We are doing cancer cell experiments in vitro. We use the trypan blue assay method for viable vs. nonviable cells. If we are 'treating' a 12 well plate or a T25 flask, how many assays do we need to do to have truly meaningful data? If we can repeat the experiments three or four times and have similar results (25% or more of cancer cells are nonviable afterwards) does this seem like enough assays (one for each experiment) to have meaningful data?
Hi Anthony,
I suppose your cancer-cells are non-adhärent cells and you take three random sample from the 12-well plate or T25 flask ? In this case I suppose 3 to 6 samples are enough to have a firm basis. Cross-check with neutral-red are also an option. I promise the test is used to dicide further how to go ahead.
all the best
Jochem
Hey Anthony,
What I would suspect if your doing something like a dose response curve biological triplicates would be required. In any case when you do your trypan exclusion test I would recommend not overly diluting before loading the hemocytometer. Generally you want a field of more less 100 cells per 4 quadrants. But i know thats not always the case, so what i would do is fill fill both sides of my hemocytometer and in the end i would average the results of 8 quadrants rather than 4. Like Jochem said it doesn't hurt to cross check your results with other cell death assays, but trypan blue exclusion is generally one of the more accepted assays for measuring cell viability. Good luck with your experiments.
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