Scientific Volume Imaging b.v.
CoLocalization analysis for 3D fluorescence microscopy is available in Huygens Essential and Huygens Professional since version 3.0, allowing you to obtain quantitative information about the amount of spatial overlap between structures in different data channels, for 3D images and 3D-time series.
As this overlap can be defined in many ways, the colocalization analyzer gives you the colocalization coefficients most commonly used in literature:
For more information about these coefficients, see Colocalization Theory.
One of the features of the colocalization analyzer is iso-colocalization object analysis. It allows you to quickly determine the properties of the different colocalization regions in your data. This is realized by visualizing the colocalization map as iso-colocalization surfaces. In the colocalization analyzer these surfaces are computed at the same time as the coefficients.
The surface objects show regions in which the degree of colocalization exceeds a certain value. By clicking on the objects local colocalization parameters are computed and reported. To relate the iso-colocalization objects to the original data the surface objects can be blended with a Red-Green MIP projection of the colocalizing channels, or with a colored MIP of another reference channel. The color range in which these objects will be displayed can be modified using a Hue Selector.
See real size image: colocScreen.png (196 kB 2008-09-15 14:53)
Notice that the Object Analyzer in the Huygens Software also provides colocalization measurements at the object level. The Colocalization Analyzer works more at the level of the whole image, despite local statistics of the colocalizing regions can be easily retrieved.
Both analyzers work, in a sense, in complementary ways.
The Object Analyzer allows you to define objects and see how much they overlap, in volume or intensity. Objects defined like this can overlap with other objects, or not.
The Colocalization Analyzer explores the whole image to search for colocalizing regions based on the usual colocalization coefficients. These regions are then segmented and treated as objects to analyze. These objects are therefore always volumes of intersection.
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