Re: Drosophila

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From:"LaCinda J. Burchell" <>
To:"Deborah Faichney (by way of Histonet)" <>
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Hello Deborah,
  Oh, I remember the days I spent playing with fruit flies! Great fun!
Here's how I've handled them with success.  I would suggest starting by
using a fixative called Prefer which is produced by a company called
Anatech  (1-800-262-8324).   I'm concerned that formalin may cause
troublesome antigen masking.  After the flies are fixed for an hour or
so I would suspend them in a liquid 1% agar made in PBS.  Be careful not
to use the agar when it is too near it's boiling point.  It should cool
to a point where it is near solidifying. Pipette off as much of the
fixative as you can, and put 200ul or so of the warm agar into the tube
containing your drosophila.  Gently mix the flies into the agar, and
then pipette the warm agar/fly batter onto a clean glass slide.  It
should set up in the shape of a soft droplet, which can then be gently
pushed off of the slide into a cassette for processing.   Paraffin will
infiltrate the agar droplet which will be clear when the paraffin is
warm, and will be an opaque white once it is embedded.  You may also
freeze the agar droplet as you would any other tissue in an OCT type
compound for frozen sectioning.  I've used agar as a suspension and
support medium for anything tiny like cells, and also for fragile things
like embryonic mouse brains.  Best of luck to you!  If you have any
questions please free to contact me.  Cindy B.
LaCinda Burchell
University of Wisconsin Madison, Med. School
Dept. of Allergy and Immunology
600 Highland Ave.  H6/367
Madison, WI 53792
FAX (608) 263-3104
Lab (608) 262-3518

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