I have been working with a ceramic based scaffold, and it is to be determined for cell viability, for which I have chosen MTT assay. Although I have tried seeding cells onto the scaffold or taking extracts of the scaffold and then treating them to the cells, I was unable to get proper results. In general, the problem faced was that the absorbance of media control is remaining higher than that of cell control, which in general shouldn't happen. It would be great if any researcher can help me sort out the issue or suggest any other method that can be followed to perform a cell viability assay.
Dear Colleague
Hi,
I propose to use XTT assay rather than utilizing MTT assay. By using XTT assay and the culture of cells within a 48 well plate, a soluble dye will be obtained. Then, you can transfer appropriate amounts of this solution (e.g. 100 uL per each microwell) to the 96 well plate and read the absorbance. Good luck
Indeed; MTT is insoluble so good for imaging, or if you want to quantify you need to extract using isopropanol with acid (protocols are around on the internet). XTT is good, but so is WST-1 or resazurin (trade name alamarBlue). And it's recommended to use media without phenol red so that doesn't interfere.
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