Parallel flow cytometer using radiofrequency multiplexing
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.
1. An apparatus comprising:
a laser; and
a frequency shifter component configured to generate a plurality of amplitude modulated frequency shifted beams of light having identical optical sensitivity.
2. The apparatus according to claim 1 , wherein the frequency shifter component comprises:
a radio-frequency source configured to generate a plurality of radio-frequency outputs.
3. The apparatus according to claim 1 , wherein the frequency shifter component comprises one or more of an acousto-optic deflector (AOD) and an acousto-optic frequency shifter (AOFS).
4. The apparatus according to claim 3 , wherein the frequency shifter component comprises an acousto-optic deflector (AOD).
5. The apparatus according to claim 4 , further comprising a radio-frequency comb generator configured to produce the plurality of frequency shifted beams of light from the laser.
6. The apparatus according to claim 5 , wherein the radio-frequency comb generator comprises a direct digital synthesizer.
7. The apparatus according to claim 5 , wherein the radio-frequency comb generator is configured to modulate the amplitude of the plurality of frequency shifted beams of light.
8. The apparatus according to claim 1 , wherein the frequency shifter component is configured to generate a local oscillator beam and the plurality of frequency shifted beams of light from the laser.
9. The apparatus according to claim 1 , wherein the frequency shifter component comprises an optical combiner configured to produce an optical interrogation beam comprising the plurality of frequency shifted beams of light.
10. The apparatus according to claim 9 , wherein the optical combiner is configured to independently control the intensity of each beam of frequency shifted light.
11. The apparatus according to claim 1 , wherein the beams of frequency shifted light are spatially distributed.
12. The apparatus according to claim 1 , further comprising a photodetector that is configured to detect fluorescence from particles irradiated by the plurality of frequency shifted beams of light.
13. The apparatus according to claim 12 , wherein the photodetector is configured to detect fluorescence from particles moving at a rate of 1 m/s through a flow channel.
14. The apparatus according to claim 12 , wherein the photodetector comprises a photomultiplier tube (PMT).
15. The apparatus according to claim 14 , wherein the photodetector is a single photomultiplier tube.
16. The apparatus according to claim 1 , wherein the laser is a continuous wave laser.
17. A method comprising irradiating with a laser a frequency shifter component to generate a plurality of amplitude modulated frequency shifted beams of light having identical optical sensitivity.
18. The method according to claim 17 , wherein the frequency shifter component comprises an acousto-optic deflector (AOD).
19. The method according to claim 18 , wherein the method comprises applying a radiofrequency drive signal to the acoustic-optic deflector with a direct digital synthesizer.
20. The method according to claim 19 , wherein the method comprises modulating the amplitude of the frequency shifted beams of light.
21. The method according to claim 19 , wherein the method comprises generating a local oscillator beam and the plurality of frequency shifted beams of light from the laser.