Daybook Analysis - Version 2.0.0 - October 2024
The co-registration accuracy is the ability of a fluorescence microscope to record images of a multi-labeled object without introducing additional shifts that are not be related to the object itself.
For any fluorescence microscope, knowledge of the co-registration accuracy, both in the lateral and axial directions, is important when color information in an image is to be used. For co-localization quantification in images of biological samples stained with several labels, the co-registration accuracy between the different channels should be known, and eventually corrected for, to prevent misinterpretation.
The “lateral co-registration accuracy” analysis provides the lateral shifts between two channels, due to the translation, rotation and scaling contributions to the co-registration inaccuracy of the system, within the entire field of view.
The “lateral co-registration accuracy” analysis is associated with the “field of rings” pattern (Pattern family B - see Figure 1). It requires images of this pattern acquired on different channels (at least two channels).
Figure 1: Example images of the “field of rings” pattern, acquired on two different channels, fulfilling the acquisition recommendations. Left: DAPI channel. Right: GFP channel. The image with the best contrast (right) should be used as the reference image, in order to improve the detection of the rings in an image with a lower contrast (left).