During the MTT assay with h357 cell line using different types of culture plates ie ELISA, PLAIN and normal culture plate, I found that when observed under microscope there was a considerable difference but there was not much difference in absorbance. Why would that be?
Dear rachana, there could be quite a few reasons. First of all MTT assay may not be sensitive in this case, you may do SRB assay, BRDU assay or thymidine incorporation assay. Secondly, small microplasma contamination also can be there which may affects during MTT assay. Be sure your cell is free from contamination.
I am quite sure that the cells were not contaminated sir... Maybe the other assays would work.. thank you
Dear Rachana
I think you should try to find the kinetic of your cell line growth and your treatment, this results may be achieved due to the cell viability or death before you add the MTT to the culture, therefore, under microscope you see the cell but they can not change the MTT and the succinct dehydrogenase enzyme in your cells dose not work!! I previously faced to same problem and it was due to the time of MTT assay (18-24 hrs).
Good luck
The attached article could maybe of help ... even if it comes very in late ..., which is better than never ...
All the best,
Robert
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