[Histonet] HIER

From:"Perry, Margaret"

We have been using the microwave HIER and have had good results, however when our microwave bits the dust I would like to have a pressure cooker method in place.  I feel the pressure cooker is more consistent for all the slides.  We are a veterinary diagnostic lab and I would like to have some idea of where to begin.  I have looked at different protocols and they often indicate HIER in a pressure cooker but do not give the details.  I currently use citrate buffer pH 6.  I put the slides in refrigerated buffer and microwave on high for 1 min 45 sec. or until the buffer just starts to boil.  I then set the microwave on 10% power for 10 minutes.  Afterward the slides are allowed to cool in the microwave for 1 hour.  We have a biocare Decloaker Chamber and I would appreciate help with the program I should use.  Do you start with cold buffer or should I prewarm it?  What temperature should I use? How long should I maintain the temperature?  How long should it be before I remove the slides?

I also am working with a new protocol that calls for heating in a steamer.  Should the temperature of the buffer be warm, cold or room temp when I start?

Thank you.

Margaret Perry HT (ASCP)
IHC Lab Manager Veterinary Science
Animal Disease Research and Diagnostic Lab
South Dakota State University
Box 2175 North Campus Drive
Brookings SD 57007

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