Re: attaching MMA thin sections to slides
Sarah,
We have good luck with 0.5 to 6 micron thin sections using the following
protocol:
1) section the 4-6 micron sections dry with a glass knife; or thinner
sections with a glass knife equipped with a boat filled with distilled
water
2) use a forceps for the dry-cut sections or a tapered wooden pick for
the wet-cut sections to place them on a dome of distilled water on the
Superfrost Plus slide
3) heat on a hot plate that is set a little less hot that that which
would make the water on the slide boil (too hot to touch). Place an
old glass knife on the hot plate so that the slide can be propped up
on it at the frosted (handling) end.
4) let the water under the section evaporate and then wait 20 seconds
before removing the slide from the hot plate and staining
5) stain with filtered toluidine blue, by placing drops of stain so
they dome over the sections and placing the slide flat on the hot plate
for 15-20 seconds. Don't let the stain boil or dry out.
6) tap off the stain and quick rinse in 3 changes of distilled water,
one change in 70% ethanol and then one more change of distilled water
and place the section back on the hot plate at an angle, over the old
glass knife until dry
7) coverslip
We have had very good luck with this protocol, in fact we have none of
the sections releasing even if we have to remove the coverslip and
restain.
Good luck,
Karen Pawlowski, Ph.D.
Research Scientist
University of Texas at Dallas
Dallas, Texas
Sarah Jones wrote:
>
> Hello all you Uncle Ben fans, hope those that need to concentrate on
> their breathing are doing well,
> I would appreciate any procedure on MMA thin sections in regards to
> attaching them to the slides. Specifically, the steps including
> sectioning through deplastinization. I've been having trouble keeping
> them on the slides with Subbed Slides and Plus Coated. How are the
> sections floated out?
> PS- I like the suggestion about using OT, thanks for the reminder. I
> think it is better to use the delete button than to get your blood
> pressure up. We don't need to lose any precious histotechs to heart
> attacks. I'm sure we would all like to become like the retired guy for
> a few years before that happens. Personally, I like a few off topic
> subjects and OT would help those that don't.
>
> Sarah Jones HT(ASCP)
> Dept. of Vet. Anatomy & Public Health
> Histology Lab
> Texas A&M University
> College Station, TX 77843-4458
> phone: 979-845-3177
> fax: 979-458-3499
> email: sjones@cvm.tamu.edu
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