Flow Cytometry Core Service Request
Fluorescence activated cell sorting (FACS) is available to our investigators and is performed on either a BD Influx or BD FACSVantage cell sorter. All sorts are performed by core personnel. Prior to any and all sorts a sorter sample questionnaire and a sort protocol questionnaire must be completed and submitted to core personnel before the sort will be scheduled. In addition all sort samples must be prepared in a serum free, EDTA containing sort buffer. Finally, all sort samples will be filtered by the sorter operator thru a 35 micron filter prior to the sort to eliminate any clumps.
A flow cytometer sorter makes it possible to isolate highly purified populations of particles having distinct characteristics. These particles are not limited to cells but may be chromosomes, microorganisms, microbeads, etc. Any parameter that is measurable on a flow cytometer can be used as a sorting criterion. Cell sorters work by rapidly vibrating a crystal connected to a flow cell, causing a stream of cells to break into uniform droplets. As each droplet separates from the stream it can be independently charged and subsequently deflected past a charged plate into a receptacle. Both sorters have software that includes signal pulse height and width measurements such that doublet discrimination and DNA analysis are available. In addition both sorters perform automatic compensation on all channels when supplied with the appropriate controls. Both the Influx and the Vantage are equipped with a sample cooling system should sort and collection temperature be an issue for your samples. For additional information on preparing cells for sorting in order to optimize your results or to view the filter configurations on the Influx and Vantage go to the sort preparation page or the filter configurations page.
The BD Influx cell sorter with five lasers (488, 640, 561, 405 and 355), 3 scatter and 19 fluorescence detectors supports up to six-way sorting as well as plate sorting. High speed sorting at fifteen to twenty thousand events per second is achievable with the Influx. In addition, the Influx has small particle detection capabilities via forward scatter which when optimized can detect particles as small as 200 nm. The Influx is housed in a Baker Bioprotect hood which allows sorting of samples rated at BSL2+ or below. The Influx runs BDFACS Sortware software.
Having four lasers (488, 561, 635 and 355), two scatter detectors and eight fluorescence detectors the Vantage is capable of a wide variety of cell sorting. The Vantage can be configured for two-way or four-way sorting as well as macro sorting, high speed sorting and single cell sorting into 96 well plates which is perfect for cloning or single cell analysis. The Vantage uses BD Diva acquisition and analysis software. Being housed in the main core lab area the Vantage is available only for sorts at or below BSL2 classification, sorting only non-primate cells that have not been transformed or have not been exposed to any potential pathogens.