U.S. patent application number 12/174336 was filed with the patent office on 2010-01-21 for use of fluid aspiration/dispensing tip as a microcentrifuge tube.
This patent application is currently assigned to Ortho-Clinical Diagnostics, Inc.. Invention is credited to David A. Heavner.
Application Number | 20100015690 12/174336 |
Document ID | / |
Family ID | 41119310 |
Filed Date | 2010-01-21 |
United States Patent
Application |
20100015690 |
Kind Code |
A1 |
Heavner; David A. |
January 21, 2010 |
USE OF FLUID ASPIRATION/DISPENSING TIP AS A MICROCENTRIFUGE
TUBE
Abstract
A fluid aspirating/dispensing member includes a sample cavity
for sample acquisition and a sealable cavity that, once sealed,
permits the separation of particles from the remainder of a fluid
sample within the sample cavity after centrifugation or other
separation means. The fluid aspirating/dispensing members, either
individually or as part of an array, increase the efficiency of
sample processing before analysis by a clinical analyzer.
Inventors: |
Heavner; David A.;
(Fairport, NY) |
Correspondence
Address: |
Hiscock & Barclay, LLP
One Park Place, 300 South State Street
Syracuse
NY
13202-2078
US
|
Assignee: |
Ortho-Clinical Diagnostics,
Inc.
Rochester
NY
|
Family ID: |
41119310 |
Appl. No.: |
12/174336 |
Filed: |
July 16, 2008 |
Current U.S.
Class: |
435/288.4 ;
435/287.1; 73/864.01 |
Current CPC
Class: |
B01L 2400/0409 20130101;
Y10T 436/2575 20150115; B01L 2300/042 20130101; Y10T 436/25375
20150115; B01L 3/5025 20130101; B01L 2400/0487 20130101; B01L
3/5021 20130101; B01L 2300/0829 20130101; B01L 3/0275 20130101;
G01N 2035/0434 20130101 |
Class at
Publication: |
435/288.4 ;
435/287.1; 73/864.01 |
International
Class: |
C12M 1/00 20060101
C12M001/00; B01L 3/00 20060101 B01L003/00 |
Claims
1. A fluid aspirating/dispensing member comprising: a) a first port
in fluid communication with an internal volume of a fluid
aspirating/dispensing member; b) a second port in fluid
communication with said internal volume, said second port being
opposite said first port; c) a sample cavity of said internal
volume disposed between said first and second ports; d) a sealable
cavity of said internal volume disposed between said second port
and said sample cavity; and e) a cap, attached adjacent to said
first port, said cap being configured to close said first port when
said cap is placed thereupon, wherein the sealing of said sealable
cavity permits the sealing of said second port to create a fluid
container, said container being configured to retain a fluid sample
in said sample cavity and to permit the separation of particles
suspended in said sample from the remainder of said sample.
2. The aspirating/dispensing member of claim 1, further configured
to be placed within a testing apparatus capable of separating
particles from the remainder of the sample.
3. The aspirating/dispensing member of claim 2, wherein said
apparatus is a clinical analyzer.
4. The aspirating/dispensing member of claim 1, wherein the
separation of the particles from the remainder of the sample
results from separation by centrifugation.
5. The aspirating/dispensing member of claim 1, wherein the walls
of said sample cavity are tapered.
6. The aspirating/dispensing member of claim 1, wherein the walls
of said sealable cavity are parallel with respect to the vertical
axis of said member.
7. The aspirating/dispensing member of claim 1, wherein said
sealable cavity is heat-sealable.
8. The aspirating/dispensing member of claim 1, wherein said cap is
removably attached.
9. The aspirating/dispensing member of claim 1, wherein the walls
of said sealable cavity are less than 2 mm apart.
10. The aspirating/dispensing member of claim 1, wherein said
sealable cavity is less than 1 cm in length.
11. The aspirating/dispensing member of claim 1, wherein said
member permits optical or visual testing of a sample within said
internal volume of said member.
12. The aspirating/dispensing member of claim 1, wherein the sample
comprises a cell suspension.
13. The aspirating/dispensing member of claim 1, wherein the sample
comprises blood.
14. The aspirating/dispensing member of claim 1, wherein the
particles are cells.
15. The aspirating/dispensing member of claim 1, further comprising
a separation barrier disposed in said internal volume of said
member.
16. The aspirating/dispensing member of claim 1, wherein the sample
further comprises reagents for particle agglutination.
17. A fluid aspirating/dispensing plate comprising an array of
fluid aspirating/dispensing members attached to a solid support,
each of said aspirating/dispensing members comprising: a) a first
port in fluid communication with an internal volume of a fluid
aspirating/dispensing member; b) a second port in fluid
communication with said internal volume, said second port being
opposite said first port; c) a sample cavity of said internal
volume disposed between said first and second ports; and d) a
sealable cavity of said internal volume disposed between said
second port and said sample cavity, wherein the sealing of said
sealable cavity of each of said aspirating/dispensing members
permits the sealing of said second ports of each of said members to
create a plurality of fluid containers, each of said containers
being configured to retain a fluid sample in said sample cavity and
to permit the separation of particles suspended in said sample from
the remainder of said sample.
18. The fluid aspirating/dispensing plate of claim 17, wherein said
solid support is planar.
19. The fluid aspirating/dispensing plate of claim 18, wherein said
support further comprises means for supporting said members.
20. The fluid aspirating/dispensing plate of claim 17, wherein said
fluid aspirating/dispensing members are reversibly attached to said
solid support.
21. The fluid aspirating/dispensing plate of claim 17, wherein said
plate comprises 96 fluid aspirating/dispensing members.
22. The fluid aspirating/dispensing plate of claim 17, said plate
being further configured to be placed within a testing apparatus
capable of separating particles from the remainder of the sample of
each said fluid aspirating/dispensing member.
23. The fluid aspirating/dispensing plate of claim 22, wherein said
testing apparatus is a clinical analyzer.
24. The fluid aspirating/dispensing plate of claim 17, wherein the
separation of the particles from the remainder of the sample
results from separation by centrifugation.
25. The fluid aspirating/dispensing plate of claim 17, wherein said
sealable cavity is heat-sealable.
26. The fluid aspirating/dispensing plate of claim 17, further
comprising a cover said cover being configured to close each of
said first ports when said cover is placed on said first ports.
27. The fluid aspirating/dispensing plate of claim 17, wherein said
each of said aspirating/dispensing members permit the optical or
visual testing of a sample within said internal volume of each of
said members.
28. The fluid aspirating/dispensing plate of claim 17, wherein said
array of aspirating/dispensing members can be aligned with the
wells of a microtiter plate.
29. The fluid aspirating/dispensing plate of claim 28, wherein each
of the wells of said microtiter plate contain a sample that is
different from each of the other wells.
30. The fluid aspirating/dispensing plate of claim 17, wherein the
sample of each of said containers comprises a cell suspension.
31. The fluid aspirating/dispensing plate of claim 17, wherein the
sample of each of said containers comprises blood.
32. The fluid aspirating/dispensing plate of claim 17, wherein the
particles are cells.
33. The fluid aspirating/dispensing plate of claim 17, each of the
fluid aspirating/dispensing members further comprise a separation
barrier disposed in said internal volume of each of said
members.
34. The fluid aspirating/dispensing plate of claim 17, wherein each
of said containers further comprises reagents for particle
agglutination.
35. A method of separating particles in a fluid sample, said method
comprising the steps of: a) loading a fluid aspirating/dispensing
member into an apparatus, said member comprising a first port, a
second port and a sample cavity in fluid communication with each of
said first and second ports; b) aspirating a sample into said
sample cavity through said second port of said fluid
aspirating/dispensing member; c) sealing said second port of said
fluid aspirating/dispensing member to create a fluid container; d)
closing said first port using a cap sized to releasably engage and
cover said first port; and e) separating particles in said sample
from the remainder of said sample, wherein the separated particles
and sample are retained within said sample cavity of said fluid
aspirating/dispensing member for detection the samples or
particles.
36. The method of claim 35, wherein said loading step includes the
step of attaching said fluid aspirating/dispensing member to a
proboscis.
37. The method of claim 35, wherein said fluid
aspirating/dispensing member is a metering tip.
38. The method of claim 35, wherein said apparatus is a clinical
analyzer.
39. The method of claim 35, wherein the separating step is
performed by centrifugation.
40. The method of claim 35, wherein the sealing step is performed
by heat sealing said second port of said member.
41. A method of separating particles in a plurality of fluid
samples, said method comprising the steps of: a) loading a fluid
aspirating/dispensing plate into an apparatus, said plate
comprising a plurality of fluid aspirating/dispensing members, each
of said members comprising a first port, a second port and a sample
cavity in fluid communication with each of said first and second
ports; b) aspirating a plurality of samples into said sample
cavities through said second ports of each of said fluid
aspirating/dispensing members; c) sealing said second ports of each
of said fluid aspirating/dispensing members to create a plurality
of fluid containers; and d) separating particles in said sample
from the remainder of the sample in each of said containers,
wherein the separated particles and sample are retained within said
sample cavity of each of said fluid aspirating/dispensing members
for detection the particles or sample.
42. The method of claim 41, wherein said loading step requires the
attachment of said fluid aspirating/dispensing plate to a plurality
of proboscis of said apparatus, said proboscis being part of a
metering mechanism.
43. The method of claim 41, wherein said fluid
aspirating/dispensing members are metering tips.
44. The method of claim 41, wherein said apparatus is a clinical
analyzer.
45. The method of claim 41, wherein the separating step is
performed by centrifugation.
46. The method of claim 41, further comprising a step, after said
aspirating step, wherein said first ports of each of said members
are closed by a lid, said lid being configured to close said first
ports of each of said member when said lid is placed on said first
ports.
47. The method of claim 41, wherein the sealing step is performed
by heat sealing said second ports of each of said members.
Description
FIELD OF THE APPLICATION
[0001] This application relates to an apparatus and a method for
sample collection and centrifugation within a single, disposable
fluid aspirating/dispensing member.
BACKGROUND OF THE INVENTION
[0002] Combinational clinical analyzers containing both "wet" and
"dry" chemistry platforms in a single apparatus for the testing of
biological samples, such as whole blood serum, are now widely used
in most modern health care facilities.
[0003] So-called "dry" chemistry systems commonly include a sample
supply, a number of sample containers, a metering/transport
mechanism, and an incubator having a plurality of test read
stations as described, for example, in U.S. Pat. No. 3,992,158, the
contents of which are hereby incorporated herein by reference in
its entirety. A typical protocol starts with the aspiration of a
known quantity of a sample into a fluid aspirating/dispensing
member. An aliquot of the sample is then dispensed onto a dry slide
element which is then loaded into an incubator. After appropriate
incubation, the amount or presence of at least one analyte in the
sample is determined using, for example, an electrometer,
reflectometer or other suitable testing devices.
[0004] So-called "wet" chemistry systems on the other hand, use a
reaction vessel, such as a cuvette, which receives predetermined
volumetric quantities of sample, reagent, and other fluids that are
appropriately metered into a reaction vessel in order to perform an
assay(s). The "wet" chemistry system commonly includes a metering
mechanism to transport a patient sample fluid from a sample supply
to the reaction vessel. After a pre-determined incubation period
during which one or more reactions occur, a measuring device, such
as an optical measuring device is used to pass a beam of light
through the reaction vessel and sample. Assays typically used in
`wet` chemistry systems include, but are not limited to,
spectrophotometric absorbance assays such as end point reaction
analysis and rate of reaction analysis, turbidimetric assays,
nephelometric assays, radiative energy attenuation assays (such as
those described in U.S. Pat. Nos. 4,496,293 and 4,743,561, the
contents of which are hereby incorporated herein by reference in
their entirety), ion capture assays, color, metric assays, and
fluorometric assays, and immunoassays, all of which are well known
in the art.
[0005] Integration of wet/dry chemistry capabilities into clinical
analyzers reduces experimental error, improves work flow and limits
the need for human intervention thereby reducing the risk of
contamination of lab personnel with human pathogens. One example of
a commercially available combination clinical analyzer employing
both wet and dry chemistry systems is the Vitros 5,1 FS Chemistry
System, which is described in further detail in U.S. Pat. No.
7,250,303 and U.S. Patent Publication No. US 2003/0026733 (each
assigned to Ortho-Clinical Diagnostics, Rochester, N.Y.), the
contents of which are incorporated herein by reference in its
entirety.
[0006] Despite the progress that has been made, these systems still
require that patient samples be first processed to remove
particulate components before presentation to a clinical analyzer.
This processing step remains cumbersome, time consuming and limits
the overall efficiency of these analyzers.
[0007] Information relevant to attempts to address this problem can
be found in U.S. Pat. Nos. D453,573; 4,933,291; 5,384,239;
5,722,553; 5,753,186; 6,001,310; 6,334,842; 6,601,725; 6,622,882;
7,064,823; the published U.S. Publication Numbers US 2001/0019842;
US 2005/0204832; US 2005/0208676; US 2007/0003443, US 2007/0017927;
International PCT application PCT/AU1992/000236 and European Pat.
No. EP743095. Each one of these references suffers, however, from
one or more of the following disadvantages: the references fail to
remedy the inefficient processing of patient samples prior to
clinical analysis and also fail to describe a fluid
aspirating/dispensing member that permits both sample collection
and centrifugation.
[0008] For the foregoing reasons, there is an unmet need in the
field to improve the efficiency of sample processing prior to
analysis by clinical analyzers.
SUMMARY OF THE APPLICATION
[0009] The invention pertains to a fluid aspirating/dispensing
member having a sealable end that can be used for both sample
collection and centrifugation. Methods are described for performing
sample collection and centrifugation using either individual fluid
members or plates comprising a plurality of fluid
aspirating/dispensing members.
[0010] According to one version, a fluid aspirating/dispensing
member is described that comprises a first port, a second port,
opposite the first port, and a cap. The cap is configured to
releasably close the first port when attached thereto. The internal
volume of the fluid aspirating/dispensing member comprises a sample
cavity between the first and second ports; and a sealable cavity
between the second port and the sample cavity. The sealing of the
sealable cavity seals the second port to create a container that is
configured to retain a fluid sample in the sample cavity and to
permit the separation of particles suspended in the sample from the
remainder of the sample.
[0011] According to another aspect, the fluid aspirating/dispensing
member is configured to be placed within a testing apparatus
capable of separating particles from the remainder of the fluid
sample. The apparatus may be a clinical analyzer. The separation of
the particles from the remainder of the sample can result, for
example, from centrifugation.
[0012] In one aspect, the walls of the sample cavity are tapered,
whereas the walls of the sealable cavity can be parallel with
respect to the vertical axis of the fluid aspirating/dispensing
member.
[0013] The sealable cavity of the fluid aspirating/dispensing
member can be heat-sealable, less than 1 cm long, with walls that
are less than 2 mm apart and parallel to the vertical axis of the
aspirating/dispensing member.
[0014] In one aspect, the cap is removably attached to the
aspirating/dispensing member.
[0015] The aspirating/dispensing member can allow for optical or
visual testing of a sample, which can be a cell suspension or
blood. Particles within the sample can be red blood cells.
[0016] The aspirating/dispensing member can further contain a
separation barrier or reagents for agglutination within the
internal volume of each member.
[0017] According to another version, a fluid aspirating/dispensing
plate is described that comprises an array of fluid
aspirating/dispensing members attached to a solid support. Each of
the aspirating/dispensing members comprises a first port and a
second port, opposite the first port. The internal volume of each
fluid aspirating/dispensing member comprises a sample cavity
between the first and second ports; and a sealable cavity between
the second port and the sample cavity. The sealing of the sealable
cavity of each of the fluid aspirating/dispensing members creates a
plurality of containers that are configured to retain a fluid
sample in the sample cavity and permit the separation of particles
suspended in the sample from the remainder of the sample.
[0018] In one aspect, the solid support is planar that can include
a means for supporting the fluid aspirating/dispensing members. The
fluid aspirating/dispensing plate can have 96 fluid
aspirating/dispensing members.
[0019] In another aspect, the fluid aspirating/dispensing members
are reversibly attached to the solid support.
[0020] In another version, the fluid aspirating/dispensing plate is
configured to be placed within a testing apparatus capable of
separating particles from the remainder of the sample of each the
fluid aspirating/dispensing member. This testing apparatus can be a
clinical analyzer.
[0021] The separation of the particles from the remainder of the
sample can result from separation by centrifugation.
[0022] The sealable cavity of the fluid aspirating/dispensing plate
can be a heat-sealable cavity.
[0023] In another version, the fluid aspirating/dispensing plate
has a cover that is configured to close each of the first ports
when the cover is placed on the first ports of the fluid
aspirating/dispensing plate.
[0024] In one aspect, the fluid aspirating/dispensing members of
the plate allow for optical or visual testing of a sample within
the internal volume of each member.
[0025] In another aspect, the array of aspirating/dispensing
members on the plate align with the wells of a microtiter plate.
Each well of the microtiter plate can retain a sample that is
different from each of the other wells in the plate, wherein the
sample can be a cell suspension or blood. Particles within the
sample can be cells.
[0026] In yet another aspect, the aspirating/dispensing members on
the plate can contain a separation barrier or reagents for
agglutination within the internal volume of each member.
[0027] According to another embodiment, a method is described for
separating particles in a fluid sample, the method comprising the
steps of (a) loading a fluid aspirating/dispensing member into an
apparatus, the member comprising a first port, a second port and a
sample cavity in fluid communication with the first and second
ports, (b) aspirating a sample into the sample cavity through the
second port of the fluid aspirating/dispensing member, (c) sealing
the second port of the fluid aspirating/dispensing member to create
a fluid container; (d) closing the first port using a cap sized to
releasably engage and cover the first port; and (e) separating
particles in the sample from the remainder of the sample, wherein
the separated particles and sample are retained within the sample
cavity of the fluid aspirating/dispensing member for detection of
the particles or sample.
[0028] The apparatus can be a clinical analyzer. The separating
step can be performed, for example, by centrifugation.
[0029] According to one aspect, the loading step requires the
attachment of the fluid aspirating/dispensing member to a proboscis
of a metering mechanism of the apparatus.
[0030] In another aspect, the fluid aspirating/dispensing member is
a metering tip.
[0031] In one aspect, the sealing step is performed by heat sealing
the second port of the fluid aspirating/dispensing member.
[0032] According to yet another embodiment, a method is described
for separating particles in a plurality of fluid samples, the
method comprising the steps of (a) loading a fluid
aspirating/dispensing plate into an apparatus, wherein the plate
comprises a plurality of fluid aspirating/dispensing members, each
of the members comprising a first port, a second port and a sample
cavity in fluid communication with each of the first and second
ports, (b) aspirating a plurality of samples into the sample
cavities through the second ports of each of the fluid
aspirating/dispensing members, (c) sealing the second ports of each
of the fluid aspirating/dispensing members to create a plurality of
fluid containers and (d) separating particles in the sample from
the remainder of the sample in each of the containers, wherein the
particles and sample are retained within the sample cavity of each
of the fluid aspirating/dispensing members for detection of the
particles or sample.
[0033] In one aspect, the apparatus is a clinical analyzer.
[0034] The separating step can be performed by centrifugation.
[0035] In one aspect the loading step requires the attachment of
the fluid aspirating/dispensing plate to a plurality of proboscis
of a metering mechanism of the apparatus.
[0036] In another aspect, the fluid aspirating/dispensing members
are metering tips.
[0037] According to another embodiment, after the aspirating step,
the first ports of each of the fluid aspirating/dispensing members
are closed by a lid that is configured to close the first ports of
each of the fluid aspirating/dispensing members when it is placed
on the first ports of the fluid aspirating/dispensing members.
[0038] The sealing step can be performed by heat sealing the second
ports of each of the fluid aspirating/dispensing members.
[0039] The previously described embodiments have many advantages,
including the ability to perform sample collection and particle
separation using either individual disposable fluid
aspirating/dispensing members or a fluid aspirating/dispensing
plate comprising a plurality of fluid aspirating/dispensing
members. The methods described herein reduce handling errors as
well as the time spent for sample processing before analysis by
clinical analyzers. Sample processing using the herein described
fluid aspirating/dispensing member is therefore faster than
conventional methods known in the art.
[0040] It should be understood that this application is not limited
to the embodiments disclosed in this Summary, and it is intended to
cover modifications and variations that are within the scope of
those of sufficient skill in the field, and as defined by the
claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0041] FIG. 1 depicts a cross-sectional view of a fluid
aspirating/dispensing member made in accordance with one
embodiment;
[0042] FIG. 2 illustrates the movement of the cap of the fluid
aspirating/dispensing member of FIG. 1;
[0043] FIGS. 3A-3G depict sequential cross-section views of the
fluid aspirating/dispensing member of FIG. 1 representing a method
of sample collection and centrifugation in accordance with a second
embodiment;
[0044] FIG. 4A depicts a view of a fluid aspirating/dispensing
plate comprising an array of fluid aspirating/dispensing members in
accordance with a third embodiment;
[0045] FIG. 4B depicts a cross-sectional view of a row of fluid
aspirating/dispensing members of the plate of FIG. 4A;
[0046] FIG. 5 illustrates the alignment of the array of fluid
aspirating/dispensing members of FIG. 4A with a microtiter
plate;
[0047] FIGS. 6A-6I depict sequential cross-section views of the
array of fluid aspirating/dispensing members of FIG. 4A
representing a method of sample collection and centrifugation in
accordance with a fourth embodiment.
DETAILED DESCRIPTION.
[0048] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of skill in the art. The following definitions are provided to help
interpret the disclosure and claims of this application. In the
event a definition in this section is not consistent with
definitions elsewhere, the definition set forth in this section
will control.
[0049] As used herein, "combinational" analyzer refers to a
clinical analyzer that includes at least two chemistry systems that
can encompass any combination of "dry" and/or "wet" chemistry
systems.
[0050] As used herein, a "clinical analyzer" refers to any
apparatus capable of analyzing clinical samples including, but not
limited to, immunodiagnostic analyzers as well as analyzers for the
automated "wet" and/or "dry" chemistry analysis of clinical
samples. "Wet" chemistry platforms typically include a
microprocessor controlling an automated fluid handling system that
aspirates and dispenses one or more samples and/or reagents into a
reaction vessel, at least one sample reservoir, optionally at least
one reagent reservoir, at least one incubator and at least one
detector, such as a spectrophotometer. Typical analyzers for use
with the fluid aspirating/dispensing member of this application are
described in further detail in U.S. Pat. Nos. 6,096,561, 7,282,372
and 7,250,303, all of which are incorporated herein by reference in
their entireties. In a preferred embodiment, the clinical analyzer
used herein refers to commercially available analyzers such as the
VITROS 5,1 FS Chemistry System manufactured by Ortho-Clinical
Diagnostics, Inc.
[0051] An element is in "fluid communication" with another element
when a fluid is able to travel from one element to the other such
as via capillary action and/or gravity. The elements may be in
direct contact, but do not need to be in direct contact; i.e.,
other elements through which said fluid can pass may be
intervening. An element is "not in fluid communication" with
another element when a fluid is not able to travel from one element
to the other via capillary action and/or gravity. Typically, the
elements are physically separated, i.e. spaced apart.
[0052] As used herein, a "fluid aspirating/dispensing member" is a
device such as a metering tip containing one or more internal
cavities in fluid communication with two or more sealable apertures
that is used for both sample collection and particle separation as
described in U.S. Pat. No. 6,797,518, the contents of which are
hereby incorporated by reference herein in its entirety. In one
embodiment, the particle separation occurs as a result of
centrifugation. A fluid aspirating/dispensing member typically
performs the aspiration or dispensing of a volumetric amount of a
sample within a clinical analyzer. In one embodiment, the fluid
aspirating/dispensing member is composed of a molded, solid
material that can be centrifuged without deformation of the
internal cavities. In another embodiment, the material does not
promote the adhesion of a biological sample to the internal walls
of the fluid aspirating/dispensing member. In another embodiment
the fluid aspirating/dispensing member is made out of a plastic
material typically by extrusion blow molding. The fluid
aspirating/dispensing member can be made from a thermoplastic
material preferably having a translucent or transparent
characteristic that allows optical testing to be performed upon the
fluid contents after aspiration therein. In one embodiment, the
fluid aspirating/dispensing member is made of a moldable plastic
such as polypropylene or polyethylene. In another embodiment, the
fluid aspirating/dispensing member is made of a homopolymer or
copolymer such as a polyallomer, wherein one of the monomers is
propylene. In yet another embodiment, the fluid
aspirating/dispensing member is made of polyethylene
terephthalate.
[0053] As used herein, a "fluid aspirating/dispensing plate" refers
to an array of multiple fluid aspirating/dispensing members
attached to a solid support. Each fluid aspirating/dispensing
member comprises one or more internal cavities in fluid
communication with two or more sealable apertures for both sample
collection and particle separation. A fluid aspirating/dispensing
plate is typically used for the aspiration or dispensing of a
volumetric amount of one or more samples within a clinical
analyzer. In one embodiment, the support is planar and rectangular
or square in shape, although the fluid aspirating/dispensing plate
described herein may have any shape. In another embodiment, the
fluid aspirating/dispensing members on the fluid
aspirating/dispensing plate are arranged in rows, each row having
the same number of fluid aspirating/dispensing members. In another
embodiment, the fluid aspirating/dispensing plate comprises a
support for the fluid aspirating/dispensing members. A person of
skill in the art will recognize that the support can be engineered
to facilitate centrifugation and robotic handling in a clinical
analyzer. In one embodiment, the fluid aspirating/dispensing
members are reversibly affixed to the solid support. In another
embodiment, the multiple fluid aspirating/dispensing members are
arrayed on a fluid aspirating/dispensing plate in such a manner so
as to align with the wells of a microtiter plate. In yet another
embodiment, the plate has a plurality of locations for the
reversible attachment of one, two, five, ten, 25, 50, 75, 100 or
more fluid aspirating/dispensing members. Each location on the
plate may or may not be filled with a fluid aspirating/dispensing
member.
[0054] As used herein, "separation of the particles from the
remainder of the sample" refers to any procedure that permits the
separation of the liquid phase of a sample from its solid
particulate phase. In one embodiment, the "separation of the
particles from the remainder of the sample" refers to the partial
or complete separation of the solid particulate phase of a sample
from its liquid phase within the sample cavity of the fluid
aspirating/dispensing member after the sealing of the fluid
aspirating/dispensing member. In another embodiment, the separation
occurs as a consequence of centrifugation. In yet another
embodiment, the separation results from a magnetic field acting on
magnetic particles that have been added to a sample.
[0055] As used herein, the term "sealable cavity" refers to a
region of the fluid aspirating/dispensing member, located between
the sample cavity and the bottom end of the aspirating/dispensing
member, which can be sealed, for example, by heat sealing or as a
result of the application of a sealant as defined herein. As a
consequence of sealing, the sample cavity is no longer in fluid
communication with the aspirating/dispensing end of the fluid
aspirating/dispensing member thus creating a fluid container for
particle separation. In one embodiment, the sealable cavity is made
of a thermoplastic melt-fusible material. In another embodiment the
thermoplastic melt-fusible material is a polyallomer or similar
thermoplastic material suitable for heat sealing as known by a
person of skill in the art.
[0056] As used herein, "heat sealing" refers to the application of
sufficient heat and pressure to the walls of the sealable cavity of
the fluid aspirating/dispensing member to cause the walls to fuse
together. Subsequent curing i.e. the hardening and solidification
of the fused thermoplastic walls creates a pressure-resistant
sealing of the sealable cavity that prohibits fluid communication
between the sample cavity and the sealable aspirating/dispensing
end of the fluid aspirating/dispensing member. A more detailed
description of heat sealing can be found in U.S. Pat. No.
3,929,943, the contents of which is hereby incorporated herein in
its entirety. In another embodiment, heat sealing is applied to the
sealable cavities of a plurality of fluid aspirating/dispensing
members arrayed on a fluid aspirating/dispensing plate. For
example, the aspirating/dispensing ends of the array of fluid
aspirating/dispensing members can be aligned and inserted into an
array of metallic molding cups that are heated to a temperature
that promotes the melting and fusion of the walls of the sealable
cavities placed therein.
[0057] As used herein, "sealing" refers to the permanent closing of
the aspirating/dispensing end of a fluid aspirating/dispensing
member. In one embodiment, sealing refers to heat sealing. In
another embodiment, sealing refers to the application of a sealant,
for example, a plastic or adhesive, that hermetically plugs the
aspirating/dispensing end of a fluid aspirating/dispensing member.
In another example, the sealant may be a bottom cap, such as a
bottom screw cap, that hermetically closes the
aspirating/dispensing end of a fluid aspirating/dispensing
member.
[0058] As used herein, a "sealant" shall mean any composition such
as an adhesive that can be used to form a connecting bond between
the walls of the sealable cavity of the fluid aspirating/dispensing
member described herein. In one embodiment the sealant is UV
curable. In another embodiment, the sealant cures at room
temperature.
[0059] As used herein, the term "polyallomer" refers to any
thermoplastic material that produces copolymers of the 1-olefins
exhibiting a degree of crystallinity normally associated only with
homopolymers. In one embodiment, the polyallomer is a random block
copolymer of propylene and ethylene. In another embodiment, the
polyallomer is a readily fusible plastic material sold by Eastman
Chemical Co. under the trade name "Tenite.RTM. Polyallomer."
[0060] As used herein, the term "cavity" refers to any
three-dimensional enclosure within the described fluid
aspirating/dispensing member. In an exemplary embodiment, one or
more cavities are in fluid communication with each other.
[0061] The term "plurality," as used herein, refers to a quantity
of two or more.
[0062] As used herein, "particles" may be cells, for example,
bacteria or red blood cells or white blood cells found in a sample.
In another embodiment, particles may be microscopic solids that are
added to a sample prior to sample processing. These particles may
be inert solids in the form of beads, beaded gels or microspheres,
although they may have any shape. Further examples of particles
include, but are not limited to, plastic particles, ceramic
particles, carbon particles, polystyrene microbeads, latex beads,
glass beads, magnetic beads, hollow glass spheres, metal particles,
particles of complex compositions, microfabricated or micromachined
particles. Inert particles may be comprised of any suitable
material, such as glass or ceramics, organic materials such as
carbon or plastic and/or one or more polymers, such as, for
example, nylon, polytetrafluoroethylene (TEFLON.TM.) or
styrene-divinylbenzene polymers. The particle size may be from
about 0.1 micron to 1000 microns. Preferably, the particle size is
from about 1 to about 10 microns. In principle, any ligand may be
covalently bound to a solid-phase matrix or particle such as
agarose beads (e.g., Sepharose Pharmacia) using known techniques,
for example as described by Heam et al., Methods in Enzymology Vol.
35:102-117 (1987). Generally, the beads are first activated by a
chemical agent, such as glutaraldehyde, carbonyldiimidizole,
cyanogen bromide hydroxysuccinimide, tosyl chloride or the like.
The chosen ligand is then covalently attached to the beads,
resulting in an extremely stable linkage of the ligand to the
support.
[0063] As used herein, "cell suspension" refers to a mixture of
cells in a liquid typically found in a sample. Cells can be
eukaryotic or prokaryotic cells. In one embodiment, the cells are
bacteria. In another embodiment, the cells are pathogenic bacteria.
In a preferred embodiment, the cells are blood cells. In another
preferred embodiment, the cells are red blood cells.
[0064] As used herein, the term "sample" refers to a material
suspected of containing at least one analyte. The sample can be
used directly as obtained from the source or following a
pretreatment to modify the character of the sample. The sample can
be derived from any biological source, such as a physiological
fluid, including, blood, plasma, saliva, ocular lens fluid,
cerebral spinal fluid, sweat, urine, milk, ascites fluid, raucous,
synovial fluid, peritoneal fluid, amniotic fluid or the like. The
sample can be pretreated before use. For example, whole blood is
typically treated with a polyanion such as heparin and the like to
inhibit blood coagulation. In another example, viscous fluids can
be diluted. Methods of treatment can also include filtration,
distillation, concentration, inactivation of interfering
components, and the addition of reagents. Besides physiological
fluids, other liquid samples can be used. In addition, a solid
material suspected of containing an analyte can be used as the
sample. In some instances it may be beneficial to modify a solid
sample to form a liquid medium or to release the analyte. In
another embodiment, a sample, as defined herein, is understood to
include one or more compounds that may be added to the sample
before or during analysis, including, but not limited to, buffers,
reagents, peptides, enzymes, ligands, ligand-binding molecules,
antibodies, particles, ligand-bound particles, magnetic particles,
and the like. A sample, as defined herein, may contain particles as
defined above.
[0065] The term "analyte," as used herein, refers to any compound
or composition or entity to be detected or measured. An analyte can
be any inorganic or organic molecule or cellular component or cell
or organism that is tested for by clinical assays known in the art.
Clinical assays may test for either the presence or absence of an
analyte. In one embodiment, an analyte has at least one epitope or
binding site or ligand. In another embodiment, an analyte can be
any substance for which there exists a naturally occurring binding
member or for which a binding member can be prepared. In yet
another embodiment, an analyte refers to a population of cells, for
example, blood cells such as white blood cells, red blood cells
(hematocrit) or platelets. Analytes may also include, but are not
limited to, metabolites, toxins, inorganic or organic compounds,
proteins, peptides, cytokines, chemokines, enzymes, amino acids,
lipids, HDL/LDL cholesterol, triglycerides, polysaccharides, blood
glucose, nucleic acids, hormones (for example, thyroid stimulating
hormone (TSH), Adrenocorticotropic hormone (ACTH), prolactin),
steroids (for example, cortisol or testosterone or estrogen),
vitamins, electrolytes, drugs, ions (for example, for pH
measurements), trace metals, microorganisms (bacteria, viruses or
parasites and the like), virus particles, metabolites of and
antibodies to any of the above substances. In another embodiment,
the analyte is C reactive protein, albumin, amylase, D-dimer,
bilirubin, alkaline phosphatase, gamma glutamyl transferase, urea,
creatine, serum iron, transferring, prostate specific antigen
(PSA), alpha fetoprotein (AFP), beta Human chorionic gonadotrophin
(bHCG), alpha 1-antitrypsin (AAT), CA-125 (also CA12.5),
carcinoembryonic antigen (CEA), fibrinogen or any other compound
which is typically tested for in a clinical sample. The term
"analyte" may include any antigenic substances, haptens,
antibodies, macromolecules and combinations thereof. In another
embodiment, the analyte is an enzyme that can be detected by
measuring enzyme activity. For example, the testing for creatine
kinase, glutamic oxaloacetic transaminase (SGOT), serum glutamic
pyruvic transaminase (SGPT) activity in blood is widely used as an
indicator of liver or heart damage.
[0066] As used herein, a "ligand" is any molecule which is capable
of binding to a ligand-binding molecule. In one embodiment, the
ligand is an epitope of an antibody. A number of ligands are also
known that bind immunoglobulin molecules and may be covalently
coupled to the particles used in this application, for example
Protein A, Protein G, Protein A/G and KappaLock..TM. (see also U.S.
Pat. No. 5,665,558, the contents of which are herein incorporated
by reference in its entirety). The ligand may bind to the isotype
of the antibody which is used or tested for or, alternatively, one
may use a bridging antibody, e.g., an IgG anti-IgM, for an IgM
antibody. Thus, an IgG anti-IgM antibody would be coupled to the
ligand as a "bridge" and an IgM antibody would bind to the IgG
anti-IgM antibody.
[0067] The term "ligand-binding," as used herein, refers to a
member of a binding pair, i.e., two different molecules wherein one
of the molecules specifically binds to the second molecule through
chemical or physical means. In addition to antigen and antibody
binding pair members, other binding pairs include, as examples
without limitation, biotin and avidin, carbohydrates and lectins,
complementary nucleotide sequences, complementary peptide
sequences, 1polysaccharides and lectins, effector and receptor
molecules, enzyme cofactors and enzymes, enzyme inhibitors and
enzymes, a peptide sequence and an antibody specific for the
sequence or the entire protein, polymeric acids and bases, dyes and
protein binders, peptides and specific protein binders (e.g.,
ribonuclease, S-peptide and ribonuclease S-protein), and the like.
Furthermore, binding pairs can include members that are analogs of
the original binding member, for example, an analyte-analog or a
binding member made by recombinant techniques or molecular
engineering. If the binding member is an immunoreactant it can be,
for example, a monoclonal or polyclonal antibody, a recombinant
protein or recombinant antibody, a chimeric antibody, a mixture(s)
or fragment(s) of the foregoing, as well as a preparation of such
antibodies, peptides and nucleotides for which suitability for use
as binding members is well known to those skilled in the art. A
ligand-binding member may be a polypeptide affinity ligand (see,
for example, U.S. Pat. No. 6,326,155, the contents of which are
hereby incorporated by reference herein in its entirety). In one
embodiment, the ligand-binding member is labeled. The label may be
selected from a fluorescent label, a chemiluminescent label or a
bioluminescent label, an enzyme-antibody construct or other similar
suitable labels known in the art.
[0068] As used herein "blood" broadly includes whole blood or any
component of whole blood, such as red blood cells, white blood
cells, plasma or serum.
[0069] As used herein, "centrifugation" refers to the rotation of
an object about an axis of rotation. Samples may be centrifuged in
a fixed angle or swing bucket rotor or any other rotor known in the
art.
[0070] As used herein, "STAT" is a medical term derived from the
Latin word "statim" which means immediately. A "STAT lane"
therefore refers to the urgent or rush processing of patient
samples.
[0071] As used herein, "emergency sample" refers to any sample that
requires immediate processing. Emergency samples typically include
those samples collected in emergency rooms or other urgent care
facilities. For example, an emergency room sample can be a blood
sample taken from a patient in an emergency room.
[0072] "Agglutination," as used herein refers to the clumping of a
suspension of cellular or particulate antigen by a reagent, usually
an antibody or other ligand-binding entities (see, for example,
U.S. Pat. Nos. 4,305,721, 5,650,068 and 5,552,064, the contents of
which are hereby incorporated herein by reference in their
entirety). In one embodiment, the reagent is Coomb's reagent.
[0073] As used herein, "Coomb's reagent" refers to a preparation of
antibodies, raised in animals, directed against one of the
following human immunoglobulin, complement or a specific
immunoglobulin e.g. anti-human IgG for use in the Coomb's test.
[0074] As used herein, "detection" refers to the detection of light
absorption or light scattering using a photodetector (see, for
example, U.S. Pat. No. 5,256,376 and published U.S. patent
application US 2004/0166551, the contents of which are hereby
incorporated herein by reference in their entirety). In one
embodiment, detection refers to the detection of bioluminescence or
chemiluminescence or fluorescence (see, for example, U.S. Pat. No.
6,596,546, the contents of which are hereby incorporated herein by
reference in its entirety).
[0075] As used herein, the term "agitating" refers to a force
acting on the contents of a fluid aspirating/dispensing member, for
example a centrifugal force or a force induced by a magnetic
field.
[0076] As used herein, a "metering device," as used herein, refers
to a component of a clinical analyzer that can reversibly attach to
a fluid aspirating/dispensing member by means of a proboscis. In
one embodiment, metering devices, controlled by an on-board
computer, coordinate the movement and/or transport of fluid
aspirating/dispensing members or a fluid aspirating/dispensing
plate within a clinical analyzer.
[0077] As used herein, the term "proboscis" refers to a component
of a clinical analyzer that attaches to one or more
aspirating/dispensing members, either individually or as part of a
fluid aspirating/dispensing plate. In one embodiment, the proboscis
is part of a metering mechanism and can be cylindrical in shape and
fits within the proboscis receptacle region of each of one or more
fluid aspirating/dispensing members in such a manner as to insert
itself against the internal wall of the proboscis receptacle region
of a fluid aspirating/dispensing member in an air-tight manner
(i.e., there is no substantial leak of air between the cylindrical
surface of the proboscis and the wall). Once the proboscis is
hermetically affixed to the proboscis receptacle region of each of
the fluid aspirating/dispensing member, the proboscis by means of
the metering mechanism, confers either a vacuum or pressure to the
internal volume of each fluid aspirating/dispensing member and
thereby drives the movement of a known volume of fluid within the
internal cavities of each fluid aspirating/dispensing member.
[0078] As used herein, the term "separation barrier" refers to a
water immiscible, typically thixotropic, gel-like or bead-like
material having a density intermediate between that found for the
light, liquid phase and the heavy, substantially particulate phases
of a sample. For example, the separation barrier is typically
disposed in the sample cavity of the fluid aspirating/dispensing
member which is then filled with the sample. Upon centrifugation,
the sample is gradient separated into its two phases and the
barrier material migrates to the interface between the phases. Upon
completion of centrifugation, the separation barrier forms a
physical and chemical barrier between the separated phases, thereby
preventing any mixing of the phases. Separation barriers can be,
for example, a mixture of silicone fluids and fine hydrophobic
silica powder, a polyester material or a hydrocarbon gel-like
material such as a polybutane or any other material known in the
art. The composition of separation barrier materials is described
in further detail in U.S. Pat. Nos. 4,190,535; 4,101,422; 4,818,418
and 4,147,628, the contents of which are hereby incorporated herein
in their entirety.
[0079] As used herein, the term "antibody" includes both polyclonal
and monoclonal antibodies; and may be an intact molecule, a
fragment thereof (such as Fv, Fd, Fab, Fab' and F(ab)' 2 fragments,
or multimers or aggregates of intact molecules and/or fragments;
and may occur in nature or be produced, e.g., by immunization,
synthesis or genetic engineering. The antibody or antigen used
herein is dependent upon the antibody or antigen that is being
tested. For example, the number of blood group antigens and thus,
antibodies to these antigens that have been identified is very
large, with more antigens and antibodies continually being
determined. The International Society of Blood Transfusion has
published a non-exclusive list of red cell antigens in Blood Group
Terminology 1990, Vox. Sang. 58:152-169 (1990 and includes, but is
not limited to, antibodies and antigens A, B, D, C, c, Cw, E, e, K,
Fya, Fyb, Jka, Jkb, S and s.
[0080] With the preceding definitions as noted herein, the
following description relates to certain preferred embodiments of
the application, and to a particular methodology for the initial
processing of patient samples prior to chemical analysis. As will
be readily apparent from the discussion, the inventive concepts
described herein can also be suitably applied to other methods that
require the efficient processing of patient samples. In addition,
such terms as "top," "bottom," "lateral," "above," "below" and the
like are also used in order to provide a convenient frame of
reference for use with the accompanying drawings. These terms,
unless stated specifically otherwise, however, are not intended to
be limiting of the present invention. A novel fluid
aspirating/dispensing member is described herein that permits both
sample collection and particle separation by centrifugation or
other separation means, thereby increasing the efficiency of sample
processing before analysis by the wet/dry chemistry components of a
combinational clinical analyzer.
[0081] According to a first embodiment, FIG. 1 shows a sealable
fluid aspirating/dispensing member 100 for sample collection and
particle separation. The fluid aspirating/dispensing member 100 is
typically composed of an injection-moldable, preferably
transparent, thermoplastic material such as polypropylene,
polyallomer, polyethylene terephthalate, co-polymers or blends of
polymers or any other suitable inert material known in the art.
Preferably, the sample being analyzed does not adhere to the walls
of the internal volume of the fluid aspirating/dispensing member
100 wherein these surfaces of the fluid aspirating/dispensing
member 100 may be treated to avoid such adherence of the sample or
reagents to the internal surfaces of the fluid
aspirating/dispensing member 100. The internal volume of a fluid
aspirating/dispensing member 100 may be from about 1 microliter to
about 2000 microliters or more. In one example, the internal volume
is from about 1 microliter to about 500 microliters. In a preferred
embodiment, the fluid aspirating/dispensing member 100 has an
internal volume of about 1 microliter to about 300 microliters. The
thickness of the wall of a fluid aspirating/dispensing member 100
is not critical provided it can withstand centrifugation without
deformation. Typically, the wall has a thickness from about 5 mm to
about 0.1 mm, from about 2 mm to about 0.5 mm or from about 1.5 mm
to about 0.75 mm. As shown in FIG. 1, the fluid
aspirating/dispensing member 100 is defined at an upper end by an
input port 104 and at a lower end by a sealable input port 108.
Between input ports 104 and 108 is an internal cavity comprising a
sample cavity 106 which is in fluid communication with input ports
104 and 108. The input port 104 is configured to attach
hermetically to the proboscis of a metering mechanism (not shown in
this view) for the movement of known volumes of air from the
internal volume of the fluid aspirating/dispensing member 100. In
one embodiment, the proboscis 320 inserts hermetically through the
input port 104 into a proboscis receptacle region 112 of the fluid
aspirating/dispensing member 100. The sealable input port 108 is
configured to permit aspiration or dispensing of fluids through the
input port 108 and into the sample cavity 106. The walls of the
sealable cavity can be thinner than those of the sample cavity 106
to facilitate heat sealing. In one version, the internal volume of
the fluid aspirating/dispensing member 100 may be pre-loaded with
reagents that are required for clinical analysis, for example,
reagents for blood agglutination or blood typing. In another
embodiment, the sample cavity of the fluid aspirating/dispensing
member 100 contains a separation barrier as defined herein.
[0082] Referring now to FIGS. 1 and 2, the herein described fluid
aspirating/dispensing member 100 is shown with an optional cap 114
connected to the upper end of the member 100 by means of a tether
116. The tether 116 may be of any length as long as it does not
interfere with sample collection or particle separation, i.e.,
centrifugation. For example, the tether 116 may be from about 0.5
cm to about 2 cm in length or from about 0.5 cm to about 1 cm in
length. By bending the tether 116 in the direction 210, the cap 114
can be reversibly inserted into the input port 104. In one version,
the cap 114 is not attached to the fluid aspirating/dispensing
member 100 (not shown) and is therefore inserted directly into
input port 104 before centrifugation. The placement of the cap 114
over input port 104 forms a hermetic seal that prevents fluids in
the sample cavity 106 from escaping through input port 104 during
centrifugation and/or handling of the fluid aspirating/dispensing
member 100. In another version, the fluid aspirating/dispensing
member 100 does not require a cap. In yet another embodiment, the
input port 108 may be sealed using a sealant as defined herein that
is cured after being inserted into the sealable cavity 110.
[0083] Referring to FIGS. 1, 2 and 3A, the herein described fluid
aspirating/dispensing member 100 includes a heat sealable cavity
110 that is disposed between the sample cavity 106 and the sealable
input port 108. Application of heat to the heat sealable cavity 110
causes the thermoplastic material of the walls to melt and fuse
thus sealing the sealable input port 108. Other methods may also be
employed to seal input port 108. For example, a sealant, such as an
adhesive, may be applied to the input port 108 and cured by cooling
or other means, for example, localized exposure to UV radiation. In
another version, the input port 108 is hermetically closed using a
bottom cap, for example, a bottom screw cap (not shown).
[0084] With the foregoing structural description of a fluid
aspirating/dispensing member 100, a method is now described with
respect to sample collection and particle separation within a fluid
aspirating/dispensing member 100 in accordance with a second
embodiment.
[0085] Referring to FIG. 3B, the fluid aspirating/dispensing member
100 is shown as attached to a metering mechanism 350 and more
specifically a proboscis 320 of a clinical analyzer. The proboscis
forms a component of a metering mechanism of the analyzer.
Appropriate robotic commands direct the metering system to align
the proboscis 320 with the input port 220 of the fluid
aspirating/dispensing member 100. After proper alignment, the
metering system hermetically inserts the proboscis 320 through the
open input port 220 into the proboscis receptacle region 112 of the
fluid aspirating/dispensing member 100. The input port 108 is then
immersed into the sample 330. The proboscis 320 of the metering
mechanism 350 creates a controlled amount of negative pressure
within the internal volume of the fluid aspirating/dispensing
member 100. The subsequent displacement of air 310 through input
port 104 promotes the movement 343 of a defined volume of sample
330 into the sample cavity 106 of the fluid aspirating/dispensing
member 100. The volume of the aspirated sample depends on the
available volume within the sample cavity 106. In one example, an
aliquot of between about 1 to about 1000 microliters of sample 330
is aspirated. In another example, an aliquot of between about 1 to
about 300 microliters of sample 330 is aspirated. In yet another
example, from about 1 to about 50 microliters of sample 330 is
aspirated into the sample cavity 106 of the fluid
aspirating/dispensing member 100.
[0086] Referring to FIG. 3C, after the aspiration of a sample 330
into the sample cavity 106 is complete, the input port 108 of the
fluid aspirating/dispensing member 100 is removed from the sample
330. The proboscis 320 of the metering mechanism 350 again creates
a controlled amount of negative pressure within the internal volume
of the fluid aspirating/dispensing member 100 that causes a small
bubble of air 347 to be aspirated through input port 108 and
displaces the aspirated sample 345 away from the input port 108 and
into the sample cavity 106 proper. By maintaining the air tight
seal between the proboscis 320 and the proboscis receptacle region
112 of the fluid aspirating/dispensing member 100, the sample is
retained within the sample cavity 106 prior to heat sealing of the
input port 108. This procedure ensures both a reliable seal and
limits any temperature rise of the aspirated sample 345 before
analysis.
[0087] Referring to FIG. 3D, the input port 104 of the fluid
aspirating/dispensing member 100 is placed in a heat sealing device
353 while the proboscis 320 remains hermetically inserted in the
proboscis receptacle region 112 of the fluid aspirating/dispensing
member 100. The thermoplastic material in the walls of the sealable
cavity 110 is then rapidly heated to its melting temperature. The
speed with which the polymer must be heated to its melt temperature
depends on the inherent viscosity of the polymer, the wall
thickness and diameter of the sealable cavity 110, as described in
detail in U.S. Pat. No. 3,929,943, incorporated herein above. The
maximum period for heating can be from about 1 to about 30 seconds
or from 5 to about 20 seconds or about 10 to about 15 seconds. The
heat sealing device can be a high-intensity radiant heater, such as
tungsten or quartz lamp. Alternatively, microwave heaters, such as
dielectric heaters or ultrasonic heaters or any other means known
in the art can be utilized. The melting temperature can be from
about 200 degrees to about 350 degrees Celsius, depending on the
thermoplastic properties of the fluid aspirating/dispensing member
100. After the walls of the heat sealable cavity 110 are heated,
they are pressed in the direction 362 by a press 357 to force the
walls together and seal the open end. In one version, the end of
the press 357 is shaped in the form of a cup 364 that fits over the
input port 108 of the fluid aspirating/dispensing member 100 and
facilitates the fusion and molding of the input port 108 into a
sealed end 355. In one version, a metallic press 357 is heated to
the appropriate melting temperature and applied directly to the
sealable input port 108. After sealing, the input port 108 is
rapidly cooled to promote the solidification of the fused
thermoplastic material thus producing a hermetically sealed fluid
container or cuvette. The press 357 and metering device 320 are
then removed.
[0088] Referring to FIGS. 3E-3G, a cap 367 can be inserted over
input port 104. The fluid aspirating/dispensing member 360 is then
robotically placed into a fixed angle or swinging bucket rotor of a
centrifuge of a clinical analyzer and rotated in the direction 370
around a vertical axis 365. The port 355 is hermetically sealed and
rendered pressure resistant by the heat sealing treatment described
above, thereby ensuring the aspirated sample 345 remains within the
sample cavity 106 during centrifugation about the vertical axis
365. As a consequence of the centrifugal force acting on the
particulate phase of the aspirated sample 345, particles within the
sample accumulate in a pellet 375 at the bottom of the sample
cavity 106 of the fluid aspirating/dispensing member 100. After
centrifugation, the closed cap 367 is removed and the sample
supernatant 380 is collected for presentation to either the "wet"
and/or "dry" chemistry components of a clinical analyzer.
[0089] The sealed fluid aspirating/dispensing member 100, may also
be made from a transparent plastic material that permits optical
testing of the fluid contents within the fluid
aspirating/dispensing member 100. Details relating to the optical
reading of the fluid contents of a sample are described in further
detail in U.S. Pat. Nos. 6,013,528 and 5,846,492, the entire
contents of each are hereby incorporated by reference. In one
example, the walls of the fluid aspirating/dispensing member can
transmit electromagnetic radiation of a wavelength required for the
excitation of a fluorescent ligand and electromagnetic radiation of
a wavelength characteristic of the subsequent emission of
fluorescence.
[0090] According to another version, the fluid
aspirating/dispensing member 100 may be preloaded with a separation
barrier material, as defined herein, that facilitates the
separation of the particulate phase from the liquid phase of a
sample during particle separation, for example, centrifugation.
[0091] A person of ordinary skill in the art will recognize that
the described embodiments can be altered in a number of ways and
still fall within the intended scope of the application. For
example, the fluid aspirating/dispensing member 100 described
herein can be used as part of a series where a second fluid
aspirating/dispensing member 100 is used to collect the supernatant
380 of centrifuged sample that was processed in a first
aspirating/dispensing member 100. For example, a sample, such as
heparinized blood from a patient, can be aspirated into the sample
cavity 106 of a first fluid aspirating/dispensing member 100. After
the sealing of the sealable cavity 106, the aliquot is centrifuged
to pellet the blood cells at the bottom of the sample cavity 106. A
second fluid aspirating/dispensing member 100 can then be used to
aspirate the serum supernatant 380 into the sample cavity 106 of
the second fluid aspirating/dispensing member 100. Sealing of the
sealable cavity 110 of the second fluid aspirating/dispensing
member 100 creates a reaction vessel that can be robotically
transported to the wet/dry components of the clinical analyzer for
additional processing. In one version, the fluid
aspirating/dispensing member 100 may be attached to a metering
mechanism 350 having a proboscis 320 that permits pre-loading of
the sample cavity with one or more reagents for clinical analysis,
for example, reagents for agglutination, i.e., anti-human
immunoglobulin (Coomb's reagent) or other reagents for blood
typing. Mixing of the sample with the reagent can be achieved by
repeated aspiration and dispensing into a suitable container
followed by the aspiration of the homogenized mixture into the
sample cavity 106 of the fluid aspirating/dispensing member 100 for
further processing.
[0092] It will be readily apparent to one of sufficient skill and
as described in greater detail that the following description is
exemplary and therefore, there is potential to use the fluid
aspirating/dispensing member 100 for the processing of, for
example, immunoassays comprising ligand-binding molecules attached
to various particles such as latex or agarose beads and the like. A
number of ligands are known that bind immunoglobulin molecules and
may be covalently coupled to the particles, for example Protein A,
Protein G, Protein A/G, Kappalock. In one embodiment, the particles
may be bound to, for example, bacteriophage expressing a ligand
binding entity (see for example, U.S. Pat. No. 6,979,534, the
contents of which are hereby incorporated by reference herein in
their entirety).
[0093] According to one version, assays used in conjunction with
the fluid aspirating/dispensing member 100 may include magnetic
particles such as magnetic beads. Magnetic particles can be
aspirated together with a sample into the sample cavity 106. After
the sealing of the sealable cavity 110, the particles are separated
from the rest of the sample as described above by simply deploying
a powerful magnet adjacent to the sample cavity in the absence of
centrifugation. Many methods are known in the art where cells can
be rendered magnetic for purposes of cell separation and the like.
For example, cells can be incubated with biotinylated antibodies or
other ligand-binding molecules that are specific for a surface
antigen, characteristic of a particular cell type. Addition of
streptavidin-conjugated magnetic beads (Invitrogen/Dynal Biotech)
then bind to the biotinylated antibodies and thereby render the
cells magnetic and hence amenable to cell separation using a
magnetic field. The description of controlled transport of magnetic
beads is disclosed in U.S. Pat. No. 7,217,561, the contents of
which is hereby incorporated herein in its entirety. Cells used in
the invention may also be tagged using labeled antibodies known in
the art. For example, the labeled antibodies may be
fluorophore-conjugated antibodies. Agglutination can be monitored
by the detection of fluorescence emitted from agglutinated
cells.
[0094] In accordance with a third embodiment, FIGS. 4A and 4B
illustrate a fluid aspirating/dispensing member assembly
configuration that is more amenable to automation and high
throughput analysis of a plurality of samples. FIG. 4A depicts a
fluid aspirating/dispensing plate 400 on which 96 of the fluid
aspirating/dispensing members 100 of FIG. 1 are arranged into 8
rows of 12 fluid aspirating/dispensing members 425. FIG. 4B
portrays a cross sectional view of the plate 400 to show row 420 of
12 fluid aspirating/dispensing members 425 attached to a solid
support 414. In one version, the attachment may be reversible. Each
of the aspirating/dispensing members 425 comprises an input port
404, proboscis receptacle region 412 attached to a solid support
414, a sample cavity 406, a sealable cavity 410 and sealable input
port 408. FIG. 5 illustrates how the sealable input ports 408 of
the fluid aspirating/dispensing plate 400 can align with the wells
510 of a microtiter plate 500.
[0095] With the foregoing structural description of a fluid
aspirating/dispensing plate 400 of fluid aspirating/dispensing
members, a method is now described with respect to sample
collection and particle separation using a fluid
aspirating/dispensing plate 400 in accordance with a fourth
embodiment.
[0096] Referring to FIG. 6A, a cross section view of the row 420 of
the fluid aspirating/dispensing members 425 of FIG. 4B is portrayed
with the sealable input ports 408 aligned with the wells 510 of a
microtiter plate 500. Appropriate robotic commands direct the
metering mechanism 603 to align the proboscises 605 with the input
ports 404 of the fluid aspirating/dispensing plate 400. After
proper alignment, the metering system hermetically inserts the
proboscises 605 through the open input ports 404 into the proboscis
receptacle regions 412 of the fluid aspirating/dispensing members
425.
[0097] Referring to FIGS. 6B and 6C, the metering mechanism then
aligns the sealable input ports 408 of the fluid
aspirating/dispensing members 425 with the wells 510 of the
microtiter plate 500 and immerses the sealable input ports 408 into
the samples 607 within each well 510. The proboscises 605 of the
metering mechanism 603 create a controlled amount of negative
pressure within the internal volume of each of the fluid
aspirating/dispensing members 425. The subsequent displacement of
air 601 through the input ports 404 promotes the movement 643 of a
defined volume of samples 607 into the sample cavities 406 of each
of the fluid aspirating/dispensing members 425. The volume of the
aspirated sample within each fluid aspirating/dispensing member 425
depends on the available volume within the sample cavities 406. In
one example, an aliquot of between 0.1 to 1000 microliters of
sample 607 is aspirated. In another example, an aliquot of between
0.1 to 300 microliters of sample 607 is aspirated. In yet another
example, from 0.1 to 50 microliters of sample 607 is aspirated into
the sample cavity 406 of each of the fluid aspirating/dispensing
members 425.
[0098] Referring now to FIG. 6D, the sealable input ports 408 are
removed from the samples 607 in the wells 510. Again the
proboscises 605 of the metering mechanism 603 exert a controlled
amount of negative pressure within the internal volume of each of
the fluid aspirating/dispensing members 425. The subsequent
displacement of air within the internal cavities of the row 420 of
fluid aspirating/dispensing members in the direction 625 causes the
movement of the aspirated sample 620 into the sample cavity 406
proper and the aspiration of a measured amount of air 610 through
the sealable input ports 408 into the sealable cavity 410 prior to
heat sealing of the sealable input ports 408. This procedure averts
the heating of the aspirated sample 620 and ensures a reliable
seal
[0099] As shown in FIG. 6E, the walls of the sealable cavities 410
are then heated to the appropriate melting temperature using the
row of heat sealing devices 640. After the walls of the heat
sealable cavities 410 are heated, they are pressed in the direction
635 by the presses 630 to force the walls together and seal the
open ends of the sealable input ports 408. In one version, the ends
of the presses 630 are shaped in the form of a cup 645 that fits
over the sealable input ports 408, thereby facilitating the fusion
and molding of the input ports 408 into the sealed ends 650,
depicted in FIG. 6F. In one version, metallic presses 630 are
heated to the appropriate melting temperature and applied directly
to the sealable input ports 408 for the melting and fusion of the
walls of the sealable cavities 410. After sealing, the sealable
input ports 408 are rapidly cooled to promote the solidification of
the fused thermoplastic material thus producing a hermetically
sealed fluid container or cuvette. The presses 630 and the
proboscises 605 are then removed.
[0100] Referring to FIG. 6G-I, an optional lid 660 is placed onto
the fluid aspirating/dispensing plate 400 and hermetically seals
the input ports 404 of the fluid aspirating/dispensing members. Lid
660 prevents the aspirated sample 620 from escaping the sample
cavity 406 of each fluid aspirating/dispensing member and prevents
cross-contamination of the aspirated samples 620. The fluid
aspirating/dispensing plate 400 is then robotically placed into a
fixed angle or swinging bucket rotor of a centrifuge and spun
around a vertical axis 365 in the direction 370 for the time
required to separate the particles within the aspirated sample from
the rest of the sample. At the conclusion of the centrifugation,
the particles within the aspirated samples 620 form pellets 670 at
the bottom of the sample cavities 406. The lid 660 is then removed
and the supernatants 680 can be collected and presented to the
wet/dry components of the clinical analyzer.
[0101] In an alternative version, the sample cavities 406 may be
preloaded with a separation barrier material, as defined herein, to
facilitate the separation of the particulate phase from the liquid
phase of the aspirated samples 620 during centrifugation. In
another version, the sample cavities 406 may be pre-loaded with
reagents, for example, reagents for agglutination or blood typing
or antibodies attached to various particles as defined herein for
immunoassays. In yet another version, the aspirated samples 620
within the fluid aspirating/dispensing members of a plate contain
magnetic particles that can be separated from the rest of the
aspirated samples 620 by placing the plate on a strong magnet
according to protocols that are well known in the art.
[0102] As noted above, the herein described processing of samples
in a plate of fluid aspirating/dispensing members is particularly
amenable to automation for the rapid processing of STAT samples in
urgent care facilities. Sample processing can be controlled by
appropriate software programs running on a dedicated computer
component of a clinical analyzer. In one version, the solid support
414 of a plate of fluid aspirating/dispensing members further
comprises appendages to facilitate robotic handling and appropriate
adapters for centrifugation.
[0103] The disclosure herein also provides for a kit format which
comprises a package unit having one or more fluid
aspirating/dispensing members of the subject disclosure and in some
embodiments includes containers of various reagents. The kit may
also contain one or more of the following items: buffers,
instructions, and positive or negative controls. Kits may include
containers of reagents mixed together in suitable proportions for
performing the methods described herein. Reagent containers
preferably contain reagents in unit quantities that obviate
measuring steps when performing the subject methods. Kits may
further comprise fluid aspirating/dispensing members pre-loaded
with reagents.
PARTS LIST FOR FIGS. 1-6
[0104] 100 fluid aspirating/dispensing member [0105] 104 input port
[0106] 106 sample cavity [0107] 108 sealable input port [0108] 110
heat sealable cavity [0109] 112 proboscis receptacle region [0110]
114 cap [0111] 116 connector [0112] 118 vertical axis [0113] 210
movement of cap [0114] 220 open input port [0115] 225 closed input
port [0116] 310 direction of movement of air [0117] 320 proboscis
[0118] 330 sample [0119] 335 direction of movement of sample [0120]
343 aspiration of sample [0121] 345 aspirated sample [0122] 347 air
space [0123] 350 metering mechanism [0124] 353 heat sealing device
[0125] 355 sealed end [0126] 357 press [0127] 360 fluid
aspirating/dispensing member in a centrifuge [0128] 362 direction
of movement of press [0129] 364 cup [0130] 365 axis of rotation
[0131] 367 closed cap [0132] 370 direction of rotation [0133] 375
pellet [0134] 380 supernatant [0135] 390 probe [0136] 400 Fluid
aspirating/dispensing plate with array of fluid
aspirating/dispensing members [0137] 404 input ports [0138] 406
sample cavities [0139] 408 sealable input ports [0140] 410 sealable
cavities [0141] 412 proboscis receptacle regions [0142] 414 solid
support [0143] 420 row of fluid aspirating/dispensing members
[0144] 425 fluid aspirating/dispensing member [0145] 500 microtiter
plate [0146] 510 wells [0147] 601 direction of movement of air
[0148] 603 array of metering mechanisms [0149] 605 array of
proboscises [0150] 607 samples [0151] 610 aspiration of air [0152]
615 array of probes [0153] 620 aspirated samples [0154] 625
direction of movement of air [0155] 630 presses [0156] 640 heat
sealer devices [0157] 643 aspiration of samples [0158] 645 cup
[0159] 650 sealed ends [0160] 660 cover [0161] 670 pellets [0162]
680 supernatants
[0163] While this invention has been particularly shown and
described with references to preferred embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
intended scope of the invention encompassed by the following
appended claims.
* * * * *