System and method of verification of a prepared sample for a flow cytometer

• US 8,507,279 B2
• Filed: 06/02/2010
• Issued: 08/13/2013
• 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: Ball, Jack T., Howes, Grant C., Rich, Collin A.
• Primary Examiner(s): Turk, Neil N
• Application Number:
  US12/792,536
• Publication Number:
  US 20110008816A1
• Time in Patent Office: 1,168 Days
• Field of Search: None
• US Class Current: 436/10
• 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 method for verifying preparation of a sample fluid drawn into an interrogation zone of a flow cytometer, comprising:

receiving the sample fluid prepared with a reference bead concentration;

drawing a sample volume of the sample fluid with the reference bead concentration into the interrogation zone with a peristaltic pump system;

analyzing the sample volume of the sample fluid in the interrogation zone to determine a parameter indicative of reference bead count;

determining an expected value of the sample volume based upon the parameter and the reference bead concentration;

determining a measured value of the sample volume of the sample fluid used to determine the expected value; and

performing an error correction action in response to a comparison between the measured value of the sample volume and the expected value of the sample volume.

2. The method of claim 1, wherein drawing a sample volume of the sample fluid into the interrogation zone with a peristaltic pump system further comprises drawing the sample volume using a sheath pump and a waste pump, wherein a pressure differential created by the sheath pump and the waste pump draws the sample fluid into the interrogation zone.

3. The method of claim 2, wherein determining a measured value of the sample volume comprises determining a measured value based on a difference between a sheath fluid volume pumped by the sheath pump and a waste fluid volume pumped by the waste pump.

4. The method of claim 3, wherein drawing a sample volume of the sample fluid into the interrogation zone includes simultaneously:

pumping the sheath fluid volume, with the sheath pump, from a sheath container into the interrogation zone and

pumping the waste fluid volume, with the waste pump, from the interrogation zone into a waste container;

wherein the interrogation zone is in fluid communication with the sheath pump, the waste pump, and a sample container containing the sample fluid, and

the sheath pump is configured to pump at a first flow rate and the waste pump is configured to pump at a second flow rate thereby creating the pressure differential to draw the sample fluid from the sample container into the interrogation zone.

5. The method of claim 3, wherein analyzing the sample volume of the sample fluid to determine a parameter indicative of reference bead count includes directly counting reference beads based upon optical detection.

6. The method of claim 1, wherein receiving the sample fluid prepared with a reference bead concentration includes receiving a blood sample with a CD4 test reagent including reference beads, and wherein analyzing the sample volume of the sample fluid includes enumerating reference beads of the reagent and CD4 cells.

7. The method of claim 1, wherein analyzing the sample volume of the sample fluid in the interrogation zone to determine a parameter indicative of reference bead count comprises directly counting reference beads based upon optical detection.

8. The method of claim 1, wherein analyzing the sample volume of the sample fluid in the interrogation zone to determine a parameter indicative of reference bead count further comprises determining a time based function of a reference bead count.

9. The method of claim 1, wherein receiving the sample fluid prepared with a reference bead concentration comprises receiving a sample fluid prepared with a first reference bead concentration and a second reference bead concentration.

10. The method of claim 9, wherein analyzing the sample volume of the sample fluid in the interrogation zone to determine a parameter indicative of reference bead count further comprises determining a ratio between a first reference bead parameter and a second reference bead parameter.

11. The method of claim 1, wherein determining a measured value of the sample volume of the sample fluid comprises calculating a difference between a waste fluid volume and a sheath fluid volume.

12. The method of claim 1, wherein determining a measured value of the sample volume of the sample fluid comprises directly measuring a volume of sample fluid drawn using a sensor.

13. The method of claim 1, wherein performing an error correction action comprises providing a user notification in response to a comparison between the measured value and the expected value of the sample volume.

14. The method of claim 13, wherein providing a user notification comprises providing an alert indicating sample preparation error.

16. The method of claim 1, wherein performing an error correction action comprises at least one of providing a user notification and performing an adjustment to a preparation of the sample fluid in response to the comparison between the measured value and the expected value of the sample volume.

15. A method for verifying preparation of a sample fluid drawn into an interrogation zone of a flow cytometer, comprising:

drawing a sample volume of the sample fluid, prepared with a reference bead concentration, into the interrogation zone with a peristaltic pump system;

analyzing the sample volume of the sample fluid in the interrogation zone to determine a parameter indicative of reference bead count;

determining an expected vale of the sample volume based on the parameter and the reference bead concentration;

determining a measured value of the sample volume of the sample fluid used to determine the expected value; and

providing a user notification in response to a comparison between the measured value and the expected value of the sample volume.