[Histonet] Processing Guinea Pig
Grantham, Andrea L - (algranth)
algranth <@t> email.arizona.edu
Mon Jun 17 10:50:09 CDT 2013
Looking at your protocol for processing the kidney and heart tissue I can't figure out why it is mushy especially if you are putting it in the cassettes so thin and it has had a chance to fix well before processing. In fact, looking at your protocol I might think that your tissue might be dried out and need to sit in cold icy water before sectioning.
At any rate, I would rinse well in running water after fixing and skip the formalin on the processor.
I'd start in 70% alcohol, maybe two changes and move on up to 80%, 2- 95%'s and 3-100%'s.
I'm not familiar with the clearing agent that you use. I use Clear Rite 3 and never have a problem - 2 changes of Clear Rite 3 and 4 paraffins.
I don't know how many cassettes you are processing but make sure there is adequate room for the reagents on the processor to move around the cassettes and that you have a good ratio of the reagents to the tissue and that the reagents are fresh. If you can increase the agitation in some of the dehydration steps it might help.
Andrea Grantham, HT (ASCP)
Senior Research Specialist
University of Arizona
Cellular and Molecular Medicine
Histology Service Laboratory
Tucson, AZ 85724
algranth <@t> email.arizona.edu<mailto:algranth <@t> email.arizona.edu>
Tel: 520.626.4415 Fax: 520.626.2097
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