cutting whole mice

From:Gayle Callis

What IS the age of the mice?  

If there is calcification, tape transfer with Instrumedics would be
helpful, and all you do is fix the sections properly with
formalin/gluteraldehyde mixture (there is a formula for Beta Gal staining)
then develop the color with Beta Gal working buffer solution containing
ferro and ferricyanide.  Roche has a lovely kit that does this or you can
do it with mass staining.  Beta Gal per se is NOT an IHC stain, the Lac Z
reporter was incorporated into the cells of the mouse, you cut the
sections, and do an enzyme histochemical reaction for beta galactosidase.
There are successful paraffin protocols available.

Also, one could do the whole mouse, looking for areas of blue, have done
this with murine lungs, once this is done, you can cut the sections post
staining.   

I have a Beta Gal "bible" that is available on the internet with all kinds
of beta gal hints, even for paraffin sectioning.



  At 03:35 PM 11/26/01 -0700, you wrote:
>fellow netters,
>i have been asked about cutting whole young mice treated with beta-gal.
>i did this at one time as frozen sections by placing a microtome in a
>chest freezer and using a tungsten carbide knife.  question:  in order
>to sucessfully do ihc for beta-gal would the sample have to be frozen or
>does it stand up to processing?
>the exact inquiry follows.
>patsy
>Dear Patsy,
>I was given your name by Nancy Shellhorn and she thought you would be
>just the person to help me!
>I work in a core lab which provides histology support to many users.  I
>have an investigator who is interested in doing whole mounts of young
>mice.  He is injecting them with Beta-Gal at birth, then wants to look
>for the stain after they have grown a little- he says they will be
>"young mice" but hasn't given me an exact age.  I believe weeks as
>opposed to months.
>Have you any experience processing entire small mice and cutting them?
>If not, can you advise me of a source (human or literature) that may be
>able to assist me?
>
>Thank You Very Much,
>Marda Jorgensen
>
>
>
>
Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)





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