I've stained the dead and live cells with the live/dead cell assay from Life Technologies, but the kit is not suitable for further fixation according to the manufacturer.
I seeded the cells on the surface of the polymeric scaffolds and I want to examine viability using dyes such as live/dead cell staining kits or DAPI/PI and fix the scaffolds for cryostat sectioning.
I'm planning PI staining prior to fixation and cryostat sectioning, but I'm not sure that PI staining would be stable through the fixation and embedding steps. Does anybody have experience on this kind of experiments?
I agree with Song and staining should not have any effect on fixation, Trypan Blue is a good alternative
Thank you Song and Brian.
However, since my cells are expressing GFP and the scaffolds that I use are not transparent, I need to use fluorescent microscopes (confocal laser microscopes). What I wanted to know is the effect of fixation on the staining and the morphology of the cells. Thanx.
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