Re: whole mouse sections

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From:"R.Wadley" <>
Date:Tue, 21 Sep 1999 11:15:18 +1000
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	Dear Mary,

	Yes, I have sectioned whole mice.  The method used was:

	The mouse was sacrificed & delivered to me frozen on dry ice.

	I removed the head, limbs & tail (bone shears are ideal for this, old
steel microtome blades are good too, but keep a close count on your
fingers!) and mounted the carcase belly up on a cryostat stub using OCT.  

	You have to be gentle as you trim into the mouse.  Cryostat temperature is
important, too cold & the bones will be too tough to section, I think we
worked at somewhere around -20C to -25C.

	When you reach the level you require you will not get good results unless
you can support the section.

	I used a Leica CM 1800 fitted with gold coated high profile disposable
blades by Sturkey.  To support the section I stuck on a piece of cold
temperature adhesive tape to the trimmed surface of the specimen, then
using the guide plate (They are useful sometimes) cut a 60 um section.
This was air dried and used to look for a radioactive tag.  I got 5 - 6
sections per blade before score marks (limb bones) become to much.

	I had silver tape, but clear tapes are available & I believe from what I
see on the Histonet that there is a histo product exactly for this task.
Thinner sections are also possible.  With clear tape you can do all your
staining as if it were a slide, then stick onto a slide (I used lantern
slides) for viewing.

	You will find that a whole mouse is almost impossible to freeze without
ice crystal formation within the body cavity.  Sectioning cold enough to
keep fat solid often means it is too cold to cut through larger bones.
Some tissues (eg bladder) which are surrounded by dense bone structures
(pelvis) may be difficult to obtain score free.


	Rob W.


At 15:24 09/20/1999 -0400, you wrote:
>Greetings Histonetters,
>	Does anyone out there in Histoland have experience with
>sectioning of whole mice?[yes, the whole animal - undissected]  Can it
>be done?  What's involved?  Details please!  One of the investigators
>here is curious and so am I. I'll compile all of the info I receive and
>give a summary. Thanks.
>Best Regards,
>Mary Vaughan HT (ASCP)
>Roswell Park Cancer Institute
>Elm + Carlton Sts.   CDC-121
>Buffalo, NY 14263

R. Wadley, B.App.Sc. M.L.S, Grad.Dip.Sc.MM
Laboratory Manager
Cellular Analysis Facility
School of Microbiology & Immunology
UNSW, New South Wales, Australia, 2052
Ph (BH) 	+61 (2) 9385 3517
Ph (AH)	+61 (2) 9555 1239
Fax 	+61 (2) 9385 1591

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