Re: [Histonet] Help-Fried tissue

From:"Anthony Reilly"

Hi Kristen
I would certainly look at replacing the alcoholic formalin with an aqueous solution as over exposure to alcohol will harden your tissue unnecessarily however I do not think that this is the main problem.  Your tissue has dried at some point and the fact that some samples have a mixture of good and bad areas would suggest it is during processing.  Other than ensuring that solution levels are adequate as previously suggested I would look at the biopsy cassettes and how they are stacked in the processor.  When using biopsy cassettes with a fine mesh there is marked reduction of the flow of solutions which will be worsened by stacking the cassettes too tightly in processor racks.  If you have not done so already I would suggest trying paper wraps (we use end papers from hairdressing suppliers) in routine cassettes which are loosely stacked into the processing racks.  Some have racks with spacers.  just a suggestion.  Good luck.
Tony Reilly

Chief Scientist
Anatomical Pathology
Pathology Queensland
Princess Alexandra Hospital
Ipswich Rd, 
Woolloongabba Q 4102
Ph: 07 32402412
Fax:07 32402930

>>> kristen arvidson  2/04/2008 10:12 pm >>>
  This is a very serious matter and we have tried everything.  So I will get right to the point...we have had this artifact on and off for several years, the tissue looks shrunken on the slide and the Paths have a very hard time diagnosing.  When embedding and cutting the pieces look smaller and are harder then their counter parts (sometimes one half of the specimen is fine while the other is destroyed in the same cassette!!).  
  So here is what we do and what we have tried to remedy the situation..
  we use recycled formalin, alcohol, and some clear-rite (BR instruments).  We use multiple sized biopsy cassettes (mesh, small hole, and slotted--by the way we do all derm).  We have tried shortening our processing times.  We just purchased two new processors (Leica ASP300S) thinking that may have been the problem and the very first day we used them we had a large number of fried tissue.  We also have two older VIP machines and it happens on and off on those as well.  All our biopsies come in alcoholic formalin.  We use paraplast.  I think I've covered everything.  We have come to some conclusions, we know that there could be air bubbles in the cassettes during processing and we've heard various things about the alcoholic formalin.  So now what?  Any other suggestions would be so helpful!!  Thank you all for your time.

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