Live / Dead assay mammalian cells - rat endothelial progenitor cells

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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Found 2 matching solutions for this experiment

Protocol tips
Add 100 µL/well (96-well plate) or 25 µL/well of CytoCalcein™
Green/Propidium Iodide dye-working solution.

Incubate the plate at room temperature or 37°C for 30 minutes to 1 hour, protected from light.
Downstream tips
Monitor the fluorescence intensity with a fluorescence plate reader (bottom read mode) at Ex/Em = 490/525 nm
Protocol tips
Add 2 µM calcein AM and 4 µM EthD-1.

Incubate the cells for 30–45 minutes at room temperature
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