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PMID: 21209361 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

A practical guide to evaluating colocalization in biological microscopy.

American journal of physiology. Cell physiology ·Vol. 300 ·No. 4 ·2011-04-00 ·Pages C723-42

Dunn KW, Kamocka MM, McDonald JH

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.

MeSH Terms
Biology/methods Fluorescent Dyes/metabolism Image Interpretation, Computer-Assisted/methods Immunohistochemistry/methods Microscopy, Fluorescence/methods Software
Chemicals
Fluorescent Dyes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Dunn Kenneth W
Dept. of Medicine, Division of Nephrology, Indiana Univ. Medical Center, Indianapolis, IN 46202, USA. kwdunn@iupui.edu
Kamocka Malgorzata M
McDonald John H
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Article Info
Journal
American journal of physiology. Cell physiology
Abbr.
Am J Physiol Cell Physiol
ISSN
1522-1563
DOI
10.1152/ajpcell.00462.2010
Email
kwdunn@iupui.edu
Published
2011-04-00
Epub
2011-00-05
Pages
C723-42
Language
English
Region
United States
NLM ID
100901225
PMCID
PMC3074624
Subset
IM
Grants
NIDDK NIH HHS · DK-51098 · United States
NIDDK NIH HHS · DK-61594 · United States
External Links
PubMed source DOI: 10.1152/ajpcell.00462.2010 PMC Full Text View journal details
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