Live-cell fluorescence imaging with extreme background suppression by plasmonic nanocoatings
(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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- author
- Schreiber, Benjamin
; Heil, Hannah S
LU
; Kamp, Martin and Heinze, Katrin G
- publishing date
- 2018-08-06
- 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}}, }