I would like to monitor cell viability during an experiment (in a quantitative way if possible) using the very same cells I'm using for the experiment. I'd be using poly IC and viruses, and would like to make sure that the potential effects are not due to cells dying, but they are actual responses by living -but stressed- cells. One option is to use a Caspase assay on a parallel set of cells, but the ideal would be to use the same cells. What are the best, most used methods to test this?
Hello,
If you have a facs (flow cytometry), it's the best option.
Good luck
Hmm... maybe just very easy by staining your cell layer after infection with crystal violet or coomassie to make sure that there is no cytopathic effect or loss of cells because of cell death (depending on cell layer size and virus).
otherwise i would do an MTT/XTT or WST-1 assay with infected and uninfected cells.
Good luck
Cell titer-glo or cell tox-glo from Promega are useful assays for this purpose. The assays measure ATP production by cells as a readout for toxicity.
Thank you all for the answers. The problem with most of the assays is that they sacrifice the cells -you cannot routinely check your treated samples, as the protocol consumes them. Perhaps the viablity stains would be the best to do. (Although they only show dead/live cells, so the dying but still live cells would not show up.)
...
Need to think hard on this. Thank you again.
Check out the PrestoBlue cell viability assay (Life Technologies). Hope this helps.
Make replicate wells of your cells and sacrifice a few wells at every time point you wish to test for viability.
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