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Hi Olaf,

we encounter the same problem on our Mira 900 with BioRad LaserSharp
1.4 - but only when scanning with certain, slower scanning speeds
like 50 lps. Other speeds: no black lines! The lines don't destroy
the acquired information, but are rather annoying. We also could not
figure out a way to solve the problem, the BioRad technicians did not
know as well.

We are using our Mira 900 with a BioRad Setup and a Nikon TE
microscope, probably different from your Leica system. Therefore:
maybe this is not a scope problem, but rather a laser problem.

In addition: no black lines with the BioRad vis lasers on our system,
but we did not test out all possible scanning speeds on the vis
lasers . I would suspect the problem in the software or the beam
controlling unit... but strange anyway.

Hope this information helps...

Cheers, Florian



>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>No, John, the horizontal lines are there with the Mira Laser as well
>as the vis lasers... it'got something to do with the scope I think.
>The best hint I got was that from Ian... I'll have to check the
>galvano-stage for vibrations.
>
>Thanks anyway for the hints on the Mira. I hope I never need them...
>;-) Our Mira is quite stable, actually. And we double check it with
>an oscilloscope.
>
>Olaf
>
>>Search the CONFOCAL archive at
>>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>>Hi all,
>>
>>We don't have the Leica, we have the Zeiss 510 with the Mira 900 (5W
>>pumped). But...
>>
>>If you see the horizontal (black)lines with Mira laser scanning and at
>>no other time, then it could be as simple as adjusting the Mira slightly
>>to make the laser stay in Mode lock.
>>
>>If you see the horizontal brighter/darker (contrasting) lines with Mira
>>laser scanning and at no other time, then it could be as simple as
>>adjusting the Mira slit slightly to remove the lines, (we don't have an
>>oscilloscope, so that is how we tweak the tuning).
>>
>>If your laser is mode locked fine but goes out of mode lock when
>>scanning starts it could be miss alignment in parts belonging to Leica.
>>We recently had this issue with our 510. (Coherent was real helpful with
>>this)
>>
>>If none of the above, turn off the Mira and cool it down along with all
>>its extra parts, including water chiller and any nitrogen purging (takes
>>about 45min) and run the scope on visible lasers checking all the
>>possible PMTs.
>>
>>If you still see the lines then you may have to call Leica.
>>
>>Hope this helps
>>
>>Jon Ekman
>>Florida State University
>>
>>On Fri, 2003-08-08 at 08:12, Olaf Selchow wrote:
>>>  Search the CONFOCAL archive at
>>>  http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>>
>>>  Hello confocal colleagues...
>>>
>>>  I have a problem with our Leica SP2 system: The images show thin
>>>  dark, horizontal stripes. Sometimes more and sometimes less. I
>>>  couldn't figure out yet when the stripes are stronger and when not.
>>>  Sometimes they seem to have gone completely. It looks a little like a
>>>  phase-problem with the line scans. But I couldn't fix it by adjusting
>>>  the phase-correction.
>>>
>>>  One more hint I got is the following: The stripes were seen the first
>>>  time after the 2Photon system (IR laser Coherent Mira 900, EOM and
>>>  all the extra bits and pieces) has been added to the standard SP2.
>>>  Unfortunately this has been before I joined the lab. So I can not
>>>  confirm this myself. Before that the images were fine (that's what
>>>  the older colleagues here in the lab say).
>>>
>>>  Independent from the 2P system I thought that the stripes could come
>>>  from some noise on the electricity supply. We're a big institute and
>>>  I don't know what other equipment is used in the building... and as I
>>>  said I couldn't correlate it with any external parameter like
>>>  daytime, weekday or something else.
>>>
>>>  The images are sufficiently good quality to interpret. But they are
>>>  no good for publication....
>>>
>>>  Has anybody had such a problem before and/or have an idea about how
>>>  to solve it?
>>>  I'd like to avaid to call (and pay) the Leica service when it's maybe
>>  > just a very easy thing that I can do myself.
>>  >
>>  > Thanks in advance for your help!
>>  >
>>  > Olaf
>>  >
>>  > --
>>  >
>>  >   --------------------------------------------
>>>  | Dr. Olaf Selchow                           |
>>>  | SFB 495 Z1 - Central Microscopy Facility   |
>>>  | Institute for Cell Biology und Immunology  |
>>>  | University of Stuttgart                    |
>>>  | Allmandring 31, 70569 Stuttgart, Germany   |
>>>  | T: +49 711 685 8209, Fax: +49 711 685 7484 |
>>>  | email: [log in to unmask]    |
>>>    --------------------------------------------
>>--
>>Jon Ekman <[log in to unmask]>
>
>
>--
>
>  --------------------------------------------
>| Dr. Olaf Selchow                           |
>| SFB 495 Z1 - Central Microscopy Facility   |
>| Institute for Cell Biology und Immunology  |
>| University of Stuttgart                    |
>| Allmandring 31, 70569 Stuttgart, Germany   |
>| T: +49 711 685 8209, Fax: +49 711 685 7484 |
>| email: [log in to unmask]    |
>  --------------------------------------------


--


----------------------------------------------------------------------------------------------
Florian Ulrich
Heisenberg Lab
Max-Planck-Institute for Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108
01307 Dresden
Germany
phone: (+49) 351 210 2689
fax:    (+49) 351 210 1489
email: [log in to unmask]