[Histonet] Could these samples have been saved?

[Roger Heyna]

We microwave process our tissues, and we've had tissues fall out of the rack after the dehydration phases and air dry in the open retort for hours. We did not continue with processing; we added them to formalin for eight hours (which could be overkill) and then started the processing from the beginning. We had no problems with cutting or staining.
 
If you look in Freida Carson's Histotechnology: A Self-Instructional Text, she includes a protocol for tissues that were well-fixed but accidentally desiccated. She suggests the following:
 
1. If the tissue has been in paraffin, blot off as much as possible with paper towels.
2. Soak the tissues overnight in a rehydrating solution: 50 mL water, 30 mL absolute alcohol, and 20 mL of 5% aqueous solution of sodium carbonate.
3. Reprocess as usual.
 
I've never done this before, and maybe others have modified it for labs without sodium carbonate.
 
Roger Heyna
Maywood, IL

>>> "Johnson, Kevin" <KJohnson <@t> med.miami.edu> 2/8/2013 12:00 PM >>>
Dear all,

During an overnight tissue processing cycle, a malfunction occurred such that the sample basket was suspended in mid-air for several hours at probably the worst spot in which to do so---after the final absolute ethanol of the dehydration series. I continued the process manually in the morning, and carried it through blocking and attempted sectioning.  However, the samples (mouse skin and fat) had been converted to uncuttable rocks.

In hindsight, should I have attempted to rehydrate and reprocess these samples in an attempt to glean even minimal information from them?  Or is there no way to unmummify a mummy?

Regards,

Kevin Johnson
University of Miami
Diabetes Research Institute
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