[Histonet] 2x3 microtomy: Adapter vs Sliding Microtome

[Anne van Binsbergen]

i  spent many many years handling  pig, mouse and human brain, cutting
sections on 2x3 slides.

the trick is to use the tap water float out before the warm water, also to
cut sections thicker than normal - cut at 10-15 and maybe even 20 microns -
trouble is at that thickness AND it being animal tissue, the sections wash
off in a flash - unless you use adhesive and cook the sections on to the
slides gently at 32C for at least overnight

i have some 20micron silver stained cerebellum on 2x3 slides here with me
right now, 20 years later they are still good as new!!

good luck
Annie



2009/12/30 Gudrun Lang <gu.lang <@t> gmx.at>

> 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
>
>
> -----Ursprüngliche Nachricht-----
> 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
>  _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



-- 
Anne van Binsbergen (Hope)
Abu Dhabi
UAE