Re: [Histonet] Thin slice embedding

From:Rene J Buesa

If I understood you well you are trying to "hold" slices of tissue flat to further section them.
  If that is the case preparing a layer of paraffin, waiting it to solidify and adding later on the tissue and another layer of paraffin (like a sandwich) did not work, and will not work.
  Try the following: get a metal flat surface the size of you tissue slices. Pour melted paraffin in a mold larger than the piece of tissue. Put the piece of tissue flat OVER the melted paraffin, and the metal flat surface HEATED above the melting point of the paraffin, PUSH the piece of tissue up to the depth you want. Gently retrieve (PULL) the piece of metal leaving the piece of tissue at the depth you selected.
  Wait until this "block" solidifies and try to section. Since the piece of tissue has not been infiltrated, just embedded, the paraffin will not hold to the tissue, but keep it in position to section.
  René J.

Shengwen Zhang  wrote:
  Hi All,

I'm trying to further cut my organotypically cultured brain slices (300 micron before culture, ~100 micron after culture). I've tried OTC compound and sucrose solutions for embedding, both didn't work well (the slices wrinkle at the tissue-medium boundary). Now I want to try paraffin. I want to keep 3-4 slices from the same brain flat and on the same plane, so I can process them together (otherwise, I would have too much work to do). I created a flat area with paraffin, after it solidified, I laid the slices, and pour another layer of paraffin. But the two layers don't stick together well enough to stand the forces during cutting. How do you embed thin slices with paraffin?

Thank you!

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