[Histonet] organotypic hippocampal cultures and immunohistochemistry


Hi Histoneters,


I have problem with immunohistochemistry on organotypic hippocampal slice
cultures (OHSC). I have stained them with OX-42(CD11b) and
glucoseoxidase-DAB development but the staining is weak and unsharp. When I
tried the same protocol on rat brain slices the staining is very nice. OHSC
are fixed for 1hour in 4% PFA then 20%sucroze and then glued on freezing
stage and cut with microtome to 30mikron slices. Does anybody have an idea
where is the bug?


Thank you very much

Best regards






Jakub Otahal MD,PhD

Department of Developemental Epileptology 

Institute of Physiology 

Czech Academy of Sciences 

Videnska 1083

142 20 Prague 4

Czech Republic

tel: +420 241062495




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