Cell fixation problem - (Oct/16/2006 )
Does anyone know how to fixed WI-38 cells, human lung fibroblasts for immunofluorescence staining? 4% paraformaldehyde didn't work very well, there are almost no cells left!
Well, i am guessing ur cells dont attach well. try using PBS with calcium and magnesium for all steps, including making PFA with the same PBS.
also u could coat ur plates or coverslips in poly-lysine before plating cells. This also can help in cells attaching well.
do u have ur cells on coverslips or still in the culture plates.
Thanks. My cells are are plate in slide flasks, after PFA I can see still some of the cells, but then during all the steps the number of cells is decreasing, so and the end the are few left.
some people also use gelatin or collagen in addition to poly-lysine or poly-ornithine so that cells stick better to the plating surface.
good Luck !!!