Re: [Histonet] frozen sections on monkey eye

From:Gayle Callis

Ethanol is antifreeze and if you have that in the tissue, you will probably 
have snap freezing issues that further sectioning problems.  This would be 
eliminated by the sucrose cryoprotection before snap freezing however.

You can try raising the cutting temperature so it is higher say at -16C to 
keep the tissues from being crunchy and harder.

If you plan to do immunohistochemistry, you should be aware that Perenyi's 
fluid is also used for decalcifying bone and it may compromise your tissue 
antigens when exposed to the nitric acid.

Another possibility is to cut the eyes with a tungsten carbide knife, much 
as one would do for undecalcified bone sections, and they do make c profile 
tungsten carbide knives at Delaware Diamond Knives.  They may even have a 
demo available to try out, before you spend the big bucks for your own 
knife.  These are designed to cut through some extremely tough tissues even 
decalcified bone frozens at times might be easier with a TC knife.

Another possibility is to use the Cryojane tape transfer system to maintain 
all parts of the eye.

At 04:31 AM 6/21/2007, you wrote:
>Hello Jennifer,
>I've worked in an eye institute before and i have done
>a lot of frozen mouse and rat eyes. Normally i'll have
>a lot of difficulties in cutting the lens as when it
>is fixed in 4% PFA, it hardens.
>I've read in one of the previous email from histonet
>that Perenyi's fluid is a good fixative for eye as
>both Nitric and chromic acids have a softening effect
>on the tissuesand ethanol is a coagulant fixative. It
>is good as it fixes retina really well with little
>separation of the layers and does not harden the lens.
>I hope this will help.
>--- "Harvey, Jennifer Lynn"
> wrote:
> >
> > I thought years of bone work I had done some of the
> > most challenging
> > histology. We move to Nashville and I took a job in
> > eye research. Was I
> > crazy??? This is worse. I need to cut frozen
> > sections on 4%PFA fixed
> > monkey eyes?  Any ideas??

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610

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