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
> To: 	CONFOCAL@LISTSERV.ACSU.BUFFALO.EDU
> Subject: 	Killing Nematodes
> 
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> 
>         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: edelmare@muohio.edu
> http://www.emf.muohio.edu
> 
> "RAM disk is NOT an installation procedure."
> 
>