I am co-culturing bacterial strains with human cells (WBCs) and there are about 3000 different bacterial strains. The idea is to carry out flow cytometry experiments with these samples (in 96 well plates).
Because of the large number of strains, I was wondering if there’s a way to fix the co-cultured samples so that I can prepare multiple plates (96 wells) and can run them at a later time.
It would be great if I could get some suggestions/ideas in regards with the above mentioned scenario.
Thanks in advance!
I wouldn't recommend staining your cells after fixation, because the process of fixing your cells can alter cell surface expression of your markers of interest, as well as possibly permeabilize your cells if you're not careful. They can be stained and then fixed, but the flourophores on the antibodies would be only so stable afterwards.
It depends on which antibodies and fluorophores you're going to use. You can either fix cells before or after staining. You should remember that fixatives can change antigen epitope structures and it may lead to loss of signal when fixation made before staining. Therefore you should check whether your antibody's binding is affected by fixation. If you fix the cells after staining, fluorophores which are conjugated to antibodies can lose signal. In this case, you should choose a fluorophore that can withstand fixation conditions. You might also find this useful: https://www.researchgate.net/post/How_to_stain_and_fix_cell_for_flow_cytometry_for_long_time
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