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Hi, Joel

Have recently undergone this operation, this question is near and 
dear to my heart!

The ultimate answer to your question rests in what they actually want 
to see.  However, in general, this sounds like one of those 
situations perfectly suited to a confocal serial section with 3D 
reconstruction.  We had a situation years ago when I was working for 
Sarastro in which a researcher wanted to image the whole cornea.  In 
his case, the most useful information came from a Z 
section.  However, more recently, when I went to the surgeon, he did 
just regular imaging to look at the cells on my cornea.. a situation 
which is very analogous to your problem.

I agree that mounting the lens cell side toward the objective makes 
the most sense.  Also, mounting in Saline (or eye drops?) will reduce 
the spherical aberration.

Good hunting! .... and let us know how it turns out!

Barbara Foster, President and Sr. Consultant
Microscopy/Microscopy Education   P: (972)924-5310   W: 
www.MicroscopyEducation.com

We are now scheduling courses through August 2012... and don't forget 
to take part in our most recent surveys.  Check 
MicroscopyEducation.com for details.

At 07:27 PM 4/19/2012, you wrote:
>Joel,
>
>Today was my first experience with the lenses.
>
>As I understand it, these are the implantable lenses used to fix 
>cataracts.  After a time in the eye, cells grow (fairly flat cells 
>in a layer that appears to be 2-3 cells thick) on the anterior 
>surface (facing out of the eye).  They mount the lens so that the 
>anterior surface is closest to the coverslip.  While I suppose 
>slicing the lens might be an option, my suspicion is that the 
>surface will always be curved in relation to the flat focal plane of 
>the confocal.
>
>The curved surface itself doesn't bother me a whole lot, but I would 
>like to think that we can nail down the sample prep to something 
>slightly less kludge-y that would give the best images 
>possible.  Based on today's observations, the part of the sample 
>that is farther away from the coverslip has less detail, most likely 
>due to spherical aberration (Thanks Jim P!) because we are imaging 
>into an aqueous solution that is beginning to be a good distance 
>from the coverslip (due to lens curvature).  I should add that my 
>objective lens options are 20x/0.7 dry, 40x/1.25 oil and 63x/1.4 
>oil.  No water lenses on this particular confocal.  I used the 20x 
>today, mostly for "field of view" reasons.
>
>Doug
>
>^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
>Douglas W. Cromey, M.S. - Assistant Scientific Investigator
>Dept. of Cellular & Molecular Medicine, University of Arizona
>1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA
>
>office:  AHSC 4212         email: [log in to unmask]
>voice:  520-626-2824       fax:  520-626-2097
>
>http://swehsc.pharmacy.arizona.edu/exppath/
>Home of: "Microscopy and Imaging Resources on the WWW"
>
>
>-----Original Message-----
>From: Confocal Microscopy List 
>[mailto:[log in to unmask]] On Behalf Of Joel B. Sheffield
>Sent: Thursday, April 19, 2012 1:56 PM
>To: [log in to unmask]
>Subject: Re: sample prep for intra-occular lenses
>
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>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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>
>This is an interesting puzzle.   Those people who are working with curved
>surfaces, such as retinas, make a series of radial cuts that allow 
>them to make flat(er) mounts.  Of course, in this case, I would 
>imagine that the lenses are also of variable thickness, so there 
>might be an additional complication.  You should be sure that the 
>cellular surface is closest to the cover slip so that the layer of 
>cells is more uniform.  Do you know on which surface of the lens the 
>cells will be found? Or are they on both?
>
>Joel
>
>
>On Thu, Apr 19, 2012 at 4:41 PM, Cromey, Douglas W - (dcromey) < 
>[log in to unmask]> wrote:
>
> > I have a new set of users for our inverted confocal that will be
> > bringing intra-occular lenses (of the kind used to fix cateracts).
> > The lenses have a thin layer of cells on the surface, which is curved.
> > Right now they are bringing them mounted in buffer as a whole mount
> > with wax to seal the coverslip edges.  They try to compress the lens
> > by putting the slide under a book after it's mounted.
> >
> > It seems like there ought to be a better way and I'm hoping that
> > someone out there has dealt with this before.
> >
> > Thanks!
> > Doug
> >
> > ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
> > Douglas W. Cromey, M.S. - Assistant Scientific Investigator Dept. of
> > Cellular & Molecular Medicine, University of Arizona
> > 1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA
> >
> > office:  AHSC 4212         email: [log in to unmask]
> > voice:  520-626-2824       fax:  520-626-2097
> >
> > http://swehsc.pharmacy.arizona.edu/exppath/
> > Home of: "Microscopy and Imaging Resources on the WWW"
> >
> >
> >
>
>
>--
>
>
>Joel B. Sheffield, Ph.D
>Department of Biology
>Temple University
>Philadelphia, PA 19122
>Voice: 215 204 8839
>e-mail: [log in to unmask]
>URL:  http://astro.temple.edu/~jbs