CA2500751A1 - Buffy coat tube and float system and method - Google Patents
Buffy coat tube and float system and method Download PDFInfo
- Publication number
- CA2500751A1 CA2500751A1 CA002500751A CA2500751A CA2500751A1 CA 2500751 A1 CA2500751 A1 CA 2500751A1 CA 002500751 A CA002500751 A CA 002500751A CA 2500751 A CA2500751 A CA 2500751A CA 2500751 A1 CA2500751 A1 CA 2500751A1
- Authority
- CA
- Canada
- Prior art keywords
- float
- support members
- tube
- main body
- body portion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims 16
- 210000003743 erythrocyte Anatomy 0.000 claims abstract 7
- 210000002381 plasma Anatomy 0.000 claims abstract 7
- 230000005484 gravity Effects 0.000 claims abstract 6
- 238000000926 separation method Methods 0.000 claims abstract 4
- 238000005119 centrifugation Methods 0.000 claims abstract 3
- 210000004369 blood Anatomy 0.000 claims 34
- 239000008280 blood Substances 0.000 claims 34
- 239000000470 constituent Substances 0.000 claims 7
- 210000004027 cell Anatomy 0.000 claims 5
- 230000000694 effects Effects 0.000 claims 3
- 239000003795 chemical substances by application Substances 0.000 claims 2
- 238000002372 labelling Methods 0.000 claims 2
- 239000000463 material Substances 0.000 claims 2
- 239000012491 analyte Substances 0.000 claims 1
- 238000005286 illumination Methods 0.000 claims 1
- 238000003384 imaging method Methods 0.000 claims 1
- 239000003446 ligand Substances 0.000 claims 1
- 230000004807 localization Effects 0.000 claims 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/505—Containers for the purpose of retaining a material to be analysed, e.g. test tubes flexible containers not provided for above
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5021—Test tubes specially adapted for centrifugation purposes
- B01L3/50215—Test tubes specially adapted for centrifugation purposes using a float to separate phases
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/81—Packaged device or kit
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/807—Apparatus included in process claim, e.g. physical support structures
- Y10S436/81—Tube, bottle, or dipstick
Abstract
A tube and float system for use in separation and axial expansion of the buf fy coat is provided. The system includes a transparent, or semi-transparent, flexible sample tube (130) and a rigid separator float (110) having a specif ic gravity intermediate that of red blood cells and plasma. The sample tube has an elongated sidewall (136) having a first cross-sectional inner diameter. T he float consists of a main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube. The main body portion and the support members of the float have a cross-sectional diameter less than that of the first cross-sectional inner diameter of the tube (138) when the sample tube is expanded, such as b y centrifugation. The main body portion of the float together with an axially aligned portion of the sidewall define an annular volume therebetween. The support members protruding from the main body portion of the float traverse said annular volume to produce one or more analysis areas. During centrifugation, the centrifugal force enlarges the diameter of the tube to permit density-based axial movement of the float in the tube. Thereafter, th e centrifugal force is reduced to cause the tube sidewall to return to its fir st diameter, thereby capturing the float and trapping the buffy coat constituen ts in the analysis area.
Claims (48)
1. A method of separating and axially expanding the buffy coat constituents of a blood sample, comprising:
introducing the blood sample into a flexible sample tube, the sample tube having an elongate sidewall having a first cross-sectional inner diameter;
introducing an elongate rigid volume-occupying float into the flexible sample tube, said rigid float having a specific gravity intermediate that of red blood cells and plasma;
said float comprising:
a main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members of the float having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas;
centrifuging the sample tube to effect a density-based separation of the blood sample into discrete layers at a rotational speed that causes a resilient enlargement of the sidewall to a second diameter in response to centrifugal force, said second diameter being sufficiently large to permit axial movement of the float in the tube;
moving said float into axial alignment with at least the buffy coat constituents of the blood sample in response to centrifugal forces produced in centrifuging the blood; and thereafter, reducing the rotational speed to cause the tube sidewall to return to said first diameter, thereby capturing the float and trapping the buffy coat constituents in the analysis area.
introducing the blood sample into a flexible sample tube, the sample tube having an elongate sidewall having a first cross-sectional inner diameter;
introducing an elongate rigid volume-occupying float into the flexible sample tube, said rigid float having a specific gravity intermediate that of red blood cells and plasma;
said float comprising:
a main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members of the float having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas;
centrifuging the sample tube to effect a density-based separation of the blood sample into discrete layers at a rotational speed that causes a resilient enlargement of the sidewall to a second diameter in response to centrifugal force, said second diameter being sufficiently large to permit axial movement of the float in the tube;
moving said float into axial alignment with at least the buffy coat constituents of the blood sample in response to centrifugal forces produced in centrifuging the blood; and thereafter, reducing the rotational speed to cause the tube sidewall to return to said first diameter, thereby capturing the float and trapping the buffy coat constituents in the analysis area.
2. The method according to claim 1, wherein said float is introduced into the blood sample tube before the blood is introduced therein.
3. The method according to claim 1, wherein said blood sample is introduced into the blood sample tube before the float is introduced therein.
4. The method according to claim 1, wherein the blood sample comprises anticoagulated whole blood.
5. The method according to claim 1, wherein the blood sample tube comprises a closed first end and an open second end adapted to receive a closure device and the float.
6. The method according to claim 1, wherein the sample tube is sized to receive a blood sample of approximately ten milliliters in volume.
7. The method according to claim 1, wherein the float is further moved into axial alignment with at least one of a portion of a separated red blood cell layer and a portion of a separated plasma layer.
8. An apparatus for separation and analysis of a target analyte in sample of anticoagulated whole blood, comprising:
an at least semi-transparent, flexible sample tube for holding the sample, the sample tube having an elongate sidewall of a first cross-sectional inner diameter;
an elongate, rigid, volume-occupying float having a specific gravity intermediate that of red blood cells and plasma, said float comprising:
a main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas;
said sidewall being resiliently radially expandable to a second diameter in response to centrifugal force, said second diameter being sufficiently large to permit axial movement of the float in the tube during centrifugation.
an at least semi-transparent, flexible sample tube for holding the sample, the sample tube having an elongate sidewall of a first cross-sectional inner diameter;
an elongate, rigid, volume-occupying float having a specific gravity intermediate that of red blood cells and plasma, said float comprising:
a main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas;
said sidewall being resiliently radially expandable to a second diameter in response to centrifugal force, said second diameter being sufficiently large to permit axial movement of the float in the tube during centrifugation.
9. The apparatus according to claim 8, wherein the blood sample tube comprises a closed first end, an open second end adapted to receive a closure device, and the float.
10. The apparatus according to claim 8, wherein the sample tube is sized to receive a blood sample of approximately ten milliliters in volume.
11. The apparatus according to claim 8, wherein the float includes opposite axial ends which are tapered in the axial direction.
12. The apparatus according to claim 8, wherein the one or more support members include one or more annular ridges.
13. The apparatus according to claim. 8, wherein the one or more support members include two annular ridges.
14. The apparatus according to claim 13, wherein the two annular ridges are disposed at opposite axial ends of the float.
15. The apparatus according to claim 8, wherein the one or more support members include three or more axially-spaced annular ridges.
16. The apparatus according to claim 8, wherein the one or more support members comprises a helical ridge.
17. The apparatus according to claim 8, wherein the one or more support members include a plurality of radially spaced-apart splines.
18. The apparatus according to claim 17 wherein, the splines are aligned parallel to an axis of the float.
19. The apparatus according to claim 17, wherein the one or more support members further include annular ridges disposed at opposite axial ends of the float.
20. The apparatus according to claim 8, wherein the one or more support members include a plurality of radially spaced-apart splines intersecting with a plurality of axially spaced-apart splines.
21 21. The apparatus according to claim 8, wherein the one or more support members include a plurality of raised protrusions spaced over the surface of the main body portion.
22. The apparatus according to claim 21, wherein the protrusions are selected from rounded bumps and faceted bumps.
23. The apparatus according to claim 8, wherein the tube is formed of a flexible transparent polymeric material and the float is formed of a hard polymeric material.
24. The apparatus according to claim 8, wherein the float has a specific gravity in the range of from about 1.029 to about 1.089.
25. A volume occupying separator float adapted for use with an associated sample tube, comprising:
a main body portion and one or more support members protruding from the main body portion to engage and support a sidewall of the sample tube, said main body portion and said support members having a cross-sectional diameter less than a first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas.
a main body portion and one or more support members protruding from the main body portion to engage and support a sidewall of the sample tube, said main body portion and said support members having a cross-sectional diameter less than a first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas.
26. The separator float according to claim 25, wherein the main body portion and the one or more support members are integrally formed.
27. The separator float according to claim 25, wherein the clearance gap has a radial extent of about 50 microns.
28. The separator float according to claim 27, wherein the sample tube is sized to receive a blood sample of approximately ten milliliters in volume.
29. The separator float according to claim 27, wherein the float includes opposite axial ends which are tapered in the axial direction.
30. The separator float according to claim 27, wherein the one or more support members include one or more annular ridges.
31. The separator float according to claim 27, wherein the one or more support members include two annular ridges.
32. The separator float according to claim 31, wherein the two annular ridges are disposed at opposite axial ends of the float.
33. The separator float according to claim 27, wherein the one or more support members include three or more axially-spaced annular ridges.
34. The separator float according to claim 27, wherein the one or more support members comprises a helical ridge.
35. The separator float according to claim 27, wherein the one or more support members include a plurality of circumferentially spaced-apart splines.
36. The separator float according to claim 35 wherein, the splines are aligned parallel to an axis of the float.
37. The separator float according to claim 35, wherein the one or more support members further include annular ridges disposed at opposite axial ends of the float.
38. The separator float according to claim 27, wherein the one or more support members include a plurality of radially spaced-apart splines intersecting with a plurality of axially spaced-apart splines.
39. The separator float according to claim 27, wherein the one or more support members include a plurality of raised protrusions spaced over the surface of the main body portion.
40. The separator float according to claim 39, wherein the protrusions are selected from rounded bumps and faceted bumps.
41. A method for detecting circulating target cells in an anticoagulated whole blood sample, comprising:
combining the blood sample with one or more labeling agents so as to differentiate target cells from other cells in the blood sample;
introducing the blood sample into an at least semi-transparent, flexible sample tube, the sample tube having an elongated sidewall of a first cross-sectional inner diameter;
inserting a volume-occupying separator float having a specific gravity intermediate that of red blood cells and plasma into the sample tube;
said separator float comprising a rigid main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas;
centrifuging the blood sample and separator float in the sample tube to effect centrifugally-motivated localization of any target cells present in the blood sample within said analysis area; and after said centrifuging, allowing the sample tube to constrict upon the separator float, and examining the blood sample present in the analysis area to identify any target cells contained therein.
combining the blood sample with one or more labeling agents so as to differentiate target cells from other cells in the blood sample;
introducing the blood sample into an at least semi-transparent, flexible sample tube, the sample tube having an elongated sidewall of a first cross-sectional inner diameter;
inserting a volume-occupying separator float having a specific gravity intermediate that of red blood cells and plasma into the sample tube;
said separator float comprising a rigid main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members traverse said annular volume to produce one or more analysis areas;
centrifuging the blood sample and separator float in the sample tube to effect centrifugally-motivated localization of any target cells present in the blood sample within said analysis area; and after said centrifuging, allowing the sample tube to constrict upon the separator float, and examining the blood sample present in the analysis area to identify any target cells contained therein.
42. The method according to claim 41, wherein the one or more labeling agents includes a fluorescently labeled ligand, and further wherein said examining step includes imaging the blood sample present in the analysis area under illumination and magnification.
43. The method according to claim 41, further comprising combining the blood sample with a stain.
44. The separated and axially expanded buffy coat constituents provided by the method of claim 1.
45. A method of separating and axially expanding the buffy coat constituents of a blood sample, comprising:
introducing the blood sample into an assembly comprising a flexible sample tube, and an elongated rigid volume-occupying float, the sample tube having an elongate sidewall having a first cross-sectional inner diameter;
the rigid float having a specific gravity intermediate that of red blood cells and plasma;
said float comprising:
a main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members of the float having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members protruding from the main body portion traverse said annular volume to produce one or more analysis areas;
centrifuging the assembly to effect a density-based separation of the blood sample in the tube into discrete layers at a rotational speed that causes a resilient enlargement of the sidewall to a second diameter in response to centrifugal force, said second diameter being sufficiently large to permit axial movement of the float in the tube;
moving said float into axial alignment with at least the buffy coat constituents of the blood sample in response to centrifugal force produced in centrifuging the blood; and thereafter, reducing the rotational speed to cause the tube sidewall to return to said first diameter, thereby capturing the float and trapping the buffy coat constituents in the analysis area.
introducing the blood sample into an assembly comprising a flexible sample tube, and an elongated rigid volume-occupying float, the sample tube having an elongate sidewall having a first cross-sectional inner diameter;
the rigid float having a specific gravity intermediate that of red blood cells and plasma;
said float comprising:
a main body portion and one or more support members protruding from the main body portion to engage and support the sidewall of the sample tube, said main body portion and said support members of the float having a cross-sectional diameter less than said first inner diameter of the tube when the sample tube is expanded, wherein said main body portion together with an axially aligned portion of said sidewall define an annular volume therebetween; and wherein said support members protruding from the main body portion traverse said annular volume to produce one or more analysis areas;
centrifuging the assembly to effect a density-based separation of the blood sample in the tube into discrete layers at a rotational speed that causes a resilient enlargement of the sidewall to a second diameter in response to centrifugal force, said second diameter being sufficiently large to permit axial movement of the float in the tube;
moving said float into axial alignment with at least the buffy coat constituents of the blood sample in response to centrifugal force produced in centrifuging the blood; and thereafter, reducing the rotational speed to cause the tube sidewall to return to said first diameter, thereby capturing the float and trapping the buffy coat constituents in the analysis area.
46. The method according to claim 45, wherein the blood sample comprises anticoagulated whole blood.
47. The method according to claim 45, wherein the sample tube is sized to receive a blood sample of approximately ten milliliters in volume.
48. The method according to claim 45, wherein the float is further moved into axial alignment with at least one of a portion of a separated red blood cell layer and a portion of a separated plasma layer.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US10/263,975 US7074577B2 (en) | 2002-10-03 | 2002-10-03 | Buffy coat tube and float system and method |
| US10/263,975 | 2002-10-03 | ||
| PCT/US2003/031205 WO2004031770A1 (en) | 2002-10-03 | 2003-10-02 | Buffy coat tube and float system and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2500751A1 true CA2500751A1 (en) | 2004-04-15 |
| CA2500751C CA2500751C (en) | 2012-01-17 |
Family
ID=32042119
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2500751A Expired - Lifetime CA2500751C (en) | 2002-10-03 | 2003-10-02 | Buffy coat tube and float system and method |
Country Status (6)
| Country | Link |
|---|---|
| US (4) | US7074577B2 (en) |
| EP (2) | EP2458381B1 (en) |
| JP (2) | JP4401963B2 (en) |
| AU (1) | AU2003277224B2 (en) |
| CA (1) | CA2500751C (en) |
| WO (1) | WO2004031770A1 (en) |
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2002
- 2002-10-03 US US10/263,975 patent/US7074577B2/en active Active
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2003
- 2003-10-02 JP JP2004542039A patent/JP4401963B2/en not_active Expired - Lifetime
- 2003-10-02 AU AU2003277224A patent/AU2003277224B2/en not_active Expired
- 2003-10-02 CA CA2500751A patent/CA2500751C/en not_active Expired - Lifetime
- 2003-10-02 EP EP12001206.7A patent/EP2458381B1/en not_active Expired - Lifetime
- 2003-10-02 WO PCT/US2003/031205 patent/WO2004031770A1/en active Application Filing
- 2003-10-02 EP EP03799393.8A patent/EP1546720B1/en not_active Expired - Lifetime
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2006
- 2006-03-07 US US11/370,635 patent/US7329534B2/en not_active Expired - Lifetime
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- 2008-02-11 US US12/029,274 patent/US7915029B2/en not_active Expired - Lifetime
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2009
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| US20040067536A1 (en) | 2004-04-08 |
| JP2006502389A (en) | 2006-01-19 |
| JP5079763B2 (en) | 2012-11-21 |
| EP2458381A3 (en) | 2012-07-25 |
| EP1546720A1 (en) | 2005-06-29 |
| EP1546720B1 (en) | 2016-11-16 |
| WO2004031770A1 (en) | 2004-04-15 |
| JP2009288254A (en) | 2009-12-10 |
| US8012742B2 (en) | 2011-09-06 |
| US7074577B2 (en) | 2006-07-11 |
| CA2500751C (en) | 2012-01-17 |
| EP2458381A2 (en) | 2012-05-30 |
| AU2003277224B2 (en) | 2010-01-21 |
| US7329534B2 (en) | 2008-02-12 |
| US7915029B2 (en) | 2011-03-29 |
| AU2003277224A1 (en) | 2004-04-23 |
| US20060154308A1 (en) | 2006-07-13 |
| EP1546720A4 (en) | 2009-10-28 |
| US20110171680A1 (en) | 2011-07-14 |
| EP2458381B1 (en) | 2018-05-30 |
| US20080128340A1 (en) | 2008-06-05 |
| JP4401963B2 (en) | 2010-01-20 |
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