Sheila:
  Deparafinization is a fundamental step for IHC. If paraffin is not completely removed it will compromise IHC reactions.
  In a recent study on deparafinizing agents it was found that octane was the best but is not practical because of health hazards and costs.
  Short of that, xylene is more than adequate but you have to make sure it completely works.
  I used to have 3 xylene stations and the slides stayed 5 minutes in each. Xylenes in the last 2 stations were moved forward after 20 slides were treated and fresh xylene was added to the last station (much like moving reagents in a tissue processor).
  Hope this will help you.
  René J.

sheila adey  wrote:
  Hello Histonetters.

I am curious about how long other labs are spending in Xylene when bringing 
tissue to water? I started here 6 years ago and never questioned the 
rehydration times but I think we may not be spending enough time in Xylene. 
Our times are 1 min in each of 2 xylenes. This week we increased it to 5 
minutes in each and I think our IHCs are staining better?
Any input?
Thanks

Sheila Adey HT, MLT
Port Huron Michigan

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