Flow Cytometry: Sorting

 

The facility houses three high end cell sorters operated by a team of experienced scientists and operators. All the sorters are housed in Class II biological safety cabinet offering an extra level of protection to both operator and sort. This enables samples of human origin to be safely sorted in accordance to international guidelines. 

The ‘flag ship’ Aria III SORP (Special Order Research Product) cell sorter is highly configurable with six independent lasers allowing up to 18 fluorescence parameters (out of a choice of 23) parameters to be acquired. The Aria 3 has the capability of resolving 4 different fluorescent proteins simultaneously from ex-vivo samples enabling high content sorting of constituently expressed genes. The Influx cell sorter has an additional feature of being able to sort six populations simultaneously, rather than the usual four, so maximising the use of precious biological material. All the cell sorters can be optimised for any experiment specific conditions and any cell type, our team work closely with all the researchers to ensure precision and specificity for all sorts.

 In addition to identifying cell populations and bulk sorting, the cell sorters can physically separate cells (sorted) for further downstream molecular studies, and sort single cells into 96 or 384 well plates including index sorting.

 

The cell sorting laboratory, all sorters are enclosed in class II cabinets 

 

 

The BD Influx (the class II enclosure has been opened so to allow a clear view of the equipment)

 

Sorting human peripheral blood mononuclear cells phenotyping CD4 and CD8 naïve and memory T cells (CD45RA andCD45RO respectively). The data below demonstrates the utility of the equipment. The top panel shows the first sort with the red gating depicting the populations of interest to be sorted. The bottom panel showing the completed sort of 98% purity of the populations of interest. 

Sorting human peripheral blood mononuclear cells and phenotyping (Data generated from samples supplied by Sara Valpione, Molecular Oncology)