I treat my cells with chemotherapeutic drugs and I want to control the chemotherapeutic stimulation on my cells. so I'm looking for a test and I heard that we can use doxorubicne on flow cytometry to see if the cells can reject the doxorubicine and use non treated cells as control.
Hi Jeremy, it would be easier to titrate in doxorubicne onto cells in a 96 well plate and check for live/dead cells.
It is difficult to detect and quantify dead and live cells using flow cy because you have to trypsinize them if they are adherent. Plus once the cells have gone through the flow cy, they are gone.
Hi, I think it might work..we used microscopy in Fission yeast
https://www.ncbi.nlm.nih.gov/pubmed/25793410
I used the Rhodamine123 fluorescent dye which is known to be a substrate for the chemoresistant cells instead of Doxorubicin in the chemoresistant cells to determine accumulation and efflux of chemotherapeutics through the flow cytometer.Please find the attached paper with the protocol.Also, the second paper contains a protocol for using doxorubicin as a fluorescent dye in the flow cytometry experiments.I followed their protocol in another experiment in which I used doxorubicin.Some papers suggest using propidium iodide dye to determine the viability of the cells after using the doxorubicin Since it stains the nuclei of living cells.And you can use the proper gating in the flow jo software to analyse the data.Hope that will be useful.
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