Fixable Viability Stain 440UV

Flow cytometric analysis of human Jurkat cells stained with BD Horizon™ Fixable Viability Stain 440UV. Cells from the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line were treated with 0.025% DMSO (Left Panels) or 5 μM camptothecin (Right Panels) for 16 hours and then stained (Bottom Panels) with BD Horizon™ Fixable Viability Stain 440UV (Cat. No. 566332) in serum-free buffer. The cells were then either left unfixed (solid line histograms) or fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized in BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) (dashed line histograms). Dead cell fluorescence is retained after fixation and permeabilization. The shift in the fluorescence of the live cells is due to a shift in auto-fluorescence post-fixation, as evidenced by the shift in the unstained (Top Panels) samples. Histograms were derived from gated events with the forward and side light-scattering characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometry System. Please note that FVS440UV is also compatible with BD Phosflow™ Perm Buffer III (Cat. No.558050) or BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725).

Preparation

Bring FVS440UV dye powder and 363 μl of fresh cell culture-grade Dimethyl Sulfoxide (DMSO; eg, Sigma D2650) to room temperature. Add 363 μl of DMSO and vortex solution well. Inspect the solution and repeat vortex until the stock dye has fully dissolved. This is the Stock Solution.

Storage

Upon arrival, store the dry dye desiccated and protected from light at -80°C until use. After reconstitution with DMSO, store the Stock Solution at -20°C in small aliquots. Do not use reconstituted dye after 90 days of storage. Please discard the dye solution after 90 days post reconstitution with DMSO.