Incucyte® Cytotox NIR dye for counting dead cells

Incucyte® Cytotox dyes are formulated to detect disruption of cell membrane integrity in real time to quantify cell death. These dyes are ideal for quantification of cells undergoing cytotoxicity, using a real-time mixing and reading method. In combination with the Incucyte® live cell analyzer and integrated software, these reagents enable reliable and accurate cytotoxicity assessment. These dyes can also be used in combination with other Incucyte® reagents for multiple readings of cytotoxicity, proliferation and apoptosis within the same well. Addition of Incucyte® Cytotox dyes to normal healthy cells does not interfere with cell growth or morphology and gives little or no intrinsic fluorescent signal.

Once cells become unhealthy, the integrity of the plasma membrane is reduced, allowing Incucyte® Cytotox dye to enter and causing a 100- to 1000-fold increase in fluorescence upon binding to deoxyribonucleic acid (DNA).

Storage and stability

Incucyte® Cytotox dyes are supplied as a solution in dimethyl sulfoxide (DMSO), providing sufficient volume to perform 500–1000 tests (1 test = 1 well of a 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. If stored correctly, the solutions will be stable for 6 - 12 months.

Recommended use

We recommend that each vial of Incucyte® Cytotox Green or Red Dye be diluted to a stock concentration of 100 µM in complete medium or PBS. It can be further diluted in complete medium for direct addition to cells seeded in a 96-well plate to give a final concentration of 250 nM (eg 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 the entire medium volume. When used in the Incucyte® device, we recommend collecting data every 2-3 hours.

Features

• detect cytotoxicity in real time with non-disruptive reagents
• reduce the loss of vulnerable cells with simple mixing and reading protocols
• collect reliable data with little or no intrinsic fluorescence
• multiplex with measurements of proliferation and apoptosis

Application

• ideal for the detection of cell membrane integrity disorders and the quantitative determination of cell death in real time
• multiple applications, such as apoptosis, proliferation and growth of single spheroids, in combination with other Incucyte® reagents and metrics