I am struggling to get any results out of MTT assay because of fluorescence compound. Which is giving false positive results. I am now planning to shift in ATP based cytotoxicity assay. Have I chosen the right method or do you have any suggestion to overcome MTT assay issue?
Hi. Well, it depends on which kind of fluorescence interference you have !?!?!?!
An easy alternative method to study cell viability might be the Propidium Iodide (PI) cell exclusion. You can use a FACS instrument and analyse PI fluorescence in FL2 or FL3, that may help with your fluo-problems!
Thanks for your response. however doing FACS scan of 96 well plate will not be feasible at my end. anyways i consider that as good alternative if provided such facility.
Hi Faraz
I do MTT assays often and compounds come in that are colored and florescent all the time. I have found it best to set your assay up as usual then on the day you plan on doing MTT assay you pellet the cells in the plate and wash them twice to remove the color or florescent compound. The effect on the cells is not changed and you can then re suspend the cells in RPMI or whatever media you are using and continue your MTT assay
Hello Angela,
Thanks for this valuable suggestion. Its implemented and there is improvement in results. Thanks again.
Hi Faraz,
Or else u can always put your control which contain everything except cells. And that way you can evaluate your results by subtracting your respective control's reading.
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