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Olaf,
Does the period of intensity fluctuation change as a function of scan
speed or scan resolution? Does moving from low zoom to high zoom have
an effect on the pattern? The answers to these questions might help to
diagnose the problem.
Best,
Karl
Olaf Selchow wrote:
> 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] |
> --------------------------------------------
--
Karl Garsha
Light Microscopy Specialist
Imaging Technology Group
Beckman Institute for Advanced Science and Technology
University of Illinois at Urbana-Champaign
405 North Mathews Avenue
Urbana, IL 61801
Office: B650J
Phone: 217.244.6292
Fax: 217.244.6219
Mobile: 217.390.1874
www.itg.uiuc.edu
|
