One day processing by hand
Frouwke- Non-fatty (parenchymal organs, skin) pieces of tissue 2x2 cm,
provided they are cut 2-4 mm in thickness, should be processed by this
70% ethanol- 45 min
95% ethanol- 45 min
100% ethanol x 4 - 45 min each
xylene x 2 - 45 min each
paraffin x 3 (no hotter than 60 deg C)- 45 min each.
Total time 8 hour 15 minutes. Processed tissues here can be cooled overnight
and remelted briefly the next morning for embedding, taking care not to
exceed 60 deg C. This "breathing" actually makes for easier microtomy,
allowing for further evaporation of minute traces of retained xylene.
Extended exposure to heated paraffin is to be avoided for IHC so this is not
a good holding point. Since you are processing by hand, and don't use
vacuum, agitation is critical. A rotating shaker might be used to agitate in
I hope this helps you.
Jeff Silverman HT HTL QIHC (ASCP)
Bay Shore, New York USA
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