Live/Dead Stain Dyes for Flow Cytometry
Understanding the degree of cell death in a flow cytometry assay is critical towards any analysis of cell population. Dead cells can non-specifically bind antibodies and exhibit higher autofluorescence than live cells. This both reduces the dynamic range and can lead to false positives in your results.
It's true that using gates based on forward and side scatter can help remove confusing substances in your sample such as debris and dead cells; however, this technique does not remove them all and the use of live/dead markers remain essential in flow cytometry to elucidate viability.
RayBright® Live dyes are amine-reactive fluorescent dyes that help researchers distinguish between live and dead cells in flow cytometry analysis. Our trademarked line of dyes bind with a much brighter signal in dead cells than in living cells. Living cells refuse the dye while dead cells are stained both on the cell surface and intracellularly. Each product is specific for your excitation laser and includes a cell staining protocol. Size options are 100 or 500 tests.
Cell Viability Dyes are supplied frozen in DMSO solution at a pre-titrated concentration. We recommend end users further determine the optimal concentration for their specific cell types.
Cell Staining Protocol
- Wash cells with PBS (free of protein, Tris and sodium azide) twice.
- Wash cells with FACS buffer once
- Resuspend cells in 100 µl of PBS containing RayBright® Live dye (1:100 ~ 1:500 dilution for 106 ~ 107 cells).
- Continue for further standard flow cytometry staining.
- Incubate the cells on ice or at 4°c for 20 minutes.
- Cells can be fixed with paraformaldehyde or methanol for intracellular staining.
