Eugene Tu, Haichuan Zhang, Mark Wang, Daniel Raymond, Joon Mo Yang, Norbert Hagen, Bob Dees, Elinore Mercer, Anita Forster, Soheil Attari, Chris Richardson, Ilona Kariv, Mirianas Chachisvillis, Manami Hara, Philippe Marchand, William Butler
A microfabricated fluorescence activated cell sorter based on an optical switch enables low-stress sorting of small cell populations (i.e. 1,000 - 100,000 cells), a regime that is not addressed by current cell sorters. The glass microchannels are packaged in a self-contained plastic cartridge that enables "chip-to-world" interconnect for ease of use.
The sample is hydrodynamically focused using pneumatic pressure driven flow control and is biased to the waste reservoir by default. A 5 mW, 488 nm semiconductor laser is used for cell detection and fluorescence excitation. A sorting event is triggered based on the fluorescence and light scatter properties of the cell. The optical switch is produced from a 20 W CW 1070 nm Ytterbium fiber laser that is controlled by an acousto-optic modulator to deflect the target cell to the target reservoir. Cells can be recovered after the sort for further manipulations. Sorting performance was evaluated using a stably transfected, GFP expressing HeLa cell line spiked at varying ratios into parental HeLa cells. Throughputs ranged from 20-100 cells/sec depending on initial cell concentrations and flow parameters. Target purities greater than 90% were obtained in most cases with overall recoveries greater than 85%. A 1% subpopulation of GFP-HeLa cells could be enriched by 63-71 fold. Similar results were demonstrated using samples from pancreas and other primary tissues. Optical switching can be scaled to fluidic networks of greater complexity by multiplexing an optical source to multiple locations on a chip, and these locations can be arbitrarily reconfigurable.
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