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Dear Olaf,

After a second careful inspection of your images I came to
the conclusion that the observed stripe pattern is most
probably caused by mechanical vibration and not light
modulation or electrical interference. My conclusion is
based on the observation of ripples on the left and right
edges of your fluorescent cell. That is, the cell's edge has
been displaced in x and/or z direction during every other
scan sweep. So I would agree with Ian's suggestion to check
Leica's Galvo-Z-stage and x-y stage.

Cheers,

Franco

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]    |
>  --------------------------------------------
>

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