Judy;

Sorry forgot to attach citation.

Hiraoka Y., Sedat J.W., Agard D.A. (1990) Determination of three-dimensional
imaging properties of a light microscope system: partial confocal behavior
in epifluorescence microscopy. Biophysical Journal 57:325-333.

Regards,

Tom



At 09:01 AM 1/16/97 -0800, you wrote:
>        Dear fellow microscopists,
>
>        Can someone enlighten me on the potential pitfalls of RI mismatch
>between coverslip and immersion oil.  I am working with someone who must
>grow his cells on Aclar coverslips (RI = 1.435).  We are mounting the
>specimens in Vectashield (RI = 1.4577) and are using Zeiss oil (RI =
>1.515). We are trying to do confocal reconstructions of fluorescently
>tagged (FL and RH) mineral aggregates in these cultures (approx. 10-20 um
>thick) to visualize their substructure.  To do this we are using 100X oil
>immersion lens at Zoom 4 on a Leica TCS-NT confocal.    What sorts of
>aberrations could I expect in the reconstructed images??  Should I use an
>oil with a lower RI?
>        A related question:  Most of the fluorescent specimens I work with
>have glass coverslips and are imaged with oil immersion objective lenses
>(consistent RI), but are mounted in Vectashield (Lower RI) or similar
>anti-photobleach medium.  What problems does this pose for confocal
>imaging??
>
>Judy Drazba, Ph.D.  ([log in to unmask])
>Confocal Microscopy Facility, NC-3
>The Cleveland Clinic Foundation
>9500 Euclid Avenue
>Cleveland, OH 44195-5001
>Office (216)445-3760
>FAX    (216)444-7927
>
>
Tom Donnelly                       206-313-4549 tel
DeltaVision Systems                206-557-1055 fax
Applied Precision, Inc.            [log in to unmask]
1040 12th Ave. N.W.                Web Site: http://www.api.com
Issaquah, WA 98027-8929