Ag Retrieval, Waterbath

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From:"Carol Bobrowitz" <>
To:"histonet" <>
Date:18 May 1999 15:22:21 -0500

                      Subject:                              Time:  2:17 PM
  OFFICE MEMO         Ag Retrieval, Waterbath               Date:  5/18/99
Our lab is strictly an IHC lab and research lab.
We use a waterbath for all Antigen Retrieval.  
I purchased a large waterbath, measurements 15" long, 14" wide and 9" high,
with a cover.
We only use distilled water in it. 
We have 2 metal Tissue Tek Cytology staining racks (room for 6 white Tissue Tek
These racks are never removed from the water bath.  
Slides are placed in the gray Tissue Tek Staining Racks (up to twenty slides). 
Written on several different white staining containers the specific antigen
retrieval and pH is listed.  (If a small number of slides need ag retrieval,
Nalgene white coplin jars are used (no more than 4 slides per coplin jar).
A thermometer is in the waterbath to maintain a 97#161#C to 100#161#C at all times.  
Two problems we had and the solutions.
1)  Evaporation: A MUST, Daily, the end of the day, the water must be filled
with distilled water over the metal staining racks.  
2)  Time waiting for the waterbath to reach 97#161#C to 100#161#C.  I purchased from
two different scientific companies timers that you can set to your needs.
The waterbath comes on at 4:00 am Monday thru Friday and is ready to use by
6:30 am and goes off at 2:00 pm.   Holiday weekends can also be set.
Since using a the waterbath instead of a microwave our staining results have
been excellent.  
One MAJOR PROBLEM I have had:  Technicians rush to start the slides and do not
pay attention to the waterbath temperature.  The slides were started below
required temperature.  The results were not consistant and very poor quality. 
TIP:  We have two MOPEC slide dryers.  Both dryers are on timers.  If slides
are cut at the end of the day,  they are ready to start at 6:30 am.  We have at
times put our floatation waterbaths on times.
I hope this information helps.
Carol Ann Bobrowitz
Medical College of Wisconsin  

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