Research Technologies Branch

Frequently Asked Questions

• Can I use the same fluorochromes for sorting that I have used for analysis on my FACS Calibur/Scan?
• Can my cells be sorted under sterile conditions so that I can put them into in vitro culture after the sort?
• I have four populations that need to be sorted and the number of cells that I have is limiting; can I sort all four populations at the same time?
• Does the number of fluorochromes that I use slow down the sort or decrease the yield?
• Can I access the Flow Cytometry Section after hours?

Can I use the same fluorochromes for sorting that I have used for analysis on my FACS calibur/scan?

Yes, all fluorochromes except for PerCP can be used on the cell sorters. PerCP is photo bleached by the higher power lasers, however. The intensity of fluorescence (signal-to-noise) will be approximately five-fold lower on the cell sorters than the bench-top analyzers. This is due to the difference between a cuvette (analyzers) and stream-in-air optics (cell sorters). Other fluorochromes can be used in addition to those traditionally used for bench-top analyzers. The laser configurations are listed in the Technology section. You can use this spectrum viewer* to determine if the dye you are interested in will work on our sorters. 

Can my cells be sorted under sterile conditions so that I can put them into in vitro culture after the sort?

Yes, the instruments are checked for sterility, but it is advisable to include antibiotics in the medium that you bring the cells in. See the Protocols section for further information on what to bring. 

I have four populations that need to be sorted and the number of cells that I have is limiting; can I sort all four populations at the same time?

One of our sorters (FACS Vantage SE-Orange) is able to sort four populations simultaneously. The other sorters are currently limited to sorting two populations.

Does the number of fluorochromes that I use slow down the sort or decrease the yield?

No there is no difference in the processing speed between one color and multiple colors.

Can I access the Flow Cytometry Section after hours?

Access to the Flow Cytometry Section's instrumentation on nights and weekends can be arranged. Your NIH ID card can be programmed to grant access to the FACScan and Calibur in Bldg. 50, room 5430 and into Bldg. 4; a key can be made available for access to the laboratory in Bldg. 4, Room B1-34.

*Note: Some of the links on this page connect to information sources outside of NIAID and are provided as a convenience for World Wide Web users. Please see the NIAID disclaimer.

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