Tip induced fluorescence quenching for nanometer optical and topographical resolution

Schulz, O.; Zhao, Zhao; Ward, Alex; Koenig, Marcelle; Koberling, Felix; Liu, Yan; Enderlein, Jörg; Ros, Robert

doi:10.1186/2192-2853-2-1

Cited by 27 publications

(19 citation statements)

References 53 publications

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“…Fitting of experimental lifetime values by using Equation(6). The unperturbed lifetime t 0 obtained as a fit parameter is 2.78 ns, which is in good agreement with the lifetime of Atto655 measured in air on glass [39]. …”

supporting

confidence: 78%

Single‐Molecule Metal‐Induced Energy Transfer (smMIET): Resolving Nanometer Distances at the Single‐Molecule Level

et al. 2014

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We present a new concept for measuring distance values of single molecules from a surface with nanometer accuracy using the energy transfer from the excited molecule to surface plasmons of a metal film. We measure the fluorescence lifetime of individual dye molecules deposited on a dielectric spacer as a function of a spacer thickness. By using our theoretical model, we convert the lifetime values into the axial distance of individual molecules. Similar to Förster resonance energy transfer (FRET), this allows emitters to be localized with nanometer accuracy, but in contrast to FRET the distance range at which efficient energy transfer takes place is an order of magnitude larger. Our technique can be potentially used as a tool for measuring intramolecular distances of biomolecules and complexes.

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supporting

confidence: 78%

Single‐Molecule Metal‐Induced Energy Transfer (smMIET): Resolving Nanometer Distances at the Single‐Molecule Level

et al. 2014

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“…Silicon probes expose only a moderate field enhancement and the dipolar coupling between probe and dye is less pronounced [4]. Even though, silicon tips can quench the fluorescence emission at close proximity [14]. Moreover, the elaborate probe design of custom-made tips demand complex fabrication processes and substantial experience [3,7,12–13 15–17].…”

Section: Introductionmentioning

confidence: 99%

Apertureless scanning near-field optical microscopy of sparsely labeled tobacco mosaic viruses and the intermediate filament desmin

Harder¹,

Dieding²,

Walhorn³

et al. 2013

Beilstein J. Nanotechnol.

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SummaryBoth fluorescence imaging and atomic force microscopy (AFM) are highly versatile and extensively used in applications ranging from nanotechnology to life sciences. In fluorescence microscopy luminescent dyes serve as position markers. Moreover, they can be used as active reporters of their local vicinity. The dipolar coupling of the tip with the incident light and the fluorophore give rise to a local field and fluorescence enhancement. AFM topographic imaging allows for resolutions down to the atomic scale. It can be operated in vacuum, under ambient conditions and in liquids. This makes it ideal for the investigation of a wide range of different samples. Furthermore an illuminated AFM cantilever tip apex exposes strongly confined non-propagating electromagnetic fields that can serve as a coupling agent for single dye molecules. Thus, combining both techniques by means of apertureless scanning near-field optical microscopy (aSNOM) enables concurrent high resolution topography and fluorescence imaging. Commonly, among the various (apertureless) SNOM approaches metallic or metallized probes are used. Here, we report on our custom-built aSNOM setup, which uses commercially available monolithic silicon AFM cantilevers. The field enhancement confined to the tip apex facilitates an optical resolution down to 20 nm. Furthermore, the use of standard mass-produced AFM cantilevers spares elaborate probe production or modification processes. We investigated tobacco mosaic viruses and the intermediate filament protein desmin. Both are mixed complexes of building blocks, which are fluorescently labeled to a low degree. The simultaneous recording of topography and fluorescence data allows for the exact localization of distinct building blocks within the superordinate structures.

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“…Because the brightness of the single molecules is too weak for high-speed imaging, averaging over longer exposure times was necessary. We imaged individual molecules of Atto 655, a fluorescence label shown to be very stable and to exhibit little blinking on glass surfaces under dry conditions (10). Two-dimensional Gaussians were fit to the single-molecule emission spots in the CSD image and in the CSD-ISM image for multiple scans.…”

Section: (Compare Figs 1 and 2c And Fourier Reweighting)mentioning

confidence: 99%

Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy

Schulz

Pieper

Clever

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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118

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We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions.

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Tip induced fluorescence quenching for nanometer optical and topographical resolution

Cited by 27 publications

References 53 publications

Single‐Molecule Metal‐Induced Energy Transfer (smMIET): Resolving Nanometer Distances at the Single‐Molecule Level

Single‐Molecule Metal‐Induced Energy Transfer (smMIET): Resolving Nanometer Distances at the Single‐Molecule Level

Apertureless scanning near-field optical microscopy of sparsely labeled tobacco mosaic viruses and the intermediate filament desmin

Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy

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