[Histonet] decapod cuticle sections

[David Waugh]

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