I am optimizing the analysis of the phosphorylated H2AX histone in cells exposed to nanoparticles using flow cytometry analysis and I observed some interferences of the nanoparticles with the analysis. I have already tested to filter the samples to eliminate the maximum of nanoparticles, what helped to reduce the interferences but not eliminate completely the problem. Do you know more alternatives to solve this problem?
Dear Fatima,
In which cell lines are you working either adherent cells or suspension cells? If It is adherent cells, I hope that the problem will not be there because we can wash the cells in the plate itself several times with PBS. If it is in suspension cells then it is difficult to isolate cells from the particles (present in the medium). The best option to solve this problem for both adherent cells or suspension cells can be by processing the another plate with cells plus particles and follow the same procedure that you have carried out, along with other samples. But here don't put primary antibody and use as negative control.
I hope that it may helps in the elimination of background signals due to nano-particles.
Regards
Sarat
What sort of interference are you talking about?
Because nanoparticle is very small and usually you can eliminate them by setting the threshold.
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