[Histonet] tissue loss

[Mark Elliott]

My question with regards to this is, when I add my slides to the 95C buffer in the water bath, the temperature of the buffer drops because of the cold/room temperature slides-do you start timing the 20 mins then, or when the temp gets back up to 95?  It takes awhile for the temp to get back to 95 (10+minutes).

Mark

>>> Rene J Buesa <rjbuesa <@t> yahoo.com> 07/13/06 11:49 AM >>>
Melissa:
  This is the way I used to do HIER:
  heat the citrate buffer in the MWoven without the slides up to boliling point (time will depend on the energy output of yout MWoven and the amount of buffer).
  After that, put the heated citrate buffer in either a water bath (95ºC) or a steamer (98ºC) and when the hot citrate has thermally equilibrated with the hot environment into which it has been put (about 5 minutes) is then when you add the slides to do the HIER during 20 minutes; after those 20 minutes you take the container with the hot citrate buffer and the slides and place it over the counter for another 20 minutes to complete HIER.
  You will not lose sections with this procedure.
  Hope this will help you
  René J.

Melissa Mazan <melissa.mazan <@t> tufts.edu> wrote:
  Hi - I'm trying a citrate buffer antigen retrieval protocol. After I 
have dewaxed the slides, I put them in citrate buffer - protocol says 
microwave on high for 25 minutes. However, when I remove the slides all 
the tissue has slid off the slide. Any suggestions? Melissa
-- 
Melissa R. Mazan, DVM, Diplomate ACVIM
Associate Professor of Large Animal Medicine
Director of Sports Medicine
Cummings School of Veterinary Medicine
Tufts University
200 Westborough Road
North Grafton, MA 01536
Tel: 508-839-5395
Fax: 508-839-7922
Email: melissa.mazan <@t> tufts.edu 


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