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Date: | Mon, 4 Feb 2008 23:48:24 -0500 |
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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
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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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