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From:"ronald l. salisbury" <ronald2@uakron.edu> (by way of histonet)
To:histonet@histosearch.com
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Hi Listers,
	I have been reading the queries about processing mouse tissue with much
interest as I am trying to process the necks from 5 day old neonatal rat
pups.  We to serial section the necks to get to the superior cervical
ganglion, which we use for immunohistochemistry.  However, our procedure
results in blocks that are hard and brittle. Soaking helps, but must be
repeated so often that the process takes too long to complete. There is
evidence of good fixation and all tissues are still in a developmental
state. We use a Tissue Tek VIP processor and we use the following
processing times: 70,80,95(2x), and 100% Flex for 1 hr. Each, limoneme
(Hemo D) 2x for 1 hr. Each and 4 changes of paraplus for 45 min. Each.
Vacuum is only applied in the paraffin stages. Any suggestions?  Ron
Salisbury, Dept. of Biology, U. of Akron, Akron, OH.




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