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Accurate and rapid background estimation in single-molecule localization microscopy using the deep neural network BGnet.


ABSTRACT: Background fluorescence, especially when it exhibits undesired spatial features, is a primary factor for reduced image quality in optical microscopy. Structured background is particularly detrimental when analyzing single-molecule images for 3-dimensional localization microscopy or single-molecule tracking. Here, we introduce BGnet, a deep neural network with a U-net-type architecture, as a general method to rapidly estimate the background underlying the image of a point source with excellent accuracy, even when point-spread function (PSF) engineering is in use to create complex PSF shapes. We trained BGnet to extract the background from images of various PSFs and show that the identification is accurate for a wide range of different interfering background structures constructed from many spatial frequencies. Furthermore, we demonstrate that the obtained background-corrected PSF images, for both simulated and experimental data, lead to a substantial improvement in localization precision. Finally, we verify that structured background estimation with BGnet results in higher quality of superresolution reconstructions of biological structures.

SUBMITTER: Mockl L 

PROVIDER: S-EPMC6955367 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Accurate and rapid background estimation in single-molecule localization microscopy using the deep neural network BGnet.

Möckl Leonhard L   Roy Anish R AR   Petrov Petar N PN   Moerner W E WE  

Proceedings of the National Academy of Sciences of the United States of America 20191223 1


Background fluorescence, especially when it exhibits undesired spatial features, is a primary factor for reduced image quality in optical microscopy. Structured background is particularly detrimental when analyzing single-molecule images for 3-dimensional localization microscopy or single-molecule tracking. Here, we introduce BGnet, a deep neural network with a U-net-type architecture, as a general method to rapidly estimate the background underlying the image of a point source with excellent ac  ...[more]

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