[Histonet] RE: need some help with frozen sections

[Andrea T. Hooper]

Dear Chris,

This is an *excellent* summary of what to do and not to do for frozen 
section IHC!

One thing though, you said: "At the end of the IHC staining 
procedure, after the chromogen step wash your slides with tap water. 
NEVER use distilled water as this  will ruin the tissue section 
completely!!!!". Why do you think this is so? In my experience, I 
never use tap water after staining - only MilliQ ddH20 - and have no 
deleterious effects on my frozen sections of varying thicknesses of 
both mouse and human tissue - both fixed with NBF and acetone.

Best regards,
Andrea


At 11:01 AM +0200 4/26/05, C.M. van der Loos wrote:
>    Eva,
>
>    Some DO's and DON'T's with immunostaining of cryostat tissue sections:
>      * After  cutting, let the sections dry overnight at room temperature
>        under a ventilator
>      * Fix  with  cold  acetone (10 min, 4C). Some people apply a "double
        acetone   fixation"  method:  2x  10  min  acetone  fixation 
with .........
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