Information limits of optical microscopy: application to fluorescently labelled tissue section

Abstract

The article demonstrates some less known principles of image build-up in diffractive microscopy and their usage in analysis unravelling the smallest localized information about the original object - an electromagnetic centroid. In fluorescence, the electromagnetic centroid is naturally at the position of the fluorophore. The usage of an information-entropic variable - a point divergence gain - is demonstrated for finding the most localized position of the object's representation, generally of the size of a voxel (3D pixel). These spatial pixels can be qualitatively classified and used for reconstruction of the 3D structures with precision comparable with electron microscopy.

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