Methods and apparatuses for sorting target particles
This disclosure provides methods and apparatuses for sorting target particles. In various embodiments, the disclosure provides a cassette for sorting target particles, methods for sorting target particles, methods of loading a microchannel for maintaining sample material viability, methods of quantifying sample material, and an optical apparatus for laser scanning and particle sorting.
1. A method of detecting and sorting target particles, the method comprising:
providing a substrate with a plurality of microchannels, wherein said microchannels comprise a sample material mixture comprising a plurality of target particles;
scanning the contents of said microchannels with an excitation beam;
detecting a fluorescence signal emitted from said microchannels wherein said fluorescence signal indicates the presence of said target particle in a microchannel; and
extracting the target particles from said microchannels with an extraction beam;
wherein scanning said microchannels with an excitation beam occurs at a rate from about 1,000,000 microchannels per second to 100,000,000 microchannels per second.
2. The method of claim 1 , wherein said target particles are extracted from said microchannels with an extraction beam occurs at a rate from about 500,000 microchannels per second to about 1,000,000 microchannels per second.
3. A method of detecting and sorting target particles, the method comprising:
providing a substrate with a plurality of microchannels, wherein said microchannels comprise a sample material mixture comprising a plurality of target particles;
scanning the contents of said microchannels with an excitation beam;
detecting a fluorescence signal emitted from said microchannels wherein said fluorescence signal indicates the presence of said target particle in a microchannel; and
extracting the target particles from microchannels with an extraction beam;
wherein extracting the target particles from said microchannels with an extraction beam occurs at a rate from about 500,000 microchannels per second to about 1,000,000 microchannels per second.
4. The method of claim 3 , where said microchannels are scanned with an excitation beam at a rate from about 1,000,000 microchannels per second to 100,000,000 microchannels per second.
5. The method of claim 1 , wherein said sample material mixture comprises a cellular suspension.
6. The method of claim 5 , wherein said sample material mixture comprises about 1×10 6 to about 100×10 9 cells.
7. The method of claim 1 , wherein scanning the contents of said microchannels comprises illumination of a microchannel with a first wavelength and detection of a second wavelength from said microchannel, wherein said second wavelength corresponds with said target particles.
8. The method of claim 7 , wherein said first and second wavelength are independently selected from about 200 nm to about 1.5 mm.
9. The method of claim 1 , wherein scanning the contents of said microchannels comprises illumination of a microchannel with a plurality of different wavelengths and detecting an emission from said microchannel wherein said emission corresponds with one or more target particles.
10. The method of claim 1 , wherein scanning the contents of said microchannels comprises illumination of a microchannel with a single wavelength and detection of a plurality of emissions from said microchannel, wherein said plurality of emissions correspond with one or more target particles.
11. The method of claim 1 , wherein scanning the contents of said microchannels comprises illumination of a microchannel with a plurality of different wavelengths and detection of a plurality of emissions from said microchannel, wherein one or more of said emissions correspond with one or more target particles.
12. The method of claim 1 , wherein said target particle is a cell.
13. The method of claim 12 , wherein said target particle is a cell with a desired phenotype.
14. The method of claim 13 , wherein greater that 95% of the extracted cells are cells of said desired phenotype.
15. The method of claim 13 , wherein greater than 95% of the extracted cells are viable as determined by evaluating the cells within 5 hours from said extracting.
16. The method of claim 13 , wherein the extracted cells are suitable for therapeutic use without the need for additional sterilization steps.
17. The method of claim 13 , wherein the extracted cells are essentially free of pathogens.
18. The method of claim 13 , wherein the extracted cells comprise less than 0.1% of pathogens.
19. The extracted cells obtained by the method of claim 13 .
20. A pharmaceutical composition comprising the extracted cells of claim 19 .