Posted by
Michael Weber-4 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/FW-A1-tp591961p591963.html
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi David,
the system measures the amount of fluorescence signal per pixel and
controls the AOTF for this pixel based on the measured value. So if
there is no light coming to the detector, laser will be restricted from
illuminating the sample. Or, if the detector gets close to saturation,
laser will be restricted as well. This should reduce photobleaching and
phototoxicity also in out-of-focus areas. That's at least how I
understand this.
"The CLEM control unit is an optional add-on system for the C1. The CLEM
unit calculates the integrated detected signal and exposure time for
each pixel in real time, performs AOM high speed shutter control and
high speed operation processing based on the amount of acquired
fluorescence signal, PMT gain and then outputs the resulting
fluorescence signal to the C1 controller’s line grabber electronics
circuit."
source:
http://www.nikoninstruments.eu/news.php?n=550I expect the quantification (measuring intensities) to be difficult
afterwards, since one needs to apply a correction. However, it's one of
the main things that makes the Nikon system interesting.
Michael
David Knecht wrote:
> Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>> The ability of using this system along with Controlled Light Exposure
>> Microscopy (CLEM) makes this system for long time live cell imaging and
>> confocal analysis. CLEM automatically monitors and varying the laser
>> illumination during time-lapse studies to reduce the risk of laser induced
>> bleaching, biochemical inconsistency and cell death.
>
> Can you clarify what this means? I don't see how you can monitor the
> imaging and vary the laser in a way that can avoid cell toxicity.
> Typically, you don't want the laser intensity to vary. Do you have any
> quantitative data on cellular toxicity during imaging in comparison to a
> spinning disk system?
>
> Dr. David Knecht
> Department of Molecular and Cell Biology
> Co-head Flow Cytometry and Confocal Microscopy Facility
> U-3125
> 91 N. Eagleville Rd.
> University of Connecticut
> Storrs, CT 06269
> 860-486-2200
> 860-486-4331 (fax)