Experimenters’ guide to colocalization studies: finding a way through indicators and quantifiers, in practice.
Methods in Cell Biology. 2014-01-01; : 395-408
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1. Methods Cell Biol. 2014;123:395-408. doi: 10.1016/B978-0-12-420138-5.00021-5.
Experimenters’ guide to colocalization studies: finding a way through indicators
and quantifiers, in practice.
Cordelières FP(1), Bolte S(2).
(1)Bordeaux Imaging Center, UMS 3420 CNRS-Université Bordeaux Segalen-US4 INSERM,
Pôle d’imagerie photonique, Institut François Magendie, Bordeaux Cedex, France.
(2)Sorbonne Universités-UPMC Univ Paris 06, Institut de Biologie Paris-Seine-CNRS
FR 3631, Cellular Imaging Facility, Paris Cedex, France.
Multicolor fluorescence microscopy helps to define the local interplay of
subcellular components in cell biological experiments. The analysis of spatial
coincidence of two or more markers is a first step in investigating the potential
interactions of molecular actors. Colocalization studies rely on image
preprocessing and further analysis; however, they are limited by optical
resolution. Once those limitations are taken into account, characterization might
be performed. In this review, we discuss two types of parameters that are aimed
at evaluating colocalization, which are indicators and quantifiers. Indicators
evaluate signal coincidence over a predefined scale, while quantifiers provide an
absolute measurement. As the image is both a collection of intensities and a
collection of objects, both approaches are applicable. Most of the available
image processing software include various colocalization options; however,
guidance for the choice of the appropriate method is rarely proposed. In this
review, we provide the reader with a basic description of the available
colocalization approaches, proposing a guideline for their use, either alone or
© 2014 Elsevier Inc. All rights reserved.
PMID: 24974039 [Indexed for MEDLINE]