A practical guide to evaluating colocalization in biological microscopy

作者：Kenneth W. Dunn, Malgorzata M. Kamocka, John H. McDonald · 发表于：American Journal of Physiology-Cell Physiology · 年份：2011 · DOI：10.1152/ajpcell.00462.2010 · 被引用次数：2125 · 研究领域：Advanced Fluorescence Microscopy Techniques、Cell Image Analysis Techniques、Advanced Biosensing Techniques and Applications

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.