Method of detaching adherent cells for flow cytometry
US-2015368620-A1 · Dec 24, 2015 · US
US9040304B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9040304-B2 |
| Application number | US-201414206832-A |
| Country | US |
| Kind code | B2 |
| Filing date | Mar 12, 2014 |
| Priority date | Mar 28, 2003 |
| Publication date | May 26, 2015 |
| Grant date | May 26, 2015 |
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A multi-channel system and methods for sorting particles according to one or more characteristics of the particles. The system includes multiple flow cytometry units, each unit can have a nozzle for producing a fluid stream containing a desired population of particles in a mixture of particles. Each of the units may be operable to sort said desired population of particles by interrogating the fluid stream with a beam of electromagnetic radiation and classifying particles based on one or more characteristics of the particles. The system also includes a common fluid delivery system for delivering sheath fluid to each flow cytometer unit for producing respective fluid streams.
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What is claimed is: 1. A multi-channel system for sorting particles according to one or more characteristics of the particles, said system comprising: multiple flow cytometry units, each unit having a nozzle to produce a fluid stream containing a desired population of particles in a mixture of particles, wherein each of said units is operable to sort said desired population of particles by interrogating the fluid stream with a beam of electromagnetic radiation and classifying particles based on one or more characteristics of the particles; and a common fluid delivery system for delivering sheath fluid to each flow cytometer unit for producing respective fluid streams. 2. The multi-channel system of claim 1 wherein the particles are cells. 3. The multi-channel system of claim 1 wherein the particles are sperm cells. 4. The multi-channel system of claim 1 wherein one or more of the multiple flow cytometry units comprises a jet-in-air droplet sorting flow cytometry unit. 5. The multi-channel system of claim 1 wherein the multiple flow cytometry units are provided on an integrated platform. 6. The multi-channel system of claim 5 wherein the integrated platform comprises a common input for controlling operation of said units and a common processor for receiving and processing information from said units. 7. The multi-channel system of claim 5 wherein said integrated platform comprises a common source of electromagnetic radiation for producing the beam of electromagnetic radiation for each unit. 8. The multi-channel system of claim 7 wherein the common source of electromagnetic radiation comprises a single laser producing a single laser beam and further comprising a beam splitting system for splitting the single laser beam into multiple beams and directing the multiple beams into optics systems of respective flow cytometry units. 9. The multi-channel system of claim 8 wherein said single laser beam comprises a plurality of pulses. 10. The multi-channel system of claim 9 and wherein one or more of said flow cytometry units comprises: a flow channel for directing a fluid stream containing sample particles through a particle interrogation location; a beam guidance system operable to direct a portion of the electromagnetic radiation in a pulse to the interrogation location; a timing circuit operable to produce a timing signal indicative of the arrival of electromagnetic radiation at the interrogation location; a detector adapted to detect electromagnetic radiation from the interrogation location and operable to output a time-varying analog signal indicative of the intensity of the detected electromagnetic radiation; an analog to digital converter adapted to receive the time-varying analog signal as input and to sample the analog signal to produce a digitized output; and an electronic processor operable to analyze the digitized output from the analog to digital converter as a function of the timing signal. 11. The multi-channel system of claim 5 , wherein the integrated platform comprises a common processor which performs at least one of: (1) receiving and processing information from the flow cytometry units in real time; and (2) receiving and processing information from the flow cytometry units to permit evaluation of the operation of one unit relative to another unit. 12. The multi-channel system of claim 5 wherein said integrated platform comprises a common processor operable to send control signals to the flow cytometry units in real time during a sorting process to adjust their operation as a function of information received by the common processor, and wherein the flow cytometry units are responsive to the control signals. 13. The multi-channel system of claim 1 wherein each flow cytometry unit comprises a sensor operable to generate a time-varying output signal indicative of at least one characteristic of the particles, and further comprising a common processor for receiving and processing information from the units and said information received by the common processor comprises the output signals from the respective sensors, and wherein the processor is operable to receive the output signals as a substantially continuous stream and to substantially continuously process the output signals in real time. 14. The multi-channel system of claim 1 wherein each flow cytometry unit comprises an epi-illumination optics system for interrogating a respective fluid stream. 15. The multi-channel system of claim 1 wherein at least one of said flow cytometry units comprises a photo-damage system. 16. The multi-channel system of claim 1 wherein at least one of said flow cytometry units comprises a fluid-switching sorting system. 17. The multi-channel system of claim 1 wherein said flow cytometry units are adapted to operate in parallel. 18. The multi-channel system of claim 1 wherein the multiple flow cytometry units comprise three or more flow cytometry units. 19. The multi-channel system of claim 1 wherein the multiple flow cytometry units comprise at least twelve flow cytometry units. 20. A multi-channel method of sorting particles according to one or more characteristics of the particles, said method comprising: delivering sheath fluid to multiple flow cytometry units from a common fluid delivery system; generating fluid streams containing sheath fluid at each of said units, each fluid stream also containing a desired population of particles within a mixture of particles; interrogating each fluid stream with a beam of electromagnetic radiation; classifying particles based on one or more characteristics of the particles; and sorting the desired population of particles within a mixture of particles. 21. The method of claim 20 wherein said particles are cells. 22. The method of claim 20 wherein said particles are sperm cells. 23. The method of claim 20 wherein at least one of said multiple flow cytometry units comprises a jet-in-air droplet sorting flow cytometry unit. 24. The method of claim 20 further comprising sharing a common source of electromagnetic radiation in the form of a single laser beam, said method further comprising splitting the single laser beam into multiple beams and directing the multiple beams into optics systems of respective flow cytometry units. 25. The method of claim 24 further comprising reflecting a percentage of beam light of the single beam toward the optics system of one of said flow cytometry units, and passing a percentage of beam light of the single beam for transmission to the optics system of another of said flow cytometry units. 26. The method of claim 20 further comprising the step of removably mounting said flow cytometry units in a common housing. 27. The method of claim 20 further comprising a common source of electromagnetic radiation in the form of a shared laser; the method further comprising the steps of: emitting a plurality of electromagnetic radiation (EMR) pulses from a laser, wherein a peak power of each pulse exceeds an average power of the laser; directing each pulse into a beam splitting and guidance system to intermittently illuminate each fluid stream and the particles contained therein by directing a portion of the energy in the pulses along a beam path from the laser to an interrogation location associated with each of the flow cytometer units; detecting EMR from at least one interrogation location; gener
Condition responsive control · CPC title
involving viable microorganisms · CPC title
Biological material, e.g. blood, urine (G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence); Haemocytometers (counting blood corpuscules distributed over a surface by scanning the surface G06M11/02) · CPC title
Sperm cells, spermatogonia · CPC title
Volumetric liquid transfer · CPC title
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