Re: cutting whole mice

From:rueggp

thanks Jamie,
that is what i thought.  not sure if they want to use the beta-gal enzyme or ihc method but either way i think the tissue needs to be frozen, in my experience anyway.
patsy

Jamie Erickson wrote:

> Patsy,
>         We've only do B-galactosidase staining on frozens. I don't know if this is what your asking but we treat mice with adenovirus that express the marker  gene B-galactosidase then freeze the tissues  (after days of injection of the virus ) for staining and using the enzymatic method (X-gal, Ferric and Ferro cyanide etc..) and detect B-gal in various tissues. I have not used the B-gal antibodies for B-gal staining, the enzymatic method is a lot easier. Hope this helps..
>
> Jamie Erickson
> Scientist II
> Wyeth/Genetics Institute
> 1 Burtt Rd.
> Andover, MA   01810
> work : (978) 247-1348
> FAX  : (978) 247-1389
> Jerickson@genetics.com
>
> >>> rueggp  11/26 5:35 PM >>>
> 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





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