BD FACSMelody

Transfected GFP cell sorting
Transfected GFP cell sorting
HEK-293 cells (ATCC® CRL-1573™) were transfected over 24 hours with AcGFP. Untransfected cells were used as a negative control to set the instrument and draw the gates (not shown). Two populations of GFP+ cells, dim and bright, could be discriminated. GFP+ bright cells were identified and sorted at 1,000 total events/ second in purity mode. Post-sort analysis revealed a purity of 98.8%.
Regulatory T-cell (Treg) sorting using a panel with minimal spectral overlap
Regulatory T-cell (Treg) sorting using a panel with minimal spectral overlap
Peripheral blood mononuclear cells (PBMCs) were isolated from human whole blood and stained with an antibody cocktail for the identification and sorting of Tregs. Lymphocytes were first gated based on light scatter, and following doublet discrimination (not shown), helper T cells were defined based on CD4 expression. From the CD4+ gate, Tregs were then identified as CD25+CD127dim/- cells and sorted at 3,500 total events/second in purity mode. Post-sort analysis revealed a cell purity of 99.7%. With each fluorochrome excited by a different laser and cross-laser excitation accounted for, spectral overlap between channels was minimal (<1%).