U.S. patent application number 10/775004 was filed with the patent office on 2004-08-12 for container for holding biologic fluid for analysis.
This patent application is currently assigned to Levine, Robert A.. Invention is credited to Wardlaw, Stephen C..
Application Number | 20040156755 10/775004 |
Document ID | / |
Family ID | 26759028 |
Filed Date | 2004-08-12 |
United States Patent
Application |
20040156755 |
Kind Code |
A1 |
Wardlaw, Stephen C. |
August 12, 2004 |
Container for holding biologic fluid for analysis
Abstract
A container for holding a biologic fluid sample for analysis is
provided which includes a chamber and a label. The chamber includes
a first wall, a transparent second wall, and a plurality of
features including features spatially located within the chamber.
The transparent second wall permits a fluid sample quiescently
residing within the chamber to be imaged through the second wall.
The plurality of features, including those spatially located within
the chamber, are operable to enable the analysis of the biologic
fluid. The label directly or indirectly contains information
regarding the features and the spatial location of the features
within the chamber. The sample is analyzed by an analytical device
that utilizes the information communicated through the label.
Inventors: |
Wardlaw, Stephen C.; (Old
Saybrook, CT) |
Correspondence
Address: |
McCormick, Paulding & Huber LLP
CityPlace II
185 Asylum Street
Hartford
CT
06103-3402
US
|
Assignee: |
Levine, Robert A.
31 Pilgrim Lane
Guilford
CT
|
Family ID: |
26759028 |
Appl. No.: |
10/775004 |
Filed: |
February 9, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10775004 |
Feb 9, 2004 |
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09256486 |
Feb 23, 1999 |
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6723290 |
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60077214 |
Mar 7, 1998 |
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Current U.S.
Class: |
422/400 |
Current CPC
Class: |
B01L 2300/089 20130101;
B01L 3/50 20130101; G01N 2035/00851 20130101; G01N 35/00732
20130101; B01L 2400/086 20130101; B01L 3/502746 20130101; G01N
35/00029 20130101; B01L 2400/0633 20130101; B01L 3/502738 20130101;
G01N 2001/282 20130101; B01L 9/56 20190801; B01L 2400/0406
20130101; B01L 3/545 20130101 |
Class at
Publication: |
422/102 |
International
Class: |
B01L 003/00 |
Claims
I claim:
1. A container for holding a biologic fluid sample for analysis,
said container comprising: a fluid holding chamber having a first
wall and a transparent second wall, and wherein the fluid holding
chamber has a mapped interior so that positions within the fluid
holding chamber are identifiable by a coordinate address; at least
one feature operable to enable a determination of the volume of a
field of the fluid sample, the at least one feature located within
the chamber at a predetermined coordinate address; and a label
attached to said container, said label operable to supply the
predetermined coordinate address within the fluid holding
chamber.
2. The container of claim 1, wherein the interior of the chamber is
orthogonally mapped.
3. The container of claim 2, wherein the at least one feature
operable to enable a determination of the volume of a field of the
fluid sample includes a through-plane thickness, and the field of
the fluid sample is located at the predetermined address.
4. The container of claim 2, wherein the at least one feature
operable to enable a determination of the volume of a field of the
fluid sample includes a geometric characteristic.
5. The container of claim 4, wherein the geometric characteristic
comprises a step of known height disposed in one or both of the
first wall and second wall.
6. The container of claim 4, wherein the geometric characteristic
comprises a cavity of known height or volume disposed in one or
both of the first wall and second wall.
7. The container of claim 4, wherein the geometric characteristic
comprises a protruberance of known height or volume disposed in one
or both of the first wall and second wall.
8. The container of claim 4, wherein the geometric characteristic
comprises an object of known volume.
9. A container for holding a biologic fluid sample for analysis,
said container comprising: a fluid holding chamber having a first
wall and a transparent second wall, and wherein the fluid holding
chamber has a mapped interior so that positions within the fluid
holding chamber are identifiable by a coordinate address; wherein
the fluid holding chamber has at least a first through-plane
thickness and and a second through-plane thickness, extending
between the first wall and the second wall, wherein the first
through-plane thickness and the second through-plane thicknesses
are each located within the chamber at predetermined coordinate
addresses, and the first through-plane thickness is greater than
the second through-plane thickness; and a label attached to the
container, which label is operable to supply the predetermined
coordinate addresses of the first through-plane thickness and the
second through-plane thickness within the fluid holding
chamber.
10. The container of claim 9, wherein the interior of the chamber
is orthogonally mapped.
11. The container of claim 10, wherein the at least a first
through-plane thickness and the second through-plane thickness are
relatively sized to enable iterative performance of an analysis of
a fluid sample quiescently residing within the fluid holding
chamber.
12. A container for holding a biologic fluid sample for analysis,
said container comprising: a fluid holding chamber having a first
wall and a transparent second wall, and wherein the fluid holding
chamber has a mapped interior so that positions within the fluid
holding chamber are identifiable by a coordinate address; wherein
the fluid holding chamber has: a first through-plane thickness
extending between the first wall and the second wall, wherein the
first through-plane thickness is located within the chamber at a
first predetermined coordinate address; and a second through-plane
thickness extending between the first wall and the second wall,
wherein the second through-plane thickness is located within the
chamber at a second predetermined coordinate addresses; a first
reagent located within the chamber at the first predetermined
coordinate address; a second reagent located within the chamber at
the second predetermined coordinate address; wherein the first
predetermined coordinate address and the second predetermined
coordinate address are separated from each other within the chamber
by a distance great enough such that an analysis of each of the
reagents with the fluid sample can be completed without
interference from the other of the reagents; and a label attached
to the container, which label is operable to supply the
predetermined coordinate addresses of the first through-plane
thickness and the second through-plane thickness within the fluid
holding chamber.
13. A container for holding a biologic fluid sample for analysis,
said container comprising: a fluid holding chamber having a first
wall and a transparent second wall, and wherein the fluid holding
chamber has a mapped interior so that positions within the fluid
holding chamber are identifiable by a coordinate address; means
operable to enable a determination of the volume of a field of the
fluid sample, the means located within the chamber at a
predetermined coordinate address; and a label attached to said
container, said label operable to supply the predetermined
coordinate address within the fluid holding chamber.
14. The container of claim 13, wherein means operable to enable a
determination of the volume of a field of the fluid sample includes
a through-plane thickness, and the field of the fluid sample is
located at the predetermined address.
15. The container of claim 13, wherein the means operable to enable
a determination of the volume of a field of the fluid sample
includes a geometric characteristic.
16. The container of claim 15, wherein the geometric characteristic
comprises a step of known height disposed in one or both of the
first wall and second wall.
17. The container of claim 15, wherein the geometric characteristic
comprises a cavity of known height or volume disposed in one or
both of the first wall and second wall.
18. The container of claim 15, wherein the geometric characteristic
comprises a protruberance of known height or volume disposed in one
or both of the first wall and second wall.
19. The container of claim 15, wherein the geometric characteristic
comprises an object of known volume.
Description
[0001] This application is a continuation of, and claims priority
under 35 U.S.C. .sctn.120, U.S. patent application Ser. No.
09/256,486 filed Feb. 23, 1999, and also claims the benefit of U.S.
Provisional Patent application serial No. 60/077,214 filed Mar. 7,
1998.
BACKGROUND OF THE INVENTION
[0002] 1. Technical Field
[0003] The present invention relates to apparatus for analyzing
biologic fluid samples in general, and to containers for holding a
biologic fluid sample during analytical procedures in
particular.
[0004] 2. Background Information
[0005] Most analytical methods for evaluating constituents within a
biologic fluid sample require that the sample be substantially
diluted prior to evaluation. A typical chemical analysis, for
example, involves placing a substantially diluted sample into a
transparent cuvette of known dimensions and constant light path for
evaluation. The cuvette can be made from glass or a hard acrylic
that is ground or otherwise manufactured to tight tolerances. The
tight tolerances, which are necessary to insure the accuracy of the
light path through the cuvette, also make the cuvette undesirably
expensive. In hematological analyses, a substantially diluted
sample is typically passed through a flow cell within an optical
flow cytometer or through an impedance orifice in an impedance type
flow cytometer. Most flow cytometers require mechanical subsystems
to dilute the sample, to control the sample flow rate through the
flow cell, and multiple sensors to evaluate the diluted sample. A
special problem associated with hematology measurements is the wide
dynamic range of particles that must be enumerated. The red blood
cells (RBC's) are the most numerous at about 4.5.times.10.sup.6
RBC's per microliter (.mu.l), followed by the platelets at about
0.25.times.10.sup.6 platelets per .mu.l, and the white blood cells
(WBC's) at about 0.05.times.10.sup.6 per .mu.l. Since all cells or
particles must be enumerated during a full analysis, the range of
cells/particles necessitates at least two dilution levels. The
ability to perform multiple dilutions undesirably adds to the
complexity of the machine. A person of skill in the art will
recognize disadvantages associated with flow cytometers including
plumbing leaks and inaccuracies due to fluid control
miscalibration. In both of the aforementioned analyses, the
operator (or the apparatus itself) must purge the biologic fluid
sample from the apparatus and thoroughly clean the apparatus to
avoid contaminating subsequent analyses. The substantial dilution
required in both analyses also increases the likelihood of error,
the complexity of the analysis, and the per analysis cost.
[0006] Other analytical methods minimize the above described
problems by employing a disposable sample analytical chamber. In
one chemical analytical method, the biologic fluid sample is placed
in a flexible sealed pouch where it remains during the analysis.
This approach avoids the need for plumbing, flow controls, and
cleaning the container, but requires a large diluent volume and is
restricted to standard measurements of light transmission. In the
above method, the light path dimensions are controlled by the
analytical instrument, which forms the flexible pouch into a
cuvette of the desired thickness at the time of measurement. Other,
similar "wet chemical", systems employ a rigid analytical cuvette
of specifically manufactured thickness. Other methods for
performing a chemical analysis on a biologic fluid sample employ
single or multiple test film substrates. The test film substrates
also avoid the problems associated with dilution, flow controls,
etc., but still require precise sample measurement and placement
and are also limited to those analyses that employ light
reflectance. The test film substrate methods are further limited by
requiring that the associated disposable always have identically
located analytical regions; if the desired information is not
present in the predetermined analytical areas, then the test film
substrate will not yield useful information. Hematological
analytical methods which employ a disposable sample analytical
chamber include the HemaCue.TM. and the QBC.TM.. The HemaCue.TM.
system is a method for measuring hemoglobin using a small cuvette.
The HemaCue.TM. method is particularly useful for its intended
purpose, but it is unable to measure particulate constituents of
whole blood. The QBC.TM. system, a registered trademark of Becton
Dickinson and Company of Franklin Lakes, N.J., USA involves placing
a hematological fluid sample within a cylindrical tube and
centrifuging the tube and sample for a given period of time. The
centrifuge process separates fluid sample constituents into layers
according to their density. A float disposed within the tube
facilitates evaluation of constituents within each layer. Specific
hematological tests may be performed in a disposable test system
employing a scannable optical window in a device produced by
Biometric Imaging. In this device, a substantially undiluted sample
of whole blood is placed into a capillary of known and constant
dimension where it is subjected to a laser scan which identifies
some sub-types of WBC's. The Biometric Imaging method is also
limited in that it is unable to measure any other constituents of
whole blood.
[0007] Serologic or immunologic analyses measure soluble substances
in blood, usually proteins such as specific immunoglobulins. These
tests are often performed by admixing the sample with a sensitized
particulate, such as latex, which will agglutinate in the presence
of the protein of interest. Another method for performing a more
quantitative immunological analysis is to use enzymatically linked
color changes, such as ELISA. All of these methods are performed on
apparatus specialized for their use.
[0008] Another common specialized test is urinalysis. The analysis
of urine is generally divided into two separate phases: the
determination of the bulk and/or chemical properties of the sample
and the analysis of particulates within the sample. These analyses
require distinctly different disciplines and are usually done
separately. There are large and complicated machines that can
perform both types of analyses, but they are extremely expensive
and require moderate maintenance and operator skill.
[0009] None of the above described analytical methods is capable of
performing hematological, chemical/immunochemical, and serologic
analyses on sample constituents within the same instrument. As a
result, it has been necessary to purchase apparatus devoted to
performing chemical analyses and apparatus devoted to performing
hematological analyses. It has also been necessary to train
technicians to operate the various types of apparatus, and to
provide laboratory space and maintenance for them. It has also been
impossible to combine hematological and chemical analyses in the
same apparatus for those analyses where it would be advantageous to
combine the analyses. In an analysis to determine anemia, for
example, it is preferable to perform both hematological analyses
(e.g., hemoglobin, hematocrit, and reticulocyte count) and chemical
or immunochemical analyses (e.g., iron or ferritin, and/or vitamin
B12 or folate determinations) on the sample. None of the above
described methods permit hematological and chemical analyses on a
single sample of blood in a single disposable sample chamber. As a
result, the laboratory technician must separate and transport the
various samples to their separate instruments which are often in
separate laboratories, thereby increasing the inefficiency of the
process as well as the potential for loss or misidentification of
the sample. Also, the results of the analyses may not be available
at the same time which increases the difficulty of interpreting the
analysis results.
[0010] What is needed is a single container for holding a biologic
fluid sample that can be used for multiple analyses including but
not limited to hematological, chemical, immunological, serological,
and urine analyses, one in which multiple analyses can be performed
on the same sample in one instrument which presents a common
operator interface, one that is operable with substantially
undiluted biologic fluid samples, one whose method of sample
introduction into the container is similar for each set of
analyses, and one that can be used effectively as a disposable.
DISCLOSURE OF THE INVENTION
[0011] It is, therefore, an object of the present invention to
provide a container for holding a biologic fluid sample which
permits analysis of multiple constituents within the same sample,
and in particular analysis of constituents residing individually or
in groups, using quantitative image analysis.
[0012] It is another object of the present invention to provide a
container for holding a biologic fluid sample for analysis which is
operable for analyses which require information related to the bulk
and/or chemical properties of the sample and those which require
information related to the particulates content of the sample.
[0013] It is another object of the present invention to provide a
container for holding a biologic fluid for analysis which is
operable in multiple analytical disciplines including but not
limited to hematology, chemical/immunochemical,
serology/immunological, and urinalysis.
[0014] It is another object of the present invention to provide a
container which can include analytical chambers of varying
dimensions whose thickness can be correlated to a spatial
coordinate in order to encompass a wide dynamic range of contained
particulates.
[0015] It is another object of the present invention to provide a
container for holding a biologic fluid sample that requires only a
single instrument to sense for information within the sample or
associated with the container and interpret the sensed information
for use in multiple analyses, thereby decreasing the training and
quality control requirements of the laboratory.
[0016] It is another object of the present invention to provide a
container for holding a biologic fluid sample for analysis that can
be used effectively as a disposable.
[0017] It is another object of the present invention to provide a
container for holding a biologic fluid sample for analysis that
does not require substantial dilution of the sample before
analysis.
[0018] It is another object of the present invention to provide a
container for holding a biologic fluid sample which is operable
with minimal quantities of blood or other biologic fluid.
[0019] It is another object of the present invention to provide a
container for holding a biologic fluid sample that facilitates safe
handling of the biologic fluid sample for the test operator.
[0020] It is another object of the present invention to provide a
container for holding a biologic fluid sample that includes an
analytical area suitable for imaging by a digital camera or other
digital imaging device/image dissector which produces output
suitable for quantitative analysis.
[0021] It is another object of the present invention to provide a
container for holding a biologic fluid sample that has the
capability of retaining an untreated or a substantially undiluted
sample prior to analysis and releasing said sample into the
analytical region when needed.
[0022] It is another object of the present invention to provide a
container for holding a biologic fluid sample that carries indicia
which provides information to the instrument of use in performing
the analysis(es) at hand.
[0023] According to the present invention, a container for holding
a biologic fluid sample for analysis is provided which includes a
chamber and a label. The chamber includes a first wall, a
transparent second wall, and a plurality of features including
features spatially located within the chamber. The transparent
second wall permits a fluid sample quiescently residing within the
chamber to be imaged through the second wall. The plurality of
features, including those spatially located within the chamber, are
operable to enable the analysis of the biologic fluid. The features
may, for example, include regions where chemical constituents are
analyzed, chambers of varying height and size (i.e., physical
characteristics) where particulates may be analyzed, and regions,
which allow the calibration or quality control of the analysis. The
features may also include information that is useful to set-up,
adjust, and/or calibrate the analytical device to the task at hand;
e.g., filter alignment, lens adjustment, etc. The label directly or
indirectly contains information regarding the features and the
spatial location of the features within the chamber. The sample is
analyzed by an analytical device that utilizes the information
communicated through the label.
[0024] The preferred analytical device for use with the present
invention container is the subject of co-pending U.S. patent
application Ser. No. 09/255,673. Briefly described, the "Apparatus
for Analyzing Substantially Undiluted Samples of Biologic Fluids"
as it is referred to, includes a Reader Module, a Transport Module,
and a Programmable Analyzer. The Reader Module includes optics
which are operable to image a field within the container, and
apparatus to access information through the label attached to the
container. The Transport Module includes apparatus for moving the
container relative to the Reader Module, or vice versa. The
Programmable Analyzer is programmed with instructions to coordinate
the operation of the Reader Module and Transport Module according
to a variety of analysis algorithms. Which analysis algorithms are
used is typically determined by reading the container label.
[0025] An advantage of the present invention container is that it
is operable for a variety of analyses including but not limited to
hematological, chemical, immunochemical, serologic, urinalysis and
immunological analyses. In addition, it is possible to perform a
multitude of those analyses on the same sample, in the same
analytical device. Some traditional analysis methods pass light
into a cuvette, and interpret the light traversing through or
emitting from the cuvette to provide analytical data. Other
methods, such as those which utilize film substrates for analyzing
sample constituents utilize light reflected from the film layer.
The data available using these types of methods is relatively
uniform and does not contain any spatial information. Thus, they
are useful for analyzing bulk properties of the sample, meaning
those properties that are distributed uniformly in solution or in
suspension, but it is impossible to derive useful data about
individual particulate materials within the sample. The absence of
spatial information limits the number of tests possible on a given
sample. If a sample is tested for optical density using the above
described cuvette, for example, the test parameters will provide
information about a particular constituent, but will not provide
the information necessary to characterize cellular contents. The
present invention container, in contrast, includes an analytical
chamber (or chambers) which includes features that enable the
Reader Module to extract both spatial information and quantitative
photometric information from the sample quiescently residing within
the chamber. The ability to analyze both types of information
allows the combination of the instrument and the disposable to
analyze a large number of different constituents, and consequently
perform a far greater number of tests.
[0026] The ability of the present container to include one or more
chambers operable for a variety of analyses enables the performance
of a select battery of tests on a particular sample. A person of
skill in the art will recognize that it is common to perform a
battery of tests on a patient sample, and that there is great
utility in being able to perform that battery on a single sample in
a single analytical device. For example, when a patient's blood
sample is being analyzed to determine the hemoglobin concentration,
it is usual to measure the hematocrit and white blood count and may
be useful to enumerate the reticulocytes. Using the present
invention, a chamber having the features of a known or determinable
through-plane thickness and appropriate reagents would be used to
measure the hemoglobin, and another chamber having the features of
a dispersed colorant would be used to measure the hematocrit and
white blood count. Another section of that chamber, or a separate
chamber, would have the features of a colorant plus a region of
very thin through-plane thickness where the reticulocytes would be
enumerated. In the case of a battery of tests for the detection of
a myocardial infarction, there could be a chamber having features
operable to determine treponin and/or myoglobin, a chamber with
features for determining potassium concentration, one that analyzed
the creatinine phosphokinase levels, etc. An advantage of the
present invention is that the chambers and features may be located
anywhere within the confines of the container so long as the
features are locatable by the analytical device, which allows for a
test or series of tests to be designed using the best chamber and
feature geometry for the particular analyses on a particular sample
without being limited to a chamber region of a predetermined shape
or size at a particular location.
[0027] The ability to perform different discipline analyses, for
example hematological and chemical analyses, is significant for
several reasons. First, the amount of equipment required to do the
same number of analyses is reduced significantly. It follows that
the cost of procuring and maintaining that equipment is similarly
reduced. Also, the personnel training required to operate the
equipment is reduced. Another reason is the versatility provided by
a device that can perform different discipline analyses. Many
clinical offices and laboratories are presently unable to justify
the office space and expense associated with available test
apparatus for each analytical discipline. With the versatile
present invention, however, it will be possible to have greater
in-house analytical ability because of the present invention's
relative minimal space requirements and low cost.
[0028] Another advantage of the present invention is that a
disposable container for holding, analyzing, and disposing of a
biologic fluid sample for analysis is provided. The present
invention container is independent of the analytical device,
inexpensive, readily loaded, and easily handled by an automated
analytical device. The disposable container utilizes a standardized
exterior configuration so that the analytical device can be set up
for a standard container, regardless of the configuration of the
chamber or chambers. In the example above where the container
includes one or more chambers having features operable to enable a
select battery of tests, the analytical device is set up to accept
the standard container and the container label indicates directly
or indirectly the tests to be perform and the feature or features
within the chamber or chambers that enable those tests. These
characteristics make the present container a desirable disposable.
As a disposable, the present invention obviates the need to clean
the sample chamber after each use and therefore the opportunity for
contamination from the prior sample. The disposable nature of the
present invention container also facilitates safe handling of the
biologic fluid sample for the test operator by minimizing contact
with all fluids.
[0029] Another advantage of the present invention container is that
it uses a relatively small volume of biologic fluid rather than a
large volume of significantly diluted biologic fluid. A person of
skill in the art will readily recognize the advantages of avoiding
the plumbing and fluid controls associated with most flow
cytometers which require relatively large volume of diluted sample,
as well as the advantages of avoiding the dilution steps, the
dilution hardware, and the need for diluent.
[0030] Another advantage of the present invention is that it can
hold an untreated or substantially undiluted sample prior to
analysis and selectively release that sample into the analytical
region when needed. As a result, those analyses which are time
dependent can be performed using the present invention.
[0031] These and other objects, features and advantages of the
present invention will become apparent in light of the detailed
description of the best mode embodiment thereof, as illustrated in
the accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0032] FIG. 1 is a diagrammatic view of the present invention
container.
[0033] FIG. 2 is a sectional view of the container shown in FIG. 1,
sectioned along line A-A.
[0034] FIG. 3 is a sectional view of the container shown in FIG. 1,
sectioned along line B-B.
[0035] FIG. 4 is the sectional view of FIG. 3, showing the valve
actuated open.
[0036] FIG. 5 is a diagrammatic view of a present invention
container having two chambers.
[0037] FIG. 6 is a diagrammatic illustration of a field within the
chamber.
[0038] FIG. 7 is a diagrammatic view of a chamber.
[0039] FIGS. 8A-8F are sectioned diagrammatic views of chambers
having a variety of features.
[0040] FIG. 9 is a diagrammatic view of a present invention
container having two chambers.
[0041] FIG. 10 is a diagrammatic view of a chamber.
BEST MODE FOR CARRYING OUT THE PRESENT INVENTION
[0042] Referring now to FIGS. 1-4, a container 10 for holding a
biologic fluid sample includes at least one chamber 12, a label 14,
a reservoir 16, a channel 18, and a valve 20. The container 10
holds a biologic sample in a manner that enables analysis of the
sample by an analytical device (not shown) as will be described
below. The container 10 embodiment shown in FIGS. 1-4 includes a
first piece 22 and a second piece 24 snapped together. The chamber
12 includes a first wall 26 disposed in the first piece 22 and a
transparent second wall 28 held between the first piece 22 and
second piece 24. An opening 27 disposed within the second piece 24
exposes the transparent second wall 28, and thereby provides visual
access into the chamber 12. In some embodiments, the first wall 26
may also be transparent thereby enabling light to pass through the
container 10 by way of the chamber 12. The chamber 12 has a
through-plane thickness ("t") at any given point. FIG. 6 shows a
diagrammatic illustration of a field within a chamber to better
illustrate the relationship between volume and through-plane
thickness. As used herein, the term "through-plane thickness"
refers to a line of sight which corresponds to the shortest
distance between the interior chamber surface 30 of the first wall
26 and the interior chamber surface 32 of the second wall 28. The
reservoir 16 typically holds 50 .mu.l of biologic fluid sample and
preferably includes a cap 34 for sealing the reservoir 16 and a
mixing element 36, such as a ball, that operates to keep the sample
in uniform suspension. The channel 18 extends between the reservoir
16 and the chamber 12. The valve 20 operates between the reservoir
16 and the chamber 12 to selectively allow passage of fluid from
the reservoir 16 to the chamber 12. As used herein, the term
"valve" includes not only a structure that includes a movable part
that selectively prevents flow, but also any structure that
selectively permits flow. The valve 20 shown in FIGS. 1 and 3-5
includes a pair of slits 38 adjacent the reservoir 16 is operated
by a rod 40 which is a part of the analytical device. The slits 38
allow the rod 40 to separate the reservoir 16 a small distance from
the first piece 22, thereby providing an opening through which
biologic fluid can pass through the channel 18 and into the chamber
12. The optimum valve 20 type will vary depending upon the
application. In those embodiments where there is more than one
chamber 12 (see FIG. 5), each chamber 12 is in communication with
the reservoir 16 via a channel 18. The reservoir 16 and valve 20
provide considerable utility for analyses where time is a
consideration as will be described below. In some instances,
however, it may be advantageous to provide a container 10 without a
reservoir 16 and/or a valve 20.
[0043] Referring to FIGS. 1,5, and 7, each container 10 includes a
plurality of features which are operable to enable the analysis of
the biologic fluid sample, some of which are located in the chamber
12. The features located within the chamber 12 are spatially
located, each having an address describable, for example, in x,y,z
coordinates. The advantage of an x,y,z type coordinate address
system is that the chamber can be mapped in an x,y,z, grid with a
locatable origin that orients the analytical device relative to the
container. The phrase "operable to enable the analysis of the
biologic fluid" is used to describe the fact that the features
either directly or indirectly provide information that enables the
analytical device to provide useful analytical information. To
explain further, most analyses require either the volume or the
through-plane thickness of the sample be known. The term "volume"
as used herein will refer to this requirement since the volume of a
given image field of view can be ascertained using the
through-plane thickness or vice versa; e.g., when the sample is
imaged using a fluorescent light source, it is the volume of the
field that provides the useful information directly since
fluorescent signal is a function of colorant per unit volume, or
when a light absorption technique is used for imaging, the volume
of the field indirectly provides the necessary useful information,
since absorption is a function of the through-plane thickness of
the field (i.e., the distance the light travels through the
sample). The through-plane thickness can be readily determined from
the sensed volume and the known field area of the analytical
device. Features disposed within the chamber can be used to
operably enable a means for determining the volume of one or more
select fields within the sample.
[0044] Referring to FIGS. 3,4, and 7, in a first embodiment of the
means for determining the volume of one or more fields within the
sample, the first wall 26 and second wall 28 of the chamber 12, or
a portion thereof, are in fixed relationship to one another and the
slope values for each wall 26,28 and a chamber 12 through-plane
thickness are known and are communicated to the analytical device
through the label 14 (the label 14 is discussed in detail below).
The possible configurations of the walls 26,28 (or a portion of the
walls 26,28) include parallel walls (i.e., slope=0) separated by a
known amount, and walls 26,28 which are at an angle toward one
another (i.e., a slope.noteq.0), separated by a known amount.
Hence, in this embodiment features that operate to enable the
analysis include a known through-plane thickness at one or more
known locations within the chamber, particularly when different
through-plane thickness' are used to facilitate the analysis. In
this embodiment, the through-plane-thickness may also be referred
to as a type of physical characteristic, and physical
characteristics are a type of feature.
[0045] A second embodiment of the means for determining the volume
of one or more select fields within the sample includes a known
quantity of sensible colorant for mixture with a known volume of
biologic fluid sample. As used herein, the term colorant is defined
as any reagent that produces a sensible signal by fluorescent
emission, or by absorption of light at a specific wavelength, that
can be quantified by the analytical device. The colorant has a
known signal magnitude to colorant concentration ratio that is
communicated to the analytical device through the label 14. The
colorant concentration is fixed by virtue of a known volume of
biologic fluid sample being added to a known quantity of colorant.
Alternatively, the signal magnitude to colorant concentration is
determinable by comparison with a second known material such as a
pad 44 of material (hereinafter referred to as a "calibration
pad"--see FIG. 1) with stable characteristics which is referenced
by the analytical device and used to calibrate the response of the
colorant. If the colorant signal is sensed in a particular field
via the analytical device, then the volume of that field can be
calculated using the magnitude of the sensed signal and the known
concentration of colorant within the sample. Features that are
operable to enable analysis of a biologic fluid with this
embodiment include chamber regions where particular analyses are
best-performed, located at known spatial locations. The chamber 12
regions where an analysis is best performed refers to those chamber
regions having physical characteristics such as a particular
through-plane thickness that allow for discrimination of particular
constituents within the sample. For example, a chamber
through-plane thickness of about 25 microns is known to be
favorable for the formation of rouleaux and lacunae within a sample
of whole blood. The absence of RBC's in the lacunae makes each
lacunae a favorable region to accurately sense colorant signal.
Hence, the feature of a chamber region(s) having an approximate
through-plane thickness at a particular location(s) within the
chamber is used by the analytical device to increase its
probability of being able to accurately determine the volume of one
or more fields in that region, which volume in turn is used
directly in the analysis at hand. In the examples hereafter, the
significance of accurately knowing the volume of a field within the
lacunae of a whole blood sample will be clearly shown. The
analytical device contains means for identifying which features,
and therefore the information available with those features, should
be used in particular analyses.
[0046] Referring to FIGS. 1 and 8A-8F, a third embodiment of the
means for determining the volume of one or more select fields
includes: 1) a quantity of colorant uniformly dispersed within the
biologic fluid; 2) one or more geometric characteristics (i.e., a
type of feature) within the chamber 12; 3) chamber 12 regions where
particular analyses are best performed; and 4) spatial information
locating the geometric characteristic(s) and optimum region(s) for
the analytical device. In this embodiment, it is not necessary to
know the amount of sensible colorant within the sample, nor the
total volume of the sample. Rather, the field volume determination
is done on a comparative basis. A field containing no geometric
characteristic is sensed and compared against a field containing a
known geometric characteristic. Examples of geometric
characteristics include, but are not limited to, a step 46 of known
height within one or both walls 26,28, a cavity 48 or protuberance
50 of known height or volume in one or both walls, or an object 52
of known volume. The known volume of the object 52, cavity 48, or
protuberance 50 (or volume which is determinable from a step of
known height and the cross-sectional area of the field which is
known to the analytical device) displaces a known volume of sample.
Since the signal from the sensible colorant is a function of sample
volume, the difference in signal sensed between the two fields is
attributable to the sample volume displaced by the geometric
characteristic. Hence, a signal to sample volume ratio can be
calculated, and applied to the whole field to ascertain the volume
of the field. Like the second embodiment of the means for
determining the volume of a sample field, the chamber 12 regions
where an analysis is best performed refers to chamber regions
having physical characteristics that allow for discrimination of
particular constituents within the sample. The spatial information
locating the geometric characteristic(s) and optimum region(s) for
the analytical device also refers to a chamber 12 coordinate system
wherein each feature within the chamber 12 has a coordinate
address. The analytical device contains means for identifying which
features, and the information available with those features, should
be used in particular analyses. In this embodiment, the features
that are operable to enable the analysis of a biologic fluid sample
include the one or more geometric characteristics and chamber
regions where particular analyses are best performed, located at
known locations within the chamber.
[0047] A fourth embodiment of the means for determining the volume
of one or more select fields includes a chamber 12 having specular
surfaces on which a virtual reflected image may be detected by the
analytical device. The specular surfaces are the two wall surfaces
30,32 in contact with the biologic fluid, or the outer surfaces if
the wall thicknesses are known. The analytical device detects the
virtual reflected image on one of the specular surfaces 30,32 and
then refocuses on the virtual reflected image formed on the second
surface 32,30. The distance the analytical device's optics must
move between the two images is the through-plane thickness of the
chamber 12 in the particular field. The label 14 communicates the
coordinate addresses of the select fields within the chamber 12 to
the analytical device.
[0048] Referring to FIG. 8E, in the second and third embodiments of
the means for determining the volume of one or more select fields,
one or both of the first or second walls 26,28 may be formed from a
flexible material that will deflect a determinable amount due to
capillary forces presented by the sample acting on the wall 26,28,
and thereby form a desirable convergent relationship between the
first wall 26 and the second wall 28.
[0049] Referring to FIG. 7, for chemical/immunochemical analyses of
a biologic fluid sample, the features operable to enable the
analysis of a biologic fluid sample include a plurality of
different chemical reagents 54, each located at a particular
coordinate address within the chamber 12, and may also include
chamber 12 regions where particular analyses are best performed and
coordinate addresses locating those optimum regions. In a first
embodiment, a known quantity of each chemical reagent 54 is
disposed at a particular coordinate address, usually in the form of
a coating bound to one of the chamber walls 26,28. When the
biologic fluid sample is introduced into the container chamber 12,
the biologic sample admixes with each reagent 54. The fluid sample
may be contiguous in those regions, but there is no appreciable
reagent mixing between adjacent regions for a period of time
because of the chamber configuration. Specifically, although the
rates of diffusion vertically and laterally are equal, the chamber
12 through-plane thickness (which may be described as a physical
characteristic of those regions) is small enough relative to the
possible lateral expanse that the chemical reagent 54 will diffuse
vertically and reach equilibrium at a much faster rate than it will
laterally. In fact, because vertical diffusion reaches equilibrium
much faster than lateral diffusion, lateral diffusion may be
considered negligible for a short period of time. The lateral
spacing between the addresses of the different chemical reagents 54
is such that during that short period of time in which lateral
reagent diffusion is negligible, useful analysis of any reaction
that may be occurring at a particular address can be performed. The
coordinate addresses of the various chemical reagents 54 enable the
analytical device to access each reagent 54 and perform meaningful
analyses. In those instances where chemical and hematological
analyses are desirable, the above described chamber configuration
can be provided in a particular region of a single chamber 12 and
other configurations provided elsewhere within that chamber 12. The
negligible lateral diffusion of the reagent 54 prevents
interference with contiguous chamber 12 regions which may be
devoted to other type analyses. Alternately, the different reagent
regions may be partially or completely isolated in subcompartments
of the chamber by means of intervening partitions 55 formed within
one or both of the chamber 12 surfaces (see FIG. 8F).
[0050] Referring to FIGS. 1 and 5, the label 14 is a mechanism for
communicating information to the analytical device. A practical
example of a label 14 is one which is machine readable and one
which is capable of communicating information including, but not
limited to: 1) type of analysis(es) to be performed; 2) information
concerning the type of features, and the coordinate addresses of
those features located within the sample chamber; 3) reagent
information; 4) lot information; 5) calibration data; etc. In one
form, the label 14 may be a magnetic strip or a bar code strip, or
the like, which directly contains all the information useful to the
analytical device in the performance of the analysis(es). This type
of label 14 is particularly useful in those instances where the
information to be communicated is limited. In those instances where
the quantity of information to be communicated is considerable, it
may be more desirable to have the label 14 direct the analytical
device to a data file (stored within the analytical device or
remotely accessible by the analytical device via modem, network
link, etc.) containing the appropriate information. In this
instance, the label 14 can be said to indirectly contain the
information by providing the necessary path to the information.
Here again, the label 14 could be a bar code or magnetic strip,
which in this case communicates a particular code that is
interpreted by the analytical device as being associated with a
certain data file. The same result could be achieved by
incorporating a physical feature 56 in the container (e.g., a
notch, a tab, etc.--see FIG. 5) that is interpretable by the
analytical device. Other labels 14 which function to communicate
information to the analytical device can be used alternatively.
[0051] The container 10 also preferably includes a human readable
label 58 to facilitate handling within the laboratory or clinic.
The human readable label 58 may include information such as the
patient's name, a sample taken date, an office address, an
appropriate warning (e.g., "Biohazard--Handle with Care"),
trademarks, etc. The sides 60 of the container 10 are suitable to
interact with a transport means (not shown) contained within the
analytical device. The transport means is operable to move the
container 10 relative to an imaging device (not shown) contained
within the analytical device.
[0052] As stated above, the considerable utility of the container
10 enables a wide variety of analyses to be performed on a single
sample, using a single analytical device. The examples given below
are offered so that a complete appreciation of the present
invention container 10 may be gained.
EXAMPLE I
Hematological Analyses
[0053] Referring to FIGS. 1 and 4, to enable an analysis of white
blood cells (WBC's) within an anticoagulated whole blood sample,
the container 10 includes approximately 0.8 micrograms (.mu.g) of a
sensible colorant disposed within the reservoir 16. EDTA is an
example of an anticoagulating agent that may be used with the
sample and a fluorescent highlighting supravital stain such as
acridine orange, basic orange-21, or the like are examples of
sensible colorants that may be added to the reservoir 16. For
purposes of evaluating WBC's, it is preferable to have a region
within the chamber 12 that has a plurality of select fields with a
through-plane thickness on the order of 20 microns in magnitude. A
chamber 12 through-plane thickness (i.e., a type of physical
characteristic) of approximately 20 microns is chosen for a couple
of reasons. First, an evaluation volume of 0.02 .mu.l, (formed by a
particular field of the chamber 12 having a cross-sectional area of
1 millimeter (mm) and a thickness of 20 microns) typically contains
50-200 WBC's which is a favorable quantity for evaluative purposes.
Second, a through-plane thickness of 20 microns provides an optimal
chamber 12 for rouleaux and lacunae formation. The coordinate
addresses of select fields are communicated to the analytical
device by way of the label 14. In the example, therefore, the
plurality of features operative to enable analysis of the biologic
fluid sample include: 1) the sensible reagent disposed within the
reservoir 16; and 2) the chamber 12 region(s) having a plurality of
select fields with a particular through-plane thickness, at known
coordinate addresses within the chamber 12.
[0054] Approximately 20 .mu.l of anticoagulated whole blood is
placed into the reservoir 16 by the operator and the cap 34
secured. The container is gently shaken until the reagent and whole
blood sample are adequately mixed. A mixing ball 36 disposed in the
reservoir 16 facilitates mixing. The container 10 is inserted into
the analytical device and the valve 20 is subsequently actuated to
release the sample into the chamber 12 by way of the channel 18.
Once the sample is distributed within the chamber 12, the sample
resides quiescently. The only sample motion within the chamber 12
will possibly be Brownian motion of the sample's formed
constituents, and that motion is non-disabling for the present
invention. Note that for simple tests such as a WBC count where
timing is not important, a sample could be deposited into the
chamber 12 directly, thereby obviating the need for a reservoir 16
and valve 20.
[0055] Immediately after the sample has been inserted into the
chamber, the sample will appear opaque when examined either with
transmitted light, or more preferably by epi-illuminated
fluorescence. The opaque appearance is caused by the red blood
cells (RBC's), which form an overlapping mass prior to the
formation of the rouleaux. After lying substantially motionless for
approximately thirty (30) seconds, within the chamber 12, the RBC's
will have spontaneously clustered into rouleaux, leaving lacunae
between the rouleaux. It is in these lacunae where the other whole
blood sample constituents (e.g., WBC's and platelets) can be found
and evaluated. If a count of WBC's is desired, a square millimeter
field of the 20 micron thick chamber 12, which contains 0.02 .mu.l
of whole blood sample, can be evaluated. A 0.02 .mu.l sample is
chosen to keep the number of WBC's reasonable (a normal whole blood
sample contains approximately 7,000 WBC's per .mu.l of sample; a
0.02 .mu.l sample of normal whole blood contains approximately 140
WBC's). A number of these fields would be evaluated until enough
cells are counted to get a number which has sufficient statistical
accuracy, which is in practice approximately 1000 cells. If
additional WBC information is sought, the WBC's (lymphocytes,
granulocytes, monocytes, etc.) can be analyzed within the sample
using an image dissector such as a CCD camera, for example, alone
or with analysis software. A differential count could be determined
from the data collected.
[0056] The above example of the utility of the present invention
container 10 in hematological analyses includes a plurality of
features operative to enable analysis of the biologic fluid sample.
In a preferred embodiment, the features not only include the
plurality of select fields with a through-plane thickness (i.e., a
type of physical characteristic) on the order of 20 microns, but
fields of slightly larger and smaller volume as well. The
larger/smaller field volumes can be created by several of the
mechanisms described above; e.g., convergent chamber walls 26,28,
or steps 46 within one or both walls 26,28, etc. A range of field
volumes is advantageous because constituent populations quite often
vary in magnitude within the biologic fluid sample. If, for
example, the WBC population within the sample was abnormally high,
a chamber 12 region having a through-plane thickness of 20 microns
may have more than an optimal number of WBC's for evaluative
techniques such as counting. Changing to a field of smaller volume
would decrease the number of WBC's and therefore facilitate the
analysis at hand. On the other hand, if the WBC population within
the sample was abnormally low, a chamber 12 region having a
through-plane thickness of 20 microns may have less than an optimal
number of WBC's for evaluative purposes. Changing to a field of
larger volume would increase the number of WBC's and likewise
facilitate the analysis at hand. The spatial locations of alternate
features (i.e., larger or smaller through-plane thickness regions
in the above example) are communicated to the analytical device
through the label 14.
EXAMPLE II
Chemical Analyses
[0057] Referring to FIG. 9, a complete blood count requires that
the RBC's be evaluated for hemoglobin content. In a first
embodiment, the hemoglobin evaluation is performed in a first
chamber 62 which is connected to the reservoir 16 by a channel 18.
At least two chemical reagents 64,66 are initially stored within
the first chamber 62. The reagents 64,66 are shown in the first
chamber 62 as independent deposits to illustrate the use of
multiple reagents. Reagents can often be combined into a single
reagent mixture stored as a single deposit. One of the chemical
reagents 64 is a lysing reagent which breaks down RBC's within the
sample and thereby releases the hemoglobin stored within the RBC's.
The other reagent 66 is a hemoglobin stabilizer that increases the
reliability of the hemoglobin evaluation. In most cases, the
hemoglobin evaluation is performed after the lysing agent has been
introduced into the sample for a given period of time, or at
particular intervals. Using the present invention, the period of
time begins when the valve 20 is actuated to permit the sample to
enter the first chamber 62 and a second chamber 68. The remaining
analyses associated with a complete blood count are performed in
the second chamber 68. In this embodiment, the features operable to
enable the analysis of the biologic fluid sample include: 1) the
chemical reagents 64,66 disposed in the first chamber 62 at known
spatial locations; 2) one or more select fields at know locations
where the chemical reagents are disposed; and 3) the valve 20
between the reservoir 16 and the chambers 64,66 that initiates the
time period. Additional features such as those described heretofore
in the "Hematological Analyses" example may be present in the
second chamber 68.
[0058] Referring to FIG. 10, in a second embodiment all of the
complete blood count analyses are performed in a single chamber 12.
The portion of the biologic fluid sample used for the hemoglobin
evaluation is contiguous with remaining portion of the fluid
sample, but that portion is preferably oriented toward one side of
the chamber 12 to minimize potential mixing of the lysing agent
with the remaining portion of the fluid sample. In addition to
orienting the hemaglobin evaluation to one side, it is also
preferable to choose a chamber 12 through-plane thickness small
enough such that vertical diffusion (and ultimate equilibrium) of
the chemical reagents 64,66 within the biologic fluid sample occurs
at a much faster rate than lateral diffusion. The difference in
diffusion rates is such that lateral diffusion may be considered
negligible for a short period of time. The lateral spacing between
the hemoglobin evaluation site and the remainder of the fluid
sample is such that during that short period of time in which
lateral reagent diffusion is negligible, the remainder of the
desired analyses can be performed without interference from the
lysing agent. In this embodiment, the same two chemical reagents
64,66 as described above are initially deposited in the hemoglobin
evaluation region of the chamber 12, and actuating the valve 20
begins the time period for the evaluation. The features operable to
enable the analysis of the biologic fluid sample are: 1) the
chemical reagents 64,66 disposed in the aforementioned chamber 12
region at known spatial locations; 2) the chamber configuration
functionally operable to separate the hemoglobin evaluation region
from the remainder of the biologic fluid sample; 3) the valve 20
between the reservoir 16 and the chamber 12 used to initiate the
time period; and 4) any features such as those described above in
the "Hematological Analyses" example.
EXAMPLE III
Urinalysis
[0059] Referring to FIG. 5, a complete urinalysis requires a
chemical analysis and a particulate analysis of the urine sample.
Chemical reagents 70 spatially located at particular coordinate
addresses within a chamber are used to colorometrically relate
information after a given period of time. The particulate analysis
involves detecting, evaluating and/or enumerating the particles
within the sample. In a first embodiment, the chemical analysis is
performed in a first chamber 72 and the particulate analysis is
performed in a separate second chamber 74. Both the first chamber
72 and the second chamber 74 are in fluid communication with the
reservoir 16. In a manner similar to that described above, the
through-plane thickness and other physical characteristics of the
first chamber 72 and the second chamber 74 are chosen to facilitate
the chemical and particulate analyses, respectively. In the first
embodiment, the features operable to enable the analysis of the
biologic fluid sample are, therefore: 1) the chemical reagents 70
disposed in the first chamber 72at a known spatial location(s); 2)
the physical features at a known spatial location(s) within the
chamber 12 chosen to facilitate the chemical analysis; and 3) the
valve 20 between the reservoir 16 and the chamber 12 used to
initiate the time period. In a second embodiment, the chemical and
particulate analyses are performed in the same chamber 12. In a
manner similar to the hemoglobin evaluation described above (see
FIG. 10), the chamber 12 region devoted to the chemical analysis is
preferably oriented to one side of the chamber 12 and the
through-plane thickness is such that interference from the chemical
reagents will be negligible if at all. The features within the
second embodiment operable to enable the analysis of the biologic
fluid sample are: 1) the chemical reagents 70 disposed in the
chamber 12at a known spatial location(s); 2) the chamber 12
configuration functionally operable to separate the chemical
evaluation region from the remainder of the biologic fluid sample;
and 3) the valve 20 between the reservoir 16 and the chamber 12
that initiates the time period.
[0060] Although this invention has been shown and described with
respect to the detailed embodiments thereof, it will be understood
by those skilled in the art that various changes in form and detail
thereof may be made without departing from the spirit and the scope
of the invention.
* * * * *