Re: [Histonet] crumbly femurs

From:"Amy Porter"

Potentially lack of rinsing after decal....we always rinse our decal 
specimens in running tap water before proceeding with any other steps. 
Maybe you needed to rinse and then cryoprotect with sucrose prior to flash 
freezing.  Just a thought.
----- Original Message ----- 
From: "Dana Marshall" 
Sent: Tuesday, November 08, 2005 3:37 PM
Subject: [Histonet] crumbly femurs

Hi everyone,
I am trying to section some mouse legs.  The bones were fixed/decalcified 
for one week then quick frozen in OCT and put at -20C where they have been 
for about 2 months.  Regardless of how thick I cut the section (5-20 
microns), whether I cut free hand, use our cryojane or the old fashioned 
scotch tape type of tape transfer, there is little/no visible bone in the 
section.  Just OCT and an empty space where bone would be.  If I cut without 
tape I can see the bone crumble as I cut.  This seems to be a catastrophic 
failure of some sort and may be beyond any type of help at this point but if 
anyone has any ideas as to why this is happening I would be happy to hear 
from you.
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