Disposable hematology cartridge

• US 6,537,501 B1
• Filed: 10/28/1999
• Issued: 03/25/2003
• Est. Priority Date: 05/18/1998
• Status: Expired due to Term

• Current Assignee: Washington University In St Louis
• Original Assignee: University of Washington
• Inventors: Morff, Robert J., Edwards, Floyd, Holl, Mark R., Klein, Gerald L.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): WRIGHT, PATRICIA KATHRYN
• Application Number:
  US09/428,801
• Time in Patent Office: 1,244 Days
• Field of Search: 422/58, 422/55, 422/61, 422/68.1, 422/81, 422/82, 422/82.05, 422/82-8-, 422/102, 422/103, 422/101, 436/43, 436/52-53, 436/164-165, 436/172, 436/177, 436/180, 738/632.1, 738/637--86, 356/73, 356/246
• US Class Current: 422/537
• CPC Class Codes: B01F 25/433 Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
• CPC Class Codes: B01F 25/4331 Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
• CPC Class Codes: B01F 25/4332 Mixers with a strong change of direction in the conduit for homogenizing the flow
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 3/502707 : characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502776 : specially adapted for focusing or laminating flows
• CPC Class Codes: B01L 3/565 : Seals in general F16L
• CPC Class Codes: G01N 15/14 : Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: Y10T 156/1064 : Partial cutting [e.g., grooving or incising]
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/118339 : with formation of a segmented stream
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25125 : Digestion or removing interfering materials
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The present invention provides an apparatus and method for storing a particle-containing liquid. The storage apparatus comprises a microfluidic convoluted flow channel having a plurality of particle capture regions. The storage channel is preferably an isotropic spatially periodic channel. Sedimented particles can be resuspended following storage. This invention further provides a microfluidic analysis cartridge having a convoluted storage channel therein. The sample analysis can use optical, electrical, pressure sensitive, or flow sensitive detection. A plurality of analysis channels can be included in a single cartridge. The analysis channels can be joined to reagent inlets for diluents, indicators or lysing agents. A mixing channel can be positioned between the reagent inlet and the analysis region to allow mixing and reaction of the reagent. The cartridge can include additional valves and pumps for flow management. The analysis cartridge can be a self-contained disposable cartridge having an integral waste storage container. This invention further provides a sheath flow assembly. The sheath flow assembly includes a sample channel and first and second sheath fluid channels positioned on either side of and converging with the sample channel. The assembly also includes upper and lower sheath fluid chambers positioned above and below and converging with the sample channel. The flow cartridges of this invention can be formed by molding, machining or etching. In a preferred embodiment they are laminated. This invention further provides a method of fabricating a laminated microfluidic flow device. In the method, flow elements are formed in rigid sheets and abutting surfaces of the sheets are bonded together.

Claims

1. A disposable fluidic hematology cartridge for analyzing a particle-containing liquid sample, comprising:

a sample inlet;

an optical measuring channel fluidically coupled with said sample inlet and having an optical measuring region;

a first valve interface positioned between said sample inlet and said optical measuring region;

a flow cytometric measuring channel fluidically coupled with said sample inlet;

a second valve interface positioned between said sample inlet and said flow cytometric measuring channel wherein said flow cytometric measuring channel further comprises a flow cytometric measuring region;

a sheath flow assembly positioned along said flow cytometric measuring channel before said flow cytometric measuring region; and

a plurality of laminate layers wherein at least one of said sample inlet, optical measuring channel, optical measuring region, first or second valve interfaces, flow cytometric measuring channel or flow cytometric measuring region is formed by at least three of said laminate layers.

2. The hematology cartridge of claim 1 wherein said optical measuring channel is formed by a first sheet attached to a second sheet having a cutout region attached to a third sheet attached to the second sheet.

3. The hematology cartridge of claim 1 wherein said optical measuring channel and said flow cytometric measuring channel are positioned in parallel.

4. The hematology cartridge of claim 1 wherein said optical measuring and said flow cytometric measuring regions each comprise a first transparent window positioned over said, measuring channels.

5. The hematology cartridge of claim 4 wherein said optical measuring and said flow cytometric measuring regions each further comprise a second transparent window, positioned under said measuring channels.

6. The hematology cartridge of claim 5 wherein the width of said optical measuring channel is increased in said optical measuring region.

7. The hematology cartridge of claim 4 wherein the optical pathlength of said optical measuring channel is increased by increasing the channel depth at the optical measuring region.

8. The hematology cartridge of claim 1 wherein said flow cytometric measuring channel is narrowed in said flow cytometric measuring region to constrict particles into single file.

9. The hematology cartridge of claim 1 wherein said sheath flow assembly comprises first and second sheath flow channels on either side of and converging with said flow cytometric measuring channel.

10. The hematology cartridge of claim 9 wherein the width of said flow cytometric measuring channel does not contract within said sheath flow assembly.

11. The hematology cartridge of claim 9 wherein said sheath flow assembly further comprises upper and lower sheath fluid chambers positioned above and below and converging with said flow cytometric measuring channel.

12. The hematology cartridge of claim 1 further comprising a first reagent inlet, positioned along said optical measuring channel before said optical measuring region.

13. The hematology cartridge of claim 12 further comprising a second reagent inlet, positioned along said flow cytometric measuring channel before said flow cytometric measuring region.

14. The hematology cartridge of claim 13 further comprising said sheath flow assembly positioned along said flow cytometric measuring channel between said second reagent inlet and said flow cytometric measuring region.

15. The hematology cartridge of claim 13 wherein each of said first and second reagent inlets comprises a syringe pump interface.

16. The hematology cartridge of claim 13 further comprising a mixing channel positioned along said flow cytometric measuring channel between said second reagent inlet and said flow cytometric measuring region.

17. The hematology cartridge of claim 16 wherein said mixing channel is a spatially periodic channel.

18. The hematology cartridge of claim 1 further comprising a waste storage container positioned downstream of said flow cytometric measuring region.

19. A method of blood analysis using the hematology cartridge of claim 1, comprising the steps of:

introducing a sample of blood into said sample inlet;

measuring the absorption of said blood in said optical measuring region;

measuring the scattering by said blood in said flow cytometric measuring region;

and using said sheath flow assembly to hydrodynamically focus said blood.

20. The method of blood analysis of claim 19 wherein said hematology cartridge further comprises a convoluted sample storage channel positioned before said flow cytometric measuring region, and wherein said method further comprises the step of storing said blood in said storage channel, whereby particles in said blood sediment in said storage channel.

21. The method of blood analysis of claim 20 further comprising the step of resuspending said particles in said blood.

22. The method of blood analysis of claim 19 wherein said hematology cartridge further comprises a first reagent inlet positioned between said sample inlet and said optical measuring region, and wherein said method further comprises the steps of introducing a cell lysing agent through said first reagent inlet and obtaining the hemoglobin content of said blood from the measured absorption.

23. The method of blood analysis of claim 22 wherein said hematology cartridge further comprises a second reagent inlet positioned between said sample inlet and said flow cytometric measuring region, and wherein said method further comprises the steps of introducing a second reagent through said second reagent inlet and characterizing the white blood cells from the measured scattering.

25. The method of blood analysis of claim 23 wherein said hematology cartridge further comprises a mixing channel positioned between said second reagent inlet and said flow cytometric measuring region, and wherein said method further comprises the step of allowing said blood and said second reagent to mix and react in said mixing channel.

26. The method of blood analysis of claim 19 wherein said hematology cartridge further comprises a waste storage container positioned downstream of said flow cytometric measuring region, and wherein said method further comprises the step of collecting said blood in said waste storage container.

27. The method of blood analysis of claim 26 further comprising the step of disposing of said hematology cartridge after use.

28. The hematology cartridge of claim 1 whereby said optical measuring channel is an absorption measuring channel and said optical measuring region is an absorption measuring region.

24. The method of blood analysis of claimed 23 wherein said step of introducing a second reagent comprises introducing a reagent for masking red blood cells and platelets.

29. A disposable fluidic hematology cartridge for analyzing a particle-containing liquid sample, comprising:

a sample inlet;

an optical measuring channel fluidically coupled with said sample inlet and having an optical measuring region;

a first valve interface positioned between said sample inlet and said optical measuring region;

a flow cytometric measuring channel fluidically coupled with said sample inlet;

a second valve interface positioned between said sample inlet and said flow cytometric measuring channel wherein said flow cytometric measuring channel further comprises a flow cytometric measuring region;

a convoluted sample storage channel positioned before said flow cytometric measuring channel; and

a plurality of laminate layers wherein at least one of said sample inlet, optical measuring channel, optical measuring region, first or second valve interfaces, flow cytometric measuring channel or flow cytometric measuring region is formed by at least three of said laminate layers,

and wherein said first valve interface is positioned between said storage channel and said optical measuring region and said second valve interface is positioned between said storage channel and said flow cytometric measuring region.

30. The hematology cartridge of claim 29 wherein said storage channel is a spatially periodic channel.

31. The hematology cartridge of claim 30 wherein said storage channel is an isotropic spatially periodic channel.