UNIT 4.19 Quantitative Colocalization Analysis of Confocal Fluorescence Microscopy Images

  1. Vadim Zinchuk1,
  2. Olga Grossenbacher-Zinchuk2

Published Online: 1 SEP 2011

DOI: 10.1002/0471143030.cb0419s52

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Zinchuk, V. and Grossenbacher-Zinchuk, O. 2011. Quantitative Colocalization Analysis of Confocal Fluorescence Microscopy Images. Current Protocols in Cell Biology. 52:4.19:4.16.1–4.16.19.

Author Information

  1. 1

    Department of Anatomy and Cell Biology, Kochi University, Faculty of Medicine, Japan

  2. 2

    Institute of Anatomy, University of Bern, Bern, Switzerland

Publication History

  1. Published Online: 1 SEP 2011
  2. Published Print: SEP 2011

This is not the most recent version of the article. View current version (3 MAR 2014)


Colocalization is an important finding in many cell biological studies. This unit describes a protocol for quantitative evaluation of images with colocalization based on the calculation of a number of specialized coefficients. First, images of double-stained sections are subjected to background correction. Then, various coefficients are calculated. Meanings of the coefficients and a guide to interpretation of their results indicating either presence or absence of colocalization are given. Success in colocalization studies depends on the quality of analyzed images, proper preparation of the images for coefficients calculations, and correct interpretation of obtained results. This protocol helps to ensure reliability of colocalization coefficients calculations. Curr. Protoc. Cell Biol. 52:4.16.1-4.16.16. © 2011 by John Wiley & Sons, Inc.


  • quantitative colocalization;
  • confocal fluorescence microscopy;
  • image analysis