Re: [Histonet] Processing protocol for VIP

From:Rene J Buesa

Hi Julie:
  If you get enough processing protocols, you will probably realize that there are as many protocols as laboratories there are; everybody has personal preferences.
  For tissue like you describe I always tried the following:
  1- process the next day they were received, after a good 24 hours fixation;
  2- pieces as thin as possible
  3- a 15 hours processing without the first fixation step in the first 2 VIP stations:
  a- 60% ethanol and 80% ethanol: 1 station x 30 min each
  b- 95% ethanol x 1 hour
  c- 100% ethanol: 2 x 1 hour + 1 x 2 hours
  d- xylene x 1 hour
  e- xylene x 2 hours
  f- 4 paraffin stations: first 3 x 1 hour each + last x 2 hours
  TOTAL = 15 hour.
  Now the balance was: clear time / dehydration time = 3/7
  or (clear + infiltration) / dehydraiton = 8/7
  You could change the times but try to maintain those ratios.
  On June/99 I changed all my procedures to mineral oil (eliminated xylene) and "difficult" specimens (breast, colon, uterus, brain) began to process even better.
  If you want I can send you that procedure.
  I hope this will help you!
  René J. 
  

"Sanders, Julie, VHACIN"  wrote:
  

Hi netters,
I'm back with more questions about processing...We received our HQIP evaluation and everything was fine except for the breast and skin sections, both of which contained lots of soft tissue and fat. The comments were about "muddy nuclear detail, and hematoxylin not crisp..." Now the chief of my lab (a clinical pathologist) is up in arms suggesting that ALL of our work is sub-par. I cannot convince him that this is a problem with the type of tissue and fixation. So...with that said...anyone willing to share their processing protocols would be greatly appreciated. We have two processors, one for biopsies only, which we do not have a problem with, and the other for large cases..colons, breast, etc., it is a 12 hour protocol. I would be interested to see what everyone else is doing. Oh, we use VIP processors.
Feel free to send to me personally if you wish, but I'm sure others on the net might be interested as well.
Thanks in advance!

Julie

Julie Sanders, BA, HT(ASCP)
Supervisor, Anatomic Pathology
VAMC, Cincinnati, Ohio
Julie.Sanders@va.gov

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