Re: [Histonet] Methacrylate cuttting
In a message dated 8/9/2005 7:31:29 AM US Mountain Standard Time,
> HELP!! To all of you this will probably be such an easy solution. My boss
> on annual leave for 3 weeks and has left me the task of practising cutting
> methacrylate sections at 3microns.
If you are using glass knives, are they the triangular type of knives, or the
longer type of "Ralph" knives? Are you using an ultramicrotome? Or are you
using a glass knife adapter on a regular rotary microtome? And can you put a
"boat" or a "trough" on the knife so it can hold a little water?
Methacrylate tends to be a little soft, and it's not uncommon to see the
sections crumple and stick to the knife edge. You could probably get better
results if you could use the sectioning techniques that are used for electron
microscopy. These involve bringing water up to the knife edge, so the sections
float onto the water surface as they are cut. You can then retrieve the sections
with a small wire loop or a hair mounted on a stick, and transfer them to a
drop of water on a glass slide.
Also, using water during sectioning will help the sections "relax" and
flatten out. A lot of the methacrylates are hydrophilic, and you can actually watch
them expand slightly as they come off the knife edge and float on the water
If you were cutting slightly thicker, say around 5-8 microns, you might be
able to *carefully* use a pair of fine-tip watchmaker's forceps or a hair and
gently guide the sections down the knife as they are cut. But this may not work
at 3 microns thickness.
This may have been discussed previously on Histonet...I can'r remember for
sure. You can go to:
and try searching the Histonet archives using words like:
Hope this helps!
Robert (Bob) Chiovetti, Ph.D.
The Microscope Works
Arizona's Microscopy Resource
132 North Elster Drive
Tucson, AZ 85710-3212 USA
Member, Arizona Small Business Association - ASBA
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