Multi-spectral filter profiling and quality control for flow cytometry

Patent metadata

Field	Value
Publication number	US-10429291-B2
Application number	US-201916247424-A
Country	US
Kind code	B2
Filing date	Jan 14, 2019
Priority date	Aug 12, 2015
Publication date	Oct 1, 2019
Grant date	Oct 1, 2019

Abstract

Disclosed is a system and method for characterizing optical filters in a flow cytometer and optionally checking the operation of detectors in the flow cytometer. In some embodiments, the system may utilize an LED board having an opening through which the fluorescence and side scatter beams, rays, or images pass and light emitting diodes around the opening that emit light having different spectral profiles. The different spectral profiles allow the system to identify the optical filters that are placed in the flow cytometer, to verify detector operation, to assist in instrumentation troubleshooting, and to provide a quantitative reference for detector comparison.

First claim

What is claimed is:

1. A method for determining filtering characteristics for one or more flow cytometry systems including a first flow cytometry system, the method comprising:
emitting, at different first times, different spectral profiles of calibration light from a calibration light source installed in the first flow cytometry system, wherein each spectral profile of calibration light has one or more intensity peaks at different wavelengths, and wherein the calibration light source is different than a first sample illumination source of the first flow cytometry system, the first sample illumination source being configured to deliver light to one or more first sample locations of the first flow cytometry system, thereby producing first emitted or scattered sample light;
directing some of the calibration light emitted at the first times along at least a portion of one or more first optical paths of the first flow cytometry system, wherein each first optical path passes through a corresponding one or more first optical filter elements of the first flow cytometry system, and wherein each first optical path directs first emitted or scattered sample light from one of the one or more first sample locations to a corresponding first detector of the first flow cytometry system when the first sample illumination source is caused to deliver light to the one or more first sample locations;
measuring, for each different spectral profile of calibration light emitted at one of the first times, the light intensity at each of the one or more first detectors, wherein each of the one or more first detectors produces first output data that is indicative of the measured light intensity of the calibration light that reaches the first detector; and
analyzing the first output data from one of the one or more first detectors produced during the emission of at least two different spectral profiles of calibration light by the calibration light source in association with the corresponding first times to determine the first filtering characteristics of the first optical filter elements along the first optical path corresponding to that first detector.

2. The method of claim 1 , further comprising:
comparing the first filtering characteristics for the first optical filter elements along the first optical path with predefined filtering characteristics associated with that first optical path; and
providing an indication via a user interface as to whether the first filtering characteristics for the first optical filter elements along the first optical path are within a threshold amount of the predefined filtering characteristics associated with that first optical path.

3. The method of claim 1 , wherein a different subset of the one or more first optical paths passes through each first optical filter element.

4. The method of claim 1 , wherein each of the one or more first optical paths pass through a pinhole aperture.

5. The method of claim 1 , wherein:
the calibration light source includes a printed circuit board having an opening therethrough and a plurality of light emitting diodes (LEDs) placed around the periphery of the opening,
the plurality of LEDs are configured to emit the calibration light, and
the one or more first optical paths pass through the opening.

6. The method of claim 1 , further comprising directing some of the calibration light emitted at the first times through the first sample location, wherein the first sample illumination source is oriented such that the light delivered by the first sample illumination source to each first sample location is directed into each first sample location along a direction that is not aligned with the one or more first optical paths.

7. The method of claim 1 , wherein the calibration light source comprises a plurality of light emitting diodes (LEDs) having different peak wavelengths, and wherein emitting different spectral profiles of calibration light at different times comprises separately illuminating different subsets of plurality of LEDs, where in each different subset has a different set of peak wavelengths.

8. The method of claim 7 , wherein each of the plurality of LEDs is configured to emit calibration light having a bandwidth in a range of 100 nm to 200 nm.

9. The method of claim 7 , wherein each of the plurality of LEDs is configured to emit calibration light having a bandwidth in a range of 10 nm to 100 nm.

10. The method of claim 1 , further comprising:
installing the calibration light source in a second flow cytometry system;
emitting, at different second times, different spectral profiles of calibration light from the calibration light source installed in the second flow cytometry system, wherein the calibration light source is different than a second sample illumination source of the second flow cytometry system, the second sample illumination source being configured to deliver light to one or more second sample locations of the second flow cytometry system, thereby producing second emitted or scattered sample light;
directing some of the calibration light emitted at the second times along at least a portion of one or more second optical paths of the second flow cytometry system, wherein each second optical path passes through a corresponding one or more second optical filter elements of the second flow cytometry system, and wherein each second optical path directs second emitted or scattered sample light from one of the one or more second sample locations to a corresponding second detector;
measuring, for each different spectral profile of calibration light emitted at one of the second times, the light intensity at each of the one or more second detectors, wherein each of the one or more second detectors produces second output data that is indicative of the measured light intensity of the calibration light that reaches the second detector; and
analyzing the second output data from one of the one or more second detectors produced during the emission of at least two different spectral profiles of calibration light by the calibration light source to determine the second filtering characteristics of the second optical filter elements along the second optical path corresponding to that second detector.

11. The method of claim 10 , further comprising:
comparing the first output data to the second output data to determine if there is a difference in optical performance between the first flow cytometry system and the second flow cytometry system for instances where the spectral profiles of calibration light emitted by the calibration light source for the first flow cytometry system and the second flow cytometry system are the same.

12. The method of claim 11 , further comprising:
adjusting the second detectors based on the difference in optical performance between the first flow cytometry system and second flow cytometry system.

13. The method of claim 1 , further comprising:
causing the calibration light source to emit, at an initial first time of the first times, first calibration light having an initial first spectral profile;
obtaining initial first output data from at least one of the one or more first detectors responsive to the at least one of the one or more first detectors receiving the first calibration light from the calibration light source at the initial first time;
causing the calibration light source to emit, at a subsequent first time after the initial first time, the first calibration light having the initial first spectral profile;
obtaining subsequent first output data from the at least one of the one or more first detectors responsive to the first detectors receiving the first calibration li

Assignees

• Bio Rad Laboratories

Inventors

• Fox Daniel Nelson
• Hunter Susan

Classifications

• G01J3/0218

  using optical fibers · CPC title

• G01N2015/1006

  for cytology · CPC title

• G01N15/1459

  the analysis being performed on a sample stream · CPC title

• G01N15/1434

  Optical arrangements · CPC title

• G01N15/1012 Primary

  Calibrating particle analysers; References therefor · CPC title