Bi-Plane Calibration in Super-Resolution Microscopy

H. Kirshner, T. Pengo, N. Olivier, D. Sage, S. Manley, M. Unser

Proceedings of the Twelfth Conference on Methods and Applications of Fluorescence Spectroscopy, Imaging and Probes (MAF'12), Strasbourg, France, September 11-14, 2011, in press.

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We consider the task of aligning the imaging detectors of a bi-plane microscope for super-resolution applications [1]. Such a microscope consists of two separate focal planes and the optical misalignment is modelled as an affine transform. In particular, u₁ = A u₂ + b accounts for translation, rotation and scaling operations (see illustration for an example of misaligned data). Here, u₁, u₂ describe a single point in space in terms of the two coordinate systems, A is a 3×3 matrix, and b is a translation vector. There are 12 misalignment parameters that need to be found, i.e. the matrix A and the vector b.

References

1. M.F. Juette, T.J. Gould, M.D. Lessard, M.J. Mlodzianoski, B.S. Nagpure, B.T. Bennett, S.T. Hess, J. Bewersdorf, "Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples," Nature Methods, vol. 5, no. 6, pp. 527-529, June 2008.