Description
• Detect live and dead cells by microscopy, plate reader, or flow cytometry
• Both live and dead cells may be quantified on a fluorescence plate reader; live cells may also be quantified on a standard colorimetric (ELISA) plate reader
The CytoSelect Cell Viability and Cytotoxicity Assay Kit provides a simple format for monitoring cell viability via metabolic activity. Live cells are detected with either MTT (colorimetric detection) or Calcein AM (fluorometric detection). Dead cells are detected by EthD-1 reagent (fluorometric). All 3 detection reagents are included, along with Saponin (a cell death initiator). Prior to the assay, cells may be treated with compounds or agents that affect cell viability. This kit is suitable for eukaryotic cells, not yeast or bacteria.
Introduction: The measurement and monitoring of cell viability is an essential technique in any laboratory focused on cell-based research. This skill allows for the optimization of cell culture conditions as well as the determination of cytokine, growth factor, or hormone activity. More importantly, the cytostatic nature of anticancer compounds in toxicology testing, the efficacy of therapeutic chemicals in drug screening, and cell-mediated cytotoxicity can all be assessed through the quantification and monitoring of cell viability and growth.
Cell viability characteristics include cellular metabolic activity and cell membrane integrity. One method for measuring metabolic activity is to incubate the cells with a tetrazolium salt such as MTT, which is cleaved into a colored formazan product by metabolically active cells. The green fluorescent viability dye Calcein AM can measure intracellular esterase activity, which is another indicator of cell viability. Live cells can convert the nonfluorescent, cell-permeable polyanionic calcein acetoxymethyl (Calcein AM) dye to the highly fluorescent calcein. The cleaved calcein remains in the cells. Ethidium homodimer (EthD-1) is an excellent marker for measuring dead cells. EthD-1 is a red fluorescent dye that can only penetrate damaged cell membranes. EthD-1 will fluoresce with a 40-fold enhancement upon binding ssDNA, dsDNA, RNA, oligonucleotides, and triplex DNA. Background fluorescence levels are very low because the dyes are virtually non-fluorescent before interacting with cells. This method of detection is more efficient, safer, less expensive, and a more sensitive method for determining cell viability or cytotoxicity compared to traditional viability assays such as 51Cr release or trypan blue exclusion