[Histonet] need tips for cross-sectioning of cortical bone

[gayle callis]

You did not say whether these are cross sections or mid sagittal?  Cross
sections are always tougher.  Key is to make sure the cortical bone is well
processed and infiltrated with a hard paraffin e.g. Tissue Prep 2 (Fisher
Scientific ala Thermo Scientific).  Try this old bonehead trick is cut tiny
V-shaped notches with razor blade or used microtome blade on the sides of
block, parallel to the blade.  You may have to do this top and bottom too.
Take care to NOT make these notches huge.  This permits the paraffin of each
section in ribbon to expand, relax onto the water bath along with the bone
section. 

Another trick is try laying section on RT 5 to 10% alcohol, pick up on
slide, then go to warm water bath, lower section slowly to flattening.  The
key here is to NOT let upper part of paraffin of a section totally release
from slide while going into warm water (at an angle) in other words, the
section flattens while paraffin portion is still attached to slide during
flattening. 

Also, change the blade frequently.  Sharpest possible edge helps, and
hopefully high profile which is more stable than low profile for decalcified
bone microtomy.  

Good luck

Gayle M. Callis
HTL/HT/MT(ASCP)



   

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jack Ratliff
Sent: Tuesday, May 11, 2010 5:03 AM
To: Connolly, Brett M
Cc: <histonet <@t> lists.utsouthwestern.edu>
Subject: Re: [Histonet] need tips for cross-sectioning of cortical bone

Why not embed in resin (MMA) and take thicker sections and then grind/ 
polish them down? If you went this route, you could then use  
flourescent labels and quantify mineral apposition rate and bone  
formation rate. Let me know if you are interested. I can help you get  
started and direct you to low cost equipment options.

Jack

On Apr 22, 2010, at 9:58 AM, "Connolly, Brett M" <brett_connolly <@t> merck.com 
 > wrote:

> A colleague is having trouble getting wrinkle-free sections of
> decalcified, paraffin embedded femur.
>
> Any tips??
>
> Thanks,
>
> Brett M. Connolly, Ph.D.
> Molecular Imaging Team Leader
> Merck & Co., Inc.
> PO Box 4, WP-44K
> West Point, PA 19486
> tel. 215-652-2501 fax. 215-993-6803
> brett_connolly <@t> merck.com
>
>
>
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