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On Mon, 16 Jul 2007, Sven Terclavers wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> I would suggest to use a polarization filter on a wide-field microscope.
> We've been using polarizing microscopy always for collagen staining, since
> for analysis it's important to distinguish the different types of collagen
> by color.  Confocal microscopy will be a very hard job (if not as good as
> impossible?) to obtain nice images of these staining, but a good polarizing
> filter will give you an amazing image of the Masson staining, which can be
> than easily (and nicely) merged with a normal image of the ammonium-silver
> staining.
>
> Sven Terclavers


One thing I did on a lark many years ago was the following:

Use eosin-stained block (which has gangbusters flourescence) to get
a single slice confocal image at very high resolution.

Take a pic using standard staining from a wide-field microscope.

Map the color from the wide-field microscope image to the structure in
the confocal image.  This was some years ago, but one problem I remember
running into was that there were areas much lighter or without structure
in the confocal slice where there was staining in the thick slice.
So (once again, this is from memory), I ended up not placing a direct
color map, but mapping only hue, or something like that.  I think I
also made an image using the confocal data as a bump map onto the
widefield image, which made a neat background image for my laptop :-)

The result was, for me, what looked like an extraordinarily high
resolution wide field image.

Of course, I was doing it because I thought it was pretty, not for
analysis purposes -- you'd have to justify any numbers or conclusions
you derived from the process...


billo