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,
> 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
> 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
> 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
> 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: email@example.com
> "RAM disk is NOT an installation procedure."
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