With colocalization analysis for fluorescence microscopy you can obtain quantitative information about the amount of spatial overlap between structures in different channels for 3D images and 3D-time series. As the spatial overlap can be defined in many ways, the Huygens Colocalization Analyzer gives you eleven different colocalization coefficients to determine spatial overlap. Learn more.

From Huygens Colocalization Analyzer 18.10 and newer, there are three options for the Colocalization Analyzer: The Wizard, Advanced, and Compact mode. All three modes have the same funcionalities only the presentation is different. The Wizard guides the users step by step, while the other two modes provide an overview of all functions and data in a single window. The Compact mode is specially designed for smaller screens.

You can do that very easily:

• Via Colocalization Analyzer: Measure the Co-localization coefficients -> Go to Object Statistics -> select Analyze all
• Via Object Analyzer: Select Experimental preset -> Colocalization

Huygens supports many different colocalization coefficients. And it can be hard to choose which one provides the right information for your experiment. All colocalization coefficents use the intensity from the 2 channels you are measuring. Each pixel contributes a certain amount towards your total coefficient, depending on the intensities in the channels you are comparing.

For a basic overview of the challenges in colocalization measurements, visit Colocalization Basics. For a comprehensive overview with examples you can visit the page Colocalization Coefficients. For the exact definition of the colocalization coefficients, visit the page on Colocalization Theory.