Hi Andi:

Depending on the size of your flies, you may be overprocessing.  We do blue
crab parts (about 3x4x4mm) and whole grass shrimp (these are split
longitudinally and fit in a regular cassette)on the following schedule:

70% EtOH    delay station
80%         20 min, vacuum
95%           "
95%           "
100           "
100           "
100           "
xylene        ", 35 degrees
xylene        ", 45 degrees
Paraplast      30 min, vac.
Paraplast      20 min, vac.
Paraplast      20 min, vac.

I prefer Paraplast + because of the crunchy nature of arthropod cuticle
(better penetration and higher melting point), however one of the techs who
uses my lab has a morbid terror of DMSO - also does not consider gloves an
essential part of routine laboratory work.

You may find the exoskeleton difficult to section, as it doesn't infiltrate
that well.

Hope this helps.

Regards -
Carol
*****************
Carol B. McCollough, HT(ASCP)
Diagnostics & Histology Laboratory Manager
Maryland Department of Natural Resources
Fisheries Service
Cooperative Oxford Laboratory
904 S. Morris Street
Oxford, MD 21654


---100--Original Message-----
From: Andrea Grantham [mailto:algranth@U.Arizona.EDU]
Sent: Thursday, July 06, 2000 3:01 PM
To: histonet@pathology.swmed.edu
Subject: processing bugs


Hello Histonetters,
I was just asked to start processing whiteflies for in situ detection of
intracellular symbiotic bacteria.
Two questions -
Is anybody processing bugs? I'm looking for a protocol.
My usual processing protocol for animal tissue is one hour each in
increasing strengths of ETOH starting with 70% and going to Paraplast Xtra.
Total processing time is 12 hours. Is this too long for insect processing?
Thanks.
Andi


.....................................................................
: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
: Sr. Research Specialist       University of Arizona               :
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