I do not know whether cytotoxicity assays like MTT works with Bacteria or fungi, you can give it a try. Morever, you can simply go for trypan blue staining where you can demarcate between the live and dead cell via trypan blue staining and distinguish human cells from bacteria or fungi by cell size. Moreover, you can also have an idea of the proportion of live:dead cells by hemocytometer counting as well.

Dear Soumya Kundu ,

Thanks for you answer!

I just did a quick search and MTT seems to be also metabolized by bacteria.

I have considered Trypan Blue staining but If it stains also bacteria and fungi, it is not appropriate for a microplate colorimetric assay.

I would need a fast and high-throughput test in microplates that permits me to differentiate the human cells from my microbial cells (bacteria & fungi), but not working on green/red fluorescence.

This technique should certainly do it for you, but it could get quite pricey - so only best if you'll be doing this assay frequently.

It measures impedance across the bottom of a culture plate as a surrogate of cell viability. (I am assuming that your primary human cells are adherent - if they're not there are workarounds).

https://www.aceabio.com/products/rtca-sp/

Hi Elliot,

Thanks for your suggestion!

Yes I will have adherent primary lung cells. I will also measure TEER and I guess it is the same idea as for impedance, as I understood.