Nearly all multi-channel images show at least some degree of chromatic aberration causing shifts in xyz, and possible scaling and rotation differences between fluorescent channels. Huygens Chromatic Aberration Corrector quickly estimates, visualizes, and corrects chromatic aberrations. Learn more

The chromatic shift, usually being the largest component of the chromatic aberration, is a misalignment across the channels of a multichannel image which can be corrected by simply shifting the channels. There are several circumstances that can lead to chromatic shifts in the microscopic images:

• Chromatic aberrations in the microscope optics.
• Misaligned beam splitters.
• Misaligned excitation lasers.
• Misaligned color filters.
• Faulty color interpolation in color cameras.
• Any other internal misalignments in the microscope.

To spot chromatic aberration there should be objects in your image that are stained with different fluorophores, these objects are then expected to overlap perfectly. The advise is to take a reference image with multicolored beads. To determine all possible aberrations (shifts, scaling, rotations and none-linear distortions) a 3D image is required with multiple beads distributed in 3D as well. For shifts a single bead is sufficient, but still multiple beads are recommended.

Chromatic aberration has a large effect on the correlation between different channels. If you want to do collocalization analyzis, or other analysis methods that depend on the exact location of objects in multiple channels, it is essential to check and if needed correct for chromatic abberation.