Methods for complete blood count measurement

• US 10,641,698 B2
• Filed: 11/21/2017
• Issued: 05/05/2020
• Est. Priority Date: 06/12/2015
• Status: Active Grant

• Current Assignee: Cytochip Inc.
• Original Assignee: Cytochip Inc.
• Inventors: Shi, Wendian, Ding, Yuzhe
• Primary Examiner(s): Sines, Brian J.
• Application Number:
  US15/819,416
• Publication Number:
  US 20180095023A1
• Time in Patent Office: 896 Days
• Field of Search: 422 681, 422502, 422503, 422504, 422554, 436 43, 436174, 436180
• CPC Class Codes: B01L 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 2300/168 Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0457 passive flow or gravitation
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 2400/088 by specific surface properties
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: G01N 15/01 : specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/06 : Investigating concentration of particle suspensions by weighing G01N5/00; investigating sedimentation of particle suspensions G01N15/04; investigating individual particles G01N15/10
• CPC Class Codes: G01N 15/075 : by optical means
• CPC Class Codes: G01N 15/1404 : Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1459 : the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1484 : microstructural devices
• CPC Class Codes: G01N 2015/012 : Red blood cells
• CPC Class Codes: G01N 2015/016 : White blood cells
• CPC Class Codes: G01N 2015/018 : Platelets
• CPC Class Codes: G01N 2015/0687 : in solutions, e.g. non volatile residue
• CPC Class Codes: G01N 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1486 : Counting the particles
• CPC Class Codes: G01N 21/05 : Flow-through cuvettes G01N21/09 takes precedence; handling fluid samples G01N1/10
• CPC Class Codes: G01N 21/53 : within a flowing fluid, e.g. smoke
• CPC Class Codes: G01N 21/6486 : Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/85 : Investigating moving fluids or granular solids
• CPC Class Codes: G01N 33/5094 : for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/726 : Devices
• CPC Class Codes: G01N 33/80 : involving blood groups or blood types or red blood cells white blood cells G01N33/56972

Abstract

The disclosure relates to devices and methods for analyzing blood cells in a sample. In various embodiments, the present disclosure provides devices and methods of performing complete blood count (CBC) testing. In various embodiments, the present disclosure provides a cartridge device and a reader instrument device, wherein the reader instrument device receives, operates, and/or actuates the cartridge device. In various embodiments, the present disclosure provides a method of using a device as disclosed herein for analyzing blood cells in a sample.

Claims

1. A device for analyzing blood cells in a sample, comprising:

a cartridge device, wherein the cartridge device comprises;

a fluidic conduit configured for receiving the sample into the cartridge device;

a chamber fluidly connected to the fluidic conduit and configured for mixing at least a portion of the sample with at least of a portion of a reagent to form one or more sample mixtures; and

a flow cell fluidly connected to the chamber and configured for forming one or more sample streams from the one or more sample mixtures without a sheath flow; and

a reader instrument device configured for receiving the cartridge device, measuring one or a plurality of signals from the sample streams in the flow cell, and analyzing the blood cells in the sample.

2. The device of claim 1, wherein the reader instrument device is configured for detecting, identifying, characterizing, quantifying, and/or numerating leukocyte cells, erythrocyte cells, or platelet cells, or a combination thereof.

3. The device of claim 1, wherein the flow cell has a width in the range of about 1-10, 10-40, 40-100, or 100-200 μ

m; and

a depth in the range of about 1-10, 10-40, 40-100, or 100-200 μ

m.

4. The device of claim 1, wherein the flow cell has a length in the range of about 1-10, 10-100, 100-1,000, 1,000-5,000 μ

m, or 5,000-10,000 μ

m.

5. The device of claim 1, wherein the flow cell comprises a transparent surface for an optical signal from the sample streams in the flow cell;

and wherein the reader instrument device is configured for measuring the optical signal.

6. The device of claim 5, wherein the optical signal comprises light scattering, light absorption, light extinction, or fluorescence, or a combination thereof.

7. The device of claim 1, wherein the chamber has a volume in the range of about 0.01-0.1 ml, 0.1-0.2 ml, 0.2-0.4 ml, 0.4-0.8 ml, 0.8-2 ml, or 2-10 ml.

8. The device of claim 1, wherein the chamber comprises a venting port configured for receiving a pneumatic pressure source.

9. The device of claim 8, wherein, when the cartridge device is in use, the chamber is so positioned that at least a portion of the fluid inside the chamber is pulled by gravity away from the venting port and/or towards the lower bottom of the chamber.

10. The device of claim 1, wherein the cartridge device further comprises a cuvette;

and wherein the reader instrument device is configured for measuring a signal from the sample mixtures in the cuvette to determine the hemoglobin concentration in the sample.

11. The device of claim 10, wherein the reader instrument device is configured for measuring a light absorption signal from the sample mixtures in the cuvette to determine the hemoglobin concentration in the sample.

12. The device of claim 1, wherein the reagent comprises an osmolality-adjusting compound for forming sample mixtures with an osmolality in the range of about 140-160, 160-180, 180-200, 200-220, 220-240, 240-260, 260-280, 280-300, 300-320, 320-340, 340-360, 360-380, or 380-400 mOsm/L.

13. The device of claim 1, wherein the reagent comprises a sphering compound for transforming erythrocyte cells from disk shape into sphere shape.

14. The device of claim 1, wherein the reagent comprises a fluorescent labeling reagent for labeling nucleic acids in blood cells in the sample mixtures;

and wherein the reader instrument device is configured for measuring a fluorescence signal from the sample streams in the flow cell.

15. The device of claim 1, wherein the reagent comprises a lysing compound for lysing erythrocyte cells in the sample mixtures to release hemoglobin.

16. The device of claim 1, wherein the cartridge device further comprises a flow sensor fluidly connected to the flow cell;

and wherein the reader instrument device is configured for measuring a sensing signal from the flow sensor when the sample streams enter the flow sensor.

17. The device of claim 16, wherein the reader instrument device is configured for using the sensing signal from the flow sensor to determine the absolute count of the blood cells in the sample.

18. The device of claim 16, wherein the measured sensing signal comprises an optical signal.

19. The device of claim 16, wherein the fluidic connection between the flow cell and the flow sensor is configured for a sample stream to have the same flow rate flowing through the flow cell and the flow sensor.

20. The device of claim 16, wherein the flow sensor comprises a fluidic channel and a sensing zone on the fluidic channel, wherein the fluidic channel is fluidly connected to the flow cell to allow the sample streams to flow through;

and wherein the reader instrument device is configured for measuring a sensing signal from the sensing zone when the sample streams enter the sensing zone.

21. The device of claim 1, further comprising a microfluidic channel fluidly connected to the chamber and a valve on the microfluidic channel, wherein the microfluidic channel has a cross section in the range of about 0.001-0.01 mm², 0.01-0.1 mm², 0.1-0.25 mm², 0.25-0.5 mm², 0.5-1 mm², 1-2 mm², or 2-10 mm².

22. The device of claim 21, wherein the valve is a passive valve that comprises one of the following structures:

(i) a patch of hydrophobic surface in a channel having a hydrophilic surface, (ii) a patch of hydrophilic surface in a channel having a hydrophobic surface, (iii) an enlargement of the channel cross section along the flow direction in a channel having a hydrophilic surface, and (iv) a contraction of the channel cross section along the flow direction in a channel having a hydrophobic surface.

23. The device of claim 1, wherein the reader instrument device is configured for applying an actuation mechanism to control the fluid transfer in the cartridge device, and wherein the actuation mechanism comprises a pneumatic pressure source.

24. The device of claim 23, wherein the applied pneumatic pressure source actuates the cartridge device to transfer the sample mixtures from the chamber into the flow cell to form the sample streams.

25. The device of claim 23, wherein the cartridge device is configured for mixing two separate portions of the sample received in the fluidic conduit with the reagent to form two separate sample mixtures.

26. The device of claim 24, wherein the cartridge device is configured for forming the two separate sample mixtures in the same chamber separately.

27. The device of claim 24, wherein the reader instrument device is configured for operating and/or actuating the cartridge device to form two separate sample streams in the same flow cell from the two separate sample mixtures.

28. The device of claim 23, wherein the reader instrument is configured for numerating leukocyte cells in one of the two sample mixtures and erythrocyte cell and/or platelet cells in the other of the two sample mixtures.

29. The device of claim 1, wherein the cartridge device comprises an inlet port fluidly connected to the fluid conduit;

and wherein the inlet port comprises a valve or an external structure to close or seal the inlet port after the sample is received into the fluid conduit.

30. The device of claim 1, wherein the fluid conduit has a fixed orientation and/or a fixed position in the cartridge device.

31. The device of claim 1, wherein the cartridge device is configured for transferring at least a portion of the reagent into the fluidic conduit to flush at least a portion of the received sample into the chamber to form a sample mixture.

32. The device of claim 1, wherein the reagent consists of a fluorescent labeling agent.

33. The device of claim 1, wherein the reagent consists essentially of a fluorescent labeling agent.

34. The device of claim 1, wherein the reagent does not contain a magnetic material.

35. The device of claim 1, wherein the flow cell is configured for forming one sample stream without branching.