Re: [Histonet] embedding in agarose then paraffin

From:Andrea Grantham

Histogel processes along with the tissue and should cut fine. I 
haven't injected it into a lung or heart but I think it might work. 
I've embedded tissues and other types of samples (like collagen 
fibers) in histogel with good results. Helps with orientation. The 
gel does not get replaced with paraffin and cuts fine. After staining 
there might be a sort of halo around the tissue where the histogel is 
but I haven't found that it interferes with the tissue when you look 
at it under the microscope.

Andi Grantham

At 03:26 PM 1/7/2008, Gayle Callis wrote:
>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
>----- Original Message ----- From: "Judith L. Williams" 
>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 
>>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
>>Histonet mailing list
>Histonet mailing list

: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
: Sr. Research Specialist       University of Arizona               :
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