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Am J Physiol Cell Physiol. 2011 Apr;300(4):C723-42. doi: 10.1152/ajpcell.00462.2010. Epub 2011 Jan 5.

A practical guide to evaluating colocalization in biological microscopy.

Author information

1
Dept. of Medicine, Division of Nephrology, Indiana Univ. Medical Center, Indianapolis, IN 46202, USA. kwdunn@iupui.edu

Abstract

Fluorescence microscopy is one of the most powerful tools for elucidating the cellular functions of proteins and other molecules. In many cases, the function of a molecule can be inferred from its association with specific intracellular compartments or molecular complexes, which is typically determined by comparing the distribution of a fluorescently labeled version of the molecule with that of a second, complementarily labeled probe. Although arguably the most common application of fluorescence microscopy in biomedical research, studies evaluating the "colocalization" of two probes are seldom quantified, despite a diversity of image analysis tools that have been specifically developed for that purpose. Here we provide a guide to analyzing colocalization in cell biology studies, emphasizing practical application of quantitative tools that are now widely available in commercial and free image analysis software.

PMID:
21209361
PMCID:
PMC3074624
DOI:
10.1152/ajpcell.00462.2010
[Indexed for MEDLINE]
Free PMC Article

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