[Histonet] embedding method for DRGs

Barone, Carol cbarone <@t> NEMOURS.ORG
Fri Mar 26 13:37:26 CDT 2010


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 everyone
hates to do them. This is the only technique I have ever used....but
always 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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