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  4. Imaging tissues and cells beyond the diffraction limit with structured illumination microscopy and Bayesian image reconstruction
 
research article

Imaging tissues and cells beyond the diffraction limit with structured illumination microscopy and Bayesian image reconstruction

Pospisil, Jakub
•
Lukes, Tomas  
•
Bendesky, Justin
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January 1, 2019
GigaScience

Background: Structured illumination microscopy (SIM) is a family of methods in optical fluorescence microscopy that can achieve both optical sectioning and super-resolution effects. SIM is a valuable method for high-resolution imaging of fixed cells or tissues labeled with conventional fluorophores, as well as for imaging the dynamics of live cells expressing fluorescent protein constructs. In SIM, one acquires a set of images with shifting illumination patterns. This set of images is subsequently treated with image analysis algorithms to produce an image with reduced out-of-focus light (optical sectioning) and/or with improved resolution (super-resolution). Findings: Five complete, freely available SIM datasets are presented including raw and analyzed data. We report methods for image acquisition and analysis using open-source software along with examples of the resulting images when processed with different methods. We processed the data using established optical sectioning SIM and super-resolution SIM methods and with newer Bayesian restoration approaches that we are developing. Conclusions: Various methods for SIM data acquisition and processing are actively being developed, but complete raw data from SIM experiments are not typically published. Publically available, high-quality raw data with examples of processed results will aid researchers when developing new methods in SIM. Biologists will also find interest in the high-resolution images of animal tissues and cells we acquired. All of the data were processed with SIMToolbox, an open-source and freely available software solution for SIM.

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Type
research article
DOI
10.1093/gigascience/giy126
Web of Science ID

WOS:000458893400001

Author(s)
Pospisil, Jakub
Lukes, Tomas  
Bendesky, Justin
Fliegel, Karel
Spendier, Kathrin
Hagen, Guy M.
Date Issued

2019-01-01

Publisher

Oxford University Press (OUP)

Published in
GigaScience
Volume

8

Issue

1

Article Number

giy126

Subjects

Multidisciplinary Sciences

•

Science & Technology - Other Topics

•

super-resolution microscopy

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simtoolbox

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structured illumination microscopy

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open-source software

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fluorescence

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bayesian methods

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lamp1

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live cell imaging

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live cells

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resolution limit

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light

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speed

Note

This article is licensed under a Creative Commons Attribution 4.0 International License

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LBEN  
Available on Infoscience
June 18, 2019
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/157608
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