Live / Dead assay mammalian cells - HepaRG human hepatoma

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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Upstream tips
Prepare a mix by adding 2 µL of 1 M calcein-AM stock solution and 3 µL of 1.5 M PI solution to 1 mL of Dulbecco’s modified Eagle’s medium supplemented with 2 mM of l-glutamine, 100 U/mL of penicillin and 100 µg/mL of streptomycin.
Protocol tips
Mix 100 µl of the staining solution with 200 µl of the cell suspension, and incubate the mixture for 15 minutes at 37C
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