Nanoscale Probing of Dielectric Interfaces with Single-Molecule Excitation Patterns and Radially Polarized Illumination

Chizhik, Alexey I.; Chizhik, Anna M.; Huss, Anja; Jäger, Regina; Meixner, Alfred J.

doi:10.1021/jz200934y

Cited by 14 publications

(10 citation statements)

References 41 publications

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“…However, this modification requires additional camera frames and thus additional photostability. θ can also be measured by altering a confocal excitation field using annular illumination or a radially polarized beam [41][42][43].…”

Section: Methodsmentioning

confidence: 99%

Enhanced DNA Imaging Using Super-Resolution Microscopy and Simultaneous Single-Molecule Orientation Measurements

Backer

Lee

Moerner

2016

Conference on Lasers and Electro-Optics

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Single-molecule orientation measurements provide unparalleled insight into a multitude of biological and polymeric systems. We report a simple, high-throughput technique for measuring the azimuthal orientation and rotational dynamics of single fluorescent molecules, which is compatible with localization microscopy. Our method involves modulating the polarization of an excitation laser, and analyzing the corresponding intensities emitted by single dye molecules and their modulation amplitudes. To demonstrate our approach, we use intercalating and groovebinding dyes to obtain super-resolved images of stretched DNA strands through binding-induced turn-on of fluorescence. By combining our image data with thousands of dye molecule orientation measurements, we develop a means of probing the structure of individual DNA strands, while also characterizing dye-DNA interactions. This approach may hold promise as a method for monitoring DNA conformation changes resulting from DNA-binding proteins.

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Section: Methodsmentioning

confidence: 99%

Enhanced DNA Imaging Using Super-Resolution Microscopy and Simultaneous Single-Molecule Orientation Measurements

Backer

Lee

Moerner

2016

Conference on Lasers and Electro-Optics

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“…Focused or defocused imaging with an azimuthally polarized laser beam (APLB), which can measure and provide the information of excitation or emission transition dipole moment (TDM) respectively [94][95][96] , has also been employed by the authors 93 to uncover the emission mechanism of carbon dots. The results are illustrated at figure 10b.…”

Section: Surface Dipole Emission Centrementioning

confidence: 99%

Novel properties and applications of carbon nanodots

2018

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“…Alas, this method does not give access to the out-of-plane angle. By scanning a radially polarized excitation focus the full orientation of the absorption dipole can be probed (35, 36), although this is not directly applicable to localization microscopy.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous orientation and 3D localization microscopy with a Vortex point spread function

Hulleman

Thorsen

Stallinga

et al. 2020

Preprint

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We have developed an engineered Point Spread Function (PSF) to enable the simultaneous estimation of dipole orientation, 3D position and degree of rotational constraint of single-molecule emitters from a single 2D focal plane. Besides giving access to orientation information, the Vortex PSF along with the vectorial PSF fitter avoids localization bias common in localization microscopy for fixed dipole emitters. We demonstrate this technique on reorienting single-molecules and using binding-activated localization microscopy on DNA intercalators, corroborating perpendicular azimuthal angles to the DNA axis for in-plane emitters but find a non-uniform distribution as a function of the polar angle. The Vortex PSF is realized by an affordable glass phase mask and has a compact footprint that can easily be combined with localization microscopy techniques on rotationally constrained emitters.

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Nanoscale Probing of Dielectric Interfaces with Single-Molecule Excitation Patterns and Radially Polarized Illumination

Cited by 14 publications

References 41 publications

Enhanced DNA Imaging Using Super-Resolution Microscopy and Simultaneous Single-Molecule Orientation Measurements

Enhanced DNA Imaging Using Super-Resolution Microscopy and Simultaneous Single-Molecule Orientation Measurements

Novel properties and applications of carbon nanodots

Simultaneous orientation and 3D localization microscopy with a Vortex point spread function

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