[Histonet] RE: need some help with frozen sections-tap vs. distilled water
Hi Andrea and Gayle,
The story of the ruined cryo's after distilled water came from my
preparations with the NSH hands-on workshop in Long Beach 2002. I was
told that the NSH would supply me with just distilled water for
everything. So I rehearsed the experiments here with distilled water
only. Next, I was confronted with the poorest IHC staining ever,
especially with respect to the nuclear counterstain/tissue morphology.
It took me days before I figured out that the final washing step with
distilled water was the real problem. I have the strong impression
that especially lymphoid tissues are far more vulnerable to "distilled
water washing" than other tissues. I don't know if this has something
to do with the quality of our distilled water. Perhaps I need to
repeat those simple experiments with the more standard MilliQ water as
I have placed a picture at www.histonet.org illustrating the
difference I found with tap water vs. distilled water (tap vs. dist.
Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center M2-230
NL-1105 AZ Amsterdam
phone: +31 20 5665631
fax: +31 20 6960389
----- Original Message -----
"Andrea T. Hooper"
Tue, 26 Apr 2005 13:11:27 -0400
"C.M. van der Loos" ,
[Histonet] RE: need some help with frozen sections
This is an *excellent* summary of what to do and not to do for frozen
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.
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