Live / Dead assay mammalian cells - mouse bone marrow-derived macrophages

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
Post infection with bacteria, add 100–150 µL of the combined LIVE/DEAD® assay
reagent.

Incubate the cells for 30–45 minutes at room temperature
Protocol tips
Add reconstituted Substrate Mix to each well of the plate.

Incubate at room temperature, protected from light, for 30 minutes
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