Live / Dead assay mammalian cells - human fibroblast tissue

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 3 matching solutions for this experiment

Upstream tips
Prepare a staining solution of 2 uM calcein AM/4 uM EthD-III by adding 5 uL of 4 mM calcein AM and 20 uL of 2 mM EthD-III to 10 mL of PBS or other serum-free buffer or medium
Protocol tips
Incubate the cells for 30-45 minutes at room temperature.
Upstream tips
Mix 1 µL of Solution A and 1 µL of Solution B in 1 mL of Staining Buffer to make a staining solution
Protocol tips
Incubate for 15 minutes at 37°C.
Protocol tips
Mix 2 µM Calcein AM and 4 µM Ethidium homodimer‐1 in PBS and incubate at 37°C for 20 min.
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