http://confocal-microscopy-list.275.s1.nabble.com/Changing-system-from-NIR-two-photon-to-UV-tp1694402p1695250.html
> Hi Esteban,
>
> Just a few thoughts, because I've never attempted such a swap of lasers: for
> DAPI, we use a 405 nm laser, and we found no need for real UV. However,
> even with this near-UV line, the light path is separate from the visible
> light paths and the near-UV optics contains correction pinhole lenses (one
> pinhole lens for each objective lens) in order to be able to align the
> visible and near-UV generated confocal images (even a slight misalignment of
> these corrective elements results in a 1 to 5 micron misalignment between
> the two images). We also image nuclei regularly and the light paths have to
> be well aligned if we want to have images of any use, thus I very much doubt
> that you would get away with a simple swap.
> I hope this helps a bit, I'm sure our more experienced list members will
> fill us in with great info soon. Best regards,
>
> Zoltan
>
> On Tue, Dec 23, 2008 at 4:47 PM, G. Esteban Fernandez <
>
[hidden email]> wrote:
>
>> Hello all,
>>
>> On our Zeiss LSM 510 META two-photon system we have a Coherent Chameleon
>> NIR laser that may need major repair. Coherent has been very good in
>> providing remote technical assistance to us but we're at a point where a
>> service visit is required and we're not under contract. Since we do more
>> shallow imaging of UV dyes like DAPI on that system that two-photon imaging,
>> we would replace the NIR laser with UV if it is more cost-effective than
>> repairing the NIR. Zeiss says a switch from NIR to UV would require a major
>> (and pricey) overhaul of the optics. I don't know what that entails but
>> it is understandable given the huge difference in wavelength. However, I
>> wonder how badly out of focus or aberrated the UV image would actually be
>> with the optics that are already in place if we put a UV laser in there in
>> place of the NIR; we do have a UV laser dichroic. Might it be good enough
>> for at least localizing nuclei? (I do know how to do non-confocal UV imaging
>> on the system) Can people advise on an NIR-to-UV swap of lasers?
>>
>> Thanks,
>> Esteban
>>
>> --
>> G. Esteban Fernandez, Ph.D.
>> Associate Director
>> Molecular Cytology Core Facility
>> University of Missouri
>> 120 Bond Life Sciences Center
>> Columbia, MO 65211
>>
>>
http://www.biotech.missouri.edu/mcc/>>
>> 573-882-4895
>> 573-884-9395 fax
>>
>
>
>
> --
>
> Zoltan Cseresnyes
> Facility manager, Imaging Suite
source to the microscope port. After that, you should be good to go and your
objectives should be fine for the imaging. "Normal" fiber optics won't work
for the 405 laser. In addition, collimators or other lenses in this path need
to be UV capable. If this was not the focus of your question, my apologies.