RE: Killing Nematodes with Fire

From:"Monson, Frederick C."

Well, you won't believe this, but in my references it says that the only way
to 'get' the 'buggers' is to warm the fixative to 60oC, place the worms in a
dish and flood them with the fixative.  Hot solutions include, ethanol (from
50-95%) and just plain water.  No suggestions for hot aldehyde solutions.
[Good for them!!!]  

According to Peter Gray in the Microtomist's Formulary and Guide (complete
citation if you need it), "the worms so treated usually straighten out.
Those that don't should be thrown away."  [Sounds like Guantanamo Bay.]  

NOTE:  P. Gray, like the rest of us, is/was primarily a vertebrate
histo-ist, so one cannot expect complete sincerity or attention to detail
when dealing with these, ............., parasites!

NOTE2:  I didn't think it would be appropriate to convey the method which
calls for fixative that is "almost bubbling",  or the one in which the worm
is placed in a depression slide in a drop of whatever and the slide held
over a Bunsen burner until the 'bugger' 'straightens out', but not until it
dries out.  Well, maybe they said, "relaxes".

Again, actual references on request - pages cost extra!

Regards and best wishes to your vermiform student,

Fred Monson

> ----------
> From: 	Richard Edelmann
> Reply To: 	edelmare@MUOHIO.EDU
> Sent: 	Thursday, January 31, 2002 11:17 AM
> Subject: 	Killing Nematodes
> Search the CONFOCAL archive at
>         I've got a student who is looking at Nematode intestinal tracks
> with CLSM
> using Syto-13 (488/509) as a general stain.  Works fine.
>         The problem comes in killing the buggers.  We originally tried
> Glutaraldehyde (killing, fixingn and good auto-fluoresencefor general
> imaging,
> eh?) but it was very slow (up 15% glut still wiggling at 60-90mins!).  The
> student has been using Sodium Azide in water to kill them but again this
> can
> take 30-40 mins!
>         Does anyone have any suggestions for killing nematodes reasonably
> quickly but not interfering with the Syto-13 DNA staining?
> Richard E. Edelmann, Ph.D.
> Electron Microscopy Facility Supervisor
> 352 Pearson Hall
> Miami University, Oxford, OH 45056
> Ph: 513.529.5712        Fax: 513.529.4243
> E-mail:
> "RAM disk is NOT an installation procedure."

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