Hello everyone,
I have a question about the MTT assay. In the attached image, why do some cells show MTT color while others don't? Some wells only display color around the edges. All cells appear intact under the microscope.
Has anyone encountered this issue and found a solution?
Thank you for your assistance.
Cell used-C20 cells
Treatment time- 48 hours
Control Seems ok, for other well I have used different concentration of Curcumin. (100uM to 0.01 uM)
Protocol-
Hello Grinsun,
As the MTT assays are normally used for identificate the viability of cells treated with any kind of reactive/conditions, is absolutelly normal that some of the wells present color and others not. MTT is a reactive that allows you to "measure" the mithocondrial activity of the cells: formazal crystals are formed due to the oxidation activity of the mithocondria on the MTT (methiltiazol). So, when the cells on the well are viable, they oxidate the MTT and the color appears, but when cells are with a low metabolic rate, or death, the color is not present.
In your case, clearly there are some cells that havent response to the treatment and have loose their viability. For the analysis of ypur result, i suggest to normalize every absorbance to the C_. This way, you will have a change fold of viability related with your treatments.
I hope this response will help you
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