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Live-cell fluorescence imaging with extreme background suppression by plasmonic nanocoatings

Schreiber, Benjamin ; Heil, Hannah S LU orcid ; Kamp, Martin and Heinze, Katrin G (2018) In Optics Express 26(16). p.21301-21313
Abstract

Fluorescence microscopy allows specific and selective imaging of biological samples. Unfortunately, unspecific background due to auto-fluorescence, scattering, and non-ideal labeling efficiency often adversely affect imaging. Surface plasmon-coupled emission (SPCE) is known to selectively mediate fluorescence that spatially originates from regions close to the metal interface. However, SPCE combined with fluorescence imaging has not been widely successful so far, most likely due to its limited photon yield, which makes it tedious to identify the exact window of the application. As the strength of SPCE based imaging is its unique sectioning capabilities. We decided to identify its clear beneficial operational regime for biological... (More)

Fluorescence microscopy allows specific and selective imaging of biological samples. Unfortunately, unspecific background due to auto-fluorescence, scattering, and non-ideal labeling efficiency often adversely affect imaging. Surface plasmon-coupled emission (SPCE) is known to selectively mediate fluorescence that spatially originates from regions close to the metal interface. However, SPCE combined with fluorescence imaging has not been widely successful so far, most likely due to its limited photon yield, which makes it tedious to identify the exact window of the application. As the strength of SPCE based imaging is its unique sectioning capabilities. We decided to identify its clear beneficial operational regime for biological settings by interrogating samples in the presence of ascending background levels. For fluorescent beads as well as live-cell imaging as examples, we show how to extend the imaging performance in extremely high photon background environments. In a common setup using plasmonic gold-coated coverslips using an objective-based total internal reflection fluorescence microscope (TIRF-M), we theoretically and experimentally characterize our fluoplasmonics (f-Pics) approach by providing general user guidance in choosing f-Pics over TIRF-M or classical wide-field (WF).

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Please use this url to cite or link to this publication:
author
; ; and
publishing date
type
Contribution to journal
publication status
published
in
Optics Express
volume
26
issue
16
pages
21301 - 21313
publisher
Optical Society of America
external identifiers
  • pmid:30119432
  • scopus:85051244392
ISSN
1094-4087
DOI
10.1364/OE.26.021301
language
English
LU publication?
no
id
64216ec3-597b-48ff-8cc8-7acb535ba145
date added to LUP
2025-04-26 12:13:42
date last changed
2025-04-29 03:31:58
@article{64216ec3-597b-48ff-8cc8-7acb535ba145,
  abstract     = {{<p>Fluorescence microscopy allows specific and selective imaging of biological samples. Unfortunately, unspecific background due to auto-fluorescence, scattering, and non-ideal labeling efficiency often adversely affect imaging. Surface plasmon-coupled emission (SPCE) is known to selectively mediate fluorescence that spatially originates from regions close to the metal interface. However, SPCE combined with fluorescence imaging has not been widely successful so far, most likely due to its limited photon yield, which makes it tedious to identify the exact window of the application. As the strength of SPCE based imaging is its unique sectioning capabilities. We decided to identify its clear beneficial operational regime for biological settings by interrogating samples in the presence of ascending background levels. For fluorescent beads as well as live-cell imaging as examples, we show how to extend the imaging performance in extremely high photon background environments. In a common setup using plasmonic gold-coated coverslips using an objective-based total internal reflection fluorescence microscope (TIRF-M), we theoretically and experimentally characterize our fluoplasmonics (f-Pics) approach by providing general user guidance in choosing f-Pics over TIRF-M or classical wide-field (WF).</p>}},
  author       = {{Schreiber, Benjamin and Heil, Hannah S and Kamp, Martin and Heinze, Katrin G}},
  issn         = {{1094-4087}},
  language     = {{eng}},
  month        = {{08}},
  number       = {{16}},
  pages        = {{21301--21313}},
  publisher    = {{Optical Society of America}},
  series       = {{Optics Express}},
  title        = {{Live-cell fluorescence imaging with extreme background suppression by plasmonic nanocoatings}},
  url          = {{http://dx.doi.org/10.1364/OE.26.021301}},
  doi          = {{10.1364/OE.26.021301}},
  volume       = {{26}},
  year         = {{2018}},
}