[Histonet] embedding in agarose then paraffin
Judith L. Williams
juditw <@t> u.washington.edu
Mon Jan 7 12:50:14 CST 2008
Hi- I will be trying to embed entire mouse lungs and heart to do a 3-D reconstruction using paraffin sections stained H&E. I am thinking that filling the lungs with agarose and then embedding it in agarose will help inflate the lungs and keep cell/tissue continuity. Then I want to slice the agarose into rather thick sections - say four per lung- and put these into cassettes for paraffin processing. Each block will then be serially sectioned and slides stained and scanned for reconstruction.
so questions are:
1. is Matrigel, Extragel or agarose (4 to 10%) best?
2. has anyone ever filled lungs, heart with melted agarose gel prior to embedding?
3. will the agarose- gel stay in the tissue throughout the paraffin processing?
4. will the gel stain differently in the blocks embedded in paraffin and stained with H&E?
I would appreciate hearing from those of you doing whole embryos or large parts that need orientation and sectioning.
Judy Williams, HT, PhD
Dept. of Comparative Medicine
University of Washington
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