When I used to do this with 1 micron sections of kidney in methyl/butyl
methacrylate I would cut the section with a 20% acetone filled trough
attached. I picked up and floated onto a water bath kept at 65 C, lined
several section up and mounted on the slide. I baked on for at least a
couple of hours.
To stain, I removed the plastic with an hour in acetone, xylene or,
preferably, chloroform at room temperature. I used toluidine blue for the
stain. H&E came out very pale, even if I used Cole's hemalum for an hour at
60 C with no differentiation.
I found that sections thicker than 1 micron were increasingly more difficult
to deal with as the section thickness increased. 3 microns was the
practical limit unless I was willing to spend a lot of time fiddling around.
However, the whole point of such sections was to cut them thinner than I
could from a paraffin block, which I also had from a second biopsy (and a
third for EM)..
----- Original Message -----
From: "Esther Peters"
Sent: Tuesday, March 11, 2008 8:31 AM
Subject: [Histonet] [Fwd: urgent H&E issues]
> I'm passing this question along from a research colleague, because I have
> never worked with plastic (I'm not sure whether she is using MMA or GMA,
> either). I tried a Google search but did not see a direct answer. We look
> forward to your assistance!
> We have been cutting some plastic here over the last few days (with a
> borrowed microtome), and are having some difficulties with the H&E
> staining. It seems that the section gets tons of little bubbles under
> the plastic (or so it seems). We do not see such bubbles when we stain
> with basic fuchsin and think maybe its due to the alcohol or xylenes used
> at the end of the H&E. We are not removing the plastic (methacrylate).
> Do you have any ideas or can track down an H&E staining procedure
> especially for plastic?
> Esther Peters, Ph.D.
> George Mason University
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