Posted by
Cromey, Douglas W - (dcromey) on
URL: http://confocal-microscopy-list.275.s1.nabble.com/sample-prep-for-intra-occular-lenses-tp7482155p7482215.html
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:
[hidden email]
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:
[hidden email]] On Behalf Of Joel B. Sheffield
Sent: Thursday, April 19, 2012 1:56 PM
To:
[hidden email]
Subject: Re: sample prep for intra-occular lenses
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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) <
[hidden email]> 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:
[hidden email]
> 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:
[hidden email]
URL:
http://astro.temple.edu/~jbs