AW: [Histonet] 2x3 microtomy: Adapter vs Sliding Microtome

[Gudrun Lang]

Have you tried to float the section first in a roomtemp. waterbath (bowl
with tapwater)? There you can flatten the wrinkles with a brush, then mount
it on the glass slide and bring it into the warm water. While gliding from
the glass you can also use the brush to pull softly on the section to
stretch it.

If the infiltrated tissue is still too soft, I suggest longer infiltration
time in xylen and paraffin to get rid of the fatty part of brain tissue.
Sometimes even cutting on next day of embedding (one day waiting) helps.
Do you cool the blocks before cutting? Long enough?

For big blocks we usually use one blade for trimming and a new one for
getting a thin section.

Hope this helps
Gudrun


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Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von R C
Gesendet: Dienstag, 29. Dezember 2009 23:16
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] 2x3 microtomy: Adapter vs Sliding Microtome

Hi. I'm working on a protocol for cutting monkey brain sections to be
mounted on 2x3 slides. I've read about utilizing a sliding microtome but in
short, have decided to use the 2x3 adapter for a standard Microm microtome.
During microtomy I've noticed many wrinkles in the sections, particularly
within the folds of the cerebellum. The wrinkles worsen as the sections
float in the water bath (temp=38).

In troubleshooting, a co-worker suggests inadequate fixation. I on the other
hand believe that the wrinkles relate to the Type R paraffin, which contains
polymers as well as the use of the adapter versus the sliding microtome.

Can anyone offer any first hand experience/guidance?

Thanks
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