SYSTEM AND METHOD OF VERIFICATION OF A PREPARED SAMPLE FOR A FLOW CYTOMETER

• US 20110008816A1
• Filed: 06/02/2010
• Published: 01/13/2011
• Est. Priority Date: 06/02/2009
• Status: Active Grant

• Current Assignee: Accuri Cytometers, Inc. (Becton, Dickinson & Co), Huawei Technologies Co., Ltd. (Huawei Investment & Holding Co., Ltd.)
• Original Assignee: Accuri Cytometers, Inc. (Becton, Dickinson & Co)
• Inventors: Howes, Grant C., Rich, Collin A., Ball, Jack T.
• Granted Patent: US 8,507,279 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: G01N 15/1012 Calibrating particle analysers; References therefor
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• 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 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1014 Constitution of reference particles
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2333/70514 CD4
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]

Abstract

A system and method for a flow cytometer system including a prepared sample fluid with reference beads; an interrogation zone that analyzes the prepared sample fluid; a peristaltic pump system that draws the sample fluid through the interrogation zone; and a processor that monitors a measured volume of sample fluid sampled by the peristaltic pump system and an expected sample volume based on data generated by the analysis of the sample fluid. A system and method is additionally described using an alternative volume sensing fluidic system.

Claims

1. A flow cytometer system for a prepared sample fluid comprising:

an interrogation zone that analyzes the prepared sample fluid;

a peristaltic pump system that draws the sample fluid through the interrogation zone; and

a processor that monitors a measured volume of sample fluid sampled by the peristaltic pump system and an expected sample volume based on data generated by the analysis of the sample fluid.

2. The system of claim 1, wherein the peristaltic pump fluidic system includes:

a sheath pump that pumps a sheath fluid into the interrogation zone;

a waste pump that pumps a waste fluid from the interrogation zone; and

a sample injection probe (SIP) through which the sheath and waste pump cooperatively draw in the sample fluid.

3. The system of claim 2, wherein data from the sheath pump and the waste pump is used to measure the volume of sample fluid sampled by the peristaltic pump system.

4. The system of claim 3, wherein the prepared sample includes a concentration of reference beads, and wherein the interrogation zone detects the number of reference beads identified in the sample fluid from which the expected volume is calculated.

5. The system of claim 3, wherein the prepared sample fluid includes a reagent for a CD₄ test.

6. A flow cytometer system for a prepared sample fluid with a concentration of reference beads comprising:

an interrogation zone that analyzes the prepared sample fluid and counts detected reference beads;

a volume sensing fluidic system that draws the sample fluid through the interrogation zone and measures the volume of sample fluid drawn through the interrogation zone; and

a processor that monitors measured volume of sample fluid and an expected sample volume based on the counted reference beads and reference bead concentration.

7. The system of claim 6, wherein the volume sensing fluid system directly measures the volume of sample fluid drawn into the interrogation zone.

8. The method of claim 7, wherein the volume sensing fluidic system includes a direct container volume sensor coupled to a sample fluid container wherein the container volume sensor senses the volume change in the sample fluid container to calculate the volume sampled by the fluidic system.

9. The method of claim 7, wherein the volume sensing fluidic system includes a direct container volume sensor coupled to a drawtube through which the sample fluid is introduced to the volume sensing fluidic system, wherein the direct container volume sensor measures sample fluid flow to calculate the volume sampled by the fluidic system.

10. The method of claim 6, wherein the volume sensing fluid system indirectly measures the volume of sample fluid drawn into the interrogation zone.

11. A method for operating a flow cytometer comprising:

preparing a sample fluid;

analyzing sample fluid including;

drawing a volume of sample fluid into an interrogation zone with a peristaltic pump system; and

inspecting the sample fluid in the interrogation zone;

determining an expected sample volume based on the inspected sample fluid;

comparing volume of sample fluid drawn into the interrogation to the expected volume; and

performing an error correction action if expected volume does not equal the actual volume.

12. The method of claim 11, wherein the step of drawing a volume of sample fluid into an interrogation zone with a peristaltic pump system further comprises a sheath pump and waste pump cooperatively drawing a sample fluid into the interrogation zone.

13. The method of claim 12, further comprising sensing the operation of at least the sheath pump or waste pump to determine volume of sample fluid drawn into the interrogation zone.

14. The method of claim 13, wherein the step of a sheath pump and waste pump cooperatively drawing a sample fluid into the interrogation zone includes simultaneously:

the sheath pump pumping a sheath fluid from a sheath container into the interrogation zone;

a waste pump pumping waste fluid from the interrogation zone into a waste container; and

wherein the flow rate of the sheath fluid is different from the flow rate of the waste fluid thereby drawing the fluid from the sample container into the interrogation zone.

15. The method of claim 13, wherein the sample fluid is prepared with reference beads, and the step of inspecting the sample fluid includes counting reference beads.

16. The method of claim 11, wherein the step of preparing a sample fluid includes preparing a blood sample with a CD₄ test reagent including reference beads, and wherein the step of inspecting the sample fluid includes counting the reference beads of the reagent and the number of CD₄ cells.

17. A method for operating a flow cytometer with a fluidic system comprising:

preparing a sample fluid with reference beads;

analyzing a sample fluid and counting the reference beads;

determining an expected sample volume from the counted reference beads;

measuring a sample fluid volume introduced into the fluidic system; and

performing an error correction action if the measured sample fluid volume does not equal the expected sample volume.

18. The method of claim 17, wherein the step of measuring the sample fluid volume includes directly measuring the volume of sample fluid.

19. The method of claim 17, wherein the step of measuring the sample fluid volume includes indirectly measuring the volume of sample fluid by measuring a sheath fluid volume and waste fluid volume from the fluidic system.

20. The method of claim 17, wherein the step of preparing a sample fluid includes preparing a blood sample with a CD₄ test reagent, and wherein the step of analyzing the sample fluid further includes counting the number of CD₄ cells.