Re: sample prep for intra-occular lenses

Posted by Sylvie Le Guyader on Apr 23, 2012; 8:19am
URL: http://confocal-microscopy-list.275.s1.nabble.com/sample-prep-for-intra-occular-lenses-tp7482155p7491036.html

Hi Doug

Try 1% low melting point agarose in culture medium. It is still liquid at 37 deg so it works
for live cells and you will have no problem with index mismatch.

Med vänlig hälsning / Best regards
 
Sylvie
 
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Sylvie Le Guyader
Live Cell Imaging Unit
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14183 Huddinge
Sweden
office: [phone]
LCI room: [phone]
mobile: [phone]

>
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> > On Thu, 2012-04-19 at 14:13 -0700, 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
> >>
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> >>