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From:"Sarah Christo" <> (by way of histonet)
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Drop the third absolute alcohol step.  Shorten the times to 15 minutes or
less.  A second 80% alcohol step would be OK.  No heat, no vacuum.  Sarah

Sarah Christo, HT (ASCP)
Texas A&M University
College of Veterinary Medicine
Dept. of Vet. Anatomy & Public Health
Histology Laboratory
College Station, TX  77868-4458
phone (409) 845-3177
fax (409) 847-8981

>>> "Philopena, Jennifer" <> 11/18 11:30 AM >>>
Hi.  I really appreciate so many enthusiastic replies.  Here's some
additional information, provided to mystify the believers.  I've used longer
processing times in the Citadel, up to 2 hrs/reagent.  I just demo'ed the
Sakura VIP processer for 2 weeks so I had the chance to try vacuum on all
steps, heat on all steps, and I varied the processing time from 20
minutes/reagent to 2 hours/reagent.  The livers and spleens are still dry.
The Citadel only has vacuum capabilities on the final paraffin step, and no
heat for the ethanol or Propar steps.  I'm processing liver pieces that
don't exceed 4mm thickness, and I process no more than 21 tissues per run
(even though the Citadel manual claims it can handle 150 cassettes).  Propar
is a blend of alkanes and propylene-based glycol ether.  Below is just a
re-hash of my initial email.  THANKS SO MUCH!!!

Shandon Citadel 2000
rat and mouse tissues (liver, spleen, kidney, heart).
consistently very dry
fix for 24 hours in 10% neutral buffered formalin
store in 70% reagent alcohol
20min in 80% reagent alcohol
2X 20min in 95% reagent alcohol
3X 20min in 100% reagent alcohol
3X 20min in Propar (Xylene substitute)
2X 20min in paraffin (Paraplast Plus

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