>Jonathan,
>
>so far noone on the list seems to have a suggestion what stain you
>could use. I was wondering if your problem is low contrast of your
>material under bright field conditions and whether that this is the
>reason why you want the counterstain. If this is the case you could
>try DIC or Phase contrast, which greatly improve contrast in
>specimen. As I understand you work with fixed material, consequently
>the time delay between subsequent images caused by switching between
>different set ups would not matter.
>Probably your microscope has a cameraport so that you can obtain
>photographs of your bright field images and match them with the
>fluorescent images gathered on the confocal.
>Is this a reasonable suggestion?

Thanks for the suggestion.

Unfortunately, we need to to use automated image analysis/3D reconstruction
techniques and need some good boundary detail

and my fluorescent microscope lacks DIC/phase (actually, I have it, but
can't use it at the same time as fluorescence - dammit - old-fashioned
system!)

Plus we use a digital camera system

But the point is well taken - if I stop down the condenser diaphragm, I may
get sufficent contrast for my purposes - I'll try it.

Thanks again,

Jonathan

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                      Jonathan Bard

Anatomy Department, Edinburgh University, Edinburgh EH8 9AG, UK

Telephone (44) (0)131.650.3107    Fax: (44) (0)131.650.6545

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