Posted by
Jeremy Adler-4 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Cosmic-ray-particles-centenary-The-Chase-2012-tp7578803p7578828.html
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Comparisons between instruments are difficult and tend of focus on
resolution but one area that tends to be overlooked is noise.
A simple noise correction/measurement that we are pushing to
accurately measure colocalization is to compare two images of the same
specimen - the difference is the noise. This can be visualized either
by subtraction or by plotting the intensities of pixels in the two
images against each other - a standard scattergram. The scattergram
makes the level of noise blindingly obvious and the method could
easily be implemented during acquisition, when users have the chance
to do something about it, but currently it is not available in any
commercial software.
Noise clearly depends on acquisition time but ultimately is limited by
photobleaching - a compeletly bleached image has had all the available
information extracted. So a method of comparing microscopes is to
measure the noise for a similar amount of photobleaching.
Quoting Arvydas Matiukas <
[hidden email]>:
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> To join, leave or search the confocal microscopy listserv, go to:
>
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>
> Dear List,
>
> I am looking for simple and quick test (and sample) to compare
> acquisition across different confocals. Specifically, to our LSM510
> there was recently added Nikon C2 , and users want to know how
> they compare by few practical parameters, e.g. signal sensitivity,
> spectral bleedthrough, bleaching, and maybe resolution .
>
> I am aware of papers by Zucker, Pawley, Cole that describe detail
> evaluation of confocal performance, and even recently measured
> some PSFs. However, I am looking for just simple and quick
> test that would allow direct visual comparison of images (simple
> analysis like getting intensity histogram is fine) acquired on different
> confocals.
>
> Please share your thoughts and/or experience.
>
> Thank you in advance,
> Arvydas
> ***************************
>
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> Department of Pharmacology
> SUNY Upstate Medical University
> 766 Irving Ave., WH 3167
> Syracuse, NY 13210
> tel.: 315-464-7997
> fax: 315-464-8014
> email:
[hidden email]
>
Jeremy Adler
IGP
Rudbeckslaboratoriet
Daghammersköljdsväg 20
751 85 Uppsala
Sweden
0046 (0)18 471 4607