UNIT 4.19 Quantitative Colocalization Analysis of Fluorescence Microscopy Images

  1. Vadim Zinchuk1,
  2. Olga Grossenbacher-Zinchuk2

Published Online: 3 MAR 2014

DOI: 10.1002/0471143030.cb0419s62

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Zinchuk, V. and Grossenbacher-Zinchuk, O. 2014. Quantitative Colocalization Analysis of Fluorescence Microscopy Images. Current Protocols in Cell Biology. 62:4.19:4.19.1–4.19.14.

Author Information

  1. 1

    Department of Neurobiology and Anatomy, Kochi University, Faculty of Medicine, Japan

  2. 2

    Institute of Anatomy, University of Bern, Bern, Switzerland

Publication History

  1. Published Online: 3 MAR 2014


Colocalization is an important finding in many cell biological studies. This unit describes a protocol for quantitative evaluation of fluorescence microscopy images with colocalization based on calculation of a number of specialized coefficients. Images of double-stained sections are first subjected to background correction, and then various coefficients are calculated. Meanings of the coefficients and a guide to interpretation of the results of calculations based on the use of linguistic variables are given. Success in colocalization studies depends on the quality of images analyzed, proper preparation of the images for coefficients calculations, and correct interpretation of results obtained. This protocol helps ensure reliability of colocalization coefficient calculations. Curr. Protoc. Cell Biol. 62:4.19.1-4.19.14. © 2014 by John Wiley & Sons, Inc.


  • quantitative colocalization;
  • fluorescence microscopy;
  • image analysis;
  • data interpretation