[Histonet] RE: need some help with frozen sections-tap vs. distilled water

From:"C.M. van der Loos"

   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
   Gayle used.

   I have placed a picture at [1]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
   Meibergdreef 9
   NL-1105 AZ Amsterdam
   The Netherlands

   phone:  +31 20 5665631
   fax:    +31 20 6960389
   e-mail: [2]c.m.vanderloos@amc.uva.nl

   ----- 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

   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,


   1. http://www.histonet.org/
   2. mailto:c.m.vanderloos@amc.uva.nl
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