http://confocal-microscopy-list.275.s1.nabble.com/405-nm-laser-via-NIR-port-on-Zeiss-510-tp6345367p6346333.html
We do "DAPI piping" via standard HBO (!) . Drawback: Pinhole has to be at max to collect enough signal, so it s not so much confocal
Advantage: Cheap (zero costs), efficient, good enough for recognizing nuclei , and it can be done in seconds without any installations
Whoops! Got mixed up regarding what laser needed to be brought in: the 405 can be piped in via fiber but it will be fairly important that the fiber is single-mode at 405nm.
> The primary dichroic is typically the largest problem. Our Nikon
> technician went through some fairly heroic efforts to install a
> dichroic into an A1 FN1. We had the filter custom made at significant
> expense for our ultrabroadband Ti:Saph laser (Octavius), so the
> possibility of damaging the filter during installation was a critical
> concern. After the technician partly disassembled the scan head in
> our lab, we ended up drafting a nice lady with very tiny fingers who
> did needlepoint and crocheting as a hobby to finally get the filter
> into place in the A1 scan head. This was something normally done at
> the factory... That said, it did work out for us and Nikon was very accommodating in helping us with the modification.
> Regarding piping the laser in to the Zeiss scope: I was assuming your
> laser could be brought in free space. If you have to go through a
> fiber then you probably won't be able to do it due to dispersion in
> the glass fiber. If you have a prechirping unit then you *might* be
> able to pull it off, but it will be tricky.
>
> Craig
>
>
>
> On Mon, May 9, 2011 at 4:06 PM, Armstrong, Brian <
[hidden email]>wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>> *****
>>
>> Craig makes a good point. The HFT700/488 or KP650 (for example) that
>> you have been using for your 2P laser will not work for a 405 line.
>> Therefore you would need to change the dichroics. This is not done in
>> the field and you would need to send your scan-head to the factory in
>> Germany. You would also need to think about how to get your laser
>> into the fiber (AOTF?), how to adjust the power (AOM is no longer
>> useful), and whether or not you have the secondary dichroics that will pass the emitted light.
>>
>> You should talk to Zeiss before considering this idea any further.
>>
>> Cheers,
>>
>>
>> Brian D Armstrong PhD
>> Assistant Research Professor
>> Light Microscopy Core
>> Beckman Research Institute
>> City of Hope
>> Dept of Neuroscience
>> 1450 E Duarte Rd
>> Duarte, CA 91010
>> 626-256-4673 x62872
>>
>>
>>
http://www.cityofhope.org/research/support/Light-Microscopy-Digital-I>> maging/Pages/default.aspx
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:
[hidden email]]
>> On Behalf Of Craig Brideau
>> Sent: Monday, May 09, 2011 2:52 PM
>> To:
[hidden email]
>> Subject: Re: 405 nm laser via NIR port on Zeiss 510
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>> *****
>>
>> For starters, what do you have for a Dichroic that could pipe in the
>> 405 and let the rest of the visible pass? Most 2p dichroics split
>> off at around
>> 680
>> or 700nm to allow the IR to pass and the emission light to reflect,
>> or vice-versa.
>>
>> Craig
>>
>>
>> On Mon, May 9, 2011 at 1:16 PM, Phillips, Thomas E.
>> <
[hidden email]>wrote:
>>
>> > *****
>> > To join, leave or search the confocal microscopy listserv, go to:
>> >
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>> > *****
>> >
>> > The NIR laser for two photon excitation on our Zeiss 510 died on
>> > us. The repair would be at least 60K. We are trying to find funds
>> > for a new
>> confocal
>> > so I don't know if it is worth repairing. The biggest practical
>> > problem
>> is
>> > that we can't do DAPI excitation on our routine preps. Has anyone
>> > tried retrofitting a 405 nm laser through the NIR port? Are there
>> > optical components/mirrors in the pathway that only work with
>> > longer
>> wavelengths?
>> > Any thoughts on this are welcome. Tom
>> >
>> >
>> > Thomas E. Phillips, Ph.D
>> > Professor of Biological Sciences
>> > Director, Molecular Cytology Core
>> > 2 Tucker Hall
>> > University of Missouri
>> > Columbia, MO 65211-7400
>> > 573-882-4712 (office)
>> > 573-882-0123 (fax)
>> >
[hidden email]<mailto:
[hidden email]>
>> >
>> >
http://www.biology.missouri.edu/faculty/phillips.html>> >
http://www.biotech.missouri.edu/mcc/>> >
>>
>>
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