Re: Leica resonant scanner-live for cell imaging

Posted by Martin Seem on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Leica-resonant-scanner-live-for-cell-imaging-tp590925p590933.html

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Hi Jim,

we tested the PE Ultraview at to separate occasions. Once with a normal
ORCA CCD-camera at the Molecular Imaging Center (MIC) in Bergen
(Norway) and once with an EM CCD camera at PEs training facility at
Seer Green right outside London. The VisiTech QLC100 was a CSU10 based
model equipped with an ORCA CCD-camera while the Andor system used an
EM CCD camera.

Thanks,
Martin
------------------------------------------------------------------------

Martin Seem
Graduate student
Group for Cell and Molecular Biology
Norwegian University of Science and Technology




Quoting James Pawley <[hidden email]>:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> >Search the CONFOCAL archive at
> >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> >Hi Benedikt,
> >
> >we acquired a Leica resonant scanner based confocal about 6 months
> >ago. Before buying this system we tested different types of
> >microscopes including three spinning disc based systems (Andor, PE
> >and Visitec QLC100), two point-scan based systems (Leica and Visitec
> >Eye) and the new Zeiss LSM5 Live system. We brought our own samples
> >(transgenic plant material) and used these to test the different
> >systems in terms of speed, photobleaching and overall image quality.
> >
> >We found the spinning disc based systems to be slightly faster than
> >the Leica system especially when acquiring square images (e.g.
> >512x512 pixels). However the speed of the Leica system increases
> >significantly when you reduce the number of lines in the Y
> >direction. You should have no problem acquiring 10 fps with a
> >resolution of 512x512 pixels resolution and about 15-20 fps with a
> >resolution of 512x128 pixels.
> >
> >In our experience the level of photobleaching is quite low when
> >using the Leica system. I would say it is comparable to the spinning
> >disc systems we tested and significantly lower than the Visitec Eye
> >system that bleached my sample (transgenic GFP5-ER plants) within
> >seconds (this system did however work nicely for calcium imaging in
> >dendrites so the level of photobleaching depends on the properties
> >of the fluorechrome).
> >
> >In the end we chose the Leica system because it was sufficiently
> >fast, had an adjustable and round pinhole making it fully confocal,
> >had low levels of photobleaching, had a very flexible bandpass
> >filter solution and could easily and relatively cheaply be upgraded
> >to a tandem scanner system (both resonant and conventional scanner
> >in the same system).
> >
> >
> >Best regards
> >Martin Seem
>
>-------------------------------------------------------------------------
> >
> >Martin Seem
> >Graduate student
> >Group for Cell and Molecular Biology
> >Norwegian University of Science and Technology
>
> Thank you for this Martin,
>
> Could you please tell us about the type of CCD cameras used with the
> various disk-scanners? EM-CCD or normal CCD?
>
> Thanks,
>
> Jim P.
> --
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