cell detachment in IHC - can I incubate cells with hoechst, heparin and PFA at the same time? (Jan/21/2010 )

Hi everybody, I'm a new member!

I have problems in IHC. I change the cell support from a glass coverslip to 8-wells labtek and now cells start detaching from the surface after fixation with PFA. I have to incubate with heparin and hoechst and I was wandering if it is possible to incubate with PFA, heparin and Hoechst 33258 at the same time in order to reduce the number of washing steps and then to lose less cells.

thanks!

use PBS with Ca ang Mg for washing steps and for diluting all IHC solutions. this should cells detaching. Also coat your plates with poly-lysine or something similar.