[Histonet] Distilled water rinse versus tap water rinsing, a curious thing

Gayle Callis gcallis <@t> montana.edu
Tue Apr 26 13:48:04 CDT 2005

Andrea and Chris,

I have seen Chris's results on rinsing.    We collaborated on this after I 
got home from visiting his lab.  I repeated his rinsing experiments in my 
lab to see if I had the same problems. I did NOT get the same results he 
observed.   Could this be caused by differences in another countries water 
and purification systems? Very curious why this happens as rinsing with 
distilled water is a very common practice.

My experience with acetone or acetone/ethyl alcohol combo fixed mouse 
frozen section IHC is the same as Andrea's, no problems or damage to 
sections after either rinsing with MilliQ, RO or tap water.  Acetone fixed 
sections are less robust than the AA combo fixed sections, but distilled 
water rinse was not a problem anyway.  We do not use NBF for murine IHC 
work here.

  11:11 AM 4/26/2005, you wrote:
>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,
>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 
> .........

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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