Global correction of optical distortions in multicolor single-molecule microscopy using Zernike polynomial gradients
K.A. Mc Cluskey (Kavli institute of nanoscience Delft, TU Delft - BN/Nynke Dekker Lab)
Edo van Veen (TU Delft - BN/Nynke Dekker Lab, Kavli institute of nanoscience Delft)
Jelmer Cnossen (TU Delft - Team Carlas Smith)
W.J. Wesselink (Kavli institute of nanoscience Delft, TU Delft - BN/Nynke Dekker Lab)
F.M. Asscher (TU Delft - BN/Nynke Dekker Lab)
S Smith (TU Delft - Delft Center for Systems and Control, TU Delft - ImPhys/Computational Imaging, TU Delft - Team Carlas Smith)
NH Dekker (TU Delft - BN/Nynke Dekker Lab, Kavli institute of nanoscience Delft)
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Abstract
Accurate image alignment is critical in multicolor single-molecule fluorescence microscopy. Global alignment using affine transformations leaves residual errors due to the nonlinearity of the distortions, which decreases the effective field of view. Subsequent local refinement demands either large amounts of reference data and processing time or specialized imaging techniques like active stabilization. Here, we present a global alignment method, S/T polynomial decomposition, that uses sums of Zernike polynomial gradients to decompose the distortion between two images, correcting both linear and nonlinear distortions simultaneously. With minimal reference data, we gain diagnostic information about the distortion and achieve a colocalization accuracy comparable to local registration methods across the entire field of view.