Live / Dead assay mammalian cells - mouse microglia

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

LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	Add 100–150 µL of the combined LIVE/DEAD® assay reagent and incubate the cells for 30–45 minutes at room temperature	Following the dye incubation, add about 10 µL of the fresh LIVE/DEAD® reagent solution or D-PBS to a clean microscope slide.

Protocol tips
Add 100–150 µL of the combined LIVE/DEAD® assay reagent and incubate the cells for 30–45 minutes at room temperature
Downstream tips
Following the dye incubation, add about 10 µL of the fresh LIVE/DEAD® reagent solution or D-PBS to a clean microscope slide.

Manufacturer protocol Publication protocol 1 Relevant paper

Zombie Violet™ Fixable Viability Kit

BioLegend

Upstream tips	Protocol tips	Downstream tips
Don’t use Tris buffer as a diluent and be sure that the PBS does not contain any other protein like BSA or FBS	Dilute Zombie Violet™ dye at 1:100-1000 in PBS and add to cells. Incubate the cells at room temperature, in the dark, for 15-30 minutes

Upstream tips
Don’t use Tris buffer as a diluent and be sure that the PBS does not contain any other protein like BSA or FBS
Protocol tips
Dilute Zombie Violet™ dye at 1:100-1000 in PBS and add to cells. Incubate the cells at room temperature, in the dark, for 15-30 minutes

Manufacturer protocol Publication protocol 1 Relevant paper

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