I need to measure cell viability in 48/24 well plates. However, since the cells are under treatment of an organic material which has some absorbance itself, I can't use/ trust the optical density of the culture. I would be appreciated if you suggest a method (not trypan blue) by which I can measure the cell viability in well plates.
Hi, you can use fluorescence imaging and/or fluorescence intensity measurement (RFU) with the live/dead assay, for example.
Dear Mahbobeh,
You can do a live pI (propidium Iodide) stain. for that you can stain your cells with pI (a 1mg/ml stock solution), use 3-5 ul of the same for around 1 lac cells, incubate for 10 minutes in dark and then trypsinize the cells (pI is a good stain and trypsinization will still maintain the fluorescence,just tc to do procedure in dark), after u get cells in solution, you can opt two approaches, go to a flow cytometer and perform live PI stain by gating in pI gate. second if u want images for cells positive for pI, go to a Image cytometer, acquire 25ul of the sample and it will give you all stats of live/dead along with photographs of cells with fluorescent red nuclei (dead cells). Hope it helps.
You can use Alamar Blue assay (http://www.ncbi.nlm.nih.gov/pubmed/24910583). You could use our protocol.
You could also use CyQuant assay (based on DNA content).
Article Evaluation of The Alamarblue Assay for Adherent Cell Irradia...
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