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Dear Simon

Perhaps you might be interested to try CyGEL?
This is a thermoreversible, hydrogel mountant designed for the imaging
of live cells / tissues.  Unusually, it's a gel when warm and a liquid
when cold which means your sample is stable on the stage.  The gel can
be supplemented with media and growth factors.  

In this case, I would recommend stretching the sample as you need and
then overlaying with CyGEL to help maintain it's integrity and provide
it with media, etc.  The gel will set and it can be chilled again to
spread it (either AFTER applying a coverslip or BEFORE covering with
oil).

It also depends whether you need to keep the tissue taught. If not, the
gel might be sufficient on its own.

You can see more information on our website.

I hope this helps.  You can always contact me directly if you want to
discuss this further.

Very best regards
Roy
 
Roy Edward
Biostatus Ltd
[log in to unmask]
www.biostatus.com
 
56 Charnwood Road, Shepshed, LEICESTER LE12 9NP U.K.
tel: +44(0)1509 558163 fax: +44(0)1509 651061
 

-----Original Message-----
From: Simon Walker [mailto:[log in to unmask]] 
Sent: 18 January 2007 13:54
Subject: Mounting peripheral nerves for imaging

Search the CONFOCAL archive at
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Dear List,

We are hoping to do some experiments using confocal microscopy to image 
fluorescently labelled organelles in live peripheral nerves.  The tissue

will be excised from mice and imaged on an inverted microscope.  The 
problem is, we are not sure how to mount the tissue for imaging.

For widefield imaging using an upright microscope the excised nerves
have 
been mounted in 35 mm dishes onto Sylgard and pinned at both ends to
keep 
the tissue taught (the pins go in to the Sylgard).  However, our
confocal 
is based around an inverted microscope and we now want to use high mag 
lenses with short working distances.  In other words, we can't use
Sylgard 
and pins.

The tissue needs to be in some kind of dish so that it can be immersed
in 
medium, the bottom of the dish needs to be cover-glass, and the tissue 
needs to be held at each end to keep it taught and flat to the bottom of

the dish.

Any ideas gratefully received.
Thanks,
Simon