Large field-of-view nanometer-sectioning microscopy by using metal-induced energy transfer and biexponential lifetime analysis

Hwang, Wonsang; Seo, Jinwon; Kim, Dong-Eun; Lee, Chang Jun; Choi, In Hong; Yoo, Kyung Hwa; Kim, Dug Young

doi:10.1038/s42003-020-01628-3

Cited by 12 publications

(8 citation statements)

References 37 publications

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“…As a simple and reliable far-eld optical technique, the SAF-based uorophore-height measurement is an interesting alternative and complement to other axial nanoscopies. For the sub-20-nm range where our technique lacks sensitivity, quenching-based optical rulers, based on plasmonics 45 , distance-dependent uorescence-lifetime modulation 46 , metal-induced energy transfer (MIET) [47][48][49] , or transparent conductive oxide (TCO-) based transparent rulers 50 can close the gap, offering nm sensitivity.…”

Section: Discussionmentioning

confidence: 99%

Characterization of nanometric thin films with far-field light

Salomon

Klimovsky

Shavit

et al. 2022

Preprint

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The fabrication and characterisation of ultra-thin, transparent films is paramount for protective layers on semiconductors, solar cells, as well as for nano-composite materials and optical coatings. Similarly, the probe volume of nano-sensors, as well the calibration of axial distances in super-resolution microscopies, all require the metrology of axial fluorophore distances. However, the reliable production and precise characterisation of such nanometric thin layers are difficult and labor-intense and they require specialized equipment and trained personnel. In our present work, we describe a simple, non-invasive, all-optical technique for simultaneously measuring the refractive index, thickness, and homogeneity of such thin films. We assemble transparent layers from My-133-MC, a biomimetic transparent polymer with a refractive index of 1.33, amenable for applications in the life sciences. All parameters characterising the films are obtained in a single measurement from the analysis of supercritical angle fluorescence radiation patterns acquired on a minimally modified inverted microscope. Results compare favorably to those obtained through a combination of atomic force and electron microscopy, surface-plasmon resonance spectroscopy and ellipsometry. To illustrate the utility of our technique, we present two applications, one in metrology and one in bio-imaging; (i), the calibration of axial fluorophore distance in a total internal reflection fluorescence geometry; and, (ii), live-cell super-resolution imaging of organelle dynamics in cortical astrocytes, an important type of brain cell. Our approach is cheap, versatile and it has obvious applications in profilometry, biophotonics, photonic devices, and optical nano-metrology.

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Section: Discussionmentioning

confidence: 99%

Characterization of nanometric thin films with far-field light

Salomon

Klimovsky

Shavit

et al. 2022

Preprint

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“…The authors of ref. [174] used the advantage of MIET that it does not require high-NA objectives for presenting a large field-of-view implementation of the method.…”

Section: Using Near-field Coupling For Axial Localizationmentioning

confidence: 99%

Super-resolution imaging: when biophysics meets nanophotonics

et al. 2021

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Probing light–matter interaction at the nanometer scale is one of the most fascinating topics of modern optics. Its importance is underlined by the large span of fields in which such accurate knowledge of light–matter interaction is needed, namely nanophotonics, quantum electrodynamics, atomic physics, biosensing, quantum computing and many more. Increasing innovations in the field of microscopy in the last decade have pushed the ability of observing such phenomena across multiple length scales, from micrometers to nanometers. In bioimaging, the advent of super-resolution single-molecule localization microscopy (SMLM) has opened a completely new perspective for the study and understanding of molecular mechanisms, with unprecedented resolution, which take place inside the cell. Since then, the field of SMLM has been continuously improving, shifting from an initial drive for pushing technological limitations to the acquisition of new knowledge. Interestingly, such developments have become also of great interest for the study of light–matter interaction in nanostructured materials, either dielectric, metallic, or hybrid metallic-dielectric. The purpose of this review is to summarize the recent advances in the field of nanophotonics that have leveraged SMLM, and conversely to show how some concepts commonly used in nanophotonics can benefit the development of new microscopy techniques for biophysics. To this aim, we will first introduce the basic concepts of SMLM and the observables that can be measured. Then, we will link them with their corresponding physical quantities of interest in biophysics and nanophotonics and we will describe state-of-the-art experiments that apply SMLM to nanophotonics. The problem of localization artifacts due to the interaction of the fluorescent emitter with a resonant medium and possible solutions will be also discussed. Then, we will show how the interaction of fluorescent emitters with plasmonic structures can be successfully employed in biology for cell profiling and membrane organization studies. We present an outlook on emerging research directions enabled by the synergy of localization microscopy and nanophotonics.

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“…Hwang and colleagues used MIET to prove a nanometer sectioning method in a large field-of-view in a confocal microscope with a low numerical aperture objective. They tested the method in human aortic endothelial imaging a large field-of-view of 120 μm × 96 μm (Hwang et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Plasmonics for advance single-molecule fluorescence spectroscopy and imaging in biology

Zaza

Simoncelli

2022

Front. Photon.

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The elucidation of complex biological processes often requires monitoring the dynamics and spatial organization of multiple distinct proteins organized on the sub-micron scale. This length scale is well below the diffraction limit of light, and as such not accessible by classical optical techniques. Further, the high molecular concentrations found in living cells, typically in the micro- to mili-molar range, preclude single-molecule detection in confocal volumes, essential to quantify affinity constants and protein-protein reaction rates in their physiological environment. To push the boundaries of the current state of the art in single-molecule fluorescence imaging and spectroscopy, plasmonic materials offer encouraging perspectives. From thin metallic films to complex nano-antenna structures, the near-field electromagnetic coupling between the electronic transitions of single emitters and plasmon resonances can be exploited to expand the toolbox of single-molecule based fluorescence imaging and spectroscopy approaches. Here, we review two of the most current and promising approaches to study biological processes with unattainable level of detail. On one side, we discuss how the reduction of the fluorescence lifetime of a molecule as it approaches a thin metallic film can be exploited to decode axial information with nanoscale precision. On the other, we review how the tremendous progress on the design of plasmonic antennas that can amplify and confine optical fields at the nanoscale, powered a revolution in fluorescence correlation spectroscopy. Besides method development, we also focus in describing the most interesting biological application of both technologies.

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Large field-of-view nanometer-sectioning microscopy by using metal-induced energy transfer and biexponential lifetime analysis

Cited by 12 publications

References 37 publications

Characterization of nanometric thin films with far-field light

Characterization of nanometric thin films with far-field light

Super-resolution imaging: when biophysics meets nanophotonics

Plasmonics for advance single-molecule fluorescence spectroscopy and imaging in biology

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