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Chromatic aberrations are highly objective/specimen dependent. The biggest concern for confocal is longitudinal chromatic aberrations as the pinhole mostly takes care of spherical aberration (by dumping photons).

My 2c

Mark



On 28/02/2014, at 8:27 am, Sergey Tauger <[log in to unmask]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
> 
> Hi Gary,
> 
> For any objective, aberrations are corrected only for first 0.5-1 micrometer and 
> only if you use coverglass of recommended thickness with accuracy of 1%. I.e. 
> if 0.17 cg is recommended, you will get identical images in terms of quality 
> analysis only for cg thickness range 168-172 micrometer. 
> 
> However, chromatic aberrations are negligible if you image not deeper than 3 
> micrometer into specimen and use standart cg, i.e 170 +/- 15. For deeper 
> imaging  you can get false negative\positive in FISH. 
> 
> If you want quantitative result for imaging depth up to 20-30 micrometer, it is 
> better to deconvolve images obtained. My favourite packages are Hyugens 
> Suite (commercial) and COSMOS (opensource), they perform most accurately 
> among the ones I tested.
> 
> Best,
> Sergey Tauger
> 
> PhD student
> Cell motility lab.
> Dept. of Biology
> Moscow State University

Mark  B. Cannell Ph.D. FRSNZ
Professor of Cardiac Cell Biology
School of Physiology &  Pharmacology
Medical Sciences Building
University of Bristol
Bristol
BS8 1TD UK

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