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