Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Hi Peng,
The Zeiss LSM 510 software says it has 80 mW. That would be split
over two lines, passed through a fiber optic cable, through a
collimator lens assembly, and into the microscope. I do not have a
light meter, so do not know how much reaches the specimen. I usually
use only the 364 nm line, except for days (lenses? specimens?) where
the 364 nm line is very dim.
Best wishes with NADH and FAD imaging.
George
At 09:49 PM 2/5/2008, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi George,
> Nice to hear from you. Do you seriously need 80mW -- when I did
>autofluorescence spectroscopy in Hong Kong I used no more than 1mW on
>sample (355/457nm), otherwise the photobleaching is very huge. I am
>planning to buy a 20mW 375nm laser, it will be bad if I end up with no
>signal. Please help me to double check the power on your 364nm line
>(it is not surprise for an argon laser to have 80mW output on
>488nm/514nm).
> It will be nice if I can use DAPI, but I plan to image the
>metabolic state with autofluorescence of NADH/FAD, through in vivo
>imaging. :) Thank you for reminding me with GFP! Long time ago we have
>an transgenic mouse that have GFP, but I need to know more about it
>since I am not a biologist.. :)
> Today is the Chinese New year, and I wish all of you a very happy 2008!
>
>Peng
>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, George McNamara <[log in to unmask]> 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)
> >
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)
|
