Live / Dead assay bacteria - Corynebacterium glutamicum

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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3 years ago

3 years 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
Remove the supernatant and resuspend the pellet of one tube in 0.3 ml of 0.85% NaCl solution and another tube in 1 ml of 0.85% NaCl.

Add 0.7 ml isopropyl alcohol into the tube with 0.3 ml of 0.85% NaCl and mix well

Incubate both samples at room temperature for 1 hour, mixing every 15 minutes.
Protocol tips
Incubate with a mixture of SYTO 9 and propidium iodide for 15 min in the dark
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