[Histonet] cryosectioning rat bone

[Patsy Ruegg]

John,

I have used the Instrumedics tape transfer system for frozen rat bones for
years and I am yet to get all of cortical bone to stay attached to the slide
and not come off with the tape.  Here is my best effort:  I infiltrate the
samples in 30% sucrose overnight before snap freezing, I use a tungsten
carbide D profile knife of course, I cut the section as thin as possible (I
shoot for 4 microns), then I really spend a lot of time and pressure rolling
the section onto the coated slide, flash it with UV about 3 times, then
before removing the tape I lay the slide flat on a block of dry ice and let
it get really cold before slowly with even pulling remove the tape starting
at a corner and pulling diagonally from corner to corner.  John Trapley
wrote a good how too article for JOH a several years ago which I contributed
to, on this.  Gayle Callis is another with experience and Diane Sterchi
developed a method using another 3M tape I believe.  This method is a good
sent for cutting mineralized bone in the cryostat but it is not perfect or
at least I have not ever gotten a completely perfect section with all the
bone intact left on the coated slide.  

Diane send me the ordering info for that tape please, if you are out there.

Patsy

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of John Baker
Sent: Thursday, October 12, 2006 11:33 AM
To: Histonet
Cc: ewaldorf <@t> engin.umich.edu
Subject: [Histonet] cryosectioning rat bone

Hi All,   Does anyone out there in histoland get good cryo-sections of 
rat femur or tibia?  I get great 5-7 micron sections of mouse bone 
using the CryoJane Tape System.  Whether longitudinal or cross the 
cortical bone will not laminate and comes off with the window.  I 
certainly don't get sections at all using the anti-roll plate.  I have 
a Hacker-Bright OTF cryostat and use t-c blades.  Any suggestions are 
greatly appreciated.  Thanks in advance.  John

John A. Baker
The University of Michigan
Orthopaedic Research Laboratories
Histology Unit
109 Zina Pitcher Place, 2218 BSRB
Ann Arbor, MI 48109-2200
734-936-1635


_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet