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)