Huygens Colocalization Analyzer

With unique 3D visualization

Huygens Colocalization Analyzer produces all the common colocalization coefficients for multichannel images. Colocalization is visually presented in a unique surface map projected on a MIP of the concerned channels. The analysis can be focused on a specific region of interest (ROI), and is more reliable when preceded by Huygens deconvolution, correction for hot/cold pixels, crosstalk, and chromatic aberration correction.

Why Huygens?

11 colocalization coefficients available

Huygens Deconvolution for more reliable results

Various thresholding options (incl. Costes)

Region of Interest tools

Unique 3D map to visualize colocalization

Object analysis for investigating local colocalization

Mis12 at kinetochore complex | Pearson = 0.458

3D widefield image of a U2OS cell. Inset: Colocalization Analyzer-produced iso-surfaces (yellow) from pixels contributing 8% to the total Pearson colocalization (0.458) between labeled Mis12 (green) and Aurora B kinase (red). Huygens Deconvolution and Chromatic Aberration Correction was used for improving the reliability of the analysis. Data from Dr. Livio Kleij en Martijn Vroomans, UMC Utrecht, The Netherlands.

Synaptic protein at dendritic spines | Overlap = 0.281

Coloc Analysis Neuron
Colocalization Analyzer-produced colocalization map (left, yellow) shows regions of colocalization between a synaptic protein (green) and a dendrite (red) at the dendritic spine heads. 3D confocal image of brain tissue was deconvolved with Huygens before analysis. Data from dr. Nawal Zabouri and Mariska Willemsen, Netherlands Institute for Neuroscience.

Coefficients in one click

Find all common colocalization coefficients in just one click.

3D iso-colocalization map

Automatically create isosurfaces showing the regions of colocalization.

Step-by-step guidance

The Colocalization Wizard guides beginners through the colocalization analysis step by step.


Everything is working smoothly. The colocalisation module is very good, the best I have tested so far.

Prof. Miguel Angel Peñalva Soto, group leader at Centro de Investigaciones Biológicas, Madrid, Spain.
I previously used ImageJ for Colocalization Analysis, but I actually find Huygens much more convenient and easier to use, moreover, it is much faster and the results are more consistent [..].

Merel Stiekema, Dept. of Molecular Cell Biology, University of Maastricht, The Netherlands.

Everything for reliable colocalization analysis

Intuitive Interface


Use in research

Berndt-Paetz et al. IC/BPS‐associated alterations of M2 and M3 muscarinic acetylcholine receptor trafficking in human detrusor.
Huygens was used for deconvolution, colocalization, and object analysis Neurourology & Urodynamics 2019; 1-10.

Stewart DJ et al. (2019) COMMD1 and PtdIns(4,5)P2 interaction maintain ATP7B copper transporter trafficking fidelity in HepG2 cells.
Colocalization is described in high detail as performed with Huygens Colocalization Analyzer
Journal of Cell Science 132: jcs231753.

For more, see Scientific Publications

Multi-channel images often suffer from chromatic aberrations and crosstalk (a.k.a. bleed-through), which should be corected before performing any quantitative analysis.

Chromatic Aberration Corrector Crosstalk corrector

More information

Colocalization Coefficients
Colocalization Basics
Colocalization Theory
Webinar Colocalization Analyzer