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Hi Glen,

in our facility we use the LEICA 63x / 1.3 glycerol immersion objective.
The working distance of 300 micrometers is very fine (opposed to 70
micrometers of our standard 63x / 1.32 oil lens). By carefully correcting
cover slip thickness with the correction collar ( using reflection signal
on the slide with xz scan) and matching refractive indices, we can obtain
fine signals (green and red fluorescence) down to a depth of 300 microns (
with PFA-fixed / triton-permeabilized samples, equilibrated in buffer /
glycerol, r.i. 1.45). Of course there are some depth limitations when using
shorter wavelength dyes (e.g DAPI) or with samples still scattering in this
embedding mixture.

If you plan to image samples of this type, I would highly recommend this
lens. I have no experiences with the ZEISS lens yet.

Guenter

Disclaimer: I have no financial interest in any of the products and / or
companies mentioned.

At 18:29 17.05.04, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>On the topic of immersion objectives, has anyone experience with high
>magnification glycerol immersion objectives, in contrast to
>multi-immersion objectives?  the MI units produce beautiful images but
>are only available at a max mag. of 25X (at least from Zeiss).  Has
>anyone used the Zeiss 100X/NA1.2 glycerol objective?  given that the RI
>of brain tissue is closer to glycerol, glycerol immersion seems a more
>reasonable approach than water immersion. A higher mag than 25X would
>be useful for widefield systems using a CCD camera.  A 63X oil
>immersion with a coverslip correction collar might be a more practical
>and affordable alternative, if it existed.
>
>Regards,
>Glen
>
>
>Glen MacDonald
>Core for Communication Research
>Virginia Merrill Bloedel Hearing Research Center
>Box 357923
>University of Washington
>Seattle, WA 98195-7923  USA
>(206) 616-4156
>[log in to unmask]
>
>************************************************************************
>******
>The box said "Requires Windows 95 or better", so I bought a Macintosh.
>************************************************************************
>******

------------------------------------------
Dr. Guenter Giese
Light Microscopy Facility
Dept. of Biomedical Optics, MPI fuer Medizinische Forschung
Jahnstr. 29, D-69120 Heidelberg, Germany
Phone (+49) 6221-486-360 (Fax: -325)
e-mail: [log in to unmask]
http://sun0.mpimf-heidelberg.mpg.de/~ggiese/lightmicro/index.html