[Histonet] organotypic hippocampal cultures and immunohistochemistry

From:=?iso-8859-2?Q?Jakub_Ot=E1hal?=

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

 

 

 

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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

jotahal@ftvs.cuni.cz

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