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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
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(Analytical Imaging Core Facility)