[Histonet] I am having trouble with testis tissue slide preps.

[William Weber]

I am having trouble with testis tissue slide preps. I am fixing and
embedding rodent testis in paraffin, however the tissue is not embedding
completely in the wax. It falls out of the block and when it stays in the
block the microtome will chunk it off.


My Protocol

1. fixation in Bouin’s fixative

2. dehydration to 100% ethanol

3. 2 1hr paraffin baths

4. paraffin embedding

- I use a little was in the mold before placing the tissue in, then I place
the cassette and fill the     mold the paraffin this whole time is molten.
I immediately move it to a cold plate to solidify

5. Sectioning

-I am able to get good ribbons, but there are holes in the ribbon where the
tissue should be. It either falls from the ribbon of is pushed back into
the block, often when it does cut a section it cuts a section much thicker
than set for (e.g. 100 microns instead of 5).


If anyone has any suggestions it would be greatly appreciated.

-- 
W.David Weber, M.Sc.
Pre-Doctoral Candidate
University of Maryland
Alt EMAIL; widawe at hotmail.com
TEL; (301)405-5737