Cell fixation problem - (Oct/16/2006 )

Hi!
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.