[Histonet] RE: Histonet Question

[Tyrrell, Jannifer]

I'm new to the Histonet website, but not new to histotechnology.

I'm trying to salvage some irreplaceable Zucker Diabetic Fatty rat tissue samples that were embedded in paraffin in May 2009. They have sat on my bench since they were embedded as we thought the entire experiment was a huge failure.  However, recent examination of some of the flash-frozen tissues has shown us that the experiment was actually a huge success.

Which leads me to my issues and question...

I needed to section these tissues (liver & pancreas) and have attempted to do so by several troubleshooting means - soaking the block in warm water then rapid cooling, place ice on the block then rapid slicing, changing the blade, using a different microtome, changing the blade angle, you name it, I think I've tried it.

Now I've got a bunch of sections that look like accordions that will not flatten, but hardly anything left in the actual blocks. These horrible sections are on slides, but not stained yet.  I only put them on slides because I NEED the tissues. But to stain the sections as they are would be a waste of the tissues and the reagents.

I saw a protocol for formol-glycerol re-hydration and have thought about using it...but now I can't get the tissue sections off of the glass slides.

ANY and all help or advice would be INCREDIBLY and GREATLY appreciated! This is the last bit of work I have to complete in my dissertation project and the data, I'm certain, will be astounding.

Jannifer Tyrrell
NIEHS/NIH-Sponsored Pre-Doctoral
Ruth Kirschstein Federal Grant Recipient
PhD Candidate
Department of Pathology
Wayne State University
School of Medicine
550 E. Canfield Ave.
111 Lande Building
Detroit, MI 48201

jtyrrell <@t> med.wayne.edu