# US 2019/0376887 A1 · Parallel Flow Cytometer Using Radiofrequency Multiplexing

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- Number: US 2019/0376887 A1
- Document ID: US20190376887A1
- Status: Published application
- Kind: A1
- Application number: 16/528,426
- Filed: 2019-07-31
- Published: 2019-12-12
- Earliest claimed: 2014-03-18
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: Bahram Jalali; Eric D. Diebold; Brandon Buckley
- Web page: https://patentgrep.com/patent/US20190376887A1

## CPC

- G01N 15/1434: Optical arrangements
- G01N 15/1459: {the analysis being performed on a sample stream}
- G01N 15/1484: {microstructural devices}
- G01N 21/64: Fluorescence; Phosphorescence
- G01N 33/537: with separation of immune complex from unbound antigen or antibody
- G01N 21/6428: {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)}
- G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)}
- G01N 21/6486: {Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence)}
- G01N 2015/1006: {for cytology}
- G01N 2015/1477: {Multiparameters}
- G01N 2021/6421: {Measuring at two or more wavelengths}
- G01N 2201/067: Electro-optic, magneto-optic, acousto-optic elements
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## 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.

## Claims

### Claim at position 1

1-31. (canceled)

### Claim 32

An apparatus comprising:
a laser; and
a frequency shifter component configured to generate a plurality of independently controlled frequency shifted beams of light.

### Claim 33 (depends on 32)

The apparatus according to claim 32, wherein the frequency shifter component comprises:
a radio-frequency source configured to generate a plurality of radio-frequency outputs.

### Claim 34 (depends on 32)

The apparatus according to claim 32, wherein the frequency shifter component comprises one or more of an acousto-optic deflector (AOD) and an acousto-optic frequency shifter (AOFS).

### Claim 35 (depends on 34)

The apparatus according to claim 34, wherein the frequency shifter component comprises an acousto-optic deflector (AOD).

### Claim 36 (depends on 35)

The apparatus according to claim 35, further comprising a radio-frequency comb generator configured to produce the plurality of independently controlled frequency shifted beams of light from the laser.

### Claim 37 (depends on 36)

The apparatus according to claim 36, wherein the radio-frequency comb generator comprises a direct digital synthesizer.

### Claim 38 (depends on 41)

The apparatus according to claim 41, wherein the radio-frequency comb generator is configured to modulate the intensity of each of the plurality of independently controlled frequency shifted beams of light.

### Claim 39 (depends on 32)

The apparatus according to claim 32, wherein the frequency shifter component is configured to generate a local oscillator beam and the plurality of independently controlled frequency shifted beams of light from the laser.

### Claim 40 (depends on 32)

The apparatus according to claim 32, wherein the frequency shifter component comprises an optical combiner configured to produce an optical interrogation beam comprising the plurality of independently controlled frequency shifted beams of light.

### Claim 41 (depends on 32)

The apparatus according to claim 32, wherein the beams of frequency shifted light are spatially distributed.

### Claim 42 (depends on 32)

The apparatus according to claim 32, further comprising a photodetector that is configured to detect fluorescence from particles irradiated by the plurality of independently controlled frequency shifted beams of light.

### Claim 43 (depends on 42)

The apparatus according to claim 42, wherein the photodetector is configured to detect fluorescence from particles moving at a rate of 1 m/s through a flow channel.

### Claim 44 (depends on 42)

The apparatus according to claim 42, wherein the photodetector comprises a photomultiplier tube (PMT).

### Claim 45 (depends on 44)

The apparatus according to claim 44, wherein the photodetector is a single photomultiplier tube.

### Claim 46 (depends on 32)

The apparatus according to claim 32, wherein the laser is a continuous wave laser.