RE: Grasshoppers

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From:"McCollough, Carol" <>
To:"''" <>,
Date:Tue, 15 Jun 1999 08:14:38 -0400
Content-Type:text/plain; charset="iso-8859-1"

-----Original Message-----
Sent: Monday, June 14, 1999 9:53 PM
Subject: Grasshoppers

   I am asking for any help I can get and give to an entomologist on
campus here.  He is looking for any special handling, processing, etc.
for grasshoppers???>>


I don't have anything specific on grasshoppers, but we do crustaceans here,
which may present some of the same problems.  We fix tissues in Davidson's
fixative for at least 18 hours.  

95 percent ethanol		330 ml
37-40 percent formaldehyde	220 ml
glacial acetic acid		115 ml
distilled water			335 ml

Other fixatives that have been used on crabs in the past are Bouin's and
Helly's, both of which have disposal problems that we prefer to avoid.

We then transfer to 70 percent ethanol for short term storage.  Our
processing schedule is all under vacuum:
70 percent ethanol	10 min
80 percent ethanol	10 min
95 percent ethanol	10 min
95 percent ethanol	10 min
100 percent ethanol	10 min
100 percent ethanol	10 min
xylene			10 min
xylene			10 min
Paraplast Plus		30 min
Paraplast Plus		20 min
(Obviously the pieces of tissue we dissect out are small.)

We embed in regular Paraplast and section at 5 microns using poly-L-lysine
coated slides.

Two references you may find helpful are:

Messick, G. (1995) Laboratory techniques to detect parasites and diseases in
blue crabs, Callinectes sapidus.  Techniques in Fish Immunology 4:187-200.

Johnson, P.T. (1980) Histology of the blue crab, Callinectes sapidus: a
model for the Decapoda.  Praeger.  440pp.

Regards -
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

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