|9808||Phosphate Buffered Saline (PBS-20X)||
The CellSimple™ Cell Health Assay Kit is a fluorescent assay designed for use with the CellSimple™ Cell Analyzer. The Cell Health Assay can determine cell viability by measuring intracellular esterase activity and plasma membrane integrity. The assay kit contains the fluorescent dyes Calcein-AM and Propidium Iodide (PI) and is therefore able to stain both live and dead cells simultaneously.
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The CellSimple™ Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.
Measures of cell viability and cytotoxicity are broadly used to study the effects of growth factors and cytokines, inhibitors and activators, and immune response signals. Calcein-AM is the acetomethoxyl form of calcein, a highly lipophilic, cell membrane permeable dye. Intracellular esterase activity converts the non-fluorescent Calcein-AM to the highly fluorescent Calcein, which is retained only within live cells (1,2). The DNA-binding agent PI is cell membrane impermeable and only enters dead cells or those with damaged cell membranes. Intracellular PI binds DNA and undergoes an approximate 40-fold enhancement in fluorescence intensity. As a result, live cells will produce a strong green fluorescence resulting from the conversion of Calcein-AM to Calcein, while dead cells produce a strong red fluorescence due to the presence of PI (3,4). These fluorescent signals can be detected using the Cell Health application on the CellSimple™ Cell Analyzer.
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