Re: cartilage

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From:lpwenk@netquest.com (Wenk, Lee & Peggy) (by way of histonet)
To:histonet <histonet@magicnet.net>
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NTT@shcc.org wrote:
>
> Hi,
>
> I know I've posted this in the past, but does anyone have a good
>technique for
> sectioning of frozens or paraffin embedded cartilage.

(I know the following works for paraffin. Have never tried it with FS.)

Have you ever tried floating the tissue out on 25% alcohol, then
transferring it to the flotation bath?

We take a blank slide (frosted end ones OK), lay it flat on
the counter, and flood it from a squirt bottle with 25% alcohol.

Then lay the tissue on the alcohol.

Carefully drain off the excess alcohol onto a paper towel by
lifting one corner of the slide and holding onto the tissue.

Then slowing lower your slide into the water bath at a slight
angle. The wrinkles/folds should be parallel to the edge of
water coming up the slide. In other words, you can put the
slide in bottom first or side first or diagonally. Just so
that the wrinkles are parallel to the surface of the water,
not perpendicular.

The difference in surface tension should put a little more
"pull" on the section, straightening out more of the
wrinkles, folds. The sections might spin around due to this
surface tension, so watch it if you have to maintain the
same orientation slide after slide. The more alcohol you drain
off, the less spinning, but also less pulling. So play around
with the amount you need drained off.

You can do this with one section, or a ribbon, as long as
it fits on the slide.

About the only difference I have ever seen between this and
slides that I did NOT float on the alcohol is that those with
the alcohol might stain a SLIGHTLY lighter shade with the
eosin. Nothing noticable, unless you were doing serial sections,
some with and some without alcohol, and then laid them
next to each other in a folder and LOOKED for the difference.
Microscopically, they look the same.

I'd slide with the subbed slides.

Hope this helps.

> CryoJane, but my boss is not will to spend the $6000. Or if anyone has
> cartilage sections that they're willing to share, I'd love to hear from you.
> It can be human or bovin (prefer human).   Or if you know of a vendor that
> sells cartilage sections, please let me know.
>
> I can section my own, but I always get a wrinkle in the middle of the
>section.
> Unfortunately, to publish our results, I need wrinkle free slides.  I am
>using
> plus charge slides.
>
> .
>
> Thanks,
> Noi

--
Peggy A. Wenk, HTL (ASCP)
Anatomic Pathology
Wm. Beaumont Hospital
3601 W. 13 Mile Rd.
Royal Oak, MI 48073-6769




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