[Histonet] JB4 users

[Pamela Marcum]

I agree with Gayle.  GMA does not work well with mouse femurs.  Not only are
the issues with infiltration a problem, even when using mixtures of ethanol
to pure GMA it still will not penetrate well  and polymerizes for to quickly
and too hot to make a good block.  I personally find it too soft for the
calcified femurs to hold in the block well.  

We don't use a kit and make our own MMA mixture so we can control the
softeners better for sectioning and the temperature.

Pamela A Marcum
University of Pennsylvania 
School of Veterinary Medicine
Comparative Orthopedic Laboratory (CORL)
382 W Street Rd
Kennett Square PA 19438
610-925-6278

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of gayle callis
Sent: Friday, January 30, 2009 11:57 AM
To: 'Denise Crowley'; Histonet
Subject: RE: [Histonet] JB4 users

GMA in general, does not infiltrate into calcified mouse femurs as well as
methyl methacrylate, and controlling polymerization is not easy with
calcified bone in GMA.  You have already tried extended infiltrations, but
didn't change the results. I suggest you change to a Technovits 9100 kit,
and do methyl methacrylate instead. Be sure to allow more time for
processing compared to paraffin or even 
GMA.  

Your microtome and tungsten carbide blade are not the problem, it is using
calcified mouse femurs with GMA. 

You can remove methyl methacrylate from sections for immunostaining or other
stains that will not penetrate this very hydrophobic plastic.  

Gayle M. Callis
HTL(ASCP)HT,MT
Bozeman MT 59175

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Denise
Crowley
Sent: Friday, January 30, 2009 6:56 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] JB4 users

We are a research core Histology facility accepting specimens from  
within the MIT community.  One of our researchers is trying to embed  
mouse femurs in JB4 for us to section on our Thermo Finesse microtome  
utilizing a tungsten carbide blade.    The tissue just crumbles out  
of the resin and we are not able to pick up any sections.  We asked  
the researcher to infiltrate longer, which she did, but the results  
are the same.  We have sectioned other tissues in JB4 with this  
instrument, so I do not believe the problem is us or the microtome.   
Any suggestions would be appreciated.

Denise Crowley
Facility Manager Histology
David H. Koch Institute for Integrative Cancer Research
Massachusetts Institute of Technology
40 Ames St. E17-427
Cambridge MA  02139
617-258-8183
dencrowl <@t> mit.edu




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