Photon signal processing for particle detection
A method is described wherein a plurality of lasers are used to irradiate particles in a flow cytometer's flow stream. In certain embodiments, a light source having a plurality of lasers configured for irradiation of a flow stream are disclosed where discrete intervals of irradiation by one or more discretely activated lasers are triggered by irradiation of one or more particles in the flow stream with one or more continuously on lasers.
1. A method of detecting particles in a flowcytometry system, comprising:
irradiating a flow stream having one or more particles by a plurality of lasers;
collecting light from a single fiber optic from the one or more particles in the flow stream irradiated by each of the plurality of lasers; and
detecting light conveyed by the single fiber optic by a single photodetector having one or more channels, wherein discrete intervals of irradiation by one or more discretely activated lasers of the plurality of lasers is triggered by irradiation of one or more particles in the flow stream by one or more continuously on lasers of the plurality of laser as registered by the single photodetector; and
calculating by a processor timing associated with each discrete interval of irradiation of the flow stream by each of the one or more discretely activated lasers, wherein the processor is accompanied by a tangible non-volatile computer-readable storage medium (memory) operably coupled to the processor.
2. The method of claim 1 , wherein each of the one or more discretely activated lasers is configured to irradiate the flow stream at a position downstream from the one or more continuously on lasers.
3. The method of claim 1 , wherein the wavelength of light from the one or more continuously on lasers is shorter than light from one or more of each of the one or more discretely activated lasers.
4. The method of claim 1 , wherein the wavelength of light from the one or more continuously on lasers is longer than wavelength of light from one or more of each of the one or more discretely activated lasers.
5. The method of claim 2 , wherein each of the one or more discretely activated lasers outputs a different wavelength of light.
6. The method of claim 2 , wherein each discrete interval of irradiation of the flow stream by each of the one or more discretely activated lasers is independently triggered by irradiation of the one or more particles in the flow stream with the one or more continuously on lasers.
7. The method of claim 1 , wherein the memory comprises instructions stored thereon, which when the instructions are executed by the processor, cause the processor to calculate timing of irradiation of the flow stream by each of the one or more discretely activated lasers by: irradiating the one or more particles in the flow stream with the one or more continuously on lasers; detecting light from the flow stream in response to irradiation of the one or more particles with the one or more continuously on lasers; and calculating a time interval between irradiation of the one or more particles by the one or more continuously on lasers and each of the one or more discretely activated lasers.
8. The method of claim 1 , wherein the method further comprises modulating each of the one or more discretely activated lasers by a modulator.
9. The method of claim 8 , wherein the step of modulating each of the one or more discretely activated lasers occurs in response to irradiation of the one or more particles by the one or more continuously on lasers.
10. The method of claim 1 , wherein the method further comprises diverting light from the one or more discretely activated lasers away from the flow stream by a modulation device positioned in the beam path between the one or more discretely activated lasers and the flow stream.
11. The method of claim 10 , wherein the modulation device functions as a beam diverter configured to direct light from the one or more discretely activated lasers to the flow stream in response to irradiation of the one or more particles by the one or more continuously on lasers.
12. The method of claim 11 , wherein the modulation device comprises an acousto-optical device.
13. The method of claim 1 , wherein the method further comprises modulating an amount of light that is conveyed from the one or more continuously on lasers to the single photodetector by an optical adjustment component.
14. The method of claim 13 , wherein the optical adjustment component is a dichroic filter.
15. The method of claim 1 , wherein the single photodetector is a photodetector array.
16. The method of claim 1 , wherein the single photodetector is selected from the group consisting of photomultiplier tubes (PMT), Geiger avalanche photodiode (APD), silicon photomultipliers (SiPM), and combinations thereof.
17. The method of claim 1 , wherein the single photodetector includes 42 channels.