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Well, I've never imaged 'real' lenses, but I did assist with a (very ambitious) PhD project to grow replacement lenses in a culture dish. Not only did we need to image them, we also had to work out an optic system to show that they did have a focus. Amazingly, they did! Not that it was good enough to give anyone very good vision, but as a proof of concept it was incredible. I would just add that if you do have to use a cover slip you may be able to buy 'water' ones with approximately the RI of water. Check the archives, because I do remember that it was discussed here some time ago. Maybe they were from Olympus?
Guy
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Mark Cannell
Sent: Friday, 20 April 2012 5:19 PM
To: [log in to unmask]
Subject: Re: sample prep for intra-occular lenses
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Having imaged real lenses, I think you need to define what you want to image (i.e. low res or high res over just a bit). A 40x dipping lens has several mm of working distance and even with a #0 coverslip in place gives quite good images. You may not be able to reach the entire lens surface, perhaps you should cut it up (carefully) .
Hope this helps.
Mark
On 19/04/2012, at 10:13 PM, Cromey, Douglas W - (dcromey) 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
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> 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
>
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