A guided tour into subcellular colocalization analysis in light microscopy.

S. BOLTE, F. P. CORDELIÈRES
J Microsc. 2006-12-01; 224(3): 213-232
DOI: 10.1111/j.1365-2818.2006.01706.x

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1. J Microsc. 2006 Dec;224(Pt 3):213-32.

A guided tour into subcellular colocalization analysis in light microscopy.

Bolte S(1), Cordelières FP.

Author information:
(1)Plateforme d’Imagerie et de Biologie Cellulaire, IFR 87 la Plante et son
Environnement, Institut des Sciences du Végétal, Avenue de la Terrasse, 91198
Gif-sur-Yvette Cedex, France.

Comment in
J Microsc. 2007 Jul;227(Pt 1):83; author reply 84-5.

It is generally accepted that the functional compartmentalization of eukaryotic
cells is reflected by the differential occurrence of proteins in their
compartments. The location and physiological function of a protein are closely
related; local information of a protein is thus crucial to understanding its role
in biological processes. The visualization of proteins residing on intracellular
structures by fluorescence microscopy has become a routine approach in cell
biology and is increasingly used to assess their colocalization with
well-characterized markers. However, image-analysis methods for colocalization
studies are a field of contention and enigma. We have therefore undertaken to
review the most currently used colocalization analysis methods, introducing the
basic optical concepts important for image acquisition and subsequent analysis.
We provide a summary of practical tips for image acquisition and treatment that
should precede proper colocalization analysis. Furthermore, we discuss the
application and feasibility of colocalization tools for various biological
colocalization situations and discuss their respective strengths and weaknesses.
We have created a novel toolbox for subcellular colocalization analysis under
ImageJ, named JACoP, that integrates current global statistic methods and a novel
object-based approach.

DOI: 10.1111/j.1365-2818.2006.01706.x
PMID: 17210054

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