[Histonet] Advice.

[Rittman, Barry R]

Ian
I did a lot of sectioning of dragonfly larvae a long time ago and tried all sorts of processing procedures.
The best way that I found to keep tissue soft enough for paraffin wax sections was to use a chloral hydrate: phenol mixture.
After fixation and dehydration to absolute ethanol, soaked in a mixture of equal parts of chloral hydrate:phenol (heated gently to allow mixing of these two).  Then used chloroform intermediary agent, then paraffin etc.
Also found that if soaked the insects in equal parts of chloroform and paraffin at least overnight at room temperature that this significantly cut down the time needed in molten paraffin wax.
I realize that chloral hydrate is not that easy to obtain (was 38 years ago that I did this) but I found this the best method.
 
Another method that has been used is to use N-butyl alcohol or isopropyl alcohol for dehydration as these have a less hardening effect. Have not used N-butyl alcohol  but have used isoproyl and this is somewhat miscible with paraffin wax.  This method was an improvement over routine wax processing but not as good as chloral hydrate:phenol softening.
Hope that this helps.

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Ian Montgomery
Sent: Thu 1/26/2006 9:29 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Advice.



         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,
Histotechnology,
IBLS Support Services,
Graham Kerr Building,
Institute of Biomedical & Life Sciences,
University of Glasgow,
Glasgow,
G12 8QQ.
Tel: 0141 339 8855
Office: 4652
Lab: 6644.
Pager: 07623 975451
e-mail: ian.montgomery <@t> bio.gla.ac.uk 


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