[Histonet] decapod cuticle sections
Rene J Buesa
rjbuesa <@t> yahoo.com
Mon Dec 29 14:52:28 CST 2008
Under separate cover I am sending some tips on floating paraffin sections that may help you.
--- On Mon, 12/29/08, David Waugh <dwaugh <@t> kent.edu> wrote:
From: David Waugh <dwaugh <@t> kent.edu>
Subject: [Histonet] decapod cuticle sections
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, December 29, 2008, 2:19 PM
OK, I'm not a histologist, I'm a paleontologist (&%**'it Jim, I
a Doctor not a....). I am trying to make paraffin sections of decalcified
decapod cuticle (from modern material, not fossil). The major problem I have
been having is that the sections keep falling off the slides during staining....
I was making my own slides with a solution from EMS
(3-aminopropyltriethoxysilane in acetone) I lost more than 1/2 of my sections...
so I got pre-coated slides (Poly-L-Lysine) from EMS. The sections still fall
off! After the section are cut I float them on warm water (48C), they never
fully flatten out...? I then slip the slides underneath to pick up the sections.
With the slides I subbed, I followed the directions and heated them on a
hotplate. The presubbed slides I just air dried. To help the sections stick, I
also tried pushing some of them down on the slide with a wet filter paper after
removal from the water bath. The slides where then de-waxed in two xylene baths
and brought to water in a series of ETOH baths. The sections seem to fall off
mostly in the ETOH baths, but sometimes in the xylene. I'm not looking for
perfection here, I just need the sections to stick. When a few sections make it
through my protocol they look OK. I'm doing this on a low budget, but keep
spending more and more on stains, slides etc. I'm getting very frustrated!
Any ideas? how long should the slides dry? heat, no heat? Thanks in advance!
Kent State University
Department of Geology
Kent, Ohio 44242
dwaugh <@t> kent.edu
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