[Histonet] embedding method for DRGs

[John Kiernan]

Dear Carol Barone,
 
You are asking the wrong group (Histonet), and almost anonymously. 
 
Your email address indicates that you work for a BIG company. Your employer should send you on a course to learn how to do microdissection of fetal and neonatal mice. You may get some free advice from listservers, but how will you know if it's any good? The skills you ask about need expert hands-on teaching, such as a few months in a lab that does that kind of work.
 
John A. Kiernan MB, ChB, PhD, DSc
Professor, Dept of Anatomy & Cell Biology        
The University of Western Ontario            
LONDON,  Canada  N6A 5C1
  Phone: (519) 679-2111 x 86822
  FAX: (519) 661-3936
  http://biostain.com
  http://publish.uwo.ca/~jkiernan/
  http://instruct.uwo.ca/anatomy/530/
= = =  
 ----- Original Message -----
From: "Barone, Carol " <cbarone <@t> NEMOURS.ORG>
Date: Friday, March 26, 2010 14:38
Subject: [Histonet] embedding method for DRGs
To: histonet <@t> lists.utsouthwestern.edu

> Histonetters- Back to these DRGs again: 
>  
> We are looking for a better way to embedd DRGs from neo-natal 
> mice, for
> cryotomy.. We normally...using a dissecting scope, remove the 
> DRG from
> the eppendorph tube with a small spatula from ...touch the drg 
> to some
> OCT (colored) ....and then touch the OCT to the bottom of a disposable
> embedding mold (peel-away)...the DRG releases and we fill the 
> mold and
> move on. 
>  
> But many times we need to "encourage" these neo-natal DRGs off the
> spatula, with another spatula to get the DRG to release into the OCT.
> Though this technique works...it is a very time consuming method 
> - and
> we occassionally do lose one or two of these things, because 
> they are so
> very very small.  We are using marker dye to help us locate 
> after they
> are on the spatula and getting them from the tube to the spautla 
> AOK, it
> is getting them from spatula to OCT that is the killer. 
>  
> We are currently drawling off the dye with the point of a 
> Kimwipe and
> touching the DRG to a frozen dot of OCT, using the cold to 
> capture and
> hold the DRG on the dot, before finishing the embedding process. 
> This is working slightly more efficiently, but....
>  
> Does anyone have a better technique to share? I am losing tech's 
> to "DRG
> blindness"!...It's like trying to embedd a speck of dust...and 
> everyonehates to do them. This is the only technique I have ever 
> used....butalways on full term mice..No problem. It works great; 
> but on the
> neo-natal mice....well you all get the picture. Too teeny
> tiny.....frustrating, and takes a two man team to do! Ever so time
> consuming to do!
>  
> PS: histo-gel doesn't not work, either.
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