Fluidic cartridge for cytometry and additional analysis

• US 10,634,602 B2
• Filed: 11/03/2017
• Issued: 04/28/2020
• 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:
  US15/803,133
• Publication Number:
  US 20180052093A1
• Time in Patent Office: 907 Days
• Field of Search: 422 50, 422 681, 422 81, 422 82, 422 8205, 422 8201, 436 43, 436 63, 436174, 436180, 436177
• CPC Class Codes: A61B 10/0096 Casings for storing test samples A61B10/0038 and A61B10/0045 take precedence; preservation of living parts of the human or animal body A01N1/02; containers for retaining a material to be analysed B01L3/50; containers for enzymology or microbiology C12M1/16; swab-sampler being part of enzymology or microbiology container C12M1/30
• CPC Class Codes: A61B 5/0071 by measuring fluorescence emission
• CPC Class Codes: B01F 31/00 Mixers with shaking, oscillating, or vibrating mechanisms
• CPC Class Codes: B01F 31/65 the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/30351 using hydrophilic/hydrophobic surfaces
• CPC Class Codes: B01F 35/71745 using pneumatic pressure, overpressure, gas or air pressure in a closed receptacle or circuit system
• 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 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• 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 15/06 : 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 15/1484 : 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 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1486 : Counting the particles
• CPC Class Codes: G01N 33/5094 : for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/56972 : White blood cells

Abstract

The disclosure relates to devices and methods for analyzing particles in a sample. In various embodiments, the present disclosure provides devices and methods for cytometry and additional analysis. 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 particles in a sample.

Claims

1. A device for analyzing target particles in a sample, comprising:

a cartridge device, wherein the cartridge device comprises;

an inlet configured for receiving the sample into the cartridge device;

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

a flow cell fluidly connected to the fluidic structure and configured for forming one or more sample streams from the one or more sample mixtures, wherein the sample streams are formed in the flow cell without a sheath flow, and wherein the flow cell comprises an optically transparent area configured for measuring an optical signal from the sample streams to detect the target particles in the sample; and

a flow sensor fluidly connected to the flow cell and configured for measuring a sensing signal from the sample streams that enter the flow sensor.

2. The device of claim 1, wherein the fluidic structure comprises one or a plurality of chambers, wherein each 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;

and wherein the fluidic structure is configured for transferring the sample mixtures from one of the chambers to the flow cell to form the sample streams.

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

m, 10-40 μ

m, 40-100 μ

m, or 100-200 μ

m and a depth in the range of about 1-10 μ

m 10-40 μ

m, 40-100 μ

m, or 100-200 μ

m; and

wherein the sample streams have a cross section of the same size as the flow cell.

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

m, 10-100 μ

m, 100-1,000 μ

m, 1,000-10,000 μ

m, or 10,000-50,000 μ

m.

5. The device of claim 1, wherein the optically transparent area on the flow cell has a transmission rate of 50-60%, 60-70%, 70-80%, 80-90%, 90-96%, or 96-99.9% for the optical signal from the sample streams, and wherein the optical signal comprises scattered light, reflected light, transmitted light, fluorescence, light absorption, light extinction, or white light image, or a combination thereof.

6. The device of claim 1, wherein the optically transparent area on the flow cell is made of a plastic material.

7. The device of claim 1, wherein the cartridge device further comprises a reagent.

8. The device of claim 1, wherein the reagent comprises a fluorescent labeling agent that selectively labels the target particles in the sample with fluorescence, and wherein the optical signal from the sample streams comprises fluorescence.

9. The device of claim 1, 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 a sensing signal is measured when the sample streams enter the sensing zone.

10. The device of claim 9, wherein the fluidic channel in the flow sensor has a channel width in the range of about 0.001-0.05 mm, 0.05-1 mm, or 1-5 mm, and a channel depth in the range of about 0.001-0.01 mm, 0.01-0.5 mm, 0.5-1 mm, or 1-2 mm.

11. The device of claim 1, wherein the sensing zone comprises an optically transparent area configured for measuring an optical signal that changes levels between the absence and presence of the sample streams in the sensing zone.

12. The device of claim 1, 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.

13. The device of claim 1, wherein the fluidic structure comprises at least one basic fluidic unit that comprises:

a chamber configured to accommodate a fluid;

a venting port connected to the chamber, wherein the venting port is connected to a pneumatic pressure source, an ambient pressure, or the atmosphere pressure;

a microfluidic channel connected to the chamber; and

a valve on the microfluidic channel.

14. The device of claim 13, 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, and wherein the microfluidic channel has a cross section of a size 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.

15. The device of claim 13, wherein the cartridge device is configured for transferring the sample mixtures from the chamber into the flow cell to form the sample streams when an external actuation mechanism is applied to the cartridge device, and wherein the external actuation mechanism comprises a pneumatic pressure source.

16. The device of claim 13, wherein, when the cartridge device is in use, the chamber is so positioned that the at least a portion of the fluid inside the chamber is pulled by gravity towards the microfluidic channel and/or away from the venting port.

17. The device of claim 13, wherein, when the cartridge device is in use, the chamber has a volume larger than the volume of the fluid accommodated therein and an air gap exists between the venting port and the fluid accommodated therein.

18. The device of claim 13, wherein the valve is a passive valve that is configured for allowing a fluid flow to pass through the microfluidic channel when a pneumatic pressure is applied to the fluid flow and stopping the fluid flow when no pneumatic pressure is applied to the fluid flow.

19. The device of claim 13, wherein the valve is a passive valve that comprises one of the following structures:

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

20. The device of claim 1, further comprising a reader instrument device, wherein the reader instrument device is configured for receiving, operating, and/or actuating the cartridge device.

21. The device of claim 20, wherein the reader instrument device is configured for measuring the optical signal at the flow cell to quantify the target particles in the sample.

22. The device of claim 20, wherein the reader instrument device is configured for measuring the optical signal at the flow cell and the sensing signal at the flow sensor to determine the concentration of the target particles in the sample.

23. A method for analyzing target particles in a sample using the device of claim 1, the method comprising:

applying the sample to the cartridge device, which is configured for collecting a predetermined sample volume into the cartridge device;

transferring the cartridge device into a reader instrument device;

mixing at least a portion of the collected sample and at least a portion of a reagent to form one or more sample mixtures inside the cartridge device;

forming one or more sample streams from the one or more sample mixtures in the flow cell, wherein the one or more sample streams are formed in the flow cell without a sheath flow;

measuring the optical signal from the one or more sample streams at the flow cell to detect the target particles in the sample streams;

using the reader instrument device to analyze the measured optical signal to quantify the target particles in the sample.

24. The method of claims 23, further comprising:

flowing the sample streams through a flow sensor that is fluidly connected to the flow cell;

measuring a sensing signal from the sample streams at the flow sensor to detect the entrance of the sample streams into the flow sensor and/or the exit of the sample streams out of the flow sensor; and

using the reader instrument device to analyze the measured optical signal and sensing signal to determine the concentration of the target particles in the sample.

25. The method of claim 23, wherein the target particles have a size in the range of 0.1-1 μ

m, 1-10 μ

m, 10-15 μ

m, 15-30 μ

m, 30-50 μ

m, or 50-100 μ

m; and

wherein the target particles have a concentration in the range of 1-100, 100-1000, 1000-5000, 5000-20,000, or 20,000-50,000 target particles per μ

l sample steam.

26. The method of claim 23, wherein the reagent comprises a fluorescent labeling agent that selectively labels the target particles in the sample with fluorescence, and wherein the optical signal from the sample streams comprises fluorescence.

27. The method of claims 23, wherein the mixing step is performed in a fluidic structure comprising one or a plurality of chambers, wherein each 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.

28. The method of claim 23, wherein the flow cell has a width in the range of about 1-10 μ

m, 10-40 μ

m, 40-100 μ

m, or 100-200 μ

m and a depth in the range of about 1-10 μ

m, 10-40 μ

m, 40-100 μ

m, or 100-200 μ

m; and

wherein the sample streams have a cross section of the same size as the flow cell.

29. The method of claim 23, wherein the sample streams in the flow cell have a flow rate in the range of 0.001-0.01, 0.01-0.1, 0.1-1, 1-50, 50-200, or 200-1000 μ

l/min when the optical signal is measured from the sample streams.

30. The method of claim 23, wherein the optical signal measured from the sample streams at the flow cell comprises scattered light, reflected light, transmitted light, fluorescence, light absorption, light extinction, or white light image, or a combination thereof.

31. The method of claim 23, 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 a sensing signal is measured when the sample streams enter the sensing zone.

32. The method of claim 31, wherein the fluidic channel in the flow sensor has a channel width in the range of about 0.001-0.05 mm, 0.05-1 mm, or 1-5 mm, and a channel depth in the range of about 0.001-0.01 mm, 0.01-0.5 mm, 0.5-1 mm, or 1-2 mm;

and wherein the sample streams in the flow cell and the flow sensor have the same flow rate.

33. The method of claim 31, wherein the sensing zone comprises an optically transparent area configured for measuring an optical signal that changes levels between the absence and presence of the sample streams in the sensing zone.

34. The method of claim 23, wherein mixing is performed in at least one basic fluidic unit that comprises:

a chamber configured to accommodate a fluid, 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;

a venting port connected to the chamber, wherein the venting port is connected to a pneumatic pressure source, an ambient pressure, or the atmosphere pressure;

a microfluidic channel connected to the chamber, wherein the microfluidic channel has a cross section of a size 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; and

a valve on the microfluidic channel.

35. The method of claim 34, wherein the sample mixtures are transferred from the chamber into the flow cell to form the sample streams when an external actuation mechanism is applied to the cartridge device, and wherein the external actuation mechanism comprises a pneumatic pressure source.

36. The method of claim 34, wherein, when the cartridge device is in use, the chamber is so positioned that the at least a portion of the fluid inside the chamber is pulled by gravity towards the microfluidic channel and/or away from the venting port.

37. The method of claim 34, wherein, when the cartridge device is in use, the chamber has a volume larger than the volume of the fluid accommodated therein and an air gap exists between the venting port and the fluid accommodated therein.

38. The method of claim 23, wherein at least two separate sample mixtures are transferred into the same flow cell to form at least two separate sample streams.

39. A method for analyzing target particles in a sample using the device of claim 1, the method comprising:

applying the sample to the cartridge device, which is configured for collecting a predetermined sample volume into the cartridge device;

transferring the cartridge device into a reader instrument device;

mixing at least a portion of the collected sample and at least a portion of a reagent to form one or more sample mixtures inside the cartridge device;

forming one or more sample streams from the one or more sample mixtures in the flow cell, wherein at least two separate sample mixtures are transferred into the flow cell to form at least two separate sample streams without a sheath flow;

measuring the optical signal from the sample streams at the flow cell to detect the target particles in the sample streams; and

using the reader instrument device to analyze the measured optical signal to quantify the target particles in the sample.

40. The method of claim 39, wherein a portion of the collected sample is mixed with a first reagent to form a first sample mixture and another portion of the collected sample is mixed with a second reagent to form a second sample mixture;

and wherein the two sample mixtures are separately transferred into the flow cell to form two separate sample streams.

41. The method of claim 40, wherein the two sample mixtures are separately formed in a chamber or separately transferred into a chamber before being separately transferred into the flow cell.

42. The method of claim 41, 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.

43. The method of claim 39, wherein the sample is collected into a fluidic conduit.

44. The method of claim 43, wherein the fluidic conduit is closed by a valve and/or sealed by an external structure after the sample is collected into the fluidic conduit.

45. The method of claim 43, wherein the fluidic conduit is configured for collecting a predetermine sample volume in the range of about 0.1-1 μ

L, 1-5 μ

L, 5-10 μ

L, 10-20 μ

L, or 20-50 μ

L.

46. The method of claim 43, wherein at least a portion of the reagent is transferred into the fluidic conduit to flush a portion of the collected sample into a chamber to form a sample mixture.

47. The method of claim 39, wherein at least one sample stream comprises white blood cells as the target particles detected in the flow cell and at least another sample stream comprises red blood cells and/or platelet cells as the target particles detected in the flow cell.

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

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

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

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