The LIVE/DEAD Viability/Cytotoxicity Assay Kit (Green/Deep Red) is a two-color fluorescent cell viability assay that simultaneously identifies live and dead cells. Live cells are labeled with calcein dye and emit a green fluorescence, while dead cells are labeled with SYTOX Deep Red dye and emit a red fluorescence. These dyes can be multiplexed with other orange/red fluorophores with compatible filter sets.

• Determines cellular viability with two-color fluorescence
• Detect live cells with GFP/FITC filter set and dead cells with Deep Red/CY5 filter set
• Multiplexable with orange/red fluorophores such as RFP, mCherry, Mitotracker Red/Orange, Lysotracker Red, TMRM, and others

Live cells are distinguished by the presence of ubiquitous intracellular esterase activity, determined by the enzymatic conversion of the virtually non-fluorescent cell-permeant Calcein AM to the intensely fluorescent calcein. Calcein is well retained within live cells, producing an intense uniform green fluorescence in live cells (ex/em ∼495/∼515 nm). Calcein can be detected with a traditional FITC/GFP filter set. SYTOX Deep Red enters cells with damaged membranes and increases its fluorescence many-fold upon binding to DNA. Dead or plasma membrane-compromised nuclei fluorescence brightly and can be detected with a CY5/Deep Red filter set (ex/em maxima: 660/682 nm).

The kit is suitable for use with fluorescence microscopes or high content analysis (HCA) fluorescence multiwall plate scanners and easily adaptable for use other fluorescence detection systems. The assay principles are general and applicable to most eukaryotic cell types, including adherent and non-adherent cells.