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J Phys Chem B. 2014 Jul 17;118(28):8313-29. doi: 10.1021/jp501778z. Epub 2014 May 12.

Extending single-molecule microscopy using optical Fourier processing.

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Institute for Computational and Mathematical Engineering and ‡Department of Chemistry, Stanford University , Stanford, California 94305, United States.


This article surveys the recent application of optical Fourier processing to the long-established but still expanding field of single-molecule imaging and microscopy. A variety of single-molecule studies can benefit from the additional image information that can be obtained by modulating the Fourier, or pupil, plane of a widefield microscope. After briefly reviewing several current applications, we present a comprehensive and computationally efficient theoretical model for simulating single-molecule fluorescence as it propagates through an imaging system. Furthermore, we describe how phase/amplitude-modulating optics inserted in the imaging pathway may be modeled, especially at the Fourier plane. Finally, we discuss selected recent applications of Fourier processing methods to measure the orientation, depth, and rotational mobility of single fluorescent molecules.

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