[Histonet] Poor quality frozen sections, feline skin.

[Julie Bilkey]

Hi Peter,

We use 20-25% sucrose on our fresh specimens for immunofluorescence (IF). We
only deal with human skin samples so I will not comment on feline skin. I
sit the specimen in three changes of sucrose for 10 minutes each. Before
embedding in the cryostat, I gently blot it on a tissue. My understanding is
that the sucrose does give support like a matrix, and it has saved dried
specimens for me in the past.

I hope this helps.

Julie Bilkey.
Sydney Skin Pathology.

On Fri, Oct 7, 2011 at 7:54 AM, Peter Bodig <pbodig <@t> histoprep.com> wrote:

> I have some feline skin samples for frozen sections that were sent to us
> because our client was not able to section them themselves. Unfortunately we
> are having the same problems trying to make good sections ourselves. The
> samples seem dry and unsupported when we try to cut them. The tissue breaks
> into small pieces and falls out or the OCT when we try to cut it. We have
> tried fixation with 10% NBF for various times up to 24 hrs prior to doing
> frozen sections to give the tissue better support, which helps, but not
> enough, and then the tissue falls off the coated slides when we try to stain
> them. Sectioning at 8µ helps as well (that is as thick as out cryostat will
> go).
>
>
>
> Does anyone have suggestions on how to get good quality frozen sections
> from feline skin?
>
>
>
> We have come up with some questions as well:
>
>
>
> 1.      Is there a way to support the tissue by infiltrating it with OCT or
> other similar media to improve our sections?
> 2.      I have read about using 30% sucrose solution for cryo protection,
> does that aide in making good quality sections or just reduce freezing
> artifact?
> 3.      Does tissue need to be fixed prior to using cryo protection?
>
>
>
> Our client will be using the sections for Senescence β-Galactosidase
> Staining.
>
>
>
> Thank you for any help you can provide,
>
>
>
>
>
> Péter Bodig
>
> Laboratory Coordinator
>
> Colorado Histo-Prep
>
> E-mail: pbodig <@t> histoprep.com
>
>
>
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