I have known that MTT assay can be used to check cell toxicity of some materials such as nanoparticles.
MTT assay data is obtained through measuring absorbance in specific wavelength. However, if the materials, especially relatively large particles could not be cleaned completely by suction, the materials could make optical problem, such as scattering.
In this case, how can I measure the cell viability or cell toxicity? Is there any other method besides MTT assay?
Hi Chansong,
Perhaps quantitate scattering by creating a serial dilution of your particles and then measuring absorbance at the same wavelength as you would use for MTT assay. That way you could determine whether scattering is a possibility, and what amount of particles would have to remain for scattering to be an issue.
It may not be something you have to worry about in the first place. In this review Article Toxicity of Nanoparticles and an Overview of Current Experim...
they say that MTT assay has been used to quantify cytotoxicity of nanoparticles. Perhaps read some more through the methods of the papers cited there.
If scattering does turn out to be an issue and you know the average size of your particles and your cells, perhaps you could select a filter to sort cells and beads before addition of the MTT to remove any particles. I would imagine that would work well as long as there's no chance that the particles are either bound to the cell surface or taken up by the cell. This is not my area of study so I'm not certain of the dynamics of the beads.
Best of luck.
Hello Chansong,
In addition/alternatively to what Sarah said, you could try a luminescent assay like Cell Titer Glo, or a competitor's equivalent. These assays quantitative cellular ATP in a luminescent reaction, and your particles might not play such a big role. However, you still might want to quantitate their effect as described above.
Hope that helps!
why don't you just count the number of living cells?
MTT has many drawbacks, e.g. it often interferes with some chemicals people apply, nano-p too; it only tells about activity of mitochondria in the cells (read the manual how MTT works).
Instead, taking a small fraction of cells stained with trypan blue (the simplest approach) can tell you of how many living cells are present vs overall number of cells.
FACS with proper markers can give you more accurate reading.
I recommend Thomas Mühlenberg answer to your problem. Try using Cell Titer glo, which involves measuring cellular ATP alternatively you can also use WST 1 assay which uses mitochondria enzyme.
Good luck !!
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