Live / Dead assay bacteria - Staphylococcus aureus

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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1 year ago

1 year ago by M. Daecher Germany

Live/dead assay Bacteria

Hello everyone! I am going to do a live/dead assay for my cells and I saw that I can use both fluorescence and absorbance as my detection method. Is there a difference in the results depending on the method? Is one method preferred over the other in certain situations?

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Protocol tips
Wash cells with PBS buffer (no Tris buffer and protein free).

Dilute Zombie Aqua™ dye at 1:100-1000 in PBS. Resuspend 1-10 x 10 cells in diluted 100 µl
Zombie Aqua™ solution.

Incubate the cells at room temperature, in the dark, for 15-30 minutes.
Protocol tips
Add 0.5 mL of dye solution containing 1 mM SYTO9 and 5 mM propidium iodide (PI).

Incubate for 20 min at room
Protocol tips
Add 500 μL of 1X Live and Dead Dye to cells.

Incubate 10 min at room temperature
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