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Steve Bagley wrote:

> I was wondering which cover slips are in general use for confocal  microscopy or has anyone reviewed cover slips that are commercially  available?
> As it is possible for a user to inadvertently introduce error into a well-corrected system just by selecting the wrong cover slip, I have been instructing people who use the microscopes to utilise the correct thickness of the cover glass (0.17 millimeters).
> Unfortunately with high quality cover glasses having a tolerance of +/- 10 micrometers, consequently the FWHM is detrimentally altered by more than a factor of two if a cover slip is on the extreme of this tolerance.
>
> Are there coverslips available with a finer degree of tolerance than the  standard +/- 10 ?

Dear Steve--

Are you imaging cells grown on coverslips?  If so, then your concern is
well-founded (at least with high-dry objectives).  However, if you're
working with material mounted on a slide, then the mounting material
under the coverglass (plus the thickness of the specimen itself) may
have as large, or larger influence than the thickness of the coverglass
itself.  With that in mind, for my slide-mounted material I always
deliberately use coverslips that are "too thin"--#1 instead of #1-1/2.
On a bad day (i.e. when a thick layer of DPX has been slopped on), I
still need to crank back the compensation collar on my 40x objective.

Good luck!

Martin Wessendorf
--
Martin Wessendorf, Ph.D.                       office:  (612) 626 0145
Assoc Prof, Dept Neuroscience                     lab:  (612) 624 2991
University of Minnesota                 Preferred FAX:  (612) 624 8118
6-145 Jackson Hall, 321 Church St. SE        Dept FAX:  (612) 626 5009
Minneapolis, MN  55455              e-mail:  [log in to unmask]