Re: Configuration problem with Zeiss LSM 510

Posted by G. Esteban Fernandez on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Configuration-problem-with-Zeiss-LSM-510-tp2303218p2303330.html

You need to go into the Maintain > Pinhole menu and optimize the X/Y positions of pinhole 3.  The message means that has never been done for that particular configuration of dichroics.  The pinhole is probably not centered directly in front of the PMT so you don't get a good image if it is closed down, but you'll probably get an image if you open the pinhole to max.

 

-Esteban

 

On Tue, Feb 10, 2009 at 9:54 AM, yee z <[hidden email]> wrote:

Dear listers,   We have a Zeiss LSM510 confocal microscopy system at our core facility. And I got some problems while writing new configuration. Here is an example: Single track configuartion for FITC, I used to use HFT 405/488/543/633,  NFT 545, NFT 490 and BP505-550. It works fine.  (Screen shot: http://i561.photobucket.com/albums/ss60/mizhzmkm/Q-1A.jpg)   But when I try to change the primary dichroic beam splitter to HFT 488/594 and keep everything else the same, the systme shows "Pinhole PH3 is not adjusted". (Screen shot: http://i561.photobucket.com/albums/ss60/mizhzmkm/Q-1B.jpg)    Also if I change the NFT 545 to NFT 595, the system also shows "Pinhole PH3 is not adjusted" (Screen shot: http://i561.photobucket.com/albums/ss60/mizhzmkm/Q-1C.jpg) I am still be able to take the images regardless the error window.   So does anyone here have the same problem? What does "Pinhole PH3 is not adjusted" mean, any reason for this?   Any suggestion will be appreciated.   Thanks,   Yi

--
G. Esteban Fernandez, Ph.D.
Associate Director
Molecular Cytology Core Facility
University of Missouri
120 Bond Life Sciences Center
Columbia, MO  65211

http://www.biotech.missouri.edu/mcc/

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