I have performed MTT assay with cell densities ranging from 1000-9000 cell and added MTT after 24h of incubation. The resulting absorbance is less than 0.75. Hence I selected higher cell densities ranging from 10000-30000 and the resulting absorbance for 30000 was 0.66.

Now my question is can I select this higher cell density to determine IC50 of drug?

OR ELSE

I should consider low cell density and incubate them for 48h or 72h to get the absorbance in range of 0.75-1.25?

Thank you

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