U.S. patent application number 12/627824 was filed with the patent office on 2010-06-03 for fluid processing device including size-changing barrier.
This patent application is currently assigned to LIFE TECHNOLOGIES CORPORATION. Invention is credited to Debjyoti Banerjee, Konrad Faulstich, Aldrich N. K. Lau, Umberto Ulmanella, Jun Xie.
Application Number | 20100133104 12/627824 |
Document ID | / |
Family ID | 36203645 |
Filed Date | 2010-06-03 |
United States Patent
Application |
20100133104 |
Kind Code |
A1 |
Banerjee; Debjyoti ; et
al. |
June 3, 2010 |
Fluid processing device including size-changing barrier
Abstract
A diagnostic device is provided that includes a plurality of
retainment regions interconnected through at least one fluid
processing passageway or separated by at least one barrier. A fluid
flow modulator can be provided in the fluid processing passageway
if a fluid processing passageway is provided. The barrier and/or
fluid flow modulator can comprise a polysaccharide, a derivative of
a polysaccharide, or a combination thereof. For example, the
barrier can comprise a chitosan material.
Inventors: |
Banerjee; Debjyoti; (College
Station, TX) ; Ulmanella; Umberto; (Foster City,
CA) ; Faulstich; Konrad; (Fremont, CA) ; Lau;
Aldrich N. K.; (Palo Alto, CA) ; Xie; Jun;
(Pasadena, CA) |
Correspondence
Address: |
LIFE TECHNOLOGIES CORPORATION;C/O INTELLEVATE
P.O. BOX 52050
MINNEAPOLIS
MN
55402
US
|
Assignee: |
LIFE TECHNOLOGIES
CORPORATION
Carlsbad
CA
|
Family ID: |
36203645 |
Appl. No.: |
12/627824 |
Filed: |
November 30, 2009 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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11252912 |
Oct 18, 2005 |
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12627824 |
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60619677 |
Oct 18, 2004 |
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60619731 |
Oct 18, 2004 |
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60619623 |
Oct 18, 2004 |
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Current U.S.
Class: |
204/450 ;
204/406; 204/409; 422/400; 73/864.63 |
Current CPC
Class: |
F16K 99/0001 20130101;
F16K 99/0036 20130101; Y10T 137/85978 20150401; B01L 3/502738
20130101; B01L 2400/0487 20130101; Y10T 137/1624 20150401; B01L
3/502746 20130101; Y10T 436/143333 20150115; B01L 2400/0415
20130101; B01L 2400/0683 20130101; B01L 2400/0661 20130101; B01L
2400/0406 20130101; B01L 2400/086 20130101; Y10S 422/901 20130101;
B01L 2400/082 20130101; B01L 2300/0864 20130101; B01L 2400/0655
20130101; B01L 2400/084 20130101; B01L 2200/0621 20130101; Y10T
137/0324 20150401; F16K 99/003 20130101; Y10T 137/8593 20150401;
F16K 99/0057 20130101; Y10T 436/2575 20150115; B01L 2300/087
20130101; B01L 2400/0481 20130101; Y10T 137/1812 20150401; B01L
2400/0677 20130101; B01L 2300/0867 20130101; Y10T 137/0318
20150401; Y10T 137/2191 20150401; Y10T 137/8376 20150401; F16K
2099/0084 20130101; B01L 2300/0816 20130101 |
Class at
Publication: |
204/450 ;
204/409; 204/406; 422/103; 422/100; 73/864.63 |
International
Class: |
G01N 27/26 20060101
G01N027/26; B01L 3/02 20060101 B01L003/02; F15C 1/04 20060101
F15C001/04; B81B 7/00 20060101 B81B007/00; G01N 1/12 20060101
G01N001/12 |
Claims
1. A fluid processing device comprising: a flow passageway; a
plurality of retainment regions, at least two of the retainment
regions each being in fluid communication with the flow passageway;
and at least one a fluid flow modulator arranged in the flow
passageway and being adapted to form, or modify the size of, a
fluid communication between the at least two retainment regions,
the at least one fluid flow modulator comprising a
polysaccharide.
2. The fluid processing device of claim 1, wherein the
polysaccharide is a water-soluble polysaccharide.
3. The fluid processing device of claim 1, wherein the at least one
fluid flow modulator comprise at least one of a fluid flow
modulator that is adapted to open or to increase the size of the
fluid communication between the at least two retainment regions,
and a fluid flow modulator that is adapted to close or to decrease
the size of the fluid communication between the at least two
retainment regions.
4. The fluid processing device of claim 1, wherein at least one of
the plurality of retainment regions comprises aqueous fluid
retained therein.
5. The fluid processing device of claim 1, wherein the flow
passageway is dimensioned sufficient to cause capillary flow of a
fluid from at least one of the at least two retainment regions
through the flow passageway.
6. The fluid processing device of claim 1, wherein the flow
passageway is dimensioned sufficient to cause electrokinetic flow
of charged components in a fluid from at least one of the at least
two retainment regions through the flow passageway.
7. The fluid processing device of claim 6, further comprising at
least two electrodes disposed in the device with the flow
passageway therebetween.
8. The fluid processing device of claim 1, further comprising: at
least one additional retainment region; at least one additional
flow passageway; and at least one valve comprising one or more of a
pressure-actuatable valve and a heat-actuatable valve arranged in
the at least one additional flow passageway, wherein the at least
one additional flow passageway is in fluid communication with the
at least one additional retainment region and at least one of the
plurality of retainment regions.
9. The fluid processing device of claim 1, further comprising at
least two electrodes disposed in the device with the flow
passageway therebetween.
10. A system comprising the fluid processing device of claim 9, a
power source, and at least two electrical leads forming electrical
connections, respectively, between the power source and the at
least two electrodes.
11. A system comprising the fluid processing device of claim 1, and
a pump, wherein the pump is arranged in fluid communication with at
least one of the flow passageway and one or more of the plurality
of retainment regions.
12. A fluid processing device comprising: a flow passageway; and at
least one valve arranged in the flow passageway, being adapted to
open to form a fluid communication through the flow passageway, the
valve being adapted to dissolve when contacted with an aqueous
solution at room temperature and having a pH of 9.5 or lower, and
the valve comprising poly(D-glucosamine), a derivative of
poly(D-glucosamine), or a combination thereof.
13. A fluid processing device comprising: a substrate; a plurality
of retainment regions formed in or on the substrate, comprising at
least a first retainment region and a second retainment region; and
a barrier at least partially separating the first retainment region
from the second retainment region, wherein said barrier comprises a
polysaccharide.
14. The fluid processing device of claim 13, further comprising: at
least one additional retainment region; at least one flow
passageway; and at least one valve comprising one or more of a
pressure-actuatable valve and a heat-actuatable valve arranged in
the at least one flow passageway, wherein the at least one flow
passageway is in fluid communication with the at least one
additional retainment region and at least one of the plurality of
retainment regions.
15. A method comprising: providing a fluid processing device
comprising at least a first retainment region and a second
retainment region, and a barrier arranged at least partially
between the first retainment region and the second retainment
region, wherein at least one of the first and second retainment
regions retains a solvent for the barrier, and the barrier
comprises a polysaccharide; and contacting the barrier with the
solvent to change the shape of the barrier.
16. The method of claim 15, wherein contacting the barrier with the
solvent comprises opening or increasing the size of a fluid
communication between the first retainment region and the second
retainment region, or closing or decreasing the size of a fluid
communication between the first retainment region and the second
retainment region.
17. The method of claim 15, wherein the fluid processing device
further comprises at least one additional retainment region, at
least one flow passageway, and at least one valve comprising one or
more of a pressure-actuatable valve arranged in the at least one
flow passageway, wherein the at least one flow passageway is in
fluid communication with the at least one additional retainment
region and at least one of the first retainment region and the
second retainment region, and the method further comprises opening
the pressure-actuatable valve.
18. The method of claim 15, wherein the fluid processing device
further comprises a flow passageway in fluid communication with the
first retainment region and the second retainment region, and the
method comprises migrating charged components in a sample from at
least one of the first retainment region and the second retainment
region through the flow passageway by electrokinetic motion.
19. The method of claim 15, further comprising creating a pressure
differential between the first retainment region and the second
retainment region, and moving, with the pressure differential, a
fluid from one of the first retainment region and the second
retainment region into the other of the first retainment region and
the second retainment region.
20. The method of claim 15, further comprising creating a magnetic
field across the first retainment region and the second retainment
region, and moving, with the magnetic field, magnetically
attractable materials from one of the first retainment region and
the second retainment region into the other of the first retainment
region and the second retainment region.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims benefit under 35 U.S.C. Section
119(e) from earlier U.S. Provisional Patent Applications Nos.
60/619,731, 60/619,677, and 60/619,623, all of which were filed
Oct. 18, 2004, and all of which are incorporated herein in their
entireties by reference.
INTRODUCTION
[0002] The present teachings relate to a diagnostic device and
method providing automatically controlled interconnection between a
plurality of retainment regions.
[0003] A portable or nonportable diagnostic device or method can
perform a set of predetermined assays by providing for controlled
interaction between various fluids initially present in separate
retainment regions. A device and method compatible with nucleic
acid sequence reactions, detecting such reactions, and capable of
processing automatically such a reaction is desirable.
SUMMARY
[0004] According to various embodiments, a diagnostic device is
provided that includes a plurality of retainment regions, as
exemplified below, with the retainment regions being interconnected
through a plurality of fluid communications, fluid processing
passageways, flow passageways, and/or channels. Herein the phrase
"retainment region" means a retainment or containment feature such
as a well, a fluid retainment region, a channel, a vial, a
compartment, another receptacle, a surface on which a material is
retained, or the like. The following discussion with regard to
retainment regions would be equally applicable to any of the
above-mentioned features or their equivalents.
[0005] The retainment regions can include a reaction retainment
region, one or more reagent retainment regions each containing
reagents, and a sample retainment region. A pressure-actuated valve
can be positioned in each of the fluid processing passageways
interconnecting the one or more reagent retainment regions with
respective intermediate retainment regions interposed between each
of the one or more reagent retainment regions and the reaction
retainment region. A barrier or fluid flow modulator, as
exemplified below with reference to a valve, can be provided in one
or more of the channels interconnecting the reagent retainment
regions with the reaction retainment region or intermediate
retainment regions, or interconnecting the intermediate retainment
regions and the reaction retainment region.
[0006] According to various embodiments, a method of performing a
set of predetermined assays is provided. The method can include
providing a plurality of retainment regions in a closed, and if
desired, disposable cuvette, with the retainment regions being
interconnected by fluid processing passageways but closed to fluid
flow to or from locations outside of the cuvette. First retainment
regions can be selectively closed off from fluid communication with
second retainment regions with which they are interconnected by
first channels including pressure-actuated valves positioned
therein. The pressure-actuated valves can comprise a burstable or
tearable diaphragm, or other frangible seal that can rupture, tear,
break, or the like, when exposed to a change in pressure, for
example, an increase or decrease in pressure. One or more third
retainment regions can be selectively closed off from fluid
communication with at least the second retainment regions with
which they are interconnected by second channels by valves
positioned in the second channels. Pressure can be applied to the
pressure actuated valves in the first channel sufficient to provide
fluid communication between the first and second retainment
regions. A sample to be tested or otherwise processed can be
introduced into the one or more third retainment regions, and fluid
communication can be established between the second retainment
regions and the one or more third retainment regions at a
controlled rate that can be a function of any one of a number of
stimuli and/or characteristics of at least one of the sample in the
one or more third retainment regions and a fluid within the second
retainment regions. The characteristics of at least one of the
sample in the one or more third retainment regions and a fluid
within the second retainment region can include, but are not
limited to water content, pH, chemical composition, temperature,
electrical charge, magnetic properties, or the like.
[0007] The closed, and if desired, disposable cuvette, can be
provided as a substrate that is fabricated from a single piece or
more than one piece. The retainment regions, interconnecting fluid
processing passageways, and/or valves can be fabricated all in the
single piece substrate, or if desired, can be fabricated in one or
more different pieces, which can then be combined to form the
cuvette.
[0008] One or more third retainment regions can be selectively
closed off from fluid communication with at least the second
retainment regions with which they are interconnected by second
channels by flow obstructions responsive to pH or other stimuli
that cause a change in volume of the flow obstructions positioned
in the second channels. Pressure can be applied to the pressure
actuated valves in the first channels sufficient to provide fluid
communication between the first and second retainment regions. A
sample to be tested or otherwise processed can be introduced into
the one or more third retainment regions, and fluid communication
can be established between the second retainment regions and the
one or more third retainment regions at a controlled rate. The
controlled rate can be a function of the change in volume of the
flow obstruction, such as shrinking or swelling of the flow
obstruction, as caused by the pH or other characteristics of at
least one of the sample in the one or more third retainment regions
and a fluid within the second retainment regions. The
characteristics of at least one of the sample in the one or more
third retainment regions and a fluid within the second, can
include, but are not limited to the water content, the pH, the
chemical composition, the temperature, electrical charge, magnetic
properties, or the like.
[0009] According to some embodiments, the device can comprise no
vent, at least one vent, or a plurality of vents, to relieve
pressure resulting from a flow of a fluid and its communication. A
vent can comprise a vent channel configured to relieve such
pressure. A vent can be provided in communication with a retainment
region, such that upon fluid flow resultant pressure is released. A
vent channel can comprise a hydrophobic vent channel that allows
air to travel through the channel but does not allow the flow of an
aqueous fluid.
[0010] According to some embodiments, a device is provided that can
comprise no vent and can be manufactured and sealed under vacuum
whereby the device can comprise a low internal gas pressure
relative to the external ambient pressure.
[0011] Additional features and advantages of various embodiments
will be set forth in part in the description that follows, and in
part will be apparent from the description, or can be learned by
practice of various embodiments. Other advantages of the various
embodiments will be realized and attained by means of the elements
and combinations exemplified in the application.
DRAWINGS
[0012] The skilled artisan will understand that the drawings,
described below, are for illustration purposes only. The drawings
are not intended to limit the scope of the present teachings in any
way.
[0013] FIGS. 1(a), 1(b), and 1(c) schematically illustrate various
stages in the operation of a valve according to various
embodiments.
[0014] FIG. 2 is a schematic illustration of a diagnostic device
according to various embodiments.
[0015] FIG. 3 is a schematic illustration of a diagnostic device
according to various embodiments.
[0016] FIGS. 4A-4J schematically illustrate various stages in the
operation of a diagnostic device according to an embodiment.
[0017] FIGS. 5A-5J schematically illustrate various stages in the
operation of a diagnostic device according to an embodiment.
[0018] FIG. 6 illustrates the assembly of a diagnostic device from
two separate pieces.
[0019] FIG. 7 illustrates the effect of misalignment of the pieces
shown in FIG. 6 for an embodiment of the device as shown in FIG.
3.
[0020] FIG. 8 illustrates the effect of misalignment of the pieces
shown in FIG. 6 for an embodiment of the device as shown in FIG.
2.
[0021] FIG. 9 illustrates an arrangement of retainment regions and
a valve according to various embodiments.
[0022] FIG. 10 illustrates an arrangement of retainment regions and
a valve according to various embodiments.
[0023] FIG. 11 illustrates an arrangement of retainment regions and
a valve according to various embodiments.
[0024] It is to be understood that both the foregoing general
description and the following detailed description are exemplary
and explanatory only and are intended to provide a further
explanation of the various embodiments of the present
teachings.
DESCRIPTION OF VARIOUS EMBODIMENTS
[0025] According to various embodiments, a diagnostic device, that
can be either portable or nonportable, is provided to perform one
or more predetermined assays as desired, for example, in nucleic
acid sequence detection technology. For a given assay, the assay
protocol can involve a set of fluid handling steps such as mixing,
incubation, washing, and the like, which are desirably performed in
a given sequence of steps and for specified time periods for
samples and reagents in specified volumes or proportions. The
device can be miniaturized to the point that it can be used as a
handheld portable diagnostic device. As shown in the exemplary
embodiments illustrated in FIG. 2 and FIG. 3, the diagnostic device
can include a plurality of fluid retainment regions, as exemplified
below with reference to retainment regions, interconnected by fluid
processing passageways. The plurality of retainment regions can be
included within a closed, disposable cuvette such that all of the
retainment regions are closed to fluid flow to or from locations
outside of the cuvette. The plurality of retainment regions in the
cuvette can include a reaction retainment region, one or more
reagent retainment regions, each containing unreacted reagents, and
a sample retainment region that can contain or receive a sample to
be reacted with the unreacted reagents. The unreacted reagents,
which can include enzymes, buffers, catalysts, or other reaction
components, can be contained in a first set of retainment regions
that are interconnected with an intermediate set of retainment
regions by fluid processing passageways containing
pressure-actuated valves. The intermediate retainment regions can
also be connected through fluid processing passageways to a
reaction retainment region, with the passageways that connect the
intermediate retainment regions to the reaction retainment region
comprising a material that is adapted to reduce in volume within
the fluid processing passageway when brought into contact with
fluids from the intermediate retainment regions, or when brought
into contact with a sample after the sample has been added to the
sample retainment region.
[0026] The term "fluid processing passageway" means any area, a
structure, or communication, that allows for fluid communication
between at least two fluid retainment regions, for example, a
channel connecting two regions. One or more fluid processing
passageways according to the present teachings can be configured or
adapted to provide capillary driven flow. One or more fluid
processing passageways according to the present teachings can be
configured or adapted to provide electrokinetic driven flow. One or
more of the fluid processing passageways according to the present
teachings can be configured or adapted to control the rate and
timing of fluid flow by varying the dimensions of the fluid
processing passageway.
[0027] The terms "fluid processing passageway," "a fluid
communication," "fluid flow channel," "fluid flow passageway,"
"flow channel," "flow control channel," and "flow control
passageway," are each used synonymous with the term "fluid
processing passageway," as herein defined.
[0028] According to various embodiments, the term "fluid" means a
gas, an aqueous fluid, a non-aqueous fluid, a vacuum, or a partial
vacuum. A gas can comprise, for example, air. For example, where
two retainment regions are separated by a fluid flow modulator, one
retainment region can comprise, for example, an aqueous or
non-aqueous fluid retained therein, while the other retainment
region can comprise a gas or a vacuum or partial vacuum, contained
therein.
[0029] The term "retainment region" means any area that can
comprise a reagent or other reaction component for a reaction where
the fluid retainment region is in fluid communication with, fully
separate from, or partially separate from, another retainment
region that can comprise another reagent or reaction component for
the reaction that is the same as or different from the first
reagent. A first retainment region can be separate from a second
retainment region, or a first retainment region can be surrounded
by a second retainment region, where the first and second
retainment regions are separated by a barrier comprising a
shaped-wall.
[0030] A retainment region can comprise any area, structure, or
form, capable of retaining a volume of fluid. A retainment region
can be used, for example, to retain, process, react, store,
incubate, transfer, purify, or the like, a fluid sample. A
retainment region can comprise a surface area, an area, a
reservoir, a recess, a fluid retainment region, a chamber, a
depression, a well, a space, or the like. According to some
embodiments, a retainment region can comprise, for example, a flat
surfaces with hydrophobic regions surrounding hydrophilic loci for
receiving, containing, retaining, or binding a sample. A retainment
region can comprise any shape, for example, round, teardrop,
square, polygon, star, irregular, ovoid, rectangular, or the like.
A retainment region or fluid processing passageway can comprise any
cross-section configuration, for example, square, round, ovoid,
irregular, trapezoid, or the like.
[0031] As used herein, the term "plurality" means "two or more."
Herein, the term "two or more" is used synonymously with the term
"plurality."
[0032] The terms "retainment region," "fluid retainment region,"
"reservoir," and "region," are used synonymously herein.
[0033] The term "reagent for reaction," means one or more reagents
or components necessary or desirable for use in one or more
reactions or processes, for example, one or more components that in
any way affect how a desired reaction can proceed. The reagent for
reaction can comprise a reactive component. However, it is not
necessary that the reagent participate in the reaction. The reagent
for reaction can comprise a non-reactive component. The reagent for
reaction can comprise a recoverable component comprising for
example, a solvent and/or a catalyst. The reagent for reaction can
comprise a promoter, accelerant, or retardant that is not necessary
for a reaction but affects the reaction, for example, affects the
rate of the reaction. The reagent for reaction can comprise one or
more of a solid reagent for reaction and a fluid reagent for
reaction. The term "reaction component" is used synonymous with the
term "reagent for reaction," as herein defined. The reagent for
reaction can comprise one or more of a fluid and a solid. A
retainment region can be pre-loaded with one or more reagents for
reaction.
[0034] The term "vent" means any configuration or structure that
relieves vacuum and/or back pressure, or equalizes pressure in a
fluid processing device. A vent can comprise a channel or a
microchannel. A vent can comprise a non-flow through vent in which
gas that is displaced by a fluid can collect. A non-flow through
vent can comprise, for example, a hydrophobic vent.
[0035] According to various embodiments, suitable reactions or
processes can comprise one or more of a sample preparation process,
a washing process, a sample purification process, a
pre-amplification process, a pre-amplified product purification
process, an amplification process, an amplified product
purification process, a separation process, a sequencing process, a
sequencing product purification process, a labeling process, a
detecting process, or the like. Processing components can comprise
sample preparation components, purification components,
pre-amplification reaction components, amplification reaction
components, sequencing reaction components, or the like. The
skilled artisan can readily select and employ suitable components
for a desired reaction or process, without undue
experimentation.
[0036] According to some embodiments, processing or reaction
components can be disposed in one or more retainment regions,
channels, or fluid processing passageways, using any methods known
in the art. For example, components can be sprayed and dried,
delivered using a diluent, injected using a capillary, a pipette,
and/or a robotic pipette, or otherwise disposed in the regions or
fluid processing passageways.
[0037] According to various embodiments, a fluid processing device
is provided that can comprise one or more fluid processing
passageways that can comprise one or more elements, for example,
one or more of a channel, a branch channel, a valve, a flow
splitter, a vent, a port, an access area, a via, a bead, a reagent
containing bead, a cover layer, a reaction component, any
combination thereof, and the like. Any element can be in fluid
communication with another element.
[0038] The term "fluid communication" means either direct fluid
communication, for example, two regions can be in fluid
communication with each other via an unobstructed fluid processing
passageway connecting the two regions or can be capable of being in
fluid communication, for example, two regions can be capable of
fluid communication with each other when they are connected via a
fluid processing passageway that can comprise a valve disposed
therein, wherein fluid communication can be established between the
two regions upon actuating the valve, for example, by dissolving a
dissolvable valve disposed in the fluid processing passageway.
[0039] The term "in fluid communication" refers to in direct fluid
communication and/or capable of direct fluid communication, unless
otherwise expressly stated. The term "in valved fluid
communication" refers to elements wherein a valve is disposed
between the elements, such that upon opening or actuating the
valve, fluid communication between the elements is established.
[0040] According to some embodiments, the term "capillary flow"
means passive flow resulting from a capillary potential gradient or
a surface potential gradient, created during device fabrication
that can direct the flow of liquid via capillary effect (surface
tension).
[0041] According to some embodiments a fluid processing device is
provided. The device can comprise a substrate that can comprise,
for example, a top or a first surface, and one or more fluid
processing passageways that can be provided in communication with
and/or can be defined by, for example, at least a portion of the
top or first surface of the substrate. The one or more fluid
processing passageways can be provided, for example, in a top or
first surface of a substrate, on a top or first surface of a
substrate, in a substrate, in a bottom or second surface of a
substrate, on a bottom or second surface of a substrate, in an edge
of a substrate, on an edge of a substrate, or any combination
thereof. A fluid processing device can comprise different levels
and layers of fluid processing passageways that can comprise, for
example, different levels and layers of channels and regions. For
example, a tiered, multi-channel device can comprise one or more
fluid processing passageways that traverse different heights or
levels in the substrate.
[0042] Throughout the application, descriptions of various
embodiments use "comprising" language; however, it will be
understood by one of skill in the art, that in some specific
instances, an embodiment can alternatively be described using the
language "consisting essentially" of or "consisting of."
[0043] For purposes of better understanding the present teachings
and in no way limiting the scope of the teachings, it will be clear
to one of skill in the art that the use of the singular includes
the plural unless specifically stated otherwise. Therefore, the
terms "a," "an" and "at least one" are used interchangeably in this
application.
[0044] Unless otherwise indicated, all numbers expressing
quantities, percentages or proportions, and other numerical values
used in the specification and claims, are to be understood as being
modified in all instances by the term "about." Accordingly, unless
indicated to the contrary, the numerical parameters set forth in
the following specification and attached claims are approximations
that may vary depending upon the desired properties sought to be
obtained. In some instances, "about" can be understood to mean a
given value .+-.5%. Therefore, for example, about 100 nl, could
mean 95-105 nl. At the very least, each numerical parameter should
at least be construed in light of the number of reported
significant digits and by applying ordinary rounding
techniques.
[0045] A user can operate the diagnostic device by injecting a
sample into the sample retainment region, prior to, at the same
time as, or subsequent to pushing a button or otherwise applying
pressure to the retainment regions that contain unreacted reagents.
For example, a user can inject a sample and then push a button or
other feature or area of the device. As an example of an assay
performed with a device according to various embodiments, a typical
ligation assay for detection of oligo-nucleotides can include
constituents comprising the sample, a ligation oligomer, ligation
reagent which can be a mixture of enzyme and buffer, a wash buffer,
and extension and detection reagents. The sample, the ligation
oligomer, and the ligation reagent can be allowed to mix and react
along with wash buffers and the extension and detection reagents in
an automatically controlled process. The process can occur after a
user has injected the sample into the sample retainment region and
has released the reagents from the unreacted reagent retainment
regions by applying pressure to those retainment regions.
[0046] According to various embodiments, a diagnostic device can be
provided that uses capillary driven flow for fluid actuation. The
flow cross-section of the fluid processing passageways
interconnecting the various retainment regions can contribute to
the rate at which the reagents and sample are mixed in the reaction
retainment region. Valves placed within the flow control fluid
processing passageways interconnecting the retainment regions can
provide automatic flow control and timing of the fluid
actuation.
[0047] According to various embodiments, a fluid flow modulator, as
exemplified below with reference to a valve in a flow control
passageway interconnecting retainment regions, can comprise a
material that dissolves when brought into contact with a fluid
having desired characteristics. Herein, the phrase "size-changing
valve" will be used interchangeably with the phrase "solute bridge
valve." The solute bridge valve can automatically control flow
through the fluid processing passageway interconnecting the
retainment regions and control the timing of fluid actuation by
exploiting the time it takes to dissolve, melt, otherwise wash-away
or reduce the volume, or swell or increase the volume, of the
material making-up the solute bridge valve.
[0048] According to various embodiments, a fluid processing device
comprises a flow passageway, a plurality of retainment regions, at
least two of the retainment regions each being in fluid
communication with the flow passageway, and a fluid flow modulator
arranged in the flow passageway. The fluid flow modulator can be
adapted to form, or modify the size of, a fluid communication
between the at least two retainment regions. The fluid flow
modulator can comprise a polysaccharide, for example, a
water-soluble polysaccharide. The fluid flow modulator can comprise
a valve that blocks fluid flow through the flow passageway. The
fluid flow modulator can be adapted to open, or to increase the
size of, the fluid communication between the at least two
retainment regions. The fluid flow modulator is adapted to close,
or to decrease the size of, the fluid communication between the at
least two retainment regions.
[0049] According to various embodiments, the term "fluid" means a
gas, an aqueous fluid, a non-aqueous fluid, a vacuum, or a partial
vacuum. A gas can comprise, for example, air. Where two retainment
regions are separated by a fluid flow modulator, one retainment
region can comprise, for example, an aqueous or non-aqueous fluid
retained therein, while the other retainment region can comprise a
gas, or a vacuum or partial vacuum, contained therein. In various
embodiments, the device can be manufactured to provide a vacuum on
one or more sides of a dissolvable valve, for example, to achieve a
pressure of from about 0.01 atmospheres (atm) to about 0.99 atm, or
from about 0.1 atm to about 0.5 atm.
[0050] According to various embodiments, at least one of the
plurality of retainment regions can comprise an aqueous fluid
retained therein. The aqueous fluid can have a pH of about 9.5 or
lower. The aqueous fluid can have a pH of about 6.5 or lower.
[0051] According to various embodiments, in the fluid processing
device, the fluid flow modulator can comprise a water-soluble
cellulose derivative. The fluid flow modulator can comprise
poly(D-glucosamine), a derivative of poly(D-glucosamine), or a
combination thereof. Poly(D-glucosamine) is the chemical name of
"chitosan" which is a common name arising from its original source,
i.e., chitin from shellfish. The terms "poly(D-glucosamine)" and
"chitosan" are used synonymously herein. The fluid flow modulator
can comprise poly(D-glucosamine) derivatized with poly(ethylene
glycol). The fluid flow modulator can comprise a graft polymer of
poly(D-glucosamine) and poly(methoxy polyethyleneglycol
(meth)acrylate). The fluid flow modulator can comprise a graft
polymer of poly(D-glucosamine) and poly(methoxy polyethyleneglycol
(meth)acrylamide). The fluid flow modulator can comprise
poly(D-glucosamine) substituted with hydrophilic groups. According
to various embodiments, the fluid flow modulator can comprise at
least one of N-[(2-hydroxy-3-trimethylammonium)propyl]
poly(D-glucosamine), N,O-carboxyalkyl poly(D-glucosamine),
N-ethoxylated poly(D-glucosamine), and N-benzylsulfonated
poly(D-glucosamine). The fluid flow modulator can comprise an
N-substituted poly(D-glucosamine) derived from a reductive
alkylation of at least one of an aldehydro sugar and a keto
sugar.
[0052] According to various embodiments, the fluid flow modulator
can comprise an N-substituted poly(D-glucosamine) derived from a
reductive alkylation of at least one of D-glucose,
N-acetylglucosamine, D-glucosamine, D-galactose, D-galactosamine,
D-fructose, D-glucoheptonic acid gamma-lactone, lactose,
cellobiose, maltose, melibiose, maltotoriose, streptomycin sulfate,
C6-aldehydro-cycloheptamylose, dextran, and combinations
thereof.
[0053] According to various embodiments, the fluid flow modulator
can comprise at least one of methylcellulose, hydroxymethyl
cellulose, ethylcellulose, hydroxypropyl methylcellulose,
quarternized hydroxyethylcellulose ethoxylate,
(2-hydroxyethyl)-beta-cyclodextrin, carboxymethylcellulose, a
sodium salt of carboxymethylcellulose, a potassium salt of
carboxymethylcellulose, m-aminobenzyloxymethyl cellulose,
cyanoethylated cellulose, diethylaminoethyl cellulose, and
diethyl-(2-hydroxypropyl)aminoethyl cellulose.
[0054] According to various embodiments, the fluid processing
device further comprises a liquid retained in at least one
additional retainment region. The liquid retained in the at least
one additional retainment region can have a pH of about 9.5 or
lower. The liquid retained in the at least one additional
retainment region can have a pH of about 6.5 or lower.
[0055] According to various embodiments, the fluid processing
device comprises a flow passageway dimensioned so that a flow of
fluid from at least one of two or more retainment regions and
through the flow passageway, can occur by capillary action. One or
more maximum dimensions of about 5 mm or less, for example, about 2
mm or less, or about 1 mm or less.
[0056] According to various embodiments, the fluid processing
device comprises a flow passageway dimensioned so that a migration
of charged components in a fluid, from at least one of the
retainment regions through the flow passageway, is capable of
migration by electrokinetic action. One or more maximum dimensions
of about 5 millimeter or less, for example, about 2 millimeters or
less, or about 1 millimeter or less.
[0057] According to various embodiments, the fluid processing
device can comprise at least two electrodes disposed in the device
with a flow passageway therebetween. A system can be provided that
includes electrical leads that can be electrically connected to the
electrodes.
[0058] According to various embodiments, the fluid processing
device can further comprise at least one additional retainment
region, at least one additional flow passageway, and at least one
pressure-actuatable valve arranged in the at least one additional
flow passageway. The additional flow passageway can be in fluid
communication with the additional retainment region and one or more
other retainment regions. The pressure-actuatable valve can
comprise a frangible diaphragm. The frangible diaphragm can
comprise a material that is insoluble in water at room temperature.
The pressure-actuatable valve can comprise a burstable valve that
is adapted to open and establish fluid communication only upon
receiving pressure of at least about 0.1 psig, for example, at
least about 0.5 psig, at least about 1 psig, or at least about 3
psig from a fluid in at least one additional retainment region. The
device can comprise a liquid retained in at least one additional
retainment region.
[0059] According to various embodiments, the fluid processing
device can comprise at least one heat-actuatable valve arranged in
at least one additional flow passageway. The at least one
additional flow passageway can be in fluid communication with at
least one additional retainment region and at least one of the
plurality of retainment regions. The heat-actuatable valve can
comprise at least one material selected from a rubber, a plastic, a
wax, a paraffin, a polyethylene glycol material, a derivative of a
polyethylene glycol material, a polysaccharide, a derivative of
polysaccharide, and combinations thereof. The heat-actuatable valve
can comprise a material that is insoluble in water at room
temperature. The heat-actuatable valve can comprise a material that
has a melting point of from about 35.degree. C. to about 95.degree.
C., for example, from about 35.degree. C. to about 70.degree. C.,
from about 35.degree. C. to about 65.degree. C., or from about
35.degree. C. to about 50.degree. C.
[0060] According to various embodiments, the fluid processing
device further comprises a liquid retained in the at least one
additional retainment region.
[0061] According to various embodiments, the fluid processing
device can comprise a liquid retained in at least one retainment
region. The fluid processing device can comprise a first reagent
for a reaction, retained in at least a first one of the plurality
of retainment regions. The fluid processing device can comprise a
second reagent for the reaction retained in at least a second one
of the plurality of retainment regions. The second reagent can be
the same as, or can differ from, the first reagent. According to
various embodiments, the fluid processing device further comprises
a plurality of flow passageways.
[0062] According to various embodiments, the fluid processing
device can comprise a plurality of fluid flow modulators, wherein
each fluid flow modulator can comprise a polysaccharide or a
derivative of a polysaccharide. Each fluid flow modulator can be
adapted to dissolve when contacted with an aqueous solution at room
temperature and having a pH of 9.5 or lower. Each fluid flow
modulator can be adapted to dissolve when contacted with an aqueous
solution at room temperature and having a pH of 6.5 or lower.
[0063] According to various embodiments, the fluid processing
device can comprise a flow passageway, and a valve arranged in the
flow passageway, being adapted to open to form a fluid
communication through the flow passageway. The valve can be adapted
to dissolve when contacted with an aqueous solution at room
temperature and having a pH of 9.5 or lower, for example, 6.5 or
lower. The valve can comprise poly(D-glucosamine), a derivative of
poly(D-glucosamine), or a combination thereof. The valve can
comprise poly(D-glucosamine) derivatized with poly(ethylene
glycol).
[0064] According to various embodiments, a system can comprise the
fluid processing device, a power source, and at least two
electrical leads forming electrical connections, respectively,
between the power source and the at least two electrodes. The
system can comprise the fluid processing device and a pump, wherein
the pump is arranged in fluid communication with at least one of
the flow passageway and one or more of the plurality of retainment
regions. The system can comprise the fluid processing device and a
magnet, wherein the magnet generates a magnetic field and said
fluid processing device is arranged at least partially within said
magnetic field.
[0065] According to various embodiments, instead of or in addition
to a fluid flow modulator, a barrier can be provided that at least
partially separates a first retainment region from a second
retainment region. The barrier can comprise a polysaccharide or a
derivative of a polysaccharide. The barrier can comprise any of the
polysaccharide or polysaccharide derivatives described above that
can be used for the fluid flow modulator.
[0066] According to various embodiments, the barrier can have any
of a variety of shapes. For example, the barrier can comprise an
outer wall having the shape of a polygon or a ring-shape.
[0067] The barriers, flow modulators, and valves described herein
can have the shapes, sizes, components, chemicals, and other
properties described with reference to the barriers, flow
modulators, and valves described in U.S. Provisional Patent
Application No. 60/619,677, to Banerjee et al., in U.S. Provisional
Patent Application No. 60/619,731, to Banerjee et al., and in U.S.
Provisional Patent Application No. 60/619,623, to Faulstich et al.,
all filed on Oct. 18, 2004, and all of which are incorporated
herein in their entireties by reference.
[0068] According to various embodiments, a method is provided that
comprises providing a fluid processing device comprising at least a
first retainment region and a second retainment region, and a
barrier arranged at least partially between the first retainment
region and the second retainment region. The barrier comprises a
polysaccharide or a derivative of a polysaccharide, and the method
can comprise contacting the barrier with a solvent, such as an
aqueous solution, suspension, mixture, or other material, to change
the shape of the barrier. Contacting the barrier with the solvent
can form, or change the size of, a fluid communication between the
first retainment region and the second retainment region.
Contacting the barrier with the solvent can open, or increase the
size of, a fluid communication between the first retainment region
and the second retainment region. Contacting the barrier with the
solvent can close, or decrease the size of, a fluid communication
between the first retainment region and the second retainment
region.
[0069] According to various embodiments, a method is provided that
comprises processing a fluid processing device that comprises at
least a first retainment region and a second retainment region, and
a barrier arranged between them. At least one of the first and
second retainment regions retains an aqueous solution. The barrier
can comprise at least one of a chitosan material, a chitosan
derivative material, and a combination thereof. The barrier is
adapted to swell or dissolve when contacted with the aqueous
solution. According to various embodiments, the method includes
contacting the barrier with the aqueous solution to dissolve at
least a portion of the barrier and form, or increase the size of, a
fluid communication between the first retainment region and the
second retainment region. The fluid processing device can comprise
at least one additional retainment region, at least one flow
passageway, and at least one pressure-actuatable valve arranged in
the at least one flow passageway. The at least one flow passageway
can be in fluid communication with the at least one additional
retainment region and at least one of the first retainment region
and the second retainment region. The method can comprise opening
the pressure-actuatable valve. The pressure-actuatable valve can
comprise a diaphragm and the method can comprise bursting the
diaphragm by applying pressure to the diaphragm. A heat-actuatable
valve can be actuated instead of, or in addition to, actuation of a
pressure-actuatable valve.
[0070] According to various embodiments, the method can comprise
migrating charged components in a sample from at least one of the
at least two retainment regions, through the flow passageway, by
electrokinetic motion. Migration of the charged components can be
accomplished by creating an electric field in the device. A system
can be provided that includes an electric field generator.
[0071] According to various embodiments, a method can comprise
creating a pressure differential between a first retainment region
and a second retainment region, and moving, with the pressure
differential, a fluid from one of the first retainment region and
the second retainment region into the other of the first retainment
region and the second retainment region. The pressure differential
can be generated by activating a pump. The pressure differential
can comprise a positive-pressure differential or negative-pressure
differential. A positive pressure means a pressure at or greater
than atmospheric pressure, i.e., 1 atm. A negative pressure means a
pressure less than atmospheric pressure, i.e. less than 1 atm.
[0072] According to various embodiments, the method can comprise
creating a magnetic field across a first retainment region and a
second retainment region, and moving, with the magnetic field,
magnetically attractable materials from one of the retainment
regions toward the other retainment region.
[0073] According to various embodiments, the method can comprise
performing a set of predetermined assays in a plurality of
retainment regions, for example, retainment regions, in a closed,
disposable device. An exemplary device is a cuvette. The retainment
regions can be interconnected by fluid processing passageways but
closed to fluid flow to or from locations outside of the cuvette.
The first retainment regions can be selectively closed-off from
fluid communication with second retainment regions through first
channels that interconnect them. Selective closing-off can be
provided by pressure-actuated valves positioned in the first
channels. The second retainment regions can be interconnected to
third retainment regions by second channels. Flow through the
second channels can be controlled by fluid flow modulators
positioned in the second channels, which can also provide selective
closing-off. The method can comprise applying pressure to a
pressure-actuated valve in a first channel sufficient to break the
valve and provide fluid communication between the first and second
retainment regions. Such a method can be used to introduce a sample
for testing or other processing into one or more third retainment
regions and/or establishing fluid communication between the second
retainment regions and one or more third retainment regions, at a
controlled rate. The controlled rate can be a function of
characteristics of at least one of, a fluid in a third retainment
region and a fluid within the second retainment regions.
[0074] According to various embodiments, a system is provided that
comprises a fluid processing device as described herein, and a
pump, wherein the pump is arranged in fluid communication with at
least one of a flow passageway and one or more retainment
regions.
[0075] A system can be provided that comprises a fluid processing
device as described herein, a power source, and at least two
electrical leads forming electrical connections, respectively,
between the power source and the at least two electrodes. A system
can be provided that comprises a fluid processing device as
described herein, and a magnet, wherein the magnet generates a
magnetic field and the fluid processing device is arranged at least
partially within the magnetic field.
[0076] Exemplary devices and methods according to various
embodiments are described below with reference to the drawings. The
present teachings are not limited to the embodiments depicted in
the drawings.
[0077] Referring to FIGS. 1(a)-1(c), a schematic illustration of
the process by which a solute bridge valve establishes fluid
communication between two passageways, according to various
embodiments, is shown. FIG. 1(a) shows two retainment regions 20,
24 containing fluids 30, 32, respectively, with the two retainment
regions 20, 24 being interconnected through a flow fluid processing
passageway 26. A solute bridge valve 22 can consist of a plug of
material that completely or partially blocks the flow fluid
processing passageway 26 and separates the fluids 30 and 32, e.g.
air, a gas, solvents, reagents, a vacuum or partial vacuum, or
other materials, in the respective retainment regions 20, 24. In
some embodiments, one or both of fluids 30 or 32, can, at least
initially, comprise a gas, a vacuum, a partial vacuum, and/or a
pressurized fluid.
[0078] FIG. 1(b) shows the size of the solute bridge valve 22
decreasing as the polysaccharide material that makes up the solute
bridge valve gradually dissolves into one or both of the fluids 30,
32 in retainment regions 20, 24. FIG. 1(c) shows that the fluids
30, 32 have come into contact with each other when the material
making up solute bridge valve 22 has completely dissolved. The time
required to completely dissolve the solute bridge valve 22 can be
determined by the cross-sectional area and/or shape of the
capillary flow passage 26 and the length and dissolvability of the
solute bridge valve 22. By controlling the material and/or size of
the solute bridge valve 22, it is also possible to control the
length of the time elapsed before the solute bridge valve has
completely dissolved to allow mixing of the fluids 30, 32.
[0079] It is desirable for the material that makes up the valve 22
to be a material that is compatible with the assay, and would not
adversely affect the assay condition. One example of a material
that could be used to make up the valve 22 is a polysaccharide or a
polysaccharide derivative, for example, a water-soluble
polysaccharide. An exemplary polysaccharide is poly(D-glucosamine)
or chitosan. Chitosan can be highly responsive, with a film of
chitosan varying in thickness in response to a pH change, by a
factor of 3. The source material for chitosan (chitin) is extremely
abundant in nature which makes it inexpensive. The chitosan valve
can also be manufactured by electrodeposition or by electrostatic
atomization or electrospray ionization deposition. The geometry of
a fluid flow modulator or valve produced using an electic technique
can be controlled by the shape of a metallic electrode used and the
process time. Chitosan is quite inert and does not affect
biological assays in general.
[0080] Chitosan can also be used for the feature that it will
disassociate into ionic components under acidic pH conditions,
spontaneously dissolving in an acidic solution up to pH of 9.5. The
pH of a dissolving solvent can be tailored to affect the rate of
dissolution of the chitosan. At more basic conditions, for example,
pH greater than 9.5, chitosan does not dissolve.
[0081] According to various embodiments, a pH responsive valve can
be made from a pH responsive material that partially or completely
separates two retainment regions, for example, retainment regions,
in a diagnostic device. The pH responsive material can be a
material that changes volume, (i.e., shrinks if its volume
decreases, or swells if its volume increases) in response to a
change in the local pH. Flow control through a fluid processing
passageway connecting the two regions can be achieved by the change
in the open cross-sectional area of the fluid processing passageway
between the two regions, subsequent to the change in volume of the
pH-responsive material. The actuation of the valve can consist of
the volumetric change of the material resulting from a change in pH
or other characteristics of a solution or other material contacting
the valve. If the material making up the valve completely blocks
the fluid processing passageway, the two regions can be completely
separated and the valve can be closed. When the pH of a liquid
within a region crosses a certain threshold value pH, to cause
shrinking of the pH-responsive material, the cross-sectional area
of the respective fluid processing passageway can open, and the
valve can be in an open state whereby fluids can flow through the
fluid processing passageway. The reverse operation is also
possible, that is, by adjusting or providing the pH of the liquid
in contact with the valve, the valve can be made to swell and
change from an open state to a closed state.
[0082] Several materials can be used to perform the above-described
function according to various embodiments. Chitosan is particularly
amendable to microfluidic applications, including bio-microfluidic
applications, as a result of several beneficial characteristics.
Chitosan can have a high responsiveness to variations in pH, with a
film of chitosan varying in thickness in response to a change in pH
by a factor of three. A valve made from a material such as chitosan
that can change volume in response to a change in pH can also be an
extremely simple valve that does not require moving parts and that
has a high level reliability. Furthermore, because the actuation of
the valve can occur as a result in a local change in pH, no
external energy sources are required. The source material used in
preparing chitosan, is extremely abundant in nature and therefore
is inexpensive.
[0083] According to various embodiments, beside pH, a change in
volume of the material can result from other characteristics of the
solution or solutions, such as temperature, water content, chemical
composition, electrical charge, magnetic properties, or the like.
If the material making up the valve completely blocks the fluid
processing passageway, the two regions can be completely
separated.
[0084] FIG. 2 shows an embodiment of an assay device that uses
capillary driven flow and solute bridge valves to control a
specific sequence of fluid actuation steps. The device shown in
FIG. 2 can be constructed as a microfluidic chip manufactured using
microfabrication techniques. According to various embodiments, the
chip can comprise a set of retainment regions and microchannels for
housing different liquids such as reagents, samples, etc., and for
mixing and reacting the various liquids. FIG. 2 shows a device
according to various embodiments having a sample retainment region
90 connected through a fluid processing passageway 80 to a reaction
retainment region 48. An intermediate retainment region or
retainment regions is/are connected through a fluid processing
passageway 170 containing a valve 70 to the reaction retainment
region 48, and a second intermediate retainment region 46 is
connected through a fluid flow passage 172 containing a valve 72 to
the reaction retainment region 48. A first reagent retainment
region 40 for containing unreacted reagents 120 can be connected
through a fluid processing passageway 160 containing a valve 60 to
one intermediate retainment region 44, while a second reagent
retainment region 42 containing unreacted reagents 124 can be
connected through a fluid processing passageway 162 containing a
valve 62 to the second intermediate retainment region 46. The
reaction retainment region 48 is connected through a fluid
processing passageway 174 containing a valve 74 to a first waste
retainment region 50, which can also be connected through another
fluid processing passageway 176 containing a valve 76 to a second
waste retainment region 52.
[0085] Reagent retainment regions 40, 42 connected by fluid
processing passageways 160, 162 with valves 60, 62 to the
intermediate retainment regions 44, 46 can be selectively separated
from the intermediate retainment regions 44, 46 by the pressure
actuated valves 60, 62 placed within the fluid processing
passageways 160, 162. According to various embodiments, the
pressure actuated valves 60, 62 within fluid processing passageways
160, 162 can be diaphragms that are burstable upon pressure being
applied to the reagent retainment regions 40, 42.
[0086] The intermediate retainment regions 44, 46 can be in turn
connected through the fluid processing passageways 170, 172
containing valves 70, 72 to the reaction retainment region 48.
Fluid communication through the fluid processing passageways 170,
172 containing valves 70, 72 can be controlled by the fluid flow
cross-sectional area of the passages 170, 172 as well as the
positioning of solute bridge valves 70, 72 as discussed above,
within the fluid processing passageways 170, 172. The solute bridge
valves 70, 72 contained within the fluid processing passageways
170, 172 can provide automatic control of the fluid communication
between the intermediate retainment regions 44, 46 and the reaction
retainment region 48 as a result of their responsiveness to stimuli
such as the chemical composition of the fluids within retainment
regions 44, 46 and within reaction retainment region 48. Each of
regions 90, 48, 40, 42, 50, and 52, can optionally comprise a vent
91, 49, 41, 43, 51, and 53, respectively.
[0087] FIG. 3 illustrates an embodiment wherein the retainment
regions and fluid processing passageways are arranged such that
solute bridge valves 70a, 72a, 74a, 76a corresponding to solute
bridge valves 70, 72, 74, 76 of the embodiment shown in FIG. 2, are
arranged in a line for ease of manufacture. Each of regions 90a,
48a, 40a, 42a, 50a, and 52a, can optionally comprise a vent 91a,
49a, 41a, 43a, 51a, and 53a, respectively.
[0088] FIGS. 4A-4J illustrate a sequence of events that can occur
during operation of a diagnostic device according to various
embodiments, such as exemplified in FIG. 2. Each of regions 90, 48,
40, 42, 50, and 52, can optionally comprise a vent 91, 49, 41, 43,
51, and 53, respectively.
[0089] FIGS. 5A-5J illustrate a sequence of events that can occur
during operation of a diagnostic device according to various
embodiments, such as exemplified in FIG. 3. Each of regions 90a,
48a, 40a, 42a, 50a, and 52a, can optionally comprise a vent 91a,
49a, 41a, 43a, 51a, and 53a, respectively.
[0090] In FIG. 4A a diagnostic device according to various
embodiments is provided with retainment regions 40, 42 prefilled
with, for example, a wash buffer 124 in retainment region 42 and a
detection reagent 120 within retainment region 40. Pressure
actuated valves 60, 62, such as burstable or tearable diaphragms,
can be provided within the fluid processing passageways 160, 162
separating retainment regions 40, 42 from intermediate retainment
regions 44, 46.
[0091] In FIG. 4B a user can apply pressure to the retainment
regions 40, 42, thereby actuating or bursting the valves 60, 62
within fluid processing passageways 160, 162 connecting to the
intermediate retainment regions 44, 46 and causing flow of the
buffers and/or reagents within the retainment regions 40, 42 into
intermediate retainment regions 44, 46.
[0092] As shown in FIG. 4C a user can inject a sample 126 into the
sample retainment region 90, which is connected to the reaction
retainment region 48 through a fluid processing passageway 80.
fluid processing passageways 170, 172 and 80 can be provided with
fluid flow modulators, exemplified below with reference to a valve,
and in particular a solute bridge valve, such as a plug of material
that can change volume when exposed to certain stimuli. The solute
bridge valves can control the fluid communication between
intermediate retainment regions 44, 46 and reaction retainment
region 48, as well as the fluid communication between the sample
retainment region 90 and the reaction retainment region 48. Fluid
flow of the sample from sample retainment region 90 into reaction
retainment region 48 can also be automatically controlled as a
result of the dimensions of the fluid processing passageway 80. The
fluid processing passageway 80 can be provided as a capillary
passage such that the sample material from sample retainment region
90 gradually wicks into the reaction retainment region 48, without
the need for a solute bridge valve to control this flow through
fluid processing passageway 80. Pressure can be relieved or
equalized via vent 91 and/or 49.
[0093] As shown in FIG. 4D, the sample material that is now in
reaction retainment region 48 contacts the solute bridge valve 72
on one side of the valve 72 in fluid processing passageway 172,
while the reagent in intermediate retainment region 46 contacts the
solute bridge valve 72 from the opposite side of the valve. One or
more of the reagent in retainment region 46 and/or the sample in
reaction retainment region 48 begin to dissolve or otherwise affect
the volume of the material making up the solute bridge valve 72.
After a certain amount of time that is automatically controlled by
the flow cross-section of passage 172 as well as by the volume of
material within the solute bridge valve 72, the solute bridge valve
72 in passage 172 allows the reagent in retainment region 46 to
gradually diffuse into the sample 126 in reaction retainment region
48, as shown in FIG. 4E. Pressure can be relieved and/or equalized
via vent 43.
[0094] The flow passage 174 leading from the reaction retainment
region 48 into a waste retainment region 50 can also be provided
with the dimensions that allow for capillary action, and a solute
bridge valve 74 that will gradually dissolve or otherwise change
volume as a result of contact with the fluid from reaction
retainment region 48. As shown in FIG. 4F, the effect of the fluid
within reaction retainment region 48 on the solute bridge valve 74
within flow passage 174 gradually opens the flow passage 174 within
which the valve 74 is positioned to allow fluid communication
between the reaction retainment region 48 and the first waste
retainment region 50. The flow of fluid from reaction region 48
into waste region 50 through fluid processing passageway 174
contributes to a capillary flow of more reagent from reagent region
42 through fluid processing passageway 162 and intermediate region
46 into reaction region 48. Pressure resulting from such flow can
be relieved via vent 43 and/or vent 49. Flow of fluid from reaction
region 48 into waste region 50, as shown in FIG. 4G, can also cause
more of sample 126 to flow from sample region 90 into reaction
region 48. Pressure resulting from the flow of fluid from reaction
region 48 into waste region 50, can be relieved via vent 49 and/or
vent 51. According to various embodiments, the relative dimensions
of the flow passages such as flow passage 80 leading from sample
retainment region 90 into reaction retainment region 48, and the
flow passage 172 within which valve 72 is positioned leading from
intermediate retainment region 46 into reaction retainment region
48 can be selected in order to contribute to a preferential flow of
fluid from the intermediate retainment region 46 into reaction
retainment region 48. A smaller flow cross-section through passage
80 than the flow cross-section through passage 172 would result in
more fluid flowing from the reagent retainment region 42 and
intermediate retainment region 46 into reaction retainment region
48 than the amount of sample flowing from sample retainment region
90 into the reaction retainment region 48.
[0095] After a predetermined amount of time, solute bridge valve 70
provided in the flow passage 170 between intermediate retainment
region 44 and reaction retainment region 48 can also begin to
dissolve, melt, or otherwise change in volume such that reagent 120
flows from reagent retainment region 40 through intermediate
retainment region 44 and into the reaction retainment region 48, as
shown in FIG. 4H. The relative cross-sectional flow areas of the
various flow passages connecting retainment regions as well as the
amount of material provided in the solute bridge valves within the
flow passages can be varied in order to control the amount of time
it takes for the reagents and other fluids within the retainment
regions to move from one retainment region to the next, thereby
providing a control of the fluid handling steps.
[0096] After more time has passed, solute bridge valve 76 in flow
passage 176 leading to a second waste retainment region 52 can
begin to dissolve, melt, or otherwise change in volume such that
fluid can flow from waste retainment region 50 into second waste
retainment region 52, as shown in FIGS. 4I and 4J. This flow can
cause more of the reagents and sample to flow from regions 44, 46
and 90 into reaction region 48. Pressure can be relieved via vent
49 and/or vent 91.
[0097] In an alternative embodiment, as exemplified in FIG. 3 and
FIGS. 5A-5J, the diagnostic device can comprise a set of retainment
regions and microchannels corresponding to the retainment regions
and microchannels of the embodiment exemplified in FIG. 2 and FIGS.
4A-4J, but with the solute bridge valves 72a, 70a, 74a, and 76a
being aligned so that they can be formed as a single, extended
length of the solute bridge valve material. The process by which
mixing of buffer and/or reagent from retainment regions 40a, 42a,
and sample from sample retainment region 90a is controlled
automatically corresponds with the process described above for the
embodiment shown in FIG. 2 and FIGS. 4A-4J.
[0098] A sample solution can be added to sample retainment region
90a, and supplied to a reaction retainment region 48a through a
capillary flow passage 80a, as shown in FIGS. 5C and 5D. Solutions,
such as reagents and/or wash buffers, can be dispensed from
retainment regions 40a, 42a by pressing on the clear layer of film
or other flexible covering over the retainment regions to cause
pressure-actuated valves 60a, 62a in passageways 160a, 162a to
burst and allow the solutions to move into intermediate retainment
regions 44a, 46a. Solution from intermediate retainment region 46a
then begins to act on solute bridge valve 72a in fluid processing
passageway 172a, opening up a passageway for the solution to enter
passage 80a and reaction retainment region 48a, as shown in FIGS.
5D, 5E, 5F and 5G. FIG. 5H illustrates solution from retainment
region 44a dissolving, or otherwise reducing the volume of solute
bridge valve 70a in passageway 170a. Solute bridge valve 74a in
passageway 174a has also dissolved in FIG. 5H to allow solution
from the reaction retainment region 48a to pass to a waste
retainment region 50a. As described above with regard to the
embodiment of FIG. 2, the flow of solution from reaction retainment
region 48a to the waste retainment region 50a creates a suction
that draws more solution from retainment region 46a through
passageway 172a and passageway 80a into the reaction retainment
region 48a, as shown in FIG. 5I. Solute bridge valve 76a in
passageway 176a then dissolves, allowing solution to flow to waste
retainment region 52a, and creating suction that draws solution
from retainment region 44a into reaction retainment region 48a, as
well as drawing additional solution from retainment region 46a and
additional sample from retainment region 90a, as shown in FIG. 5J.
For example, vents 41 and 43 are needed in 40 and 42, respectively,
(FIG. 4A) in order to allow 120 and 124 to flow into 44 and 46,
respectively. Likewise, vent 91 is needed in communication with
region 90. A vent 49 can be provided in communication with regions
48 (FIG. 4D) such that 120, 124 and 126 can flow into it. Without a
vent, the air trapped in 48 can prevent any inflow of liquid. The
same can apply to regions 50 and 52.
[0099] The arrangement of retainment regions, passageways and
valves of the various embodiments exemplified in FIG. 3 provides
for ease of manufacturing. As shown in FIG. 3 and FIGS. 5A-5J, the
solute bridge valves 70a, 72a, 74a, and 76a, can be aligned with
each other such that the solute bridge valves can be formed as one
length of material. The diagnostic device 130 shown in FIG. 6
exemplifies an embodiment wherein the solute bridge valves are
formed as one extended length of solute bridge valve material 270
in a substrate 134 separate from a substrate 132, within which
various retainment regions such as retainment regions 140 and 142
are formed. When the two substrates 132 and 134 are sandwiched
together to form the device 130, the length of solute bridge valve
material 270 must connect to passageways 260, 262, 264, and 266, as
shown in FIG. 7, which passageways are connected to various
retainment regions. As illustrated in FIG. 7, even if the two
substrates 132, 134 are not perfectly aligned, the length of solute
bridge valve material 270 will still connect with the passageways
260, 262, 264, and 266. FIG. 8 illustrates a situation wherein
separate substrates for solute bridge valves 370, 372 and
passageways 360, 362, 364, and 366, are not perfectly aligned when
forming a device as exemplified in the embodiment of FIG. 2. In
this situation the solute bridge valves 370, 372 may not connect to
passageways 360, 362, and 364, 366, as desired.
[0100] Referring to FIG. 9, and according to various embodiments,
retainment regions 444, 446, and 448, can be formed in a substrate
440, with retainment region 444 interconnected with retainment
region 448 through a passage 470, and retainment region 446
interconnected with retainment region 448 through a passage 472.
Solution such as reagents and/or wash buffers can be retained in
the retainment regions 444 and 446 by a flexible sheet of material
460 applied over the top surface of substrate 440 and adhered to
the top surface by an adhesive layer 462. Pressure actuated valves
470a, 472a can be positioned in the passageways 470, 472 such that
pressure applied to the solutions in retainment regions 444, 446 by
pressing down on the flexible sheet 460 over the respective
retainment regions will dispense the solutions through passages
470, 472 into the retainment region 448. A barrier 450 can define
an inner retainment region 430 and provide an automatically
controlled interaction between the solution or solutions 442 in the
retainment region 448 and a solution retained in the inner
retainment region 430. If desired, the barrier 450 can be formed
from a solute material such as chitosan that will gradually
dissolve and thereby control the interaction between the solution
or solutions 442 in outer retainment region 448 and a solution
retained in the inner retainment region 430.
[0101] According to various embodiments, and as exemplified by the
embodiment shown in FIG. 10, a device can comprise a substrate 540
having a retainment region 542 formed in the substrate and covered
by a sheet 560 that is adhered to the top surface of the substrate
540 by an adhesive 562. An inner retainment region 530 can be
defined within the retainment region 542 by a barrier 550 that can
act as a fluid flow modulator between a solution in the outer
retainment region 542 and a solution in the inner retainment region
530. The barrier 550 can comprise a portion 552 made from a soluble
material, and a portion 554 made from an insoluble material to
provide a further degree of automatic control of the interaction
between the solutions or other ingredients in retainment regions
542 and 530.
[0102] According to various embodiments, and as exemplified in the
embodiment shown in FIG. 11, a substrate 700 can be provided with a
retainment region 740 connected through a passage 770 having a
pressure actuated valve 772 to a second retainment region 742.
Similarly, another retainment region 740a can be connected through
a passage 770a having a pressure actuated valve 772a to the second
retainment region 742. A star-shaped or otherwise polygonal
retainment region 730 can be defined inside of the second
retainment region 742 by a barrier 750. All of the retainment
regions can be covered by a sheet 760 adhered to the top surface of
substrate 700 by an adhesive layer 762. A solution within
retainment region 740 can be dispensed through passage 770 by
applying pressure to the sheet 760 over the retainment region 740
to force the liquid past the pressure actuated valve 772 into the
second retainment region 742. The barrier 750 can comprise a
material such as chitosan that gradually dissolves or melts in
response to a stimuli such as characteristics of the solution that
has been introduced to the second retainment region 742, thereby
acting as a fluid flow modulator and providing an automatically
controlled interaction between the solution in second retainment
region 742 and the solution in retainment region 730.
[0103] According to various embodiments, further control of the
fluid handling steps can be provided by including various solute
structures within the fluid processing passageways and/or the
retainment regions. The solute structures can be selected to
dissolve over a finite amount of time and change the flow
properties of the fluidic circuit. As an example, raised structures
(such as pillars of different aspect ratios) made from solute
material (such as chitosan) can be fabricated by photolithography
inside the various retainment regions and/or flow passageways. The
incorporation of these structures can cause the flow paths to have
different capillarity and can cause capillary suction pressures of
different magnitudes in different parts of the fluidic circuit. The
structures can also introduce additional flow resistance, with a
variation in the flow resistance depending on the dissolution of
the solute structures.
[0104] In one example, an array of pillars made of chitosan could
be fabricated inside of the waste retainment regions 50, 52, in the
embodiment of FIG. 2, or 50a, 52a in the embodiment of FIG. 3,
which could, for example, cause higher suction pressure in waste
retainment region 50 or 50a by capillary action as compared to the
suction pressure into reaction retainment region 48 or 48a. Over
time, the solute structures within waste retainment region 50 or
50a would dissolve in the liquid, which could result in the
capillary suction pressure into retainment region 50 or 50a
reducing over time. Subsequent dissolution of a solute within flow
passage 176 between waste retainment region 50 and waste retainment
region 52, for example, could then result in the liquid in waste
retainment region 50 being pulled into waste retainment region 52
as a result of a larger capillary suction pressure in waste
retainment region 52 caused by solute structures in retainment
region 52. The pulling can be as a result of a larger capillary
suction pressure in waste retainment region 52 caused by solute
structures in retainment region 52. The chitosan cannot operate
until 74 and 76 in FIG. 2 and 170a, 172a and 174a in FIG. 3 are
open. The flow of 120, 125 and 126 in FIG. 2 into 44, 46, and 48,
respectively, relies on capillary effect alone, and does not rely
on vacuum created by the chitosan in 50 or 52 because neither 76
nor 74 are open. Chitosan can facilitate fluid flow from 48 into 50
and/or 52 without a vent. The flow of 120, 124 or 126 into 44, 46,
or 48, respectively, relies on capillary effect that requires air
vents to prevent pressure build up.
[0105] Those skilled in the art can appreciate from the foregoing
description that the present teachings can be implemented in a
variety of forms. Therefore, while these teachings have been
described in connection with particular embodiments and examples
thereof, the true scope of the present teachings should not be so
limited. Various changes and modifications can be made without
departing from the scope of the teachings herein.
* * * * *