[Histonet] embedding in agarose then paraffin

[Gayle Callis]

There was a superb poster presented at an NSH meeting where they used 
Histogel injected into a heart for slicing with a matrix slicing device, 
then placing the slices into cassettes for processing.    They used Histogel 
(from Richard Allan aka ThermoFisher Scientifc(?) which kept the heart 
distended for precise slicing.   If you fill the lungs via a v shaped cut on 
top of trachea, using an 18 or larger guage needle and syringe filled with 
Histogel, then you should be able to can distend the lungs, but need to use 
only 2.5 ml or so.  After filling, ans watch while you do this - overfilling 
with blow up alveoli - you should be able to slice a firm Histogel filled 
lung, and fix the slices appropriately.

I don't know what happens to the gel after processing, but hopefully the gel 
will be replaced with paraffin, or at least infiltrated well to maintain 
gross morphology.

Is the whole embryo question a separate question?   And what do you mean by 
large parts, whole lungs, whole legs, whole livers?????

Gayle M. Callis
HT/HTL/MT(ASCP)


----- Original Message ----- 
From: "Judith L. Williams" <juditw <@t> u.washington.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, January 07, 2008 11:50 AM
Subject: [Histonet] embedding in agarose then paraffin


>
> 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.  Thank you!
>
> Judy Williams, HT, PhD
> Dept. of Comparative Medicine
> University of Washington
>
>
>
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