[Histonet] Successful frozen sections on bone

Gayle Callis gcallis <@t> montana.edu
Mon Aug 30 09:03:06 CDT 2004

An addition to Patsy's suggestions, another trick is to flash with UV 
twice, we have even done 3 times.  The capacitor in power source needs to 
build up adequate charge (I hope I said this correctly) , so do not flash 
rapidly, do it slowly so you get the "flash".  When the green light is on, 
you have the go signal to flash.

Also remove the tape VERY slowly, and diagonally from one corner of slide 
towards the other, and make sure the slide is cold when you do this.

We use 1/2X slides, with success, if you have a great deal of problem with 
polymer slides then try 1 x or even 4X.  The latter are really gooey but 
with double UV exposure, 1/2X worked nicely.

  neAt 09:19 AM 8/28/2004, you wrote:
>I agree with Gayle on this, I have cut whole rat tibia/femurs without decal
>using the Instrumedics tape system and the sections are pretty darn good, it
>is sometimes difficult to get the tape off the slide leaving all the bone
>section behind, I expose the section to the UV light and then put it on dry
>ice for 10 min. or so before trying to remove the tape, you are still going
>to get some of the cortex left on the tape but this is the best I could do
>(my experience).
>Patsy Ruegg
>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu
>[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Gayle
>Sent: Friday, August 27, 2004 9:33 AM
>To: Lowen, Nancy; Histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] Successful frozen sections on bone
>The ONLY way we have successful cryosections of any kind of bone is using
>the Instrumedics CryoJane tape transfer system. Check out their website for
>information, how it works.
>Bone frozen sections are destroyed by disposable or c profile steel blade,
>tungsten carbide blades are ideal as you are finding out. However, I have
>never had a bone frozen section cut with a TC knife which remains intact
>unless the tape transfer method is used to maintains bone
>integrity.  Cutting temperature must be -28C up to -32C or so.
>Your tungsten carbide knife edge must be in perfectly, sharp condition even
>with tape transfer method.
>For us, cryosectioning bone without the Cryojane has been a waste of
>time.  Although a pricey bit of equipment,  it has paid for itself many
>times over with good bone frozen sections.
>I am going to attach a publication to you privately so you can see
>At 10:02 AM 8/27/2004, you wrote:
> >I am doing frozen sections on fresh, undecalcified mouse femurs.  My
> >sections seem to be losing all of the marrow and many are torn .  I have
> >tried disposable cryostat knives as well as the tungsten carbide, but the
> >sections are still not good.  If anyone else is doing this procedure, could
> >you share any tips or suggestions on your technique, cutting temperatures,
> >or overall procedure. I have tried different temperatures, but none of them
> >seem  to give me the sections I want.  Any help would
> >
> >be greatly appreciated.
> >
> >Nancy.Lowen <@t> med.va.gov
> >
> >
> >
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>Gayle Callis
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
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