Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Peng,
We have an Ar UV laser on our MRC1024 which is coupled through the side port
of a Nikon TE300 microscope (actually into the scanhead which is at the
sideport). This is (was) standard Bio-Rad design, so going through the
sideport should be no problem. I haven't actually measured the light losses
though ...
Best regards,
Kevin
Kevin Braeckmans, Ph.D.
Lab. General Biochemistry and Physical Pharmacy
Ghent University
Harelbekestraat 72
9000 Ghent
Belgium
Tel: +32 (0)9 264.80.78
Fax: +32 (0)9 264.81.89
> -----Oorspronkelijk bericht-----
> Van: Confocal Microscopy List [mailto:[log in to unmask]]
> Namens Peng Xi
> Verzonden: woensdag 6 februari 2008 3:38
> Aan: [log in to unmask]
> Onderwerp: Re: 350nm laser delivered into a microscope
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi Michael,
> Thank you for your reply! I think if it is heavily absorbed when
> using the side port, I'll use the mercury lamp port. I have
> successfullly modified a Nikon TE200 to a two-photon microscope with
> the mercury lamp port, to minimize the dispersion from the reflective
> prism. But still, I think it is much convenient to use the side-port
> for my confocal construction, if the absorption is not an issue... :)
> Thank you!
>
> Best regards,
> Peng Xi
> Associate Professor
> Institute for Laser Medicine and Biophotonics
> Shanghai Jiao Tong University
> 800 Dongchuan Rd.
> Shanghai 200240, China
> Tel: (86) 21-3420-4076
> http://biophotonics.sjtu.edu.cn/
>
>
>
> On 2/5/08, Michael Weber <[log in to unmask]> wrote:
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > Hey Peng,
> >
> > another way of bringing the laser to the object plane would be not
> > through the microscope ports, but directly below the objective via a
> > dichromatic mirror for UV. For example people in Czech Republic
> (Brno)
> > build a little laser coupling device that one can screw between
> > objective and turret. This way you can get around the spectral
> response
> > issues with the microscope prisms.
> >
> > Direct coupling of the laser or scanning system to the microscope
> would
> > allow you to work around fiber coupling issues. Then you probably
> need
> > to use existing ports - maybe one can change the prisms to good
> > dichromatic mirrors?
> >
> > ---
> >
> > Beside that, I would also be interested in news about 350nm diode
> > lasers. We have a setup with a (nearly dead) Enterprise laser and
> where
> > looking for diode lasers to get rid of the heavy and hot beast. But
> the
> > information we got so far from companies and independent people was
> that
> > the current generation of UV diode lasers has too low quality (long-
> term
> > stability, beam shape etc.) for setups like confocals.
> >
> > cheers,
> > Michael
> >
> >
> > George McNamara wrote:
> > > Search the CONFOCAL archive at
> > > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> > >
> > > Hi Peng,
> > >
> > > I estimate between 5 and 10% of confocal systems have a 351/364 nm
> laser
> > > like the 80 mW Coherent Enterprise on the Zeiss LSM 510 in our core
> (I
> > > suspect not a whole lot of the 80 ends up at the specimen for most
> > > objective lenses. Our 63x/1.4 NA lens is downright dim). We have an
> > > Axiovert 200M with the scanhead on the side port. Inside the scan
> head's
> > > two fiber input ports are collimator lenses to adjust the focus for
> UV
> > > and visible light respectively (or vis and IR for the 510
> multiphoton
> > > systems). Keep an eye on http://www.zeiss.com/sensitivity for the
> next
> > > generation.
> > >
> > > My experience has been that most autofluorescence is pretty similar
> from
> > > UV through green excitation (elastin being a partial exception). In
> the
> > > past I found that a CFP filter set worked nicely to estimate the
> > > autofluorescence component of a GFP set (both off the shelf Chroma
> > > filters).
> > >
> > > I recommend you focus more on the 364 nm laser line, if you are
> using a
> > > UV Argon ion laser that has both. I also recommend using UV to
> image
> > > moderate to high concentration DAPI, rather than low SNR dim
> > > autofluorescence.
> > >
> > > If you are using a solid state laser for 350 nm, please let me know
> what
> > > kind so I can consider replacing my Enterprise.
> > >
> > > George
> > >
> > >
> > > At 07:51 AM 2/4/2008, you wrote:
> > >> Search the CONFOCAL archive at
> > >> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> > >>
> > >> Hi,
> > >> Has anyone have the experience of delivering 350nm laser in to
> a
> > >> microscope? I'm planning to use this line for autofluorescence
> > >> excitation with a Nikon microscope, I'll use the side-port to
> deliver
> > >> the excitation to construct a custom-made confocal setup, but I
> don't
> > >> know the absorption ratio. Currently I'm worrying about the
> absorption
> > >> from the reflective prism; if the absorption is very large I'll
> > >> consider to use the back port.
> > >> Thank you for any input!
> > >>
> > >> Best,
> > >> Peng Xi
> > >> Associate Professor
> > >> Institute for Laser Medicine and Biophotonics
> > >> Shanghai Jiao Tong University
> > >> 800 Dongchuan Rd.
> > >> Shanghai 200240, China
> > >> Tel: (86) 21-3420-4076
> > >> http://biophotonics.sjtu.edu.cn/
> > >
> > >
> > >
> > >
> > >
> > >
> > > George McNamara, Ph.D.
> > > University of Miami, Miller School of Medicine
> > > Image Core
> > > Miami, FL 33010
> > > [log in to unmask]
> > > [log in to unmask]
> > > 305-243-8436 office
> > > http://home.earthlink.net/~pubspectra/
> > > http://home.earthlink.net/~geomcnamara/
> > > http://www.sylvester.org/research/SR_lab_analytical.asp?ana=desc
> > > (Analytical Imaging Core Facility)
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