Re: [Histonet] paraffin mouse femur peroist detaching from slide

From:"Gayle Callis"


Do you mean the dense cortical bone that surrounds the bone marrow?  The 
periosteum is a connective tissue that lines the outside surfaces of 
cortical bone, next to muscles.   You may have over-dried the sections in 
the oven.  A good way to dry decalcified, paraffin embedded bone sections is 
pick up on superfrost Plus charge slides, drain for a short time in the 
vertical position, then lay slides FLAT on a 37C to 40C slide warmer, and 
dry overnight.   A 37C oven is fine as long as the slides are flat and no on 
edge.  Metal slide trays help.   Longer drying is even better -  we let our 
decalcified bone sections dry for several days.  I have one bonehead friend 
who would not do any staining for two weeks, let her bone sections dry flat 
that long.  56C may be a bit too hot, and dries out the bone and cartilage 
even more after the processing., contributing to tissue liftoff.

What kind of retrieval did you use?  If the retrieval has any kind of 
mechanical manipulation of the section, then it may fall off e.g. boiling 
buffers in a microwave.  Decalcified bone sections need gentle handling, 
through deparaffinization, rehydration and all other steps for IHC, in 
particular any HIER before doing the staining. I am familiar with how the 
Ventana machine dispenses reagents but if not gentle enough,  this could be 
a mechanical stress on the bone section.

Gayle M. Callis
Bozeman MT 59715

We have more section lift
----- Original Message ----- 
From: "mesruh turkekul" 
Sent: Monday, April 28, 2008 10:59 AM
Subject: [Histonet] paraffin mouse femur peroist detaching from slide

> Dear Friends,
> I have fixed and decalcified  mouse femur with EDTA and paraffin embedded.
> The sectioning was perfect and also the H&E looked very good. I used super
> frost/plus slides and baked the sections at 56C  1 hour.
> After IHC on Ventana machines the periost was gone but the bone marrow was
> perfectly stable on the sections.
> Any suggestions?
> Mesru Turkekul
> New York, NY
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