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October 2007

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From:
Robert Zucker <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Fri, 5 Oct 2007 17:38:28 -0400
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Craig
The efficiency of detection is much less in the spectral mode due to the
design of the multianode detector. In addition as Kurt Thorn said the
light is split into a number of different bandpass channels instead of
being detected by only one channel which also limits the detection in
each channel.
Nikon compensates for this decreased light by increasing the pixel dwell
time. However photons are photons. With less photons you will get a
noisier image.
The spectral detection of any confocal system using a multianode
detector will not be as good as a PMT designed for good sensitivity and
low light detection. It will produce nosier images but if there is
enough light you will be able to determine a valuable spectrum which can
be used for scientific experiments and to manipulate images. Like
everything with confocal microscopy there is trade-offs and no perfect
system.
Best wishes.
Bob

Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
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             Craig Brideau                                              
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             RV.BUFFALO.EDU>          Re: Nikon C1si?                   
                                                                        
                                                                        
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I was wondering if you saw this decreased throughput in spectral mode
only, or also in conventional detection mode?  The spectral mode works
with a multi-anode PMT, which has a lower quantum efficiency than a
conventional PMT.  From what I have seen of the design, it also has
the option to use a conventional PMT system with the device, and I am
wondering if you have also tried that mode of operation?

Thanks,

Craig

On 10/4/07, Robert Zucker <[log in to unmask]> wrote:
> One of the major limitations that we have seen with this unit and also
> with  the Zeiss meta 510 is the decreased light throughput . This
> creates images that are noisier than conventional confocal
microscopes.
> In our hands it appears you will need to have a bright sample to make
> the spectral system work properly. ,

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