immunocytochem: is it ok to use H2O instead of PBS for washing - Problem with PBS crystallization upon drying (Jan/11/2007 )

Hi,

I've started to work on ICC and have been observing that if PBS starts to dry on the coverslip (or slide), it quickly forms a layer of salt crystals. I've tried to ensure that drying is minimal from rinsing to staining to mounting, but if I'm not careful enough, the crystal will form. Has anyone tried using H2O instead of PBS, w/o problem? Or is there any additional steps to ensure drying is at minimum?

Appreciate your help.

Cheers

For ICC, I use PBS for all the initial washes, making sure they dont dry out. But the final wash after secondary antibody, I wash once in water, so that when they dry out, no crystals are formed.

Thanks Malik. I'll try. I have high respect now for people working on ICC. Ha! ICC sure is not easy. In the future, I think I'll need more help from you guys.

Bests