We are using the WST-1 assay to look at cell viability; however, when we add the dye to one of our cell lines, we are not getting accurate results. Under the microscope we see an increase of cell death due to the drug we are using, but the WST absorbance readings (420nm) give us 95-100% cell viability still. What could be happening? This has not happened with our other cell lines.
It is really interesting. In my experience with this assay, I noticed that different cell lines metabolize WST-1 at different rate, and some cell lines need to be incubated even for 3-4 hours. Also, do you use any compound in your assays, are those compounds colorless? Do you always have a positive and negative controls?
could you tell us more about the protocol you used?
Could be as pointed out by Vilma that your cell can metabolize WST-1 quite fast, then you would need to reduce the incubation time.
We have made similar observations with certain treatments we were doing on some cell lines. I would recommend using an alternate cell death assay (neutral red uptake, Cell-Titer Glo or even measuring cell death in supernatants using G6PD release assay).
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