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dc.contributor.author
Roth, Julian
dc.contributor.author
Mehl, Johanna
dc.contributor.author
Rohrbach, Alexander
dc.date.accessioned
2020-07-31T12:35:22Z
dc.date.available
2020-07-23T05:43:52Z
dc.date.available
2020-07-31T12:35:22Z
dc.date.issued
2020-07
dc.identifier.issn
2156-7085
dc.identifier.other
10.1364/BOE.391561
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/428101
dc.description.abstract
Fluorescence microscopy is the standard imaging technique to investigate the structures and dynamics of living cells. However, increasing the spatial resolution comes at the cost of temporal resolution and vice versa. In addition, the number of images that can be taken in sufficiently high quality is limited by fluorescence bleaching. Hence, super-resolved imaging at several Hertz of low fluorescent biological samples is still a big challenge and, especially in structured illumination microscopy (SIM), is often visible as imaging artifacts. In this paper, we present a TIRF-SIM system based on scan-mirrors and a Michelson interferometer, which generates images at 110 nm spatial resolution and up to 8 Hz temporal resolution. High resolution becomes possible by optimizing the illumination interference contrast, even for low fluorescent, moving samples. We provide a framework and guidelines on how the modulation contrast, which depends on laser coherence, polarization, beam displacement or sample movements, can be mapped over the entire field of view. In addition, we characterize the influence of the signal-to-noise ratio and the Wiener filtering on the quality of reconstructed SIM images, both in real and frequency space. Our results are supported by theoretical descriptions containing the parameters leading to image artifacts. This study aims to help microscopists to better understand and adjust optical parameters for structured illumination, thereby leading to more trustworthy measurements and analyses of biological dynamics. © 2020 Optical Society of America.
en_US
dc.language.iso
en
en_US
dc.publisher
Optical Society of America
en_US
dc.title
Fast TIRF-SIM imaging of dynamic, low-fluorescent biological samples
en_US
dc.type
Journal Article
dc.date.published
2020-06-26
ethz.journal.title
Biomedical Optics Express
ethz.journal.volume
11
en_US
ethz.journal.issue
7
en_US
ethz.journal.abbreviated
Biomed. opt. express
ethz.pages.start
4008
en_US
ethz.pages.end
4026
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
Washington, DC
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02070 - Dep. Gesundheitswiss. und Technologie / Dep. of Health Sciences and Technology::02540 - Institut für Translationale Medizin / Institute of Translational Medicine::03640 - Vogel, Viola / Vogel, Viola
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02070 - Dep. Gesundheitswiss. und Technologie / Dep. of Health Sciences and Technology::02540 - Institut für Translationale Medizin / Institute of Translational Medicine::03640 - Vogel, Viola / Vogel, Viola
ethz.date.deposited
2020-07-23T05:44:05Z
ethz.source
SCOPUS
ethz.eth
yes
en_US
ethz.availability
Metadata only
en_US
ethz.rosetta.installDate
2020-07-31T12:35:38Z
ethz.rosetta.lastUpdated
2021-02-15T15:46:43Z
ethz.rosetta.versionExported
true
ethz.COinS
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