[Histonet] decapod cuticle sections

[Rene J Buesa]

Under separate cover I am sending some tips on floating paraffin sections that may help you.
René J.

--- 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!
-David


Kent State University
Department of Geology
Kent, Ohio 44242
dwaugh <@t> kent.edu
www.personal.kent.edu/~dwaugh



_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet