Incucyte® Cytotox Dye for Counting Dead Cells

Incucyte^® Cytotox Dyes are formulated to detect cell membraned integrity disruption in real time to quantify cell death. These dyes are ideally suited to the mix-and-read, real-time quantification of cells undergoing cytotoxicity. When combined with Incucyte^® Live-Cell Analysis Systems and integrated software, these reagents allow for reliable, accurate evaluation of cytotoxicity. These dyes can also be used in conjunction with other Incucyte^® reagents for multiplexed readouts of cytotoxicity, proliferation and apoptosis within the same well. Addition of the Incucyte^® Cytotox Dyes to normal healthy cells is non-perturbing to cell growth or morphology and yields little or no intrinsic fluorescent signal.

Once cells become unhealthy, the plasma membrane integrity diminishes, allowing entry of the Incucyte^® Cytotox Dye and yielding a 100- to 1000-fold increase in fluorescence upon binding to deoxyribonucleic acid (DNA). With the Incucyte^® integrated analysis software, fluorescent objects can be quantified, and background fluorescence minimized.

Presentation, Storage and Stability

Incucyte^® Cytotox Dyes are supplied as a solution in dimethyl sulfoxide (DMSO), providing sufficient quantity for performing 500–1000 tests (1 test = 1 well of 96-well microtiter plate). Upon receipt, the solution should be stored at -20°C or 4°C, depending on the reagent, and protected from light. When stored as described, the stock solutions will be stable for 6 - 12 months.

Recommended Use

We recommend that each vial of Incucyte^® Cytotox Green or Red Dye is diluted to a stock concentration of 100 µM in full media or PBS. This may then be diluted further in full media for direct addition to cells seeded in a 96-well plate to yield a final concentration of 250 nM (e.g., 1:4000 dilution of the original green or red dye).

Incucyte^® Cytotox NIR Dye should be diluted to a final concentration of 0.6 µM (1:000 dilution) in full culture media for use.

When used in an Incucyte^® Live-Cell Analysis System, we recommend data collection every 2-3 hours. Additional protocol details can be found in the Product Guide.