cell cryosection - (Aug/20/2009 )
i culture HEK cells with a compound with fluorescence and want to do cryosection to check if the compound appears in intracellular. i never do this before. can anybody help me do this experiment? thanks very much.
You don't have to do cryosection. If you culture the cells on glass cell culture slides, you can wash, fix and stain (for example with DAPI and Phalloidin) the cells directly. Or you could simply look at the culture dish with a microscope, no need to fix etc.
unfortunately, the compound can also bind to cell membrane so you cannot see intracellular under microscopy.
Do you have access to a confocal microscope?
Scan the cells across z-axes you can tell if the drug/compound is within the cells.