Parallel flow cytometer using radiofrequency multiplexing

Patent metadata

Field	Value
Publication number	US-12055476-B2
Application number	US-202318214300-A
Country	US
Kind code	B2
Filing date	Jun 26, 2023
Priority date	Mar 18, 2014
Publication date	Aug 6, 2024
Grant date	Aug 6, 2024

Abstract

An imaging flow cytometry apparatus and method which allows registering multiple locations across a cell, and/or across multiple flow channels, in parallel using radio-frequency-tagged emission (FIRE) coupled with a parallel optical detection scheme toward increasing analysis throughput. An optical source is modulated by multiple RF frequencies to produce an optical interrogation beam having a spatially distributed beat frequency. This beam is directed to one or more focused streams of cells whose responsive fluorescence, in different frequencies, is registered in parallel by an optical detector.

First claim

What is claimed is:

1. An apparatus comprising:
a flow cell comprising one or more microfluidic channels;
a light beam generator configured to generate a frequency shifted beam of light for irradiating a particle in a sample propagated in a flow stream in the one or more microfluidic channels;
a light detection system comprising a photodetector configured to detect light from the irradiated particle in the sample; and
a processor comprising memory operably coupled to the processor wherein the memory comprises instructions stored thereon, which when executed by the processor, cause the processor to:
generate one or more waveforms in response to the detected light; and
form an image of the particle from the generated waveforms.

2. The apparatus according to claim 1 , wherein the flow cell comprises a plurality of microfluidic channels.

3. The apparatus according to claim 1 , wherein the light beam generator comprises a laser.

4. The apparatus according to claim 1 , wherein the light beam generator comprises an acousto-optic device.

5. The apparatus according to claim 4 , wherein the acousto-optic device comprises an acousto-optic deflector (AOD).

6. The apparatus according to claim 5 , wherein the light beam generator further comprises a direct digital synthesizer configured to apply a radiofrequency signal to the acousto-optic deflector.

7. The apparatus according to claim 1 , wherein the photodetector comprises a photomultiplier tube (PMT).

8. The apparatus according to claim 1 , wherein the light detection system comprises a plurality of photomultiplier tubes.

9. The apparatus according to claim 1 , wherein the light detection system is configured to detect fluorescence from the sample.

10. The apparatus according to claim 9 , wherein the light detection system is configured to detect fluorescence from the sample in a plurality of pixels across the irradiated particle.

11. The apparatus according to claim 1 , wherein the system comprises an analog-to-digital converter (ADC) for generating the one or more waveforms by digitizing data signals from the photodetector.

12. The apparatus according to claim 11 , wherein the memory comprises instructions for forming the image of the particle from the digitized data signals.

13. The apparatus according to claim 1 , wherein the memory includes instructions for applying a Fourier transform to the generated waveforms.

14. A method comprising:
generating a frequency shifted beam of light;
irradiating with the frequency shifted beam of light a particle in a sample in a flow stream propagated through one or more microfluidic channels of a flow cell;
detecting light from the irradiated particle in the sample with a light detection system comprising a photodetector;
generating one or more waveforms in response to the detected light; and
forming an image of the particle from the generated waveforms.

15. The method according to claim 13 , wherein the flow cell comprises a plurality of microfluidic channels.

16. The method according to claim 13 , wherein the frequency shifted beam of light is generated by irradiating an acousto-optic device with a laser.

17. The method according to claim 13 , wherein method comprises detecting fluorescence from the particle in the sample with the photodetector.

18. The method according to claim 16 , wherein the fluorescence is detected from the sample in a plurality of pixels across the irradiated particle.

19. The method according to claim 1 , wherein the method comprises generating the one or more waveforms by digitizing data signals from the light detection system.

20. The method according to claim 18 , wherein the method comprises forming the image of the particle from the digitized data signals.

Assignees

• Univ California

Inventors

• Jalali Bahram
• Diebold Eric D
• Buckley Brandon

Classifications

• G01N2201/067

  Electro-optic, magneto-optic, acousto-optic elements · CPC title

• G01N2021/6421

  Measuring at two or more wavelengths · CPC title

• G01N21/6486

  Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence) · CPC title

• G01N21/6458

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title