biological fluid 16 as discussed herein. In some
embodiments, multiple fluid receiving devices 30 may be arranged
both in parallel and in series so as to allow simultaneous,
multi-stage processing of multiple biological fluids 16. The
microfluidic channels 31 may be fabricated from polymeric
materials, including but not limited to polymethylmethacrylate,
cylic olefin copolymer (COC), polycarbonate, polydimethylsiloxane
(PDMS), SU-8 photoresist, and the like. As previously discussed,
the microfluidic channels 31 may comprise various topographies,
geometries, and patterns, and thus should not be construed as
limited by the exemplary topographies, geometries, and patterns
shown in the exemplary figures.
[0135] The microfluidic channels 31 may be arranged in a
multilayer, 2-dimensional or 3-dimensional configuration. Moreover,
various channel geometries may be employed including, for example,
curvilinear and spiral geometries. Various channel patterns in
addition to parallel patterns may be suitable including, for
example, knot, basket-weave, and braided patterns. By way of
example and without limitation, the microfluidic channels 31 may be
straight, curved, or helical. Where multiple microfluidic channels
31 are utilized, the plurality of microfluidic channels 31 may be
arranged in parallel. In other embodiments, one or more
microfluidic channels 31 may cross paths without being fluidly
interconnected.
[0136] The microfluidic channels 31 may be formed of various
materials including polymers, such as silicon, glass or polymers,
e.g., polydimethylsiloxane (PDMS). In some embodiments, the
microfluidic channels 31 may be stacked on top of each other. The
positioning, orientation, and arrangement of the microfluidic
channels 31 may vary and thus should not be construed as limited by
the exemplary figures or description herein.
[0137] As shown in FIG. 8, another exemplary embodiment of the
fluid receiving device 30 may comprise one or more microwell arrays
32. Each microwell array 32 will generally comprise an array of
microwells into which the biological fluid 16 may be transferred
for scanning by the scanner 70 using the various techniques
discussed herein. The shape, size, and density of the individual
wells of the microwell array 32 used herewith may vary in different
embodiments.
[0138] The number of microwell arrays 32 utilized for scanning each
biological fluid 16, or for scanning multiple biological fluids 16,
may vary in different embodiments. In some embodiments, a single
microwell array 32 may be utilized such as shown in FIG. 8. In
other embodiments, multiple microwell arrays 32 may be utilized,
such as for scanning multiple biological fluids 16 simultaneously
or in-turn.
[0139] By way of example, multiple microwell arrays 32 may be
arranged in parallel so as to allow for multiple biological fluids
16, either from the same or different biological fluid sources 17,
to be simultaneously scanned and then routed accordingly. In this
manner, multiple types of biological fluids 16 from the same or
multiple biological fluid sources 17 may be processed
simultaneously. By way of example, a first biological fluid 16 may
be processed in a first microwell array 32 and a second biological
fluid 16 may be processed in a second microwell array 32.
Additional microwell arrays 32 may similarly be operated in
parallel to suit any number of biological fluids 16. Some
embodiments could combine the use of microwell arrays 32 and
microfluidic channels 31.
[0140] As a further example, multiple microwell arrays 32 may be
arranged in series such that a single biological fluid 16, or
multiple biological fluids 16, may be scanned multiple times in
multiple stages. For example, a first microwell array 32 may
process a biological fluid 16 a first time, and then the biological
fluid 16 may be transferred to a second microwell array 32 in
series with the first microwell array 32 so as to process the
biological fluid 16 a second time. Subsequent microwell arrays 32
may also be added, allowing for multi-stage separation of the
biological fluid 16 as discussed herein. In some embodiments,
multiple microwell arrays 32 may be arranged both in parallel and
in series so as to allow simultaneous, multi-stage processing of
multiple biological fluids 16.
[0141] It should also be appreciated that the type of microwell
array 32 may vary in different embodiments. The microwell array 32
may comprise single-use wells (e.g. parylene valves) or multi-use
wells (e.g., piezo locks). The number of microwells included in
each microwell array 32 may vary in different embodiments to suit
different applications. Further, the manner in which the biological
fluid 16 is introduced into the microwell array 32 may vary in
different embodiments. By way of example and without limitation,
the biological fluid 16 may be introduced into the microwell array
32 utilizing pumps, valves, microfluidic devices, pipettes, or
various combinations thereof.
[0142] As best shown in FIG. 36A, another exemplary embodiment of a
fluid receiving device 30 may comprise a droplet generator 34 in
which cells are encapsulated within droplets 39 such that the
droplets 39 may be scanned by the scanner 70. Such an embodiment
may rely on the use of droplets 39 to compartmentalize cells in
nanoliter scale compartments. Similar systems have been previously
used as reaction chambers for transcriptomic analysis. Utilizing
the systems and methods described herein, cells may be encapsulated
into droplets 39. Those droplets 39 may then be scanned by the
scanner 70 to differentiate between healthy cells (e.g., normal
blood cells) or unhealthy cells (e.g., cancer cells). The generated
droplets 39 may comprise various shapes and sizes. The generated
droplets 39 may be spherical or non-spherical (e.g., oblong).
[0143] In an embodiment utilizing droplet sorting such as shown in
FIG. 36A, a defined microfluidic channel-cross design, such as a
droplet generator 34, may be utilized. Using the droplet generator
34, two or more immiscible phase channels 35, 36 will generally
meet at an angle to generate droplets 39. A first exemplary
immiscible phase may comprise a dispersed phase channel 35 through
which an aqueous solution containing cells 18 may be routed. A
second exemplary immiscible phase channel may comprise one or more
continuous phase channels 36, through which oil may be routed.
[0144] Continuing to reference FIG. 36A, it can be seen that the
dispersed phase channel 35 includes an aqueous solution containing
cells 18 from the biological fluid 16. The continuous phase channel
36 includes oil or oils which are immiscible with the aqueous
solution. The dispersed phase channel 35 and continuous phase
channel 36 meet at a juncture 37 at an angle such that droplets are
generated which compartmentalize the cells to be scanned. In the
embodiment shown in FIG. 36A, multiple continuous phase channels 36
are utilized.
[0145] As can be seen in FIG. 36A, the juncture 37 may comprise a
four-way juncture 37 having a single dispersed phase channel 35, a
pair of continuous phase channels 36, and a scanning channel 38
wherein the droplets 39 containing compartmentalized cells 18 are
scanned. It should be appreciated, however, that such a
configuration of the juncture 37 and associated channels 35, 36, 38
may vary in different embodiments. For example, in some
embodiments, only a single continuous phase channel 36 may be
utilized.
[0146] The angle at which the dispersed phase channel 35 and
continuous phase channel(s) 36 converge at the junction 37 may vary
in different embodiments. In the exemplary embodiment shown in the
figures, a pair of continuous phase channels 36 meets at right
angles with a single dispersed phase channel 35. However, various
other angles may be utilized in different embodiments to suit
different applications. Further, the angle of the scanning channel
38 with respect to the dispersed phase channel 35 and continuous
phase channel(s) 36 may also vary in different embodiments.
[0147] The flow rates of the respective phase channels 35, 36
define the throughput of the system. The flow rate within the
respective phase channels 35, 36 may vary in different embodiments.
Generally, the flow rate of the dispersed phase channel 35,
generally containing cells 18 from the biological fluid 16 in an
aqueous media, such as an aqueous solution, will be greater than
the flow rate of the continuous phase channel 36, generally
containing an oil. However, different flow rates may be utilized to
suit different biological fluids 16, scanners 70, or other
considerations. Further, the rate of flow through the scanning
channel 38 may vary in different embodiments. In some embodiments,
the scanning channel 38 may include a channel lock such as an inlet
valve 33 so as to pause or reduce flow rate while cells 18 are
being scanned.
[0148] The size of the droplet generator 34 and the ratio of the
dispersed phase channel 35 when compared with the continuous phase
channel 36 will generally define the size of generated droplets 39
exiting the juncture 37. Thus, the size of the droplet generator
34, and the ratio of the sizes of the dispersed phase channel 35
and continuous phase channel 36, may vary in different embodiments
to suit different sizes of generated droplets 39 to be scanned by
the scanner 70. Accordingly, the size of the droplet generator 34
and ratio between the respective phase channels 35, 36 should not
be construed as the exemplary embodiment shown in FIG. 36A.
[0149] Cells 18 of the biological fluid 16 are routed first through
the dispersed phase channel 35 in an aqueous media such as an
aqueous solution. The aqueous media containing the cells 18 from
the biological fluid 16 passes through the juncture 37, at which
point oil from the continuous phase channel(s) 36 is introduced to
encapsulate the cells 18 in droplets 39, with the oil being
immiscible with the aqueous solution so as to generate the droplets
39. FIG. 36A illustrates that oil is introduced at a right angle
from two separate continuous phase channels 36. It should be
appreciated that oil may be introduced at various other angles, and
more or less continuous phase channels 36 may be utilized than are
shown in the exemplary embodiment shown in the figures.
[0150] In an exemplary embodiment, individual droplets 39 exit the
juncture 37 through a scanning channel 38 to pass a scanner 70 such
as a digital holographic microscope. The scanner 70 is configured
to scan each droplet 39 as it passes out of the juncture 37 and
through the scanning channel 38. The scanner 70 may be configured
to scan each droplet 39 within the scanning channel 38 separately
in-turn, or may be configured to scan multiple droplets 39
simultaneously. In some embodiments, multiple droplets 39 may be
scanned simultaneously within the scanning channel 38 to determine
if any of the droplets 39 encapsulate cells 18 that are undesirable
or malignant; with the droplets 39 being sorted by diverting the
flow of the droplets 39 according to the scan results.
[0151] The results from the scanner 70 are then analyzed, such as
by a control unit 80, to determine if the encapsulated cells 18
within each droplet 39 are healthy or unhealthy (e.g., malignant).
If the cells 18 are healthy, those cells 18 may be separated from
other cells 18 (e.g., malignant cells) by use of one of a pair of
valves 40. In the exemplary embodiment shown in FIG. 36A, it can be
seen that a first valve 40 leads to a return path 60 for healthy
cells and a second valve 40 leads to an isolation path 50 for all
other cells 18 (unidentified cells 18 or malignant cells 18).
[0152] When a healthy cell 18 is scanned by the scanner 70, the
first valve 40 is opened and the second valve 40 is closed. When an
undesirable or malignant cell 18 is scanned by the scanner 70, the
second valve 40 is opened and the first valve 40 is closed. The
first valve 40 may lead to a return path 60 such that healthy cells
18 may be returned to the patient 12 for therapeutic purposes, or
may lead to an isolation path 50 to be sequestered for diagnostic
purposes. The second valve 40 may lead to an isolation path 50 to
be sequestered.
[0153] It should also be appreciated that the fluid receiving
device 30 may comprise one or more inlet valves 33 such as channel
locks adapted to pause flow of the biological fluid 16 into of the
fluid receiving device 30. Such inlet valves 33 may comprise
valves, locks, or other structures which are adapted to block flow
of the biological fluid 16 at certain times. For example, one or
more inlet valves 33 of the fluid receiving device 30 may be
engaged to pause flow of the biological fluid 16 into the fluid
receiving device 30 while scanning is being performed by the
scanner 70.
D. Scanning Techniques
[0154] As described and shown herein, a wide range of scanning
techniques may be utilized to scan a biological fluid 16 and
identify any constituent biological elements thereof, such as but
not limited to white blood cells, red blood cells, CTCs, platelets,
host cells, cell-free plasma, pathogens, and the like. The types of
biological fluids 16 that can be scanned include a wide range of
biological fluids 16, such as but not limited to blood, lymphatic
fluid, cerebrospinal fluid, sweat, urine, pericardial fluid,
stools, saliva, and the like. In some embodiments, multiple types
of biological fluids 16 may be scanned either together or in turn.
For example, both blood and cerebrospinal fluids may be
simultaneously or sequentially scanned by the same fluid receiving
device 30.
[0155] Exemplary techniques for scanning may include Phase contrast
microscopy (PCM), DIC Microscopy, Hoffman modulation, polarized
light microscopy, digital holographic microscopy, confocal scanning
optic microscopy (CSOM), or laser scanning optic microscopy (SOM)
to measure voxel fluorescence, bright-field microscopy, dark-field
illumination, Raman spectrometry to measure Raman Scattering,
Optical interferometry to measure optical interference, total
internal reflection fluorescence microscopy to measure evanescent
effect, planar waveguides for refractive index detection, photonic
crystal biosensors for measure of biomolecules on cell surfaces,
and light property modulation detections such as surface plasmon
resonance (SPR) detection.
[0156] With respect to digital holographic microscopy, digital
holography is used to record a wave front diffracted from an object
by a light source 72. Utilizing the interference of light from the
light source 72, both amplitude and phase information of an object
wave 77 may be recorded to produce a hologram containing the
information of the object wave 77. A three-dimensional image may
then be reconstructed from the hologram by the control unit 80.
[0157] In an embodiment utilizing digital holographic microscopy,
the scanner 70 may include a light source 72 as previously
discussed. The light source 72 may comprise various types of
illuminating devices, such as but not limited to a laser such as a
monochromatic laser. A pair of laser light waves is generated from
the light source 72 by dividing the laser beam with a beam splitter
74 such that one of the split light waves illuminates the
biological fluid 16. The light diffracted from the biological fluid
16 forms an object wave 77, which illuminates the scanner 70 and is
collected by the microscope objective 73. The remaining laser light
wave is directly detected by the microscope objective 73 of the
scanner 70 to serve as a reference wave 78. The object and
reference waves 77, 78 interfere with each other at the scanner 70
to form an interference fringe image which is scanned by an image
sensor 76.
[0158] Continuing to reference digital holographic microscopy, the
object and reference wave 77, 78 fronts may be joined by the beam
splitter 74 such that the object and reference wave 77, 78 fronts
interfere and create a hologram which is detected by an image
sensor 76. The control unit 80 may then process the digital
hologram, with the control unit 80 functioning as a digital lens to
calculate a viewable image of the object wave 77 front utilizing a
numerical reconstruction algorithm.
[0159] While a microscope objective 73 may be used to collect the
object wave 77 front, it should be appreciated that the microscope
objective 73 is only used to collect light waves and not to form an
image. Thus, the microscope objective 73 may comprise a simple
lens, or may be omitted entirely in some embodiments. The
interference pattern (hologram) may thus be recorded in such
embodiments by a digital image sensor 76.
[0160] Digital holographic microscopy may be utilized to observe
living cells within the biological fluid 16. From the recorded
interference pattern of such living cells, the intensity and phase
shift across various points of the cells may be numerically
computed by the control unit 80. The control unit 80 may thus
measure the phase delay images of biological cells within the
biological fluid 16 to provide quantitative information about the
morphological properties (e.g., cellular dry mass, surface texture,
shape, etc.) of individual cells within the biological fluid
16.
[0161] In an exemplary embodiment, the systems and methods
described herein may utilize these quantitative indicators of
morphological properties in an algorithm to distinguish between
cell types within the biological fluid 16. By way of example and
without limitation, the control unit 80 may be adapted to extract
parameters such as cell thickness, cell area, cell volume, cell dry
mass, the phase shift across the cell, surface roughness and
texture, cell shape, elongation, convexity, luminance, circularity,
solidity, and the like.
[0162] Various types of digital holography may be utilized with the
systems and methods described herein, including but not limited to
off-axis Fresnel, Fourier, image plane, in-line, Gabor, and
phase-shifting digital holography. FIG. 31 illustrates an off-axis
embodiment. By utilizing digital holographic microscopy, the
control unit 80 may differentiate between the various constituents
13 within a biological fluid 16 sample for further processing
utilizing the systems and methods described herein.
[0163] It should be appreciated that multiple laser wavelengths may
be utilized when scanning the biological fluid 16 with digital
holographic microscopy. It has been shown that the refraction
amount increases as the wavelength of light decreases. Thus,
shorter wavelengths of light (e.g., violet and blue) are more
slowed and consequently experience more bending than longer
wavelengths of light (e.g., orange and red).
[0164] Since the morphological parameters in digital holographic
microscopy are dependent upon the wavelength of the laser used, an
exemplary embodiment of a scanning technique relying upon digital
holographic microscopy may utilize multiple lasers each having
different wavelengths. By way of example, the analysis may be
initially conducted using a light source at a first wavelength. If
the sample of cells requires additional confirmation, the light
source may be switched to a different wavelength.
E. Ex Vivo Testing
[0165] The systems and methods described herein may be utilized for
ex vivo testing of various drugs and treatments on a patient 12
specific basis. While established tissue culture cell lines are
often used for in vitro drug sensitivity assays, such cell lines
are not truly representative of the cellular heterogeneity
evidenced during metastasis and recurrence in specific patients 12.
Thus, it is would be far more desirable to test such drugs and
treatments on patient-derived CTCs and CTC-clusters.
[0166] Patient-derived CTCs and CTC-clusters offer greater
precision in predicting outcomes for a particular patient 12, as
they represent the heterogeneity profile for that particular
patient 12 at that particular time. Critically, patient-derived
CTCs and CTC-clusters may exhibit enhanced resistance to
chemotherapy during stages of relapse.
[0167] Using the systems and methods described herein, sufficient
CTCs and CTC-clusters may be extracted to permit both genomic and
transcriptomic profiling. This allows for "direct-to-drug" ex vivo
testing of treatments and drug agents to identify an optimal course
of therapy for a particular patient 12 at any particular time
during the progress of treatment.
[0168] FIG. 34 illustrates an exemplary embodiment in which a
portion of CTCs which have been identified and separated by the
systems and methods described herein may be profiled for
heterogeneity. As can be seen, CTCs and CTC-clusters may undergo
genomic and transcriptomic profiling to determine potentially
relevant drugs or treatments. After identifying potential drugs or
treatments based upon the isolated CTCs and CTC-clusters from a
particular patient 12, relevant drug targets may be identified. The
drugs or treatments may then be tested on that particular patient's
cancer cells ex vivo to identify optimal pathways.
[0169] In this manner, drugs and treatments may be optimized for
each patient 12 based upon the cancer cells identified and
separated by the methods and systems described herein to allow for
precision drug selection that is unique to each patient 12 in
consideration of the cancer cells unique to that patient 12 and in
consideration of the course of treatment to that point for that
patient 12.
F. Multi-Stage Separation
[0170] As previously discussed, the systems and methods described
herein may be utilized for multi-stage separation of non-healthy or
malignant cells from healthy cells. FIG. 35 illustrates an
exemplary embodiment of such a multi-stage separation system in
which multiple stages are utilized to separate CTCs and
CTC-clusters from a biological fluid 16. FIG. 36B illustrates
another exemplary embodiment in which an isolation path 50 is split
into additional channels for repeated, additional enrichment
through scanning.
[0171] As shown in FIG. 35, the first stage of the multi-stage
separation methodology involves receiving a biological fluid 16
from the patient 12. The biological fluid 16 may enter the
multi-stage separation directly from the patient 12. At the first
stage, passive inertial sorting, with or without the use of buffer
fluids, may be utilized to separate red blood cells and
CTC-clusters from the biological fluid 16. Red blood cells
separated at this first stage may be returned to the biological
fluid source 17 and CTC-clusters may be isolated for further
processing.
[0172] Continuing to reference FIG. 35, CTC-clusters and red blood
cells are separated from the biological fluid 16 in the first stage
by inertial sorting. The remaining cells, which will generally
include additional red blood cells, white blood cells, and CTCs,
will then be passed onto the second stage. Such remaining cells may
be passed onto a fluid receiving device 30 for a high-throughput
optical sort utilizing neural network algorithms. The second stage
will thus be utilized to further separate the cells of the
biological fluid 16. Any separated red and white blood cells may
then be returned to the patient 12, with identified CTCs and any
remaining white blood cells being passed on to a third stage.
[0173] As shown in FIG. 35, the third stage may comprise precision
optical sorting through use of an additional fluid receiving device
30 that is in series with the fluid receiving device 30 of the
second stage. Interpretable machine learning classifiers may be
utilized to separate the remaining white blood cells, thus leaving
only CTCs which are separated at the third stage. In this manner,
CTC-clusters and CTCs may be individually separated from a
biological fluid 16, with white and red blood cells from the
biological fluid 16 being returned to the patient 12. The
individually separated CTC-clusters and CTCs may then be retained
for further processing, such as ex vivo testing, therapeutics, or
diagnostics. Additional stages may also be added as-needed for
further enrichment.
G. Optical Filtration of Subsets of Healthy Cells
[0174] The methods and systems described herein may be utilized to
filter specific subsets of healthy cells (e.g., T-cells) from other
types of cells. In such an embodiment, the library of desirable
constituents 15 (e.g., healthy cells) may be configured to exclude
specific subsets of healthy cells (e.g., T-cells) so as to allow
the biological fluid filtration system 10 to filter such subtypes
of healthy cells. As previously indicated, such methods may be
applied for human or veterinary uses. The biological fluid 16 may
be scanned for subsets of healthy cells utilizing fluid receiving
devices 30 such as but not limited to microfluidic channels 31,
microwell arrays 32, and droplet generators 34.
[0175] In the course of some therapies and treatments, such as
Chimeric antigen receptor (CAR) T-cell therapy, T-cells engineered
with chimeric antigen receptors may be utilized for cancer therapy.
As is known in the art, T-cells are a type of white blood cell
which develops in the thymus gland and play a central role in the
body's immune response. CAR-T immunotherapy is utilized to modify
T-cells to recognize cancer cells in order to more effectively
target and destroy them.
[0176] T-cells are harvested, genetically altered, and then the
resulting CAR-T cells are infused into the patient 12 to attack
cancer cells. Such CAR-T cells may be derived from T-cells in the
patient's 12 own blood (autologous) or derived from the T-cells of
another healthy donor (allogenic). Once isolated, these T-cells are
genetically engineered to express a specific CAR, which programs
the T-cells to target an antigen that is present on the surface of
tumors. In other therapies, other forms of white blood cells (such
as natural killer or "NK" cells) are similarly engineered to fight
cancers.
[0177] In an exemplary embodiment of a biological fluid filtration
system 10 for filtration of subsets of healthy cells, the
biological fluid source 17 may be an aphaeretic extract from a
therapeutic apheresis or leukapheresis machine containing white
blood cells including T-cells. The systems and methods described
herein may be utilized to extract just the T-cells by recognizing
all healthy cells except for the T-cells, and then filtering out
the undesirable cells (the T-cells). As a further example, other
types of healthy cells may be omitted from recognition, such as NK
cells, so that those healthy cells may be filtered out in a similar
manner.
[0178] FIG. 37 illustrates an exemplary method of filtering out
T-cells from a biological fluid 16 containing other types of
healthy cells. It should be appreciated that the biological fluid
source 17 may be human or veterinary and may include such
biological fluids 16 as blood, lymphatic fluid, cerebrospinal
fluid, sweat, urine, pericardial fluid, stools, and saliva.
[0179] As shown in FIG. 37, an optional pre-filtration session may
be conducted to create a reference data 91 in which a sample of
biological fluid 16, from the patient 12 or others, is
pre-processed by optically scanning the sample with a scanner 70
and recognizing certain biological fluid constituents 13 such as
certain types of cells within the biological fluid 16 using an
algorithm run by a control unit 80. The optical scan of the
subsample allows for a software algorithm run by the control unit
80 to analyze the scanned data from the subsample and recognize any
subsets of healthy blood cells (e.g., red blood cells, white blood
cells) while not recognizing certain subsets of healthy blood cells
(e.g. T-cells). Such pre-processing may be utilized to create a
control group for recognizing different types of cells during the
processing phase.
[0180] Continuing to reference FIG. 37, it can be seen that in an
exemplary embodiment a patient 12 may undergo leukapheresis. The
leukapheresis extract from the patient containing healthy cells is
channeled through a fluid receiving device 30 such as microfluidic
channels, microwell arrays, or droplet generators and the extract
is scanned by the scanner 70. The control unit 80 then performs
image processing and detection of the contents within the fluid
receiving device 30, including use of any pre-processing findings
if pre-processing had previously occurred.
[0181] The control unit 80 will analyze the results of the scan
from the scanner 70 to determine where to route the biological
fluid 16. If cells other than recognized healthy cells are present,
the biological fluid 16 may be routed along a reprocessing path 62
for the contents to be processed for extraction of any undesirable
cells (e.g., T-cells). If only recognized healthy cells are
present, the contents may be routed along an isolation path 50 to
be discarded or retained for further use without cell
extraction.
H. Presorting of Biological Fluids
[0182] FIG. 38 illustrates an exemplary method of presorting and
then optically filtering a portion of a biological fluid 16 of
biological fluid constituents 13 such as pathogens or CTCs. As
shown in FIG. 38, an optional pre-filtration session may be
conducted to create a reference data 91 in which a sample of
biological fluid 16, from the patient 12 or others, is
pre-processed by optically scanning the sample with a scanner 70
and recognizing the biological fluid constituents 13 such as cells
within the biological fluid 16 using an algorithm run by a control
unit 80.
[0183] Continuing to reference FIG. 38, it can be seen that
biological fluid 16 is first drawn from the biological fluid source
17 during a filtration session. The biological fluid 16 may
comprise various fluids as discussed herein, such as but not
limited to blood, lymphatic fluid, CSF, sweat, urine, pericardial
fluid, stools, and saliva. The biological fluid source 17 may
comprise a human or an animal. After the biological fluid 16 is
drawn from the biological fluid source 17, the biological fluid 16
may be presorted, such as by using a microfluidic separation module
100.
[0184] The presorting of the biological fluid 16 may be utilized to
separate desirable constituents 15 from the biological fluid 16
prior to further processing. The manner of presorting utilized by
the microfluidic separate module 100 may vary in different
embodiments, and may include without limitation the use of inertial
sorting, centrifugal sorting, microfluidic sorting, and the like.
Characteristics utilized during presorting may include size,
density, inertial hydrodynamic, antigen binding affinity, motility,
centrifugation, electrical charge, electric dipole moment, or
magnetism.
[0185] The presorting of the biological fluid 16 by the
microfluidic sorting module 100 allows for the initial separation
of biological fluid constituents 13 without optical scanning. Any
such biological fluid constituents 13, such as CTC, may be
immediately routed along a return path 60 back to the biological
fluid source 17, or may be sequestered along an isolation path 50
for further processing. After the presorting step is completed, a
mixture of undesirable constituents 14 and desirable constituents
15 may then be transferred to a fluid receiving device 30 for
further processing.
[0186] Continuing to reference FIG. 38, it can be seen the mixture
of undesirable constituents 14 and desirable constituents 15 is
optically scanned on the fluid receiving device 30 by a scanner 70.
The control unit 80 then performs image processing and detection to
identify the contents and differentiate between undesirable
constituents 14 and desirable constituents 15. If undesirable
constituents 14 are detected in the sample, the sample is not
returned to the biological fluid source 17, but may instead be sent
along an isolation path 50 for sequestration and optional
diagnostics of the contents to identify pathogens, CTC,
CTC-clusters, host cells, cell free plasma, and the like.
[0187] Any samples comprising only undesirable constituents 14
after the presorting stage may be immediately directed along an
isolation path 50 and not returned to the biological fluid source
17. Such samples may be sequestered along the isolation path 50 for
optional diagnostics and/or therapeutics as discussed herein.
[0188] FIG. 39 illustrates another exemplary method of presorting
and then optically filtering a portion of a biological fluid 16
which, by utilizing multiple outlet ports, may be utilized to
sequester and separately hold isolated biological fluid
constituents 13 such as CTCs, CTC-clusters, white blood cells,
cell-free plasma, and the like. The sequestered outputs may then be
reprocessed single or multiple times using the systems and methods
described herein, without the aphaeretic components, to further
enrich the isolation of such biological fluid constituents 13.
[0189] As shown in FIG. 39, an optional pre-filtration session may
be conducted to create a reference data 91 in which a sample of
biological fluid 16, from the patient 12 or others, is
pre-processed by optically scanning the sample with a scanner 70
and recognizing the biological fluid constituents 13 such as cells
within the biological fluid 16 using an algorithm run by a control
unit 80. Biological fluid 16 is first drawn from the patient 12. If
necessary, an anti-coagulant may be applied to the biological
fluid. A fluid pump may draw the biological fluid 16 through a
fluid pressure sensor and pre-filter pressure sensor prior to
entering a pre-sorting module. The pre-sorting module utilizes the
techniques and/or characteristics described herein, such as but not
limited to inertial, size-based, centrifugal, dielectric, and/or
acoustic to separate biological fluid constituents 13 from the
biological fluid 16. In some embodiments, the use of additional
buffers or dilution liquids or reagents may be employed in the
pre-sorting module. In such examples, the embodiment might include
ports and reservoirs to insert, collect, and/or replenish such
fluids.
[0190] A first set of biological fluid constituents 13, such as red
blood cells, plasma, and small cells, may be transferred to a fluid
chamber to temporarily hold filtered plasma and healthy cells.
Separated large CTC-clusters may be retained and sequestered for
further processing such as diagnostics. Separated white blood cells
and small CTCs may be transferred into one or more fluid receiving
devices 30 such as microfluidic channels 31 arranged in parallel.
Each fluid receiving device 30 is independently scanned, and its
contents then undergo image processing and content detection by the
control unit 80, which may utilize findings from any optional
pre-filtration session.
[0191] If undesirable constituents 14 such as cells other than
healthy cells are detected, the contents may be re-routed back to
the fluid receiving devices 30 by a reprocessing path 62 for
further enrichment of CTCs. If no cells other than healthy cells
are detected, the contents may be combined with the filtered plasma
and healthy cells, pass through a fluid pump and pressure sensor,
undergo air bubble removal, and returned to the biological fluid
source 17 by the return path 60.
[0192] FIG. 40 illustrates yet another method of presorting and
then optically filtering a portion of a biological fluid 16. Such
embodiments may include the use of diagnostic systems to separate
pathogens or CTCs from biological fluid 16 samples for downstream
diagnostic (e.g., genomic, transcriptomic, metabolomics, drug
sensitive, drug resistance, etc.) characterization. FIG. 40
illustrates such a diagnostic embodiment without a return path 60
back to the biological fluid source 17.
[0193] As shown in FIG. 40, an optional pre-filtration session may
be conducted to create a reference data 91 in which a sample of
biological fluid 16, from the patient 12 or others, is
pre-processed by optically scanning the sample with a scanner 70
and recognizing the biological fluid constituents 13 such as cells
within the biological fluid 16 using an algorithm run by a control
unit 80. At the filtration session, biological fluid 16 is drawn
from the source and entered into a pre-sorting module to separate
biological fluid constituents 13 (e.g., inertial, size-based,
centrifuge, dielectric, acoustic, etc.). Any separated red blood
cells, plasma, and small cells are transferred to a fluid chamber
to hold plasma and healthy cells. Any separated large CTC-clusters
may be sequestered for further processing.
[0194] Samples including separated white blood cells and small CTCs
may be transferred by inertial focusing into one or more parallel
fluid receiving devices 30. Fluid flow is temporarily halted, and
each fluid receiving device 30 is scanned by a scanner 70. The
control unit 80 conducts image processing and content detection of
each such sample in-turn.
[0195] If cells other than healthy cells are present, such as
extracted CTCs along with potentially some healthy cells, the
sample may be rerouted back to the fluid receiving devices 30 by a
reprocessing path 62 for further enrichment of CTCs. Enriched
contents may be routed along an isolation path 50 for diagnostic
processing (e.g., genomic, transcriptomic, metabolomics, drug
sensitivity, drug resistance, etc.) along with any
previously-separated red blood cells, plasma, small cells, and
separated large CTC-clusters.
[0196] FIG. 41 illustrates another embodiment of a method of
presorting and then optically filtering a portion of a biological
fluid 16, with healthy cells being returned back to the biological
fluid source 17. An optional pre-filtration session may be
conducted to create a reference data 91 in which a sample of
biological fluid 16, from the patient 12 or others, is
pre-processed by optically scanning the sample with a scanner 70
and recognizing the biological fluid constituents 13 such as cells
within the biological fluid 16 using an algorithm run by a control
unit 80.
[0197] During the filtration session, biological fluid 16 is drawn
from the biological fluid source 17. Anti-coagulants may be applied
if necessary to prevent clotting within the system. The biological
fluid 16 is then pumped by a fluid pump past a fluid pressure
sensor and a pre-filter pressure sensor. The biological fluid 16
then enters the pre-sorting module to separate biological fluid
constituents 13 in any of the manners previously discussed. As with
previous embodiments, healthy cells such as separated red blood
cells, plasma, and small cells may be transferred to a fluid
chamber to be temporarily held. Malignant cells such as separated
large CTC-clusters may be transferred along an isolation path 50 to
be sequestered for diagnostic processing.
[0198] Samples with both healthy and malignant cells, such as
separated white blood cells and small CTCs, may be inertial focused
into one or more parallel fluid receiving devices 30 such as
microfluidic channels 31. Once within the one or more fluid
receiving devices 30, such samples will be held in the fluid
receiving device 30 as optical scans are performed by one or more
scanners 70. In an embodiment in which multiple scanners 70 are
utilized, each of the samples may be scanned simultaneously. In an
embodiment in which a single scanner 70 is utilized, each of the
samples may be held in place while each sample is sequentially
scanned.
[0199] It should also be appreciated, with this embodiment and with
the others described herein, that a single fluid receiving device
30 may be utilized to scan multiple distinct samples at different
times. In such embodiments, a first sample will be transferred to
the fluid receiving device 30, scanned, and the results will be
transferred to the control unit 80. Upon completion of analysis of
that sample, fluid flow may be resumed to transfer another sample
onto the fluid receiving device 30 for scanning. These steps may be
repeated until all of the samples have been scanned.
[0200] The results of each optical scan (e.g., images or other cell
characteristics) are transferred to the control unit 80 for image
processing and content detection. This step may be repeated until
all of the samples with both healthy and malignant cells have been
scanned and processed as described above. If healthy cells are
detected among unhealthy cells (e.g., CTCs), the sample may be
returned to the fluid receiving device 30 along a reprocessing path
62 for further scanning and processing. Alternatively or after
multiple such scans and enrichment, the sample may be sequestered
for diagnostic processing using any of the methods previously
discussed herein.
[0201] If only healthy cells are detected in a sample (e.g., only
white blood cells and no CTCs), the sample may be transferred via a
fluid pump and pressure sensor back to the patient 12 or biological
fluid source 17 along a return path 60. In some embodiments, any
filtered plasma and healthy cells in the fluid chamber may
similarly be returned to the biological fluid source 17, either
with the white blood cells or separately. Air bubbles will also be
removed prior to return to the biological fluid source 17. In this
manner, the embodiment shown in FIG. 41 allows for only confirmed
healthy cells to be returned to the patient 12, with any other
cells (e.g., malignant cells or undesirable cells) being
sequestered for disposal or for diagnostic processing.
[0202] In another exemplary embodiment such as shown in FIG. 42,
non-aphaeretic methods may be utilized for the removal of tumor
cell contaminants from autologous stem-cell transplant products.
Autologous stem cell transplants are generally used in patients 12
who need to undergo high doses of chemotherapy and radiation to
cure their disease. These treatments can be toxic and thus damage
the bone marrow. An autologous stem cell transplant helps to
replace the damaged bone marrow, but it is often reported that the
process to collect stem cells from the patient may lead to
contamination of such products with tumor cells. Using the method
shown in FIG. 42, the biological fluid filtration system 10 may be
used to prevent or remove such contamination of stem cells.
[0203] As shown in FIG. 42, a pre-filtration session may be
conducted to recognize healthy cells. During the filtration
session, biological fluid 16 such as leukapheresis product from the
biological fluid source 17 is transferred to a presorting module to
separate fluid constituents by any of the methods described herein.
As with the other embodiments, the pre-sorting may separate healthy
cells such as red blood cells, plasma, and small cells which are
transferred to a fluid chamber for transplantation. Malignant cells
such as large CTC-clusters may be sequestered for diagnostic
processing using any of the methods previously described.
[0204] With respect to the remaining samples containing both
healthy and malignant or undesirable cells, inertial focusing may
be utilized to transfer such samples onto one or more fluid
receiving devices 30. Each sample is then scanned (either
simultaneously using multiple fluid receiving devices 30 in
parallel or sequentially using a single fluid receiving device 30
or multiple fluid receiving devices 30 in series) and the results
transferred to the control unit 80 for image processing and content
detection.
[0205] If cells other than healthy cells are detected, such samples
may be rerouted back to the fluid receiving device 30 along a
reprocessing path 62 for further processing or may be sequestered
for diagnostic processing along an isolation path 50 using any of
the methods described herein. If only healthy cells are detected,
such samples may be transferred to the fluid chamber with the
healthy cells from the pre-sorting to be transplanted back to the
patient 12.
[0206] FIG. 43 illustrates an embodiment in which multiple
pre-sorting techniques are utilized in conjunction with optical
filtration in a biological fluid filtration system 10. Biological
fluids 16 are drawn from a biological fluid source 17 such as an
animal or human. A passive inertial sort may be utilized to
separate any CTC-clusters which may be sequestered for disposal or
diagnostics. After the initial inertial sort, centrifugation may be
utilized to remove white blood cells and CTCs (e.g.,
leukapheresis).
[0207] During leukapheresis, some red blood cells and plasma may be
returned to the patient 12. The leukapheresis product containing
white blood cells and CTCs may be transferred to a third stage for
optical filtration using the methods described herein to separate
the CTCs for diagnostic processing. Thus, the biological fluid 16
is pre-filtered across multiple stages prior to optical filtration.
As a further example, blood could first be filtered using
microfluidic inertial mechanisms to sort out large CTC-clusters.
The remainder undergoes therapeutic apheresis to separate white
blood cells and CTCs. That mix of cells is then filtered using the
optical filtration techniques discussed herein.
I. Body-Worn Aphaeretic Optical Filtration Device
[0208] While the various embodiments previously discussed are
typically conducted in clinical or lab settings, certain
embodiments may allow for the systems and methods described herein
to be utilized in a body-worn configuration outside of a clinical
or lab setting.
[0209] FIGS. 44A, 44B, and 45 illustrate an exemplary embodiment of
such a body-worn device 120. The body-worn device 120 may utilize
optical components, including one or more fluid receiving devices
30 and one or more scanners 70, to filter undesirable constituents
14 from various biological fluids 16, such as blood, lymphatic
fluid, cerebrospinal fluid, sweat, urine, pericardial fluid,
stools, and saliva. The biological fluid source 17 may be
veterinary or human.
[0210] The body-worn device 120 will generally comprise a housing
121 which houses all of the power, optical, microfluidic,
computational, and communication components necessary for the
patient 12 utilizing the body-worn device 120 to freely move about.
The body-worn device 120 will generally include a power source 122
for powering the various components of the body-worn device 120.
The power source 122 may comprise various types of batteries,
including disposable and rechargeable batteries. In some
embodiments, the power source 122 may comprise solar cells.
[0211] The body-worn device 120 may also include various indicators
132 adapted to convey various information to the user of the
body-worn device 120. For example, indicators 132 may be utilized
to indicate the power status (on or off) of the body-worn device
120, the charge remaining in the power source 122, the remaining
volume available in the cartridge 126 (e.g., whether the cartridge
is full or nearing full), and the like. The indicators 132 may be
visual (e.g., lights) and/or audible (e.g., alarms). For example,
an audible and/or visual alarm may be activated when the cartridge
126 is nearing full, or if the power source 122 is running out of
charge.
[0212] FIG. 44B illustrates an exemplary embodiment of a body-worn
device 120 which includes an anti-coagulant insert 134 which
includes anti-coagulant which is applied to the biological fluid 16
within the body-worn device 120 to prevent clotting or coagulation
of the biological fluid 16 within the body-worn device 120 as
discussed herein. A removable anti-coagulant insert 134 of such
anti-coagulant may be removably connected to the body-worn device
120 such that the anti-coagulant within the anti-coagulant insert
134 may be easily replenished as-needed. Also shown in FIG. 44B is
a buffer fluid insert 136 which may be utilized to collect or
replenish optional buffer fluids. Such buffer fluids such as
dilution fluids may be utilized for pre-sorting within the
body-worn device 120 as discussed herein.
[0213] As shown in FIG. 45, the body-worn device 120 will generally
comprise an inlet 123 and an outlet 124. The inlet 123 and outlet
124 of the body-worn device 120 are generally adapted to be
connected to intravenous (IV) or catheter ports 129a, 129b of a
catheter 128 which is inserted into the body of the patient 12 to
receive a biological fluid 16 from a biological fluid source 17. It
should be appreciated that the catheter 128 may be installed at
various locations on a patient's 12 body, and thus FIGS. 44A and
44B should not be construed as limiting in that regard. The
body-worn device 120 may be worn adjacent to the catheter 128
entering the body, or more distant from that position. Thus, the
length of the catheter 128 may vary widely in different
embodiments.
[0214] In some embodiments, the catheter 128 may be fluidly
connected to the vascular system of a patient 12 such that the
vascular system of the patient 12 acts as a biological fluid source
17 for a biological fluid 16 comprised of blood. In other
embodiments, the catheter 128 may be fluidly connected to the
nervous system of a patient 12 such that the nervous system of the
patient 12 acts as a biological fluid source 17 for a biological
fluid 16 comprised of cerebrospinal fluid. In other embodiments,
the catheter 128 may be fluidly connected to the lymphatic system
of a patient 12 such that the lymphatic system of the patient 12
acts as a biological fluid source 17 for a biological fluid 16
comprised of lymphatic fluid.
[0215] By way of example, biological fluid 16 comprised of blood
may be accessed through arteriovenous grafts, fistulas, or
catheters 128 commonly used in aphaeretic treatments such as
dialysis. Access to other types of biological fluid 16 such as
cerebrospinal fluid could be through the lumbar, peritoneum, or the
ventricles in the skull. Thus, it should be appreciated that the
catheter 128 may be positioned at various locations on the body of
the patient 12, such as but not limited to the head, arms, chest,
legs, hands, feet, and the like.
[0216] Continuing to reference FIG. 45, it can be seen that the
body-worn device 120 may comprise one or more pumps 125 for
controlling flow of the biological fluid 16 entering and exiting
the body-worn device 120. The pump 125 may include a pressure
sensor for monitoring the pressure of the biological fluid 16. The
pump 125 may be configured to run continuously or only at certain
times or depending upon certain conditions. The pump 125 may be
operated by the control unit 80.
[0217] As an example, the pump 125 may activate to draw a sample
comprising a set volume of biological fluid 16 onto the fluid
receiving device 30. Upon the set volume of biological fluid 16
being transferred to the fluid receiving device 30, the pump 125
may deactivate during the scanning process. After scanning the
sample of the biological fluid 16, the pump 125 may activate again
so as to direct the scanned sample of the biological fluid 16
either to a removable cartridge 126 or back to the biological fluid
source 17 via the outlet 124. In some embodiments, the pump 125 may
be adapted to automatically deactivate under certain conditions,
such as upon pressure being detected as being above or below
certain thresholds.
[0218] The biological fluid 16 will generally be routed from the
inlet 123 into a microfluidic fluid receiving device 30 such as is
described herein. It should be appreciated that multiple fluid
receiving devices 30 may be utilized in the body-worn device 120 in
certain embodiments, either in parallel or in series. When the
biological fluid 16 is in the fluid receiving device 30, the
scanner 70 will scan the contents of the fluid receiving device 30.
An anti-coagulant may be applied to the biological fluid 16 prior
to entering the fluid receiving device 30 so as to prevent the
biological fluid 16 from coagulating while in the body-worn device
120.
[0219] One or more scanners 70 are directed towards the fluid
receiving device 30 to scan the biological fluid 16 within the
fluid receiving device 30. The scanner 70 will generally be
communicatively connected to a control unit 80. The control unit 80
may be located within the housing 121 of the body-worn device 120,
such as by use of a microprocessor, microcontroller,
system-on-a-chip, or the like. In some embodiments, the scanner 70
may be communicatively connected to a remote control unit 80, such
as through use of a communications network. By way of example, the
scanner 70 may be communicatively connected to a control unit 80 by
Bluetooth, Wi-Fi, radio waves, or various other communications
methods known in the art.
[0220] The results of the optical scan of the biological fluid 16
within the body-worn device 120 by the scanner 70 are generally
transferred to the control unit 80 for processing and detection of
the biological fluid constituents 13. The data from the scanner 70
is compared to the reference data 91 and analyzed by the control
unit 80 to classify the biological fluid constituents 13 within the
biological fluid 16. One or more valves 127 may be utilized to
direct the biological fluid 16 along at least two different paths
depending upon the analysis of the biological fluid 16 by the
control unit 80.
[0221] If only healthy, desirable constituents 15 are detected, the
one or more valves 127 may direct the biological fluid 16 along a
return path 60 to be returned to the biological fluid source 17.
More specifically, the biological fluid 16 may exit the body-worn
device 120 through its outlet 124 and returned to the biological
fluid source 17 by the catheter 128.
[0222] If there are undesired constituents 14 or undesired
constituents such as malignant cells, the one or more valves 127
may direct the biological fluid 16 along an isolation path 50 to a
cartridge 126. The cartridge 126 may be removable and, in some
embodiments, may be disposable. The cartridge 126 generally
includes a cavity within which the biological fluid 16 may be
stored and sequestered.
[0223] The cartridge 126 is generally removably connected to the
housing 121 of the body-worn device 120. However, in some
embodiments, the cartridge 126 may instead be fixedly connected to
the housing 121 and instead include an access port through which
the biological fluid 16 may be drained from the cartridge 126
as-needed. Thus, the valve(s) 127 of the body-worn device 120 will
generally have one inlet and a pair of outlets. The inlet of the
valve(s) 127 is fluidly connected to the fluid receiving device 30.
A first outlet of the valve(s) 127 is fluidly connected to the
cartridge 126 and a second outlet of the valve(s) 127 is fluidly
connected to the outlet 124 of the body-worn device 120.
[0224] The cartridge 126, which may be replaceable and/or
disposable, may include built-in sensors to identify the protein
expression of the cells. Data from such built-in sensors may be
transferred to the control unit 80 for further processing. Thus,
the filtered contents of the cartridge may be diagnostically
profiled by the control unit 80.
[0225] FIG. 46 illustrates an exemplary method of filtering a
biological fluid 16 utilizing a body-worn device 120. Prior to
filtration, the catheter 128 will generally be inserted within the
patient 12 and fluidly connected to the biological fluid source 17.
The catheter 128 will be fluidly connected to the inlet 123 of the
body-worn device 120 and, in some embodiments, to the outlet 124 of
the body-worn device 120. Thus, the catheter 128 may include a pair
of intravenous (IV) or catheter ports 129a, 129b, such as an outlet
catheter port 129a and an inlet catheter port 129b. In such
embodiments, the outlet catheter port 129a is fluidly connected to
the outlet 124 of the catheter 128 and the inlet catheter port 129b
is fluidly connected to the inlet 123 of the catheter 128 such as
shown in FIGS. 44A and 44B.
[0226] Biological fluid 16 is drawn from the biological fluid
source 17 through the catheter 128, entering the body-worn device
120 through its inlet 123. Anti-coagulant may be applied to the
biological fluid 16 as-needed from the anti-coagulant insert 134.
The biological fluid 16 is drawn into the fluid receiving device 30
by the pump 125, which may be controlled by the control unit 80.
The scanner 70 then scans the biological fluid 16 within the fluid
receiving device 30, and the resulting data is transferred to the
control unit 80 for analysis and detection of the biological fluid
constituents 13 within the biological fluid 16.
[0227] The manner by which the biological fluid 16 is scanned by
the scanner 70 and analyzed by the control unit 80 may vary as
described herein. If a pre-filtration session was performed, the
data collected therefrom may be utilized by the control unit 80 in
detecting and identifying the biological fluid constituents 13 of
the biological fluid 16.
[0228] In some embodiments, the control unit 80 may be configured
to maintain a count of detected biological fluid constituents 13,
including healthy cells, malignant cells, and unidentified cells.
The control unit 80 may also be configured to keep track of the
total count of cells analyzed, the total volume of biological fluid
16 analyzed, cells detected per volume of biological fluid 16, and
other data.
[0229] Any biological fluid 16 samples scanned by the scanner 70
and determined by the control unit 80 to include undesirable
constituents 14 are directed by the valve(s) 127 along an isolation
path 50 into the cartridge 126. Any biological fluid 16 samples
scanned by the scanner 70 and determined by the control unit 80 to
include only healthy, desirable constituents 15 is instead routed
by the valve(s) 127 along a return path 60 out of the body-worn
device 120 via its outlet 124 to be returned to the biological
fluid source 17 by the catheter 128.
[0230] FIG. 47 illustrates a method of filtering a biological fluid
16 with a body-worn device 120 including a presorting stage. As
shown in FIG. 47, the biological fluid 16 is drawn from the
biological fluid source 17 into the body-worn device 120 via its
inlet 123 to enter a pre-sorting stage. The presorting stage sorts
the biological fluid 16 by various methods described herein (e.g.,
inertial, size, density, immunoaffinity, magnetic, dielectric). In
some embodiments, the presorting stage may include a miniature
centrifuge within the body-worn device 120. In some embodiments,
the presorting stage may utilize a buffer fluid insert 136 from
which buffer fluids and/or dilution fluids may be retrieved for
presorting.
[0231] After the presorting stage, any desirable constituents 15
may be returned along a return path 60 through the outlet 124 of
the body-worn device 120 to the biological fluid source 17 via the
catheter 128. Any undesirable constituents 14, or unhealthy cells,
may be transferred to the cartridge 126 along an isolation path 50.
Any sample of biological fluid 16 after the presorting stage which
includes a mixture of recognized fluid constituents 13 and
undesirable constituents 14, such as a mixture of healthy cells and
unhealthy cells, are transferred to the fluid receiving device 30
to be optically scanned by the scanner 70 and processed by the
control unit 80 in the manners described elsewhere herein.
[0232] FIG. 48 illustrates a method of filtering a biological fluid
16 within a body-worn device 120 which includes a drug infuser 130
in the return path 60. As shown in FIG. 48, a drug infuser 130 may
be positioned along the return path 60 between the valve(s) 127 and
the outlet 124 of the body-worn device 120. Drugs or treatments may
be infused by the drug infuser 130 with the filtered biological
fluid 16 prior to returning to the biological fluid source 17
through the outlet 124 of the body-worn device 120 via the catheter
128. Various drugs or treatments may be utilized, and the body-worn
device 120 may be configured such that the drugs or treatments may
be routinely re-filled or switched as-needed. In other example
embodiments, the body-worn device 120 may employ additional buffers
or dilution fluids to presort the biological fluid 16.
Additionally, in some embodiments, the body-worn device 120 may
employ anti-coagulants to prevent clotting.
[0233] FIG. 53 illustrates an exemplary method utilized for a
multi-stage body-worn filtration system in which the body-worn
device 120 could utilize optical filtration methods to filter and
sequester samples of biological fluids 16 (e.g., blood) that
contain CTCs and CTC-clusters. Such samples may be stored in a
replaceable cartridge 126 that may then be further processed to
enrich and extract CTCs and CTC-clusters.
[0234] As shown in FIG. 53, biological fluid 16 from a biological
fluid source 17 may be treated with optional anti-coagulant 49
prior to entering a microfluidic separation module 100 for optional
presorting. Any undesirable constituents 14 are immediately
transferred to an isolation path 50. Any samples containing both
undesirable constituents 14 and desirable constituents 15 are
transferred to a fluid receiving device 30 and scanned by a scanner
70. The resulting scanned data 90 is processed by the control unit
80. If any undesirable constituents 14 are detected, the sample may
be transferred along an isolation path 50 to be stored in a
cartridge 126 for off-line enrichment to extract CTCs. If only
desirable constituents 15 are detected, the sample may be returned
to the biological fluid source 17 by a return path 60.
[0235] FIG. 44C illustrates an exemplary portable device 140 which
is portable (as opposed to body-worn) and may be utilized in a
non-aphaeretic setting (e.g., to process cerebrospinal fluid,
saliva, or urine to extract unhealthy cells such as CTCs). As shown
in FIG. 44C, the portable device 140 will generally include a
biological fluid inlet 142 through which biological fluid 16 may be
introduced into the portable device 140. An optional buffer fluid
inlet 143 may be utilized to introduce various buffer or dilation
fluids for presorting of the biological fluid 16. The buffer fluid
may be replaced or replenished as needed. Such an embodiment may
also include anti-coagulants if utilized to process fluids such as
blood.
[0236] Continuing to reference FIG. 44C, the portable device 140
may include indicators 144 (e.g., audible or visual) which provide
various information about the operability of the portable device
140 (e.g., on/off, flow blockage, etc.). Internally to the portable
device 140, a fluid receiving device 30 and scanner 70 are provided
for scanning the biological fluid 16 within the portable device
140. Although not shown, the portable device 140 will generally
include an internal power source, though the portable device 140
may be externally powered (e.g., by a wall socket) in some
embodiments. The portable device 140 may include its own internal
control unit or may be communicatively connected to a remote
control unit for processing scanned data 90.
[0237] As shown in FIG. 44C, the portable device 140 may include a
waste fluid outlet 145 through which waste fluids and other
disposables may be removed from the portable device 140. The
portable device 140 will also generally include a removable
cartridge 146 which stores any filtered cells. The cartridge 146
may be emptied and replaced as-needed to process additional
biological fluids 16.
J. Closed-Loop Filtration, Treatment, and Monitoring
[0238] FIGS. 49 and 50 illustrate an exemplary embodiment of
utilizing a biological fluid filtration system 10 for closed-loop
filtration, treatment, and monitoring of undesirable constituents
14 such as harmful cells. In such an embodiment, undesirable
constituents 14 such as CTCs or CTC-clusters are filtered using the
optical filtration methods and systems described herein, with a
drug or treatment (e.g., chemotherapeutic agents, combination of
drugs, immunotherapy treatments, etc.) being introduced in the
return path 60 to check for its therapeutic effect.
[0239] In an example embodiment, the ratio of undesirable
constituents 14 within a certain volume of biological fluid 16 may
be monitored continuously or sporadically to determine any changes
post-administration of a certain drug or treatment. A decreasing
ratio of undesirable constituents 14 such as viable (living) CTCs
within a biological fluid 16 during aphaeresis may be an indication
that a certain drug or treatment is effective.
[0240] For example, various optical scanning methods described
herein (e.g., digital holographic microscopy) may be utilized to
count undesirable constituents 14 per given volume of processed
biological fluid 16. In some embodiments, staining or fluorescence
may be utilized. As with the other systems and methods described
herein, the closed-loop filtration, treatment, and monitoring may
be utilized in connection with humans or animals.
[0241] FIG. 49 illustrates an exemplary method of filtering,
monitoring, and treating harmful cells within a biological fluid
16. As with the other systems and methods described herein, a
pre-filtration session may be performed to improve the accuracy of
the control unit 80 in analyzing the biological fluid 16 and
identifying any biological fluid constituents 13. Data from the
pre-filtration session may be utilized in whole or in part in
development of the reference data 91 which is used by the control
unit 80 to analyze biological fluid 16 and identify any biological
fluid constituents 13 as discussed herein. The reference data 91
could include images and/or data of biological fluid constituents
13 from the same patient or from other individuals.
[0242] As shown in FIG. 49, biological fluid 16 is drawn from a
biological fluid source 17. Presorting may be applied as discussed
herein, with any undesirable constituents 14 being sequestered
along an isolation path 50 immediately after presorting. The
remaining sample of biological fluid 16 is then optically scanned
by the scanner 70 and the resulting data undergoes analysis by the
control unit 80 to identify any biological fluid constituents
13.
[0243] Any samples of biological fluid 16 containing anything other
than desirable constituents 15 are transferred along the isolation
path 50 to be sequestered along any undesirable constituents 14
from the presorting (if performed). All such undesirable
constituents 14 are monitored such as by counting the number of
viable CTCs or other harmful cells per volume of processed
biological fluid 16.
[0244] FIG. 50 illustrates an exemplary method of filtering,
monitoring, and treating harmful cells within a biological fluid 16
which includes presorting of cells prior to optical scanning,
diagnostic profiling, and ex vivo drug testing of undesirable
constituents 14 to identify an optimal drug or treatment to be
administered in the return path 60. In an exemplary embodiment, a
drug or treatment may be tested on filtered cells by measuring
changes to their cellular properties using optical techniques
(e.g., digital holographic microscopy).
[0245] As an example embodiment, prior to drug administration, a
portion of the filtered cells may be processed for genomic data
which aids in identifying suitable drug targets. Those drugs are
tested, potentially at various concentrations, on the remaining
portion of the filtered cells to determine which drug or treatment
has the most intended therapeutic effect so as to validate the
optimal drug or treatment choice. Subsequently, the optimal drug or
treatment is then introduced in the return path 60.
[0246] With reference to FIG. 50, it can be seen that a biological
fluid 16 is drawn from a biological fluid source 17, presorted,
scanned, and analyzed as described herein. Contents which are
sequestered along the isolation path 50, such as undesirable
constituents 14, undergo diagnostics such as cell counting (e.g.,
of viable CTCs), profiling (e.g., histochemistry, genomic
transcriptomic, etc.), and the like. The diagnostics are utilized
to determine potential drugs or treatments.
[0247] After determining potential drugs or treatments based on
diagnostics of undesirable constituents 14 sequestered after
presorting and optical filtration, optional drug sensitivity and
resistance assay may be performed. Based upon the results of the
assay, optimal drugs or treatments, including dosing, may be
selected. Such optimal drugs or treatments are then introduced in
the return path 60 to be returned to the biological fluid source
17.
[0248] Such an embodiment may be utilized to process multiple
biological fluids 16 from a biological fluid source 17 at the same
time. Different types of filtered cells may be held separately so
that their counts, diagnostic profiling, and drug testing may be
conducted separately. Optical scanning could include single or
multiple parallel fluid receiving devices 30. In some embodiments,
the foregoing methods may be performed on a sample of biological
fluid 16.
K. Methods of Biological Fluid Filtration
[0249] Generally, the methods of biological fluid filtration are
directed to the removal of undesirable constituents 14 which may
comprise, for example, disease-related biological or chemical
entities, from a biological fluid 16 by optically inspecting the
constituents 13 in the biological fluid 16 and filtering those that
are not recognized to be among the desirable constituents 15 of the
biological fluid 16. The methods of biological fluid filtration may
be applied in a therapeutic context, a diagnostic context, or both,
as described herein.
[0250] Generally, the methods involve directing a biological fluid
16 to a fluid receiving device 30, optically scanning the
biological fluid 16 within the fluid receiving device 30 by a
scanner 70 to generate the scanned data 90 of the biological fluid
16, comparing the scanned data 90 of the biological fluid 16 with
the reference data 91 by the control unit 80; returning the
biological fluid 16 to the biological fluid source 17 if the
scanned data 90 of the biological fluid 16 includes only desirable
constituents 15 exhibiting criteria that sufficiently match with
any of the desirable constituents 15 of the reference data 91 by
the control unit 80; and isolating the biological fluid 16 from the
biological fluid source 17 if the scanned data 90 of the biological
fluid 16 includes one or more constituents 13 not exhibiting
criteria that sufficiently match with any of the desirable
constituents 15 of the reference data 91 by the control unit 80.
The biological fluid filtration system 10 described herein may be
used to carry out the methods.
[0251] In one embodiment, the method of biological fluid filtration
includes pre-processing of the biological fluid 16 to separate
particular cellular constituents of the biological fluid 16 for
promotion to the fluid receiving device 30, as described above.
Methods may also optionally include additional filtration
processes, including antigen based filtration, described above.
[0252] In an alternative embodiment, the method includes
pre-processing of a fluid sample of the biological fluid source 17
to obtain the reference data 91 differentiating cell image data
characteristics of desirable constituents 15 to more precisely
tailor the reference data 91 the biological fluid system, including
any heterogeneity among its desirable constituents 15. Such
pre-processing may rely upon machine learning and/or artificial
intelligence models in order to more accurately and efficiently
differentiate between undesirable constituents 14 and desirable
constituents 15.
[0253] In an alternative embodiment, the reference data 91 is
developed based on relevant normative subpopulation data concerning
the desirable constituents 15 of the biological fluid system.
L. Non-Aphaeretic Filtration Methods
[0254] It should be appreciated that, while many of the embodiments
described herein utilize aphaeretic systems, additional embodiments
may be non-aphaeretic. One such exemplary embodiment is shown in
FIG. 42, in which a non-aphaeretic implementation is utilized for
the removal of tumor cell contaminants from analogous stem-cell
transplant products. Another exemplary embodiment is shown in FIG.
44C, in which a portable device 140 may be utilized for
non-aphaeretic filtration.
[0255] As shown in FIG. 42, biological fluid 16 comprised of a
leukapheresis product may be collected from a biological fluid
source 17. Pre-sorting may be performed by a microfluidic
separation module 100 so as to separate the leukapheresis product
into three primary divisions: (1) separated red blood cells,
plasma, and small cells; (2) separated white blood cells and small
CTCs; and (3) separated large CTC-clusters. In some embodiments,
pre-sorting may involve the use of chemical agents and/or buffer
solutions for red blood cell and platelet separation or lysis.
[0256] The first division, comprised of separated red blood cells,
plasma, and small cells, may be transferred to a fluid chamber to
hold filtered plasma and healthy cells for transplantation. The
third division, comprised of large CTC-clusters, may be transferred
for diagnostic processing (e.g., genomic, transcriptomic,
metabolomics, drug sensitivity and resistance) of CTCs,
CTC-clusters, and cell-free plasma.
[0257] With respect to the second division, comprised of separated
white blood cells and small CTCs, such contents may be inertial
focused into one or more parallel fluid receiving devices 30 (e.g.,
microfluidic channels 31, microwell arrays 32, and/or droplet
generators 34). Such contents may be scanned simultaneously or
sequentially by the scanner 70. The control unit 80 will then
analyze each sample to determine if cells other than healthy cells
are present (e.g., undesirable constituents 14).
[0258] If no cells other than healthy cells are present in a given
sample, that sample may be transferred to a fluid chamber along
with the first division of separated red blood cells, plasma, and
small cells. If cells other than healthy cells are present in a
given sample, that sample may be further enriched (e.g., by
returning to be reprocessed by a reprocessing path 62) or may be
sequestered along with the third division of separated large
CTC-clusters for diagnostic processing.
M. Operation of Illustrative Embodiments
[0259] The systems and methods described herein may be utilized for
therapeutic purposes, diagnostic purposes, or both. It should be
appreciated that the methods and systems used to isolate biological
fluids 16 containing undesirable constituents 14 may vary in
different embodiments. Further, the methods and systems utilized
for processing the isolated biological fluids 16 containing
undesirable constituents 14 may vary in different embodiments
depending upon whether such isolated biological fluids 16 are to be
used for therapeutic purposes, diagnostic purposes, or some
combination of both.
[0260] i. Therapeutic Systems and Methods.
[0261] As shown throughout the figures and discussed herein, the
systems and methods described herein may be utilized for various
therapeutic purposes. For example, therapeutic embodiments in which
the undesirable constituents 14 are CTCs or CTC-clusters may be
useful to reduce metastatic disease. As another example,
therapeutic embodiments in which the undesirable constituents 14
are pathogens may be useful to treat infection, including
sepsis.
[0262] ii. Aphaeretic Scanning and Filtration of Whole Blood Fluid
with CTCs on a Microfluidic Channel Platform with Reference Image
Dataset of Sampled Blood from Representative Individuals Other Than
the Patient.
[0263] One illustrative embodiment is a system and method for
aphaeretic scanning and filtration of whole blood to remove CTCs
from a patient's circulatory system. As illustrated in FIGS. 12 and
27, whole blood is pumped from a patient 12 via a receiver path 20
to a fluid receiving device 30 comprising a batch of parallel
microfluidic channels 31. Each channel 31 is optically scanned by a
scanner 70 using DIC Microscopy, DHM or other appropriate imaging
techniques to derive a scanned data 90 of the cells for each
microfluidic channel 31.
[0264] The scanned data 90 is transferred to a control unit 80,
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells (i.e.,
erythrocytes, leukocytes and platelets) in the scanned data 90.
More particularly, the algorithm 82 is designed to recognize
patterns in the reference data 91 characteristic of each type of
healthy blood cell and process each scanned data 90 transferred to
the control unit 80 to determine whether those patterns are
recognized in discreet image data obtained for each cell.
[0265] In an exemplary embodiment, the reference data 91 is
obtained through DIC Microscopy, digital holographic microscopy, or
other appropriate imaging techniques of blood samples taken from a
representative sample of individuals other than the patient. Raw
data is computer-processed to identify patterns in the images
and/or data that reflect characteristics of each type of healthy
blood cell, including, for example, cell size, shape, texture,
phase deviations, solidity and luminance. Data reflecting those
patterns is uploaded to the control unit 80 and stored to memory as
the reference data 91.
[0266] With reference to FIG. 29, if all cells in a scanned
microfluidic chamber are recognized by the algorithm 82 as healthy
blood cells, then the image processing software program generates a
first control signal instruction 83 to the control unit 80 to relay
a control signal to the valve 40 to route channel contents to the
return path 60. If, on the other hand, one or more of cells of the
scanned microfluidic chamber 31 are not recognized by the algorithm
82, then the image processing software program generates a second
control signal instruction 84 to the control unit 80 to relay a
control signal 81 to the valve 40 to route channel contents to the
isolation path 50. Filtered blood in the return path 60 is pumped
back to the patient's circulatory system. CTC-rich fluid routed to
the isolation path 50 is sequestered and optionally stored for
further processing for diagnostic or therapeutic purposes.
[0267] iii. Aphaeretic Filtration of Leukocyte-Rich Blood Fluid
with Small CTCs on a Microfluidic Channel Platform with Reference
Image Dataset of Sampled Blood from Representative Individuals
Other than the Patient.
[0268] One illustrative embodiment is a system and method for
aphaeretic scanning and filtration of Leukocyte-Rich Blood Fluid to
remove CTCs from a patient's circulatory system. In this example,
which is illustrated in FIGS. 21 and 27, whole blood is pumped from
the patient 12 into a receiver path 20 that directs flow of the
whole blood to a microfluidic separation module 100, which
pre-sorts whole blood using appropriate sorting techniques, e.g.,
Dean flow fractionation or dielectric sorting, into three
components: (1) fluid containing primarily healthy erythrocytes
(RBCs) and platelets, (2) fluid largely containing a mixture of
leukocytes (WBCs) and small CTCs, and (3) fluid containing large
CTCs and CTC-clusters. The fluid containing a mixture of leukocytes
and CTCs are promoted to fluid receiving device 30 comprising a
batch of parallel microfluidic channels 31. Each channel is
optically scanned using DIC Microscopy, DHM, or other appropriate
imaging techniques to derive a scanned data 90 of the cells for
each microfluidic channel 31.
[0269] The scanned data 90 is transferred to a control unit 80,
which follows an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. More particularly, the algorithm 82 is designed to
recognize a pattern in a reference data 91 characteristic of
healthy blood cells and process each scanned data 90 transferred to
the control unit 80 to determine whether that pattern is recognized
in discreet image data obtained for each cell. In the example, the
reference data 91 is obtained through DIC Microscopy or DHM of
blood samples taken from a representative sample of individuals
other than the patient.
[0270] With reference to FIG. 29, if all cells in a scanned
microfluidic channel 31 are recognized by the algorithm 82 as
healthy blood cells, then the image processing software program
generates a first control signal instruction 83 to the control unit
80 to relay a control signal 81 to the valve 40 to route channel
contents to the return path 60. If, on the other hand, one or more
of cells of the scanned microfluidic channel are not recognized by
the algorithm 82, then the image processing software program
generates a second control signal instruction 84 to the control
unit 80 to relay a control signal to the valve 40 to route channel
contents to the isolation path 50. Leukocytes routed to the return
path 60 are then recombined with the pre-sorted RBCs, plasma and
platelets and pumped back to the patient's circulatory system.
CTC-rich fluid routed to the isolation path 50 is sequestered and
optionally stored for further processing for diagnostic or
therapeutic purposes.
[0271] iv. Aphaeretic Scanning and Filtration of Whole Blood Fluid
with CTCs on a Microfluidic Channel Platform with Reference Image
Data of Sampled Blood from Patient.
[0272] One illustrative embodiment is a system and method or
aphaeretic scanning and filtration of whole blood to remove CTCs
from a patient's circulatory system. As illustrated in FIGS. 12 and
28, whole blood is pumped from a patient 12 via a receiver path 20
to a fluid receiving device 30 comprising a batch of parallel
microfluidic channels 31. Each channel 31 is optically scanned by a
scanner 70, such as by using DIC Microscopy, DHM, or other
appropriate imaging techniques, to derive a scanned data 90 of the
cells for each microfluidic channel 31.
[0273] The scanned data 90 is transferred to a control unit 80,
which follows an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. In the example, the reference data 91 is obtained
through DIC Microscopy, DHM, or other appropriate imaging
techniques of blood samples taken from the patient 12 prior to the
filtration session. The algorithm 82 is designed to recognize
patient-specific patterns in the reference data 91 characteristic
of each type of healthy blood cell and process each scanned data 90
transferred to the control unit 80 to determine whether those
patterns are recognized in discrete image data obtained for each
cell.
[0274] With reference to FIG. 29, if all cells in a scanned
microfluidic channel 31 are recognized by the algorithm 82 as
healthy blood cells, then the image processing software program
generates a first control signal instruction 83 to the control unit
80 to relay a control signal to the valve 40 to route channel
contents to the return path 60 If, on the other hand, one or more
of cells of the scanned microfluidic channel are not recognized by
the algorithm 82, then the image processing software program
generates a second control signal instruction 84 to the control
unit 80 to relay a control signal to the valve 40 to route channel
contents to the isolation path 50. Filtered blood in the return
path 60 is pumped back to the patient's circulatory system.
CTC-rich fluid routed to the isolation path 50 is sequestered and
optionally stored for further processing for diagnostic or
therapeutic purposes.
[0275] v. Aphaeretic Filtration of Leukocyte-Rich Blood Fluid with
Small CTCs on a Microfluidic Channel Platform with Reference Image
Data of Sampled Blood from Patient.
[0276] One illustrative embodiment is a system and method for
aphaeretic scanning and filtration of leukocyte-rich blood fluid to
remove CTCs from a patient's circulatory system. In this example,
as illustrated in FIGS. 17 and 28, whole blood is pumped from the
patient into a receiver path 20 that directs flow of the whole
blood to a microfluidic separation module 100, which pre-sorts
whole blood using appropriate sorting techniques, e.g., Dean flow
fractionation or dielectric sorting, into three components: (1)
fluid containing primarily healthy erythrocytes (RBCs) and
platelets, (2) fluid largely containing a mixture of leukocytes
(WBCs) and small CTCs, and (3) fluid containing large CTCs and
CTC-clusters. The fluid containing a mixture of leukocytes and CTCs
are promoted to fluid receiving device 30 comprising a batch of
parallel microfluidic channels 31. Each channel is optically
scanned by a scanner 70 which may, for example, use DIC Microscopy,
DHM, or other appropriate imaging techniques to derive a scanned
data 90 of the cells for each microfluidic channel 31.
[0277] The scanned data 90 is transferred to a control unit 80,
which follows an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. In the example, a reference data 91 is obtained
through DIC Microscopy, DHM, or other appropriate imaging
techniques of blood samples taken from the patient prior to the
filtration session. The algorithm 82 is designed to recognize a
pattern in the reference data 91 characteristic of the patient's
healthy cells and process each scanned data 90 transferred to the
control unit 80 to determine whether that pattern is recognized in
discreet image data obtained for each cell.
[0278] With reference to FIG. 29, if all cells in a scanned
microfluidic channel 31 are recognized by the algorithm 82 as
healthy cells, then the image processing software program generates
a first control signal instruction 83 to the control unit 80 to
relay a control signal 81 to the valve 40 to route channel contents
to the return path 60. If, on the other hand, one or more of cells
of the scanned microfluidic channel are not recognized by the
algorithm 82, then the image processing software program generates
a second control signal instruction 84 to the control unit 80 to
relay a control signal 81 to the valve 40 to route channel contents
to the isolation path 50. Leukocytes routed to the return path 60
are then recombined with the pre-sorted RBCs, plasma and platelets
and pumped back to the patient's circulatory system. CTC-rich fluid
routed to the isolation path 50 is sequestered and optionally
stored for further processing for diagnostic or therapeutic
purposes.
[0279] vi. Aphaeretic Scanning and Filtration of Whole Blood Fluid
with Pathogens on a Microfluidic Channel Platform.
[0280] An illustrative embodiment is a system and method for
aphaeretic scanning and filtration of whole blood to remove
pathogens from a patient's circulatory system. As shown in FIGS. 15
and 27-28, whole blood is pumped from a patient 12 via a receiver
path 20 to a fluid receiving device 30 comprising a batch of
parallel microfluidic channels 31. Each channel is optically
scanned using DIC Microscopy, DHM, or other appropriate imaging
techniques to derive a scanned data 90 of the cells for each
microfluidic channel 31.
[0281] The scanned data is transferred to a control unit 80, which
utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. More particularly, the algorithm 82 is designed to
recognize patterns in reference image data of the reference data 91
characteristic of each type of healthy blood cell and process each
scanned data 90 transferred to the control unit 80 to determine
whether those patterns are recognized in discreet image data
obtained for each cell. The reference data 91 is obtained through
DIC Microscopy, DHM, or other appropriate imaging techniques of
either samples taken from the patient's blood or samples taken from
a representative sample of individuals other than the patient.
[0282] With reference to FIG. 29, if all cells in a scanned
microfluidic channel 31 are recognized by the algorithm 82 as
healthy blood cells, then the image processing software program
generates a first control signal instruction 83 to the control unit
80 to relay a control signal 81 to the valve 40 to route channel
contents to the return path 60. If, on the other hand, one or more
of cells of the scanned microfluidic channel 31 are not recognized
by the algorithm, then the image processing software program
generates a second control signal instruction 84 to the control
unit 80 to relay a control signal 81 to the valve 40 to relay a
control signal to the valve 40 to route channel contents to the
isolation path 50. Filtered blood in the return path 60 is pumped
back to the patient's circulatory system. Pathogen-infected fluid
in the isolation path 50 is sequestered and optionally stored for
further processing for diagnostic or therapeutic purposes
[0283] vii. Aphaeretic Filtration of Pre-Sorted Blood Fluid with
Pathogens on Microfluidic Channel Platform.
[0284] An illustrative embodiment is a system and method for
aphaeretic filtration of Pre-Sorted Blood Fluid to remove pathogens
from a patient's circulatory system. In this example, as shown in
FIGS. 15 and 27-28, whole blood is pumped from the patient 12 into
a receiver path 20 that directs flow of the whole blood to a
microfluidic separation module 100, which pre-sorts whole blood
using appropriate sorting techniques, e.g., Dean flow fractionation
or dielectric sorting, into three components: (1) fluid containing
only healthy blood cells, (2) fluid containing a mixture of healthy
blood cells and pathogens, and (3) fluid containing only pathogens.
The fluid containing a mixture of blood cells and pathogens are
promoted to a fluid receiving device 30 comprising a batch of
parallel microfluidic channels 31. Each channel is optically
scanned using by a scanner 70 which uses DIC Microscopy, DHM, or
other appropriate imaging techniques to derive a scanned data 90 of
cells for each channel.
[0285] The scanned data 90 is transferred to a control unit 80,
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. More particularly, the algorithm 82 is designed to
recognize a patterns in a reference data 91 characteristic of each
type of healthy blood cell and process each scanned data 90
transferred to the control unit to determine whether that pattern
is recognized in discreet image data obtained for each cell. The
scanned data 90 may be obtained through DIC Microscopy, DHM, or
other appropriate imaging techniques of either samples taken from
the patient's blood or samples taken from a representative sample
of individuals other than the patient.
[0286] With reference to FIG. 29, if all cells in a scanned
microfluidic channel 31 are recognized by the algorithm as healthy
blood cells, then the image processing software program generates a
first control signal instruction 83 to the control unit 80 to relay
a control signal 81 to a valve 40 to route the channel's contents
to the return path 60. If, on the other hand, one or more of cells
of the scanned channel 31 are not recognized by the algorithm 82,
then the image processing software program generates a second
control signal instruction 84 to the control unit 80 to relay a
control signal 81 to the valve 40 to route the channel's contents
to the isolation path 50. Blood cells routed to the return path 60
are then recombined with the pre-sorted healthy blood cells and
pumped back to the patient's circulatory system. Pathogen-infected
fluid routed to the isolation path 50, along with the pre-sorted
fluid containing only pathogens, are sequestered and optionally
stored for further processing for diagnostic or therapeutic
purposes.
[0287] viii. Aphaeretic Scanning and Filtration of Whole Blood
Fluid with CTCs on a Microwell Array.
[0288] An illustrative embodiment is a system and method for
aphaeretic scanning and filtration of whole blood to remove CTCs
from a patient's circulatory system using a microwell array 32. As
shown in FIGS. 8-11, and 27-28, whole blood is pumped from a
patient 12 via a receiver path 20 to a fluid receiving device 30
comprising a microwell array 32. Each microwell of the microwell
array 32 is optically scanned with a scanner 70 using DIC
Microscopy, DHM, or other appropriate imaging techniques to derive
a scanned data 90 of the cells for each microwell of the microwell
array 32. Micro-wells may be periodically disrupted, such as by
shaking or stirring, and multiple scans taken, to ensure that the
scanned data 90 captures the entirety of the contents of the
microwells.
[0289] The scanned data 90 is transferred to a control unit 80,
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. More particularly, the algorithm is designed to
recognize patterns in reference image data of the reference data 91
characteristic of each type of healthy blood cell and process each
scanned data 90 transferred to the control unit 80 to determine
whether those patterns are recognized in discreet image data from
the reference data 91 obtained for each cell. The reference data 91
is obtained through DIC Microscopy, DHM, or other appropriate
imaging techniques of blood samples taken from either from a sample
of blood from the patient 12 or a representative sample of
individuals other than the patient 12.
[0290] With reference to FIG. 29, if all cells in a scanned
microwell of the microwell array 32 are recognized by the algorithm
as healthy blood cells, then the image processing software program
generates a first control signal instruction 83 to the control unit
80 to relay a control signal 81 to the valve 40 at the base of the
microwell to route the contents of the microwell to the return path
60 If, on the other hand, one or more of cells of the scanned
microwell are not recognized by the algorithm, then the image
processing software program generates a second control signal
instruction 84 to the control unit 80 to relay a control signal 81
to the valve 40 to route the contents of the microwell to the
isolation path 50. Filtered blood in the return path 60 is pumped
back to the patient's circulatory system. CTC-rich fluid routed to
the isolation path 50 is sequestered and optionally stored for
further processing for diagnostic or therapeutic purposes. In some
example embodiments, pipettes could be utilized to extract contents
of a microwell. In other embodiments, multiple wells in an array
could share a single valve 40.
[0291] ix. Aphaeretic Filtration of Leukocyte-Rich Blood Fluid with
Small CTCs on a Microwell Array.
[0292] An illustrative embodiment is a system and method for
aphaeretic scanning and filtration of leukocyte-rich blood fluid to
remove CTCs from a patient's circulatory system using a microwell
array 32. With reference to FIGS. 8-10 and 27-28, whole blood is
pumped from the patient 12 into a receiver path 20 that directs
flow of the whole blood to a microfluidic separation module 100,
which pre-sorts whole blood using appropriate sorting techniques,
e.g., Dean flow fractionation or dielectric sorting, into three
components: (1) fluid containing only healthy erythrocytes (RBCs)
and platelets, (2) fluid largely containing a mixture of leukocytes
(WBCs) and small CTCs, and (3) fluid containing large CTCs and
CTC-clusters. The fluid containing a mixture of leukocytes and CTCs
are promoted to a fluid receiving device 30 comprising a microwell
array 32. Each microwell is optically scanned by a scanner 70,
which may use DIC Microscopy, DHM, or other appropriate imaging
techniques to derive a scanned data 90 of the cells for each
microwell.
[0293] The scanned data 90 is transferred to a control unit 80,
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. More particularly, the algorithm 82 is designed to
recognize a pattern in a reference data 91 characteristic of
healthy blood cells and process each scanned data 90 transferred to
the control unit 80 to determine whether that pattern is recognized
in discreet image data obtained for each cell. The reference data
91 is obtained through DIC Microscopy of a sample of the patient's
blood or blood samples taken from a representative sample of
individuals other than the patient 12.
[0294] With reference to FIG. 29, if all cells in a scanned
microwell are recognized by the algorithm 82 as healthy blood
cells, then the image processing software program generates a first
control signal instruction 83 to the control unit 80 to relay a
control signal 81 to the valve 40 to route the contents of the
microwell to the return path 60. If, on the other hand, one or more
of cells of the scanned microwell are not recognized by the
algorithm 82, then the image processing software program generates
a second control signal instruction 84 to the control unit 80 to
relay a control signal 81 to the valve 40 to route the contents of
the microwell to isolation path 50. Leukocytes routed to the return
path 60 are then recombined with the pre-sorted RBCs, plasma and
platelets and pumped back to the patient's circulatory system.
CTC-rich fluid routed to the isolation path 50 is sequestered and
optionally stored for further processing for diagnostic or
therapeutic purposes. In some example embodiments, pipettes could
be utilized to extract contents of a microwell. In other
embodiments, multiple wells in an array could share a single valve
40.
[0295] x. Aphaeretic Filtration of Leukocyte-Rich Blood Fluid with
Small CTCs on a Microfluidic Channel Platform with Reference Image
Data for CTCs.
[0296] An illustrative embodiment is a system and method for
aphaeretic scanning and filtration of Leukocyte-Rich Blood Fluid to
remove CTCs from a patient's circulatory system using reference
data 91 of healthy blood cells and CTCs. With reference to FIG. 20,
a biological fluid 16 comprised of whole blood is pumped from the
patient 12 into a receiver path 20 that directs flow of the whole
blood to a microfluidic separation module 100, which pre-sorts
whole blood using appropriate sorting techniques, e.g., Dean flow
fractionation or dielectric sorting, into three components: (1)
fluid containing only healthy erythrocytes (RBCs) and platelets,
(2) fluid largely containing a mixture of leukocytes (WBCs) and
small CTCs, and (3) fluid containing large CTCs and CTC-clusters.
The fluid 16 containing a mixture of leukocytes and CTCs are
promoted to a fluid receiving device 30 comprising a batch of
parallel microfluidic channels 31. Each channel 31 is optically
scanned with a scanner 70 using DIC Microscopy, DHM, or other
appropriate imaging techniques to derive a scanned data 90 of the
cells for each microfluidic channel 31.
[0297] The scanned data 90 is transferred to a control unit 80,
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells and CTCs in
the scanned data 90. More particularly, the algorithm 82 is
designed to recognize a pattern in a reference data 91
characteristic of healthy blood cells and CTCs and process each
scanned data 90 transferred to the control unit 80 to determine
whether that pattern is recognized in discreet image data obtained
for each cell. The reference data 91, including image data of
recognized CTCs, is obtained through DIC Microscopy, DHM, or other
methods of blood samples containing CTCs taken from either from a
sample of blood from the patient or a representative sample of
individuals other than the patient. Image data from the reference
data 91 may also include scanned images of CTCs appropriately
cultured in medium or other fluids.
[0298] With reference to FIG. 29, if all cells in a scanned
microwell are recognized by the algorithm 82 as all healthy blood
cells, then the image processing software program generates a first
control signal instruction 83 to the control unit 80 to relay a
control signal 81 to the valve 40 to route the contents of the
channel to the return path 60. If, on the other hand, one or more
of cells of the scanned microwell are recognized as CTCs or not
recognized by the algorithm 82, then the image processing software
program generates a second control signal instruction 84 to the
control unit 80 to relay a control signal 81 to valve 40 to route
the contents of the channel to isolation path 50. Leukocytes routed
to the return path 60 are then recombined with the pre-sorted RBCs,
plasma and platelets and pumped back to the patient's circulatory
system. CTC-rich fluid routed to the isolation path 50 is
sequestered and optionally stored for further processing for
diagnostic or therapeutic purposes.
[0299] xi. Aphaeretic Scanning and Filtration of Whole Blood Fluid
with CTCs on a Microfluidic Channel Platform with Reference Image
Data for CTCs for Validation.
[0300] An illustrative embodiment is a system and method for
aphaeretic scanning and filtration of whole blood to remove CTCs
from a patient's 12 circulatory system using reference image data
from a reference data 91 for both healthy blood cells and CTCs.
With reference to FIG. 20, whole blood is pumped from a patient 12
via a receiver path 20 to a fluid receiving device 30 comprising a
batch of parallel microfluidic channels 31. Each channel is
optically scanned by a scanner 70 using DIC Microscopy, DHM, or
other appropriate imaging techniques to derive a scanned data 90 of
the cells for each microfluidic channel 31.
[0301] The scanned data 90 is transferred to a control unit 80,
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90 and a certifying algorithm 85 designed to recognize
CTCs in the scanned data 90. In the example, a reference data 91 is
obtained through DIC Microscopy, DHM, or other appropriate imaging
techniques of a sample of blood samples containing CTCs taken from
either from a sample of blood from the patient 12 or a
representative sample of individuals other than the patient 12.
Image data of the reference data 91 may also include scanned images
of CTCs appropriately cultured in medium or other fluids.
[0302] With reference to FIG. 29, the algorithm 82 is designed to
recognize patterns in the reference data 91 characteristic of CTCs
and each type of healthy blood cell and process each scanned data
90 transferred to the control unit 80 to determine whether those
patterns are recognized in discreet image data obtained for each
cell and generate one of two control signal instruction to the
control unit: a first control signal instruction 83 routing the
channel's contents to the return path 60 if every cell in the
channel is recognized as a healthy blood cell, and a second control
instruction 84 routing the channel's contends determined to contain
CTCs to the isolation path 50 if one or more cells in the channel
is not recognized as a healthy blood cell.
[0303] The certifying algorithm 85 is designed to generate a
certification output before a first control instruction 83 can be
executed by the control unit 80. The certifying algorithm 85 is
designed to recognize a pattern in the reference data 91
characteristic of the CTCs and verify that the CTC pattern is not
recognized in the scanned data 90 on which a generated first
control signal instruction 83 is based. If the verification
condition is met, the certifying algorithm 85 generates a
certification output and the control unit 80 is given a go
instruction to execute the first control signal instruction 83. If
the verification condition is not met, the certifying algorithm 85
generates an error output, prompting the image processing software
program to replace the first control signal instruction 83 with a
second control signal instruction 84 executed by the control unit
80. Filtered blood in return path is pumped back to the patient's
circulatory system. CTC-rich fluid routed to the isolation path 50
is sequestered and may optionally be further processed for
diagnostic or therapeutic purposes.
[0304] In some embodiments, the biological fluid 16 within the
fluid receiving device 30 may be scanned multiple times to verify
that the biological fluid 16 within the fluid receiving device 30
is indeed healthy prior to returning to the biological fluid source
17 via the return path 60. In such an embodiment, after an initial
scan of the biological fluid 16 by the scanner 70 fails to detect
any undesirable constituents 14, the biological fluid 16 will be
re-scanned, either by the same scanner 70 or by a different scanner
70 in embodiments with multiple scanners 70, to verify and confirm
the absence of any undesirable constituents 14 within the
biological fluid 16. The number of times that the biological fluid
16 is scanned in such a verification algorithm may vary in
different embodiments.
[0305] xii. Aphaeretic Filtration of Leukocyte-Rich Blood Fluid
with Small CTCs on a Microfluidic Channel Platform with Reference
Image Data for CTCs for Validation.
[0306] An illustrative embodiment is a system and method for
aphaeretic scanning and filtration of Leukocyte-Rich Blood Fluid to
remove CTCs from a patient's circulatory system using reference
image data from a reference data 91 of healthy blood cells and
CTCs. With reference to FIGS. 20 and 27-28, biological fluid 16
comprised of whole blood is pumped from the patient 12 into a
receiver path 20 that directs flow of the whole blood to a
microfluidic separation module 100, which pre-sorts whole blood
using appropriate sorting techniques, e.g., Dean flow fractionation
or dielectric sorting, into three components: (1) fluid containing
primarily healthy erythrocytes (RBCs) and platelets, (2) fluid
largely containing a mixture of leukocytes (WBCs) and small CTCs,
and (3) fluid containing large CTCs and CTC-clusters. The fluid
containing a mixture of leukocytes and CTCs are promoted to a fluid
receiving device 30 comprising a batch of parallel microfluidic
channels 31. Each channel is optically scanned by a scanner 70
using DIC Microscopy, DHM, or other appropriate imaging techniques
to derive a scanned data 90 of the cells for each microfluidic
channel 31.
[0307] The scanned data 90 is transferred to a control unit 80,
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90 and a certifying algorithm 85 designed to recognize
CTCs in the scanned data 90. In the example, a reference data 91 is
obtained through DIC Microscopy, DHM, or other appropriate imaging
techniques of blood samples containing CTCs taken from either from
a sample of blood from the patient 12 or a representative sample of
individuals other than the patient 12. The reference data 91 may
also include scanned images of CTCs appropriately cultured in
medium or other fluids.
[0308] With reference to FIG. 29, the algorithm 82 is designed to
recognize patterns in the reference data 91 characteristic of
healthy blood cells and process each scanned data 90 transferred to
the control unit 80 to determine whether those patterns are
recognized in discreet image data obtained for each cell and
generate one of two control signal instruction to the control unit:
a first control signal instruction 83 to route channel contents to
the return path 60 if every cell in the channel is recognized as
healthy blood cell, and a second control instruction 84 to route
channel contends determined to contain CTCs to the isolation path
50 if one or more cells in the channel is not recognized as a
healthy blood cell. The certifying algorithm 85 is designed to
generate a certification output before a first control instruction
can be executed by the control unit 80.
[0309] The certifying algorithm 85 is designed to recognize a
pattern in the reference data 91 characteristic of CTCs and verify
that the CTC pattern is not recognized in a scanned data 90 on
which a generated first control signal instruction 83 is based. If
the verification condition is met, the certifying algorithm 85
generates a certification output and the control unit 80 is given a
go instruction to execute the first control signal instruction 83.
If the verification condition is not met, the certifying algorithm
generates an error output, prompting the image processing software
program to replace the first control signal instruction 83 with a
second control signal instruction 84 executed by the control unit
80. Filtered blood in return path is pumped back to the patient's
circulatory system. CTC-rich fluid routed to the isolation path 50
is sequestered and may optionally be further processed for
diagnostic or therapeutic purposes.
[0310] In some embodiments, the biological fluid 16 within the
fluid receiving device 30 may be scanned multiple times to verify
that the biological fluid 16 within the fluid receiving device 30
is indeed healthy prior to returning to the biological fluid source
17 via the return path 60. In such an embodiment, after an initial
scan of the biological fluid 16 by the scanner 70 fails to detect
any undesirable constituents 14, the biological fluid 16 will be
re-scanned, either by the same scanner 70 or by a different scanner
70 in embodiments with multiple scanners 70, to verify and confirm
the absence of any undesirable constituents 14 within the
biological fluid 16. The number of times that the biological fluid
16 is scanned in such a verification algorithm may vary in
different embodiments.
[0311] xiii. Aphaeretic Scanning and Filtration of Blood with CTCs
on a Microfluidic Channel Platform Including Reference Image Data
to Limit False Positives.
[0312] An illustrative embodiment is a system and method for
aphaeretic scanning and filtration of blood fluid to remove
undesirable CTCs or pathogens from a patient's circulatory system
using reference image data that includes recognized patterns for
optic artifacts and designated pathogens not intended for
filtration to limit erroneous second control signal instructions 84
based on false positives.
[0313] In the example, as shown in FIGS. 16 and 27-28, reference
data 91 for healthy blood cells is obtained through DIC Microscopy,
DHM, or other appropriate imaging techniques, as described herein.
The reference data 91 also includes image data of known optic
artifacts and scanned data 90 of known non-target blood borne
pathogens. The image processing software program of the control
unit 80 of this embodiment comprises an algorithm 82 designed to
recognize patterns in the reference data 91 characteristic of each
type of healthy blood cell and to recognize data patterns of optic
artifacts and designated pathogens in reference data 91 to avoid
interpreting these data patterns as undesirable CTCs.
[0314] In operation, blood fluid on the fluid receiving device 30
is optically scanned, as described herein, to obtain a scanned data
90 of the cells in the scanned biological fluid 16. The scanned
data 90 is transferred to the control unit 80, and the algorithm 82
processes the scanned data 90 to determine whether patterns for
healthy blood cells, optic artifacts, and designated pathogens in
the reference data 91 is recognized in discreet image data of the
scanned data 90 for each cell. As with other embodiments, the image
processing software program generates a first control signal
instruction 83 if all cells are recognized by the algorithm 82, or
a second control signal instruction 84 if one or more cells is not
recognized. Because recognized pattern data of optic artifacts and
designated pathogens is incorporated into the reference data 91,
the presence of optic artifacts in the scanned data 90 or of
non-target pathogens in the scanned biological fluid will not cause
algorithm 82 to trigger the second control signal instruction
84.
[0315] It is preferable to maximize the amount of healthy blood
cells returned to the patient 12 and, relatedly, to limit the
amount of healthy blood cells lost during filtration. The
biological fluid filtration system 10 operates most efficiently
when scanned cells routed to the return path based on a first
control signal instruction 83 are all healthy blood cells and when
scanned cells routed to the isolation path 50 based on a first
control signal instruction 83 comprise at least one undesirable CTC
that triggered the instruction. Image data from optical artifacts
or blood borne pathogens are often picked up by optic scans, and so
have the potential to trigger a first control signal instruction
83. The present embodiment mitigates this issue by adding image
data from optical artifacts or designated pathogens to the
reference data 91 recognized algorithm 82.
[0316] xiv. Diagnostic System and Methods.
[0317] As shown throughout the figures and discussed herein, the
systems and methods described herein may be utilized for various
diagnostic purposes. For example, undesirable constituents 14 may
be sequestered and subject to a variety of in vitro diagnostic
tools for purposes of identification, prognosis, and/or treatment
determinations, within a number of methodological categories,
including microscopy, immunology-based assaying, culturing, in
vitro testing, drug sensitivity and resistance testing, and genomic
testing.
[0318] xv. Scanning and Filtration of Blood Sample for Diagnosis of
a Pathogenic Infection.
[0319] FIG. 14 is a block diagram of a diagnostic system and method
of an embodiment adapted to remove undesirable, indeterminate
pathogens from a biological fluid 16 and processing the removed
pathogens for diagnostics.
[0320] In the example, a blood sample is loaded into the fluid
receiving device 30, which comprises a batch of parallel
microfluidic channels 31 adapted to receive the blood sample. Each
channel 31 is optically scanned by a scanner 70 using DIC
Microscopy, DHM, or other appropriate imaging techniques to derive
a scanned data 90 of the cells for each microfluidic channel
31.
[0321] The scanned data 90 is transferred to a control unit 80
which follows an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. In the example, reference data 91 is obtained
through DIC Microscopy, DHM, or other appropriate imaging
techniques of blood samples taken from the patient 12 or from a
representative sample of individuals other than the patient 12
prior to the diagnostic session. The algorithm 82 is designed to
recognize a pattern in the reference data 91 and process each
scanned data 90 transferred to the control unit 80 to determine
whether that pattern is recognized in discreet image data from the
scanned data 90 obtained for each cell.
[0322] If all cells in a scanned microfluidic channel 31 are
recognized by the algorithm 82 as healthy blood cells, the control
unit 80 causes the valve 40 to direct contents of the channel 31
for disposal. If, on the other one or more of cells of the scanned
microfluidic channel 31 are not recognized by the algorithm 82, the
control unit 80 causes the valve 40 to direct the fluid 16 for
collection and diagnostic processing.
[0323] The control unit 80 is also adapted collect and store
catalogued image data for a group of catalogued pathogens, and the
algorithm 82 is further adapted to recognize patterns in the
reference data 91 characteristic of catalogued pathogens, and
process a scanned data 90 to search those patterns in the scanned
data 90, and identify any catalogued pathogen whose pattern is
recognized in the scanned data 90. The control unit 80 is further
adapted to store scanned data 90 to the catalogued image data,
assign the scan image data to a catalogued pathogen whose pattern
is recognized in the scan image data or to a pathogen later
identified in further diagnostic procedures, and the algorithm 82
is further adapted to recognize the scanned data 90 for use in
connection in with diagnostic procedures.
[0324] xvi. Scanning and Filtration of Blood Sample for Diagnosis
of a Cancer.
[0325] FIG. 14 also illustrates an embodiment diagnostic system and
method adapted to remove undesirable indeterminate CTCs or
CTC-clusters from a biological fluid 16 and process the removed
CTCs or CTC-clusters for diagnostics. Such an embodiment could be
utilized to detect the presence of cancer in an individual.
[0326] In the example, a blood sample is loaded into the fluid
receiving device 30, which comprises a batch of parallel
microfluidic channels 31 adapted to receive the blood sample. Each
channel 31 is optically scanned by a scanner 70 using DIC
Microscopy, DHM, or other appropriate imaging techniques to derive
a scanned data 90 of the cells for each microfluidic channel
31.
[0327] The scanned data 90 is transferred to a control unit 80
which utilizes an image processing software program comprising an
algorithm 82 designed to recognize healthy blood cells in the
scanned data 90. In the example, a reference data 91 is obtained
through DIC Microscopy, DHM, or other appropriate imaging
techniques of blood samples taken from the patient 12 or from a
representative sample of individuals other than the patient 12
prior to the diagnostic session. The algorithm 82 is designed to
recognize a pattern in the reference data 91 characteristic of
healthy blood cells and process each scanned data 90 transferred to
the control unit 80 to determine whether that pattern is recognized
in discreet image data from the scanned data 90 obtained from each
cell.
[0328] If all cells in a scanned microfluidic channel 31 are
recognized by the algorithm 82 as healthy blood cells, the control
unit 80 causes the valve 40 to direct contents of the channel 31
for disposal. If, on the other hand, one or more of cells of the
scanned microfluidic channel 31 are not recognized by the algorithm
82, the control unit 80 causes the valve 40 to direct the fluid 16
for collection and diagnostic processing.
[0329] The control unit 80 is also adapted collect and store
catalogued image data for a group of catalogued CTCs, and the
algorithm 82 is further adapted to recognize patterns in the
reference data 91 characteristic of catalogued CTCs, and process a
scanned data 90 to search those patterns in the scanned data 90,
and identify any catalogued CTCs whose pattern is recognized in the
scanned data 90. The control unit 80 is further adapted to store
scan image data to the scanned data 90, assign the scanned data 90
to a catalogued CTC whose pattern is recognized in the scanned data
90 or to a CTC identified in further diagnostic procedures, e.g.,
PCR, and the algorithm 82 is further adapted to recognize the
scanned data 90 for use in connection with diagnostic
procedures.
[0330] xvii. Scanning and Filtration of Leukapheresis Extract.
[0331] FIG. 32 illustrates an exemplary embodiment of scanning and
filtering a leukapheresis extract from a patient 12 undergoing
leukapheresis. As shown in FIG. 32, biological fluid 16 comprised
of a leukapheresis extract from the patient 12 containing healthy
cells and tumor cells is extracted from the patient 12. The
leukapheresis extract may be presorted using a microfluidic
separation module 100 in which healthy cells (e.g., utilizing RBC
lysis, microfluidic sorting, etc.) are separated from the
leukapheresis extract.
[0332] The remaining sample of the leukapheresis extract after
presorting is directed into a fluid receiving device 30 (e.g.,
microfluidic channels 31, microwell arrays 32, and/or droplet
generators 34) and then scanned by the scanner 70. The scanned data
90 from the scanner 70 is transferred to the control unit 80 which,
utilizing a reference data 91, detects the contents of the
leukapheresis extract.
[0333] If cells other than healthy cells are detected within the
scanned leukapheresis extract sample, the sample may be optionally
returned by a reprocessing path 62 for further enrichment of
non-healthy cells. Such samples may also be processed for
diagnostics. If only healthy cells are detected within the scanned
leukapheresis extract sample, the sample may be directed along an
isolation path 50 and not be processed for diagnostics.
[0334] FIG. 37 illustrates another exemplary method of scanning and
filtering a leukapheresis extract. As shown in FIG. 37, the
presorting stage comprised of a microfluidic separation module 100
may be utilized for optional red blood cell and platelet separation
using microfluidic or biochemical methods. After scanning by the
scanner 70 on the fluid receiving device 30, any samples containing
unrecognized cells may be processed for extraction of such
unrecognized cells (e.g., T-cell extraction) and may optionally be
reprocessed by redirecting back to the fluid receiving device 30
via a reprocessing path 62. Any samples containing only recognized
healthy cells may be directed along an isolation path 50 not to be
processed for cell extraction.
[0335] xviii. Scanning and Filtration of Stem Cell Extract.
[0336] FIG. 33 illustrates an exemplary embodiment of scanning and
filtering a stem cell extract from a patient 12 undergoing
procedures for removal of stem cells. As shown in FIG. 33,
biological fluid 16 comprised of a stem cell extract is extracted
from the patient 12 containing healthy cells and any tumor cells.
The stem cell extract may optionally be directed to a microfluidic
separation module 100 for separation of red blood cells and
platelets using microfluidic or biochemical methods.
[0337] The remaining stem cell extract after the optional
presorting stage is directed to the fluid receiving device 30 in
which it is optically scanned by a scanner 70. The scanned data 90
from the scanner 70 is transferred to the control unit 80 which,
utilizing a reference data 91, detects the contents of the stem
cell extract.
[0338] If cells other than healthy cells (e.g., tumor cells,
pathogens, etc.) are detected in the stem cell extract sample, the
sample may be optionally directed along a reprocessing path 62 for
optional reprocessing for further enrichment of non-healthy cells.
Any such stem cell extract samples (whether further enriched or
not) may then be processed for diagnostics. Stem cell extract
samples containing only healthy cells (e.g., not including tumor
cells or pathogens) may be directed along an isolation path 50 to
be utilized for stem cell transplant.
[0339] xix. Scanning and Filtration of Dialyzed Blood.
[0340] FIG. 51 illustrates an exemplary embodiment of scanning and
filtering dialyzed blood from a dialysis unit 48. In such an
embodiment, the optical filtration unit is connected to a dialysis
unit 48 (i.e., a dialysis machine), where blood (either pre- or
post-dialysis) is sent to the optical filtration unit and healthy
blood is returned back to the dialysis unit 48.
[0341] As shown in FIG. 51, biological fluid 16 from the biological
fluid source 17 is first undergoes dialysis within a dialysis unit
48. The resulting biological fluid 16 which has undergone dialysis
(e.g., dialyzed blood) is then transferred to a fluid receiving
device 30 and optically scanned by a scanner 70. The resulting
scanned data 90 is processed by the control unit 80. If only
desirable constituents 15 are detected, the sample may be returned
back to the dialysis unit 48 by a return path 60. If undesirable
constituents 14 are detected, the sample may be isolated along an
isolation path 50 and are not returned to the dialysis unit 48.
Optionally, diagnostics may be performed on the contents (e.g.,
pathogens, CTCs, CTC-clusters, host cells, cell free plasma, etc.)
that have been isolated.
[0342] xx. Scanning and Filtration of Blood Bag Contents.
[0343] FIG. 52 illustrates an exemplary embodiment of scanning a
biological fluid 16 comprised of blood from a biological fluid
source 17 comprised of a blood bag. In such an embodiment, the
contents of a blood bag from a patient 12 are processed to filter
CTCs and CTC-clusters. The remaining contents may then be separated
using a variety of methods (e.g., microfluidic, centrifugation,
biochemical, etc.) to separate white blood cells and CTCs. Those
cells may then be processed via optofluidic filtration techniques
described herein to separate CTCs.
[0344] As shown in FIG. 52, a biological fluid source 17 comprised
of a blood bag may be utilized. The biological fluids 16 within the
blood bag may be transferred to a microfluidic separation module
100 for presorting of large CTC-clusters. Any such separated large
CTC-clusters may be transferred along an isolation path 50 for
diagnostics (e.g., count, viability, genomic, transcriptomic,
molecular, morphological analyses).
[0345] The microfluidic separation module 100 may also separate
white blood cells and CTCs by using various methods such as
centrifugation, microfluidic sorting, biochemical methods, etc.).
Other methods such as RBC-lysis could also be used to separate red
blood cells from the blood. Additional buffers or reagents could
also be utilized in the presorting process. The resulting presorted
contents are then transferred to a fluid receiving device 30 to be
optically scanned by a scanner 70. The resulting scanned data 90 is
processed by the control unit 80 for detection and identification
of the contents of the biological fluid 16.
[0346] Any samples including undesirable constituents 14 may be
transferred along a reprocessing path 62 to be reprocessed or may
be isolated along an isolation path for CTC and CTC-cluster
diagnostics (e.g., count, viability, genomic, transcriptomic,
molecular, morphological analyses). Any samples which include only
desirable constituents 15 may be isolated separately and not
processed for diagnostics.
N. Example Embodiments
[0347] An example embodiment of a biological fluid filtration
system 10 may comprise a receiver path 20 adapted to receive a
biological fluid 16 from a biological fluid source 17. A fluid
receiving device 30 is fluidly connected to the receiver path 20 so
as to receive the biological fluid 16 from the receiver path
20.
[0348] A valve 40 may comprise an inlet 41 and a first outlet 42,
wherein the inlet 41 of the valve 40 is fluidly connected to the
fluid receiving device 30. An isolation path 50 may comprise an
inlet end, wherein the inlet end of the isolation path 50 is
connected to the first outlet 42 of the valve 40. A scanner 70 may
be oriented toward the fluid receiving device 30, with the scanner
70 being adapted to optically scan constituents 13 of the
biological fluid 16 within the fluid receiving device 30 so as to
derive a scanned data 90 of the constituents 13. A control unit 80
may be communicatively connected to the scanner 70 so as to receive
the scanned data 90 from the scanner 70, wherein the control unit
80 is operatively connected to the valve 40.
[0349] The control unit 80 is adapted to compare the scanned data
90 to a reference data 91. The reference data 91 may comprise
images and/or characteristics of desirable constituents 15 such as
healthy cells. The control unit 80 is adapted to switch the valve
40 so as to direct the biological fluid 16 to the biological fluid
source 17 by a return path 60 if the scanned data 90 includes only
desirable constituents 15 meeting the criteria of desirable
constituents 15. The control unit 80 is adapted to switch the valve
40 so as to direct the biological fluid 16 to the isolation path 50
if the scanned data 90 of the biological fluid 16 includes one or
more undesirable constituents 14 not meeting the criteria of
desirable constituents 15. In another exemplary embodiment of a
biological fluid filtration system 10, the reference data 91
includes image data representative of desirable constituents
15.
[0350] In another exemplary embodiment of a biological fluid
filtration system 10, the reference data 91 further comprises
criteria of optic artifacts, and the control unit 80 is adapted to
switch the valve 40 so as to direct the biological fluid 16 to the
biological fluid source 17 by the return path 60 if the scanned
data 90 of the biological fluid 16 includes only desirable
constituents 15 meeting the criteria of optic artifacts or the
criteria of desirable constituents 15.
[0351] In another exemplary embodiment of a biological fluid
filtration system 10, the reference data 91 further comprises
recognized criteria of designated pathogens. The control unit 80 is
adapted to switch the valve 40 so as to direct the biological fluid
16 to the biological fluid source 17 by the return path 60 if the
scanned data 90 of the biological fluid 16 includes only desirable
constituents 15 meeting the criteria of designated pathogens or the
criteria of desirable constituents 15.
[0352] The reference data 91 may include image data and/or
characteristics representative of healthy blood constituents. The
reference data 91 may include image data and/or characteristics
representative of erythrocytes, leukocytes, thrombocytes, and the
like.
[0353] In another exemplary embodiment, the biological fluid
filtration system 10, comprises a microfluidic separation module
100, wherein the microfluidic separation module 100 is adapted to
receive blood from the receiver path 20, generate a primarily
leukocyte-rich fluid by separating leukocytes from other desirable
constituents 15 of the blood, and advance the leukocyte-rich fluid
with any undesirable components to the fluid receiving device 30.
It should also be appreciated that, in embodiments in which a
biological fluid 16 other than blood is being scanned, the
components which are separated during presorting will by definition
be comprised of different components than with blood. For example,
in embodiments in which the biological fluid 16 is comprised of
saliva, urine, cerebrospinal fluid, lymphatic fluids, or
leukapheresis extracts, other constituents of the biological fluid
16 may be separated than the exemplary components (leukocytes,
etc.) of blood. As another example, other components of the
biological fluid 16 may be separated in sepsis-like use cases.
[0354] The one or more undesirable constituents 14 may be comprised
of circulating tumor cells and pathogens. The fluid receiving
device 30 may be comprised of one or more microfluidic channels 31.
The fluid receiving device 30 may be comprised of a plurality of
microwells, such as a plurality of microwells arranged in a
microwell array 32. The scanner 70 may be adapted to scan each of
the plurality of microwells of the microwell array 32.
[0355] The fluid receiving device 30 may be comprised of a
microfluidic droplet generator 34. In such an embodiment, a droplet
generator 34 may be utilized to encapsulate cells 18 into droplets
39, with each of the droplets 39 being scanned by the scanner 70.
The droplet generator 34 may include a dispersed phase channel 35
and one or more continuous phase channels 36 which converge at a
juncture 37 in which the cells 18 are encapsulated into the
droplets 39. The droplets 39 including the encapsulated cells 18
may then be routed through a scanning channel 38 in which each such
droplet 39 or groups of droplets 39 are scanned by the scanner
70.
[0356] The valve 40 may comprise a plurality of well valves 40 in
one-to-one fluid communication with the plurality of microwells of
the microwell array 32. The control unit 80 is operatively
connected to the well valve 40 of each of the plurality of
microwells of the microwell array 32. In some example embodiments,
pipettes could be utilized to extract contents of a microwell. In
other embodiments, multiple wells in an array 32 could share a
single valve 40.
[0357] In another exemplary embodiment of a biological fluid
filtration system 10, the fluid receiving device 30 is comprised of
a microfluidic channel 31. In some embodiments, the fluid receiving
device 30 is comprised of a plurality of microfluidic channels 31.
The plurality of microfluidic channels 31 may be arranged in
parallel or in series. The scanner 70 may be adapted to scan each
of the plurality of microfluidic channels 31. The valve 40 may
comprise a plurality of channel valves 40 in one-to-one fluid
communication with the plurality of microfluidic channels 31. The
control unit 80 may be operatively connected to the channel valve
40 of each of the plurality of microfluidic channels 31. The valve
40 may comprise a second outlet 43 in fluid communication with the
return path 60.
[0358] The return path 60 may comprise a return channel that is
fluidly connected to the biological fluid source 17 such that
filtered biological fluid 16 may be returned to the patient 12. In
some embodiments, a drug infuser 130 may be fluidly connected to
the return path 60 so as to introduce various drugs or treatments
in the return path 60 to treat the patient 12. The drug infuser 130
may comprise a reservoir containing a volume of a drug or treatment
that is fluidly connected to the return path 60. The drug infuser
130 may utilize valves, pumps, sensors, and the like to control the
dosage infused within the return path 60.
[0359] In some embodiments, a reprocessing path 62 may be included
so as to allow reprocessing of biological fluids 16 multiple times.
In such an embodiment, the reprocessing path 62 may be fluidly
connected back to the fluid receiving device 30 such that processed
biological fluid 16 may be redirected back to the fluid receiving
device 30 for additional processing. It should be appreciated that
the reprocessing path 62 may be included in addition to the return
path 60, such that certain samples of biological fluid 16 may be
returned to the patient 12, and certain other samples of biological
fluid 16 may instead be returned to the fluid receiving device 30
for additional scanning.
[0360] In another exemplary embodiment, a method of filtering a
biological fluid 16 using the biological fluid filtration system 10
comprises the steps of: directing the biological fluid 16 to the
fluid receiving device 30; optically scanning the biological fluid
16 within the fluid receiving device 30 by the scanner 70 to
generate the scanned data 90 of the biological fluid 16; comparing
the scanned data 90 of the biological fluid 16 with the reference
data 91 by the control unit 80; returning the biological fluid 16
to the biological fluid source 17 if the scanned data 90 includes
only desirable constituents 15 meeting the criteria of desirable
constituents 15; and isolating the biological fluid 16 from the
biological fluid source 17 if the scanned data 90 of the biological
fluid 16 includes one or more undesirable constituents 14 not
meeting the criteria of desirable constituents 15.
[0361] The method may comprise a therapeutic method and the
biological fluid source 17 may comprise an individual patient 12
having a wide range of conditions, such as but not limited to
cancer or infection with various pathogens. The method may further
comprise obtaining a sample of biological fluid 16 of the
individual patient 12, wherein the reference data 91 is generated
from the sample.
[0362] In another exemplary embodiment, the method may further
comprise obtaining samples of biological fluid 16 of one or more
individuals other than the individual patient 12, wherein the
reference data 91 is generated from the samples of biological fluid
16 of the one or more individuals other than the individual patient
12. The method may further comprise the step of performing
diagnostics on the undesirable constituents 14.
[0363] In another exemplary embodiment, a method of filtering a
biological fluid 16 comprises the steps of: receiving the
biological fluid 16 from a biological fluid source 17 by a receiver
path 20; transferring the biological fluid 16 from the receiver
path 20 to a fluid receiving device 30; optically scanning the
biological fluid 16 within the fluid receiving device 30 by a
scanner 70 so as to create a scanned data 90 of the biological
fluid 16; comparing the scanned data 90 of the biological fluid 16
with a reference data 91 by a control unit 80, wherein the
reference data 91 comprises criteria including images and/or
morphological parameters of desirable constituents 15; returning
the biological fluid 16 to the biological fluid source 17 by a
return path 60 if the scanned data 90 of the biological fluid 16
includes only desirable constituents 15 meeting the criteria of
desirable constituents 15; and isolating the biological fluid 16
from the biological fluid source 17 by an isolation path 50 if the
scanned data 90 of the biological fluid 16 includes one or more
undesirable constituents 14 not meeting the criteria of desirable
constituents 15. The reference data 91 may include data
representative of desirable constituents 15.
[0364] In another exemplary embodiment of the method of filtering a
biological fluid 16 using the biological fluid filtration system
10, the reference data 91 includes criteria of optic artifacts, and
further steps may be performed comprising returning the biological
fluid 16 to the biological fluid source 17 by a return path 60 if
the scanned data 90 of the biological fluid 16 includes only
desirable constituents 15 meeting the criteria of optic artifacts
or the criteria of desirable constituents 15, and isolating the
biological fluid 16 from the biological fluid source 17 by an
isolation path 50 if the scanned data 90 of the biological fluid 16
includes one or more undesirable constituents 14 not meeting the
criteria of optic artifacts or the criteria of desirable
constituents 15.
[0365] In another exemplary embodiment of the method of filtering a
biological fluid 16 using the biological fluid filtration system
10, the reference data 91 includes criteria including images and/or
morphological parametric data representative of designated
pathogens, and further steps may be performed comprising returning
the biological fluid 16 to the biological fluid source 17 by a
return path 60 if the scanned data 90 of the biological fluid 16
includes only desirable constituents 15 meeting the criteria of
optic artifacts/pathogens or the criteria of desirable constituents
15, and isolating the biological fluid 16 from the biological fluid
source 17 by an isolation path 50 if the scanned data 90 of the
biological fluid 16 includes one or more undesirable constituents
14 not meeting the criteria of optic artifacts/pathogens or the
criteria of desirable constituents 15.
[0366] The reference data 91 may include image data or
morphological characteristics representative of healthy blood
constituents. By way of example and without limitation, the
reference data 91 may include data representative of erythrocytes,
leukocytes, or thrombocytes.
[0367] In another exemplary embodiment of the method of filtering a
biological fluid 16 using the biological fluid filtration system
10, the method may further comprise the steps of: sorting the
biological fluid 16 from the receiver path 20 to separate
leukocytes from other desirable constituents 15 of the biological
fluid 16 to generate a leukocyte-rich fluid, and transferring the
leukocyte-rich fluid to the fluid receiving device 30. The one or
more undesirable constituents 14 may be comprised of circulating
tumor cells, clusters thereof, or various other pathogens.
[0368] In another exemplary embodiment of the method of filtering a
biological fluid 16 using the biological fluid filtration system
10, each of the plurality of microwells of the microwell array 32
may comprise a well valve 40 having a first port fluidly connected
to the isolation path 50 and a second port fluidly connected to the
return path 60. In other embodiments, each of the plurality of
microwells of the microwell array 32 is fluidly connected via a
well valve 40 having a single port, with the port being fluidly and
selectively connected to both the return path 60 and the isolation
path 50.
[0369] The control unit 80 may be operatively connected to the well
valve 40 of each of the plurality of microwells of the microwell
array 32. The fluid receiving device 30 may be comprised of a
microfluidic channel 31 or a plurality of microfluidic channels 31.
The plurality of microfluidic channels 31 may be arranged in a
parallel series. The scanner 70 may be adapted to scan each of the
plurality of microfluidic channels 31. In some example embodiments,
pipettes could be utilized to extract contents of a microwell. In
other embodiments, multiple wells in an array could share a single
valve 40.
[0370] In another exemplary embodiment of the method of filtering a
biological fluid 16 using the biological fluid filtration system
10, each of the plurality of microfluidic channels 31 may comprise
a channel valve 40 having a first port fluidly connected to the
isolation path 50 and a second port fluidly connected to the return
path 60. The control unit 80 may be operatively connected to the
channel valve 40 of each of the plurality of microfluidic channels
31.
[0371] In another exemplary embodiment, a method of filtering a
biological fluid 16 comprises the steps of: receiving the
biological fluid 16 from a biological fluid source 17; separating
the biological fluid 16 into a first portion, a second portion, and
a third portion, the first portion comprising only desirable
constituents 15, the second portion comprising a mix of undesirable
and desirable constituents 14, 15, and the third portion comprising
only undesirable constituents 14; returning the first portion of
the biological fluid 16 to the biological fluid source 17 by a
return path 60; isolating the third portion of the biological fluid
16 from the biological fluid source 17 by an isolation path 50;
transferring the second portion of the biological fluid 16 to a
fluid receiving device 30; optically scanning the second portion of
the biological fluid 16 within the fluid receiving device 30 by a
scanner 70 so as to create a scanned data 90 of the second portion
of the biological fluid 16; comparing the scanned data 90 of the
second portion of the biological fluid 16 with a reference data 91
by a control unit 80, the reference data 91 comprising criteria of
desirable constituents 15; returning the second portion of the
biological fluid 16 to the biological fluid source 17 by the return
path 60 if the scanned data 90 of the biological fluid 16 includes
only desirable constituents 15 meeting the criteria of desirable
constituents 15; and isolating the second portion of the biological
fluid 16 from the biological fluid source 17 by the isolation path
50 if the scanned data 90 of the biological fluid 16 includes one
or more constituents 13 not meeting the criteria of desirable
constituents 15. The step of separating the biological fluid 16 may
be comprised of dielectric sorting, Dean flow fractionation, or
hemodynamic properties.
[0372] In another exemplary embodiment of the method of filtering a
biological fluid 16 using the biological fluid filtration system
10, multiple scanners 70 may be used simultaneously to optically
scan the fluid receiving device 30. In such an embodiment,
different areas, such as different microwells of a microwell array
32, different microfluidic channels 31, or different droplets 39 of
a droplet generator 34, may be optically scanned by a plurality of
scanners 70 rather than a singular scanner 70.
[0373] In another exemplary embodiment of the method of filtering a
biological fluid 16 using the biological fluid filtration system
10, the fluid receiving device 30 may be scanned multiple times by
the scanner 70 before transfer to the return path 60. Such an
embodiment will utilize multiple scans to ensure and verify that
the biological fluid 16 being scanned does not include any
undesirable constituents 14 prior to be released to the return path
60. Such an embodiment will reduce the risks of inaccurate scans by
verifying through multiple scans prior to releasing the biological
fluid 16.
[0374] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. Although
methods and materials similar to or equivalent to those described
herein can be used in the practice or testing of the biological
fluid filtration system, suitable methods and materials are
described above. All patent applications, patents, and printed
publications cited herein are incorporated herein by reference in
their entireties, except for any definitions, subject matter
disclaimers or disavowals, and except to the extent that the
incorporated material is inconsistent with the express disclosure
herein, in which case the language in this disclosure controls. The
biological fluid filtration system may be embodied in other
specific forms without departing from the spirit or essential
attributes thereof, and it is therefore desired that the present
embodiment be considered in all respects as illustrative and not
restrictive. Any headings utilized within the description are for
convenience only and have no legal or limiting effect.
* * * * *