In Situ Hybridization query.

Dunowska, Magda via methods%40net.bio.net (by M.Dunowska from massey.ac.nz)
Wed Mar 23 04:07:06 EST 2011


Hi  Methods @   I am assisting a colleague develop an in situ hybidization (ISH) method for herpes virus in Oysters. ........Relax,  the question is more generic : )   We cut and mounted 5 um sections of Formalin fixed paraffin embedded tissue to superfrost plus slides. They were accidentally dewaxed and stained with H&E..  The sections are from a limited  supply of tissue and we are trying to figure how to preserve the sections as the probe is not ready to go quite yet. So far we have taken out the majority of the stain using a short acid alcohol step and left the slides in water.  I figured the ISH protocol would have to risk this procedure as we are using either blue black  (NBT BCIP )  or red (vector fast red) for the chromogen.
How long can you leave the slides in water ? or different aqueous medium? And beyond that time, is a solvent transition / dehydration back to paraffin feasible and likely to do more harm than good ?  Suggestions pls

cheers  M



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