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, Sort Protocol Questionnaire and demonstration of a sortable population via FACS analysis 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.