[Histonet] (no subject)

[Judith L. Williams]

Hi to Ian- Great advice and links from Phil- having worked on the second most abundant form of meiofauna- the copepods- and done light level, EM and confocal on them - fixation is a problem for sure.
there are some elegant morphology studies from the 70;s and again - most are EM.
good luck - try looking up meiofauna morphology on google or nematode morphology for the basics.
best!
Judy at U. of Washington



On Wed, 13 Feb 2008, Philip Oshel wrote:

> Ian,
>
> A couple of useful web sites:
> http://www.wormatlas.org/
> and Dave Hall's lab at Albert Einstein Coll. of Medicine:
> http://wormem.aecom.yu.edu/
> Jay Campbell at U Wisconsin has done a bunch of C. elegans EM.
> jmcampbe <@t> wisc.edu
> Mind, most of the people I know are either doing EM or confocal on whole-mount 
> or squish-mount worms. Not light microscopic sections, so I'm not sure how 
> useful these contacts will be.
> The big trick for fixation & processing at the EM level is high-pressure 
> freezing followed by freeze-substitution for embedding.
> For "regular" light histology & sectioning, I'd suggest lopping off the head & 
> tail & maybe bisecting the body, then embed in JB-4 or similar resin and cut 
> with a glass knife.
>
> Paraffin embedding and steel knives won't cut it.
>
> Phil
>
>
>>             When the select few are sitting in the nuclear bunkers the
>> dominant species on the planet will be nematodes. I've just started a
>> project for my Zoologist colleagues studying these wee devils and I'm
>> convinced they are the mega-organism. Formaldehyde, not a problem, couple of
>> weeks later, give them a rinse and away they swim. Osmium tetroxide, "are
>> they trying to annoy me with this slightly noxious compound." Managing to
>> fix them is hard enough but processing for sectioning, a nightmare. Does
>> anyone have experience processing these beasts? Hints and tips would be very
>> welcome.
>> 
>> Ian.
>> 
>> Dr. Ian Montgomery,
>> Histotechnology,
>> I.B.L.S. Support Unit,
>> Thomson Building,
>> University of Glasgow,
>> G12 8QQ.
> -- 
> Philip Oshel
> Microscopy Facility Supervisor
> Biology Department
> 024C Brooks Hall
> Central Michigan University
> Mt. Pleasant, MI 48859
> (989) 774-3576
>
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