Methods for detecting events in a flow cytometer

Patent metadata

Field	Value
Publication number	US-10024780-B2
Application number	US-201615379846-A
Country	US
Kind code	B2
Filing date	Dec 15, 2016
Priority date	Dec 18, 2015
Publication date	Jul 17, 2018
Grant date	Jul 17, 2018

Abstract

Aspects of the present disclosure include methods for detecting events in a flow cytometer. Also provided are methods of detecting cells in a flow cytometer. Other aspects of the present disclosure include methods for determining a level of contamination in a flow cell. Computer-readable media and systems, e.g., for practicing the methods summarized above, are also provided.

First claim

What is claimed is:

1. A method for detecting events in a flow cytometer, comprising:
flowing particles through a flow cell of the flow cytometer;
optically interrogating the particles flowing through the flow cell;
extracting putative event features;
time-stamping putative events;
determining a time difference between a putative previous event and a putative current event;
comparing the time difference to a threshold duration, wherein:
if the time difference is greater than the threshold duration, storing the putative current event as a current event; and
if the time difference is less than the threshold duration, comparing a peak height feature of the putative current event and a peak height feature of the putative previous event to a threshold peak height, wherein:
if the peak height feature of the putative current event is less than the threshold peak height, discarding the putative current event; and
if the peak height feature of the putative previous event is greater than the threshold peak height, storing the putative previous event as a previous event.

2. The method according to claim 1 , wherein an event is distinguished from a signal selected from the group consisting of: optical system side-lobes, fluidics drift, baseline drift, flow cell contamination, and combinations thereof.

3. The method according to claim 1 , wherein flowing particles through the flow cell comprises flowing the particles at a sheath pressure of 9 psi or greater.

4. The method according to claim 1 , wherein comparing a peak height feature of the putative current event and a peak height feature of the putative previous event to a threshold peak height comprises determining whether the peak heights of the putative current event and putative previous event are less than 2× a threshold peak height.

5. The method according to claim 1 , wherein the threshold duration is 2.5 μs or less.

6. The method according to claim 1 , wherein optically interrogating the particles comprises exciting the cells using a laser.

7. The method according to claim 1 , wherein the particles are cells and an event is a cell event.

8. The method according to claim 5 , wherein the threshold duration is 2 μs.

9. The method according to claim 6 , wherein the beam height of the laser is 5 μm or greater.

10. The method according to claim 9 , wherein the beam height of the laser is 8 μm.

11. The method according to claim 7 , wherein the cells are from a blood sample.

12. The method according to claim 11 , wherein the cell event is a small cell event.

13. The method according to claim 12 , wherein the small cell event is a platelet event.

14. The method according to claim 12 , wherein the small cell event is a microorganism cell event or a cell debris event.

Assignees

• Abbott Lab
• Abbott Laboratories Diagnostics Div

Inventors

• Shah Amish P
• Hulett Iii Frederic M

Classifications

• G01N2015/1488

  Methods for deciding · CPC title

• G01N2015/1461

  Coincidence detecting; Circuits therefor · CPC title

• G01N2021/157

  Monitoring by optical means · CPC title

• G01N2015/1006

  for cytology · CPC title

• G01N15/1459

  the analysis being performed on a sample stream · CPC title