BD Biosciences FACSAria Fusion Cell Sorter

The BD Biosciences FACSAria Fusion™ is a high-end Special Order Research Product (SORP) cell sorter configured with 5 high power lasers (UV 355 nm, Violet 405 nm, Blue 488 nm, Yellow-Green 561 nm, Red 637 nm), 18 fluorescent detectors and 2 light scatter detectors. The BD Biosciences FACSAria Fusion can sort up to 4 populations simultaneously. Samples can be sorted into 12 x 75 mm FACS tubes or 1.5 ml Eppendorf tubes for 4-way sorts, and into 15 ml centrifugation tubes or 50 ml centrifugation tubes for 2-way sorts. In addition, the FACSAria can deposit single cells into 96- or 384-well plates. The optional index sorting function ensures that a single-sorted cell has the desired phenotype. This system maintains sample sterility and can sort up to ~25,000 events per second depending on cell type and nozzle size (70, 85, 100, and 130 um). The FACSAria Fusion is operated using FACSDiva 9.1v software. FCS file versions 2.0, 3.0, and 3.1 can be exported for data analysis.

Sample Preparation for Cell Sorting (FACSAria Fusion) 
 
For sorting, a minimum of 10 x 106 cells is usually required; the optimal concentration is between 3-5 x 106 cells/ml. Cells ready for sorting should be resuspended and then collected in the media they are "happiest" in, which includes either commercially available FACS buffers, PBS, or cell culture media such as RPMI or DMEM. Realize that the presence of phenol-red in the culture media may affect the background florescence slightly; however, in general this is not a problem. 
 
In the case of adherent or inherently sticky cells, it will help to add 1mM EDTA to the sorting buffer. Adding EDTA should prevent the formation of cell clumps, which will be aborted by the machine and not sorted. Also, addition of a buffering agent like HEPES (pH 7.0) to the sorting and collection buffers will help buffer the pH of the solution and aid in cell viability as the cells exit the sorter. Bring the collection buffer of choice as well as extra sorting buffer in case the cells need to be diluted.