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CytoFLEX SRT Beckman Coulter cell sorter

Access: with an Operator or training required (1-2 experimental session)

General Description

CytoFLEX SRT is a fixed alignment benchtop cell sorter. It uses four high-powered lasers: violet (405nm, 80mW), blue (488nm, 50mW), yellow-green (561nm, 30mW) and red (638nm, 100mW). The system is equipped with forward (FSC) and side (SSC) scatter detectors and 15 fluorescence detectors:

Violet 405 (450/50, 525/40, 610/20, 660/10, 780/60)
Blue 488 (525/40, 690/50)
Yellow-green 561 (585/42, 610/20, 675/30, 710/50, 780/60)
Red 638 (610/20, 712/25, 780/60)

The tube holders for 1.5 ml, 5 ml and 15 ml are provided. There is a multi-well plate sorting module for a variety of plates including 384 format and slides. Index sorting is capable for single-cell applications. CytoFLEX SRT can sort up to four populations simultaneously. The is one 100 µm integrated nozzles available.

Sample preparation

Prepare cells suspension in filtered buffer (PBS with 2% BSA + 2.5 mM EDTA) but if necessary, in any other medium without phenol red and FCS.
The sample should have particles density of approximately 5-10 million/ml. If less cells are available put, then into minimal volume of 500 µl.
Negative controls and individual positive controls of your staining are required to set sorting gates appropriately.

Tips

To improve cells viability, make all preparation steps on ice, unless otherwise stated in your protocol.
To exclude dead cells from the analysis or sorting it is recommended to use DNA-binding dyes as a marker of dead cells.
DNA released from dead cells during cells dissociation causes stickiness. To reduce cells aggregation in your samples, incubate the cells in the presence of DNase I (100 ug/ml with 5 mM MgCl2).