Dark-field microscopy for characterization of single molecule dynamicsin vitroandin vivo

2019 ◽  
Vol 11 (21) ◽  
pp. 2778-2784
Author(s):  
Bingquan Wang ◽  
Dan Sun ◽  
Ce Zhang ◽  
Kaige Wang ◽  
Jintao Bai
Keyword(s):  
Single Molecule ◽  
Scattered Light ◽  
Dark Field ◽  
Fluorescent Labeling ◽  
Single Molecule Detection ◽  
Dark Field Microscopy

Dark-field microscopy directly detects scattered light from a sample, and therefore requires no fluorescent labeling for single molecule detection.

scholarly journals Dark-field microscopy enhance visibility of CD31 endothelial staining

2020 ◽  
Vol 64 (3) ◽  
Author(s):  
Eva Jennische ◽  
Stefan Lange ◽  
Ragnar Hultborn
Keyword(s):  
Phase Contrast ◽  
Scattered Light ◽  
Dark Field ◽  
Light Scatter ◽  
Endothelial Lining ◽  
Microscopy Technique ◽  
Dark Field Microscopy ◽  
Dark Field Illumination ◽  
Normal Human ◽  
Intense Light

A simple dark field microscopy technique was used for visualization of blood vessels in normal human renal tissues and carcinoma. Phase contrast condenser ring apt for high power objectives was combined with a 10x objective in order to create a dark field illumination of the specimens examined. The endothelial lining of the vessels had been stained by using CD31 monoclonal antibodies combined with conventional peroxidase immunohistochemistry. The final DAB addition used for this technique induced an intense light scatter in the dark field microscope. This scattered light originating from the endothelial lining made the walls of the bright vessels easily detectable from the dark background.

Internalization of hyaluronan by chondrocytes occurs via receptor-mediated endocytosis

1993 ◽  
Vol 106 (1) ◽  
pp. 365-375 ◽  
Author(s):  
Q. Hua ◽  
C.B. Knudson ◽  
W. Knudson
Keyword(s):  
Cell Surface ◽  
Articular Chondrocytes ◽  
Dark Field ◽  
Cartilage Tissue ◽  
Receptor Mediated Endocytosis ◽  
Dark Field Microscopy ◽  
Bovine Articular Chondrocytes ◽  
Hyaluronan Receptors ◽  
Hyaluronan Receptor

Several studies have suggested that chondrocytes must have the capacity to internalize and degrade extracellular hyaluronan. In the present study we show direct evidence that hyaluronan is, in fact, endocytosed by chondrocytes and that the endocytosis is mediated via cell surface CD44/hyaluronan receptors. Cultures of bovine articular chondrocytes as well as rat chondrosarcoma chondrocytes were incubated with either fluorescein- or 3H-labeled hyaluronan. Intense binding and accumulation of labeled hyaluronan was visualized by fluorescence microscopy or bright-field/dark-field microscopy following autoradiography. Cell surface hyaluronan was removed with either trypsin or Streptomyces hyaluronidase in order to distinguish and quantify intracellular endocytosed hyaluronan. Labeled hyaluronan was visualized within small discrete intracellular vesicles distributed throughout the cytoplasm. Binding and endocytosis of fluorescein- or 3H-labeled hyaluronan was totally blocked by the addition of excess unlabeled hyaluronan or hyaluronan hexasaccharides, competitive inhibitors of hyaluronan/hyaluronan receptor interactions. Binding and endocytosis was also blocked by the addition of anti-CD44 monoclonal antibodies. Characterization of endocytosed 3H-labeled hyaluronan demonstrated that a significant portion of the hyaluronan was degraded by both the bovine articular and rat chondrosarcoma chondrocytes. Interestingly, a higher proportion of bound hyaluronan was internalized by the bovine chondrocytes. Therefore, hyaluronan receptor-mediated endocytosis and degradation of hyaluronan may provide a critical link to the maintenance and homeostasis of cartilage tissue.

Engineering and Characterization of Resonant Optical Antennas

2010 ◽  
Vol 1248 ◽  
Author(s):  
Matthias D. Wissert ◽  
Andreas W. Schell ◽  
Konstantin S. Ilin ◽  
M. Siegel ◽  
U. Lemmer ◽  
...  
Keyword(s):  
Electron Beam Lithography ◽  
Dark Field ◽  
Optical Antennas ◽  
Dipole Antennas ◽  
Split Ring ◽  
Dark Field Microscopy ◽  
Scattering Spectra ◽  
Coupled Structures ◽  
Spectral Redshift

AbstractResonant optical dipole antennas, consisting either of two arms coupled by a small gap or of a single, uncoupled arm only, are fabricated by the application of electron beam lithography and gold evaporation. Using dark-field microscopy, scattering spectra of structures with varied antenna arm length and varied gap size are obtained. The results show not only a spectral redshift for coupled structures compared to single arm structures, but also that the far-field scattering intensity is significantly higher for two arm structures with gap. In addition to the dipole structures, first fabrication results on quadrupole antennas and split-ring antennas are presented, offering novel pathways for an enhancement of the optical response function.

scholarly journals A new optical method for characterizing single molecule interactions based on dark field microscopy

2007 ◽  
Author(s):  
Heidelinde R. C. Dietrich ◽  
Bart J. Vermolen ◽  
Bernd Rieger ◽  
Ian T. Young ◽  
Yuval Garini
Keyword(s):  
Single Molecule ◽  
Optical Method ◽  
Dark Field ◽  
Dark Field Microscopy ◽  
Molecule Interactions

Dark-Field Hyperspectral Imaging (DF-HSI) Modalities for Characterization of Single Molecule and Cellular Processes

2020 ◽  
pp. 231-262
Author(s):  
Nishir Mehta ◽  
Sushant Sahu ◽  
Shahensha Shaik ◽  
Ram Devireddy ◽  
Manas Ranjan Gartia
Keyword(s):  
Hyperspectral Imaging ◽  
Single Molecule ◽  
Dark Field ◽  
Cellular Processes

scholarly journals Plasmonic Photomobile Polymer Films

Crystals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 660
Author(s):  
Riccardo Castagna ◽  
Massimo Rippa ◽  
Francesco Simoni ◽  
Fulvia Villani ◽  
Giuseppe Nenna ◽  
...  
Keyword(s):  
Polymer Films ◽  
Localized Surface Plasmon Resonance ◽  
Dark Field ◽  
Localized Surface Plasmon ◽  
Force Microscopy ◽  
Atomic Force ◽  
Plasmonic Nanostructure ◽  
Dark Field Microscopy ◽  
Scanning Electron

In this work, we introduce the approaches currently followed to realize photomobile polymer films and remark on the main features of the system based on a biphasic structure recently proposed. We describe a method of making a plasmonic nanostructure on the surface of photomobile films. The characterization of the photomobile film is performed by means of Dark Field Microscopy (DFM), Scanning Electron Microscopy (SEM), and Atomic Force Microscopy (AFM). Preliminary observations of the light-induced effects on the Localized Surface Plasmon Resonance are also reported.

Homogeneous amplified single-molecule detection: Characterization of key parameters

2007 ◽  
Vol 368 (2) ◽  
pp. 230-238 ◽  
Author(s):  
Jonas Melin ◽  
Jonas Jarvius ◽  
Jenny Göransson ◽  
Mats Nilsson
Keyword(s):  
Single Molecule ◽  
Single Molecule Detection

scholarly journals Fabrication and characterization of Au dimer antennas on glass pillars with enhanced plasmonic response

Nanophotonics ◽  
2017 ◽  
Vol 7 (2) ◽  
pp. 497-505 ◽  
Author(s):  
Pedram Sadeghi ◽  
Kaiyu Wu ◽  
Tomas Rindzevicius ◽  
Anja Boisen ◽  
Silvan Schmid
Keyword(s):  
Resonance Wavelength ◽  
Dark Field ◽  
Plasmonic Resonance ◽  
High Uniformity ◽  
Dark Field Microscopy ◽  
Scattering Spectra ◽  
Scattering Response ◽  
Difference Time ◽  
Microscopy Images

AbstractWe report on the fabrication and dark-field spectroscopy characterization of Au dimer nanoantennas placed on top of SiO2 nanopillars. The reported process enables the fabrication of nanopillar dimers with gaps down to 15 nm and heights up to 1 μm. A clear dependence of the plasmonic resonance position on the dimer gap is observed for smaller pillar heights, showing the high uniformity and reproducibility of the process. It is shown how increasing the height of nanopillars significantly affects the recorded elastic scattering spectra from Au nanoantennas. The results are compared to finite-difference time-domain (FDTD) and finite-element method (FEM) simulations. Additionally, measured spectra are accompanied by dark-field microscopy images of the dimers, showing the pronounced change in color. Placing nanoantennas on nanopillars with a height comparable to the in-plane dimer dimensions results in an enhancement of the scattering response, which can be understood through reduced interaction of the near-fields with the substrate. When increasing the pillar height further, scattering by the pillars themselves manifests itself as a strong tail at lower wavelengths. Additionally, strong directional scattering is expected as a result of the interface between the nanoantennas and nanopillars, which is taken into account in simulations. For pillars of height close to the plasmonic resonance wavelength, the scattering spectra become more complex due to additional scattering peaks as a result of larger geometrical nonuniformities.

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