Re: [Histonet] Advice.
Hmmm...well, some years back I was working on a butterfly heads project
for a brain
cell count study. Never worked on insects before, so it didn't take long
before I noticed
the head of the butterfly is covered with chitin layers and I had to
section the entire head.
Here's what I did. I got beautiful paraffin sections stained w/Nissl
> fixed cut head in Carnoy's fixative - overnight @ 4C (there are many
the literature. Possible could also use glutaraldehyde containing
fixative - sure these
would be fine for LM too)
> dehydrate a bit more with ethanol alcohol 2x - 30' each
> soak (tissue) in a mixture of equal parts of chloral hydrate:phenol
to soften the entire
*Chloral hydrate:phenol made by weighing equal parts & then gently
molten. Once molten can store for several weeks. Sections cut beautiful.
Tissues were left in this mixture for a minimum of 2 hours to overnight
(perhaps even up
to a couple days).
> clear in chloroform 3x - over 2 hours to overnight (I did 1 hour each).
> place in cedarwood oil - 2 days (change oil 3x during period).
> then do routine paraffin.
*When sectioning, I remember adding a bit of glycerin in the ice water
to soak the blocks a
Ian, I hope this helps.
Maria Bartola Mejia
Smith-Kettewell Eye Research Institute
San Francisco, CA 94115
Ian Montgomery wrote:
> With cuts in staff I've been asked if I can teach the
> histology portion of a zoology class. Problem, one of the tissue I'll
> be using is insect abdomen with its chitin exoskeleton. Any hints and
> tips regarding this type of material, processing, embedding,
> sectioning, that sort of thing. Or, is it Mollifix time again?
> Dr. Ian Montgomery,
> IBLS Support Services,
> Graham Kerr Building,
> Institute of Biomedical & Life Sciences,
> University of Glasgow,
> G12 8QQ.
> Tel: 0141 339 8855
> Office: 4652
> Lab: 6644.
> Pager: 07623 975451
> e-mail: firstname.lastname@example.org
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