Re: Drosophila

<< Previous Message | Next Message >>
From:Connie McManus <conmac@cc.usu.edu>
To:Deborah Faichney <d.a.faichney@stir.ac.uk>(by way of Histonet), HistoNet@pathology.swmed.edu
Reply-To:
Content-Type:text/plain; charset="us-ascii"

Deborah,

Insects have a tough chitinous exoskeleton that must be cut into for good
fixation and processing to happen.  I've done TEM on Drosophila before and
this is how I did it...

Expose the live insects to CO2 gas for about 5 minutes to kill them.  Then,
you need to get a micro scalpel (I'm not joking).  If you don't have these
(they are very expensive) they can be made with a small shard from a double
edged razor blade mounted on the end of  a wooden applicator stick and
secured with fine copper wire stripped out of some scrap electrical wire.
Seal this with dental wax and ... Wah Lah..you have a micro scalpel. Place
the insect on double sided sticky tape in a glass petri dish, place under a
dissecting microscope, then make a very small incision in the abdomen.  Fix
and process under vacuum or microwave. The alcohols will dissolve the
adhesive on the tape.  The cellophane can be removed and discarded.  Also,
add 1% DMSO to all reagents (including the fixative) up to the absolute ETOH.
Hope this made sense.  If not, e-mail me and I'll try to explain in better
detail.

Connie McManus

At 03:25 PM 05/19/2000 -0500, Deborah Faichney wrote:
>Hello all
>
>Any ideas for fixation, processing etc of Drosophila?  Can you use and get
>good results with formalin?
>
>Most of our work is fish related, but a student wishes to do IHC and IFAT
>for detection of Rickettsia Like Organisms.  She has these techniques sorted
>for the fish part but not the Drosophila.
>
>Any hints 'n' tips would be gratefully received.
>
>Debbie Faichney
>I
>Institute of Aquaculture
>University of Stirling
>Stirling
>Scotland
>
>
>
>
Connie McManus
Veterinary Diagnostics Lab
Utah State University
Logan, UT
USA



<< Previous Message | Next Message >>