I need to do treatment of Mtt assay for 24, 48 and 72 hours treatment of my compound.
First method,
Monday - seed the cells
Tuesday- treatment for 72 hours
wednesday - treatment for 48 hours
thursday- treatment for 24 hours
friday - run for Mtt for all 3 plates (72,48,24 hours)
2nd method,
Monday- seed the cells
Tuesday- treatment for all 3 plates (24 , 48, 72 hours)
wednesday- Run Mtt for cells treatment for 24 hours
thursday- run mtt for 48 hours
friday- run mtt for 72 hours
which of these methods are preferabl? does the first method is wrong?
Hello Atielah Nabila,
I would prefer the second method. This is because all the three plates for the three time points (24, 48 and 72 hours) would be treated at the same time and uniformity would be maintained.
If I were to use the first method, I would have to prepare the compound dilutions separately on each day, and this is bound to introduce variation.
So, by using the second method, I would prepare the required quantity of working solution from the stock needed for all the three plates, and add the compound to all the three plates at the same time. In this way, I would have treated all the three plates for the three points similarly leaving no room for variation.
I would suggest whenever you perform any cell-based assay avoid introducing extra steps. The first method is not wrong, but I wouldn't consider the results as reliable. The second method is preferable by many.
Best.
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