title: A practical guide to evaluating colocalization in biological microscopy.
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.
authors.affiliation: Dept. of Medicine, Division of Nephrology, Indiana Univ. Medical Center, Indianapolis, IN 46202, USA. kwdunn@iupui.edu
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Dunn
authors.forename: Malgorzata M
authors.initials: MM
authors.lastname: Kamocka
authors.forename: John H
authors.initials: JH
authors.lastname: McDonald
mesh_terms: D001695:Biology / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D012984:Software
chemical_list: D005456:Fluorescent Dyes