Live / Dead assay bacteria - Pseudomonas aeruginosa

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.

Start discussion

Found 1 discussion for this experiment

Discussion

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?

Share your thoughts or question with experts in your field by adding a discussion!

Found 3 matching solutions for this experiment

Protocol tips
Add 50 μl of 2.5 μM SYTO9 to the wells.

Incubate on the orbital shaker for 15 minutes in the dark.
Downstream tips
Detect using a 488/20 nm excitation filter (for both SYTO9 and PI) and a 528/20 nm (SYTO9 emission wavelength) and 645/40 nm (PI emission wavelength) emission filter.
Upstream tips
Prior to use, mix 1 µl of solution A and 1 µl of solution B in 1ml of Staining Buffe
Protocol tips
Resuspend to 1 ml Staining Solution.

Incubate for 15 minutes at 37°C
Protocol tips
Incubate the cells in an incubator (37°C, 5% CO2, 90% humidity
Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!
Become shareholder Discussions About us Contact Privacy Terms