High definition aperture probes for near-field optical microscopy fabricated by focused ion beam milling

NARCIS (Netherlands)

Veerman, J.A.; Otter, A.M.; Kuipers, L.; van Hulst, N.F.

1998-01-01

We have improved the optical characteristics of aluminum-coated fiber probes used in near-field scanning optical microscopy by milling with a focused ion beam. This treatment produces a flat-end face free of aluminum grains, containing a well- defined circularly-symmetric aperture with controllable

Near-field-optical-microscopy studies of micro-modifications caused by femtosecond laser irradiation in lithium niobate crystals

International Nuclear Information System (INIS)

Lamela, J.; Jaque, D.; Rodenas, A.; Jaque, F.; Torchia, G.A.; Vazquez, J.R.; Mendez, C.; Roso, L.

2008-01-01

Near-field-optical-microscopy has been used to study the micro-modifications caused by femtosecond laser pulses focused at the surface and in the volume of lithium niobate crystals. We have found experimental evidence of the existence, close to femtosecond ablation craters, of periodic modifications in the surface reflectivity. In addition, the potential application of near-field-optical microscopy for the spatial location of permanent modifications caused by femtosecond pulses focused inside lithium niobate crystals has been also demonstrated. (orig.)

Quantitative Image Restoration in Bright Field Optical Microscopy.

Science.gov (United States)

Gutiérrez-Medina, Braulio; Sánchez Miranda, Manuel de Jesús

2017-11-07

Bright field (BF) optical microscopy is regarded as a poor method to observe unstained biological samples due to intrinsic low image contrast. We introduce quantitative image restoration in bright field (QRBF), a digital image processing method that restores out-of-focus BF images of unstained cells. Our procedure is based on deconvolution, using a point spread function modeled from theory. By comparing with reference images of bacteria observed in fluorescence, we show that QRBF faithfully recovers shape and enables quantify size of individual cells, even from a single input image. We applied QRBF in a high-throughput image cytometer to assess shape changes in Escherichia coli during hyperosmotic shock, finding size heterogeneity. We demonstrate that QRBF is also applicable to eukaryotic cells (yeast). Altogether, digital restoration emerges as a straightforward alternative to methods designed to generate contrast in BF imaging for quantitative analysis. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Scanning near-field optical microscopy of quantum dots in photonic crystal cavities

Energy Technology Data Exchange (ETDEWEB)

Skacel, Matthias; Fiore, Andrea [COBRA Research Institute, Technical University Eindhoven, Den Dolech 2, 5600 MB Eindhoven (Netherlands); Prancardi, Marco; Gerardino, Annamaria [Institute of Photonics and Nanotechnology, CNR, via del Cineto Romano 42, 00156 Roma (Italy); Alloing, Blandine; Li Lianhe, E-mail: m.s.skacel@tue.n [Institute of Photonics and Quantum Electronics, EPFL, CH-1015 Lausanne (Switzerland)

2010-09-01

Nanophotonic devices are of major interest for research and future quantum communication applications. Due to their nanometer feature size the resolution limit of far-field microscopy poses a limitation on the characterization of their optical properties. A method to overcome the resolution limit is the Scanning Near-Field Optical Microscope (SNOM). By approaching a fiber tip into the close vicinity of the sample the optical emission in the near-field regime is collected. This way of collecting the light is not affected by the diffraction limit. We employ a low temperature SNOM to investigate the photoluminescence of InAs QDs emitting at 1300nm wavelength embedded in photonic crystal cavities. At each location of an image scan the tip is stopped and a spectrum is acquired. We then plot maps of the photoluminescence for each wavelength. With this instrument it is now possible to directly observe the coupling of QDs to photonic crystal cavities both spectrally and spatially. We show first results of photoluminescence mapping of InAs QDs in photonic crystal cavities.

Particles and waves in electron optics and microscopy

CERN Document Server

Pozzi, Giulio

2016-01-01

Advances in Imaging and Electron Physics merges two long-running serials, Advances in Electronics and Electron Physics and Advances in Optical and Electron Microscopy. The series features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science, digital image processing, electromagnetic wave propagation, electron microscopy, and the computing methods used in all these domains. * Contains contributions from leading authorities on the subject matter* Informs and updates all the latest developments in the field of imaging and electron physics* Provides practitioners interested in microscopy, optics, image processing, mathematical morphology, electromagnetic fields, electron, and ion emission with a valuable resource* Features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science, and digital image pro...

Near-field optical microscopy of localized excitations on rough surfaces: influence of a probe

DEFF Research Database (Denmark)

Bozhevolnyi, Sergey I.

1999-01-01

Starting from the general principles of near-field optical microscopy. I consider the influence of a probe when being used to image localized dipolar excitations and suggest a way of evaluating the perturbation thus introduced. Using the rigorous microscopic (electric) point-dipole description, I...

Wide-field optical detection of nanoparticles using on-chip microscopy and self-assembled nanolenses

Science.gov (United States)

Mudanyali, Onur; McLeod, Euan; Luo, Wei; Greenbaum, Alon; Coskun, Ahmet F.; Hennequin, Yves; Allier, Cédric P.; Ozcan, Aydogan

2013-03-01

The direct observation of nanoscale objects is a challenging task for optical microscopy because the scattering from an individual nanoparticle is typically weak at optical wavelengths. Electron microscopy therefore remains one of the gold standard visualization methods for nanoparticles, despite its high cost, limited throughput and restricted field-of-view. Here, we describe a high-throughput, on-chip detection scheme that uses biocompatible wetting films to self-assemble aspheric liquid nanolenses around individual nanoparticles to enhance the contrast between the scattered and background light. We model the effect of the nanolens as a spatial phase mask centred on the particle and show that the holographic diffraction pattern of this effective phase mask allows detection of sub-100 nm particles across a large field-of-view of >20 mm2. As a proof-of-concept demonstration, we report on-chip detection of individual polystyrene nanoparticles, adenoviruses and influenza A (H1N1) viral particles.

Near-Field Optical Microscopy of Fractal Structures

DEFF Research Database (Denmark)

Coello, Victor; Bozhevolnyi, Sergey I.

1999-01-01

Using a photon scanning tunnelling microscope combined with a shear-force feedback system, we image both topographical and near-field optical images (at the wavelengths of 633 and 594 nm) of silver colloid fractals. Near-field optical imaging is calibrated with a standing evanescent wave pattern...

Application of carbon nanotubes to topographical resolution enhancement of tapered fiber scanning near field optical microscopy probes

Science.gov (United States)

Huntington, S. T.; Jarvis, S. P.

2003-05-01

Scanning near field optical microscopy (SNOM) probes are typically tapered optical fibers with metallic coatings. The tip diameters are generally in excess of 300 nm and thus provide poor topographical resolution. Here we report on the attachment multiwalled carbon nanotubes to the probes in order to substantially enhance the topographical resolution, without adversely affecting the optical resolution.

Fluorescence in situ hybridization on human metaphase chromosomes detected by near-field scanning optical microscopy

NARCIS (Netherlands)

Moers, M.H.P.; Moers, M.H.P.; Kalle, W.H.J.; Kalle, W.H.J.; Ruiter, A.G.T.; Wiegant, J.C.A.G.; Raap, A.K.; Greve, Jan; de Grooth, B.G.; van Hulst, N.F.

1996-01-01

Fluorescence in situ hybridization o­n human metaphase chromosomes is detected by near-field scanning optical microscopy. This combination of cytochemical and scanning probe techniques enables the localization and identification of several fluorescently labelled genomic DNA fragments o­n a single

Application of super-resolution optical microscopy in biology

International Nuclear Information System (INIS)

Mao Xiuhai; Du Jiancong; Huang Qing; Fan Chunhai; Deng Suhui

2013-01-01

Background: A noninvasive, real-time far-field optical microscopy is needed to study the dynamic function inside cells and proteins. However, the resolution limit of traditional optical microscope is about 200 nm due to the diffraction limit of light. So, it's hard to directly observe the subcellular structures. Over the past several years of microscopy development, the diffraction limit of fluorescence microscopy has been overcome and its resolution limit is about tens of nanometers. Methods: To overcome the diffraction limit of light, many super-resolution fluoresce microscopes, including stimulated emission of depletion microscopy (STED), photoactivation localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), have been developed. Conclusions: These methods have been applied in cell biology, microbiology and neurobiology, and the technology of super-resolution provides a new insight into the life science. (authors)

Fabrication of a novel nano-probe slide for near-field optical microscopy

International Nuclear Information System (INIS)

Yim, Sang-Youp; Jeang, Eun-Hee; Lee, Jae-Hoon; Park, Seung-Han; Cho, Kyu-Man

2004-01-01

A novel probe structure, which can act as a planar nano-probe slide for near-field microscopy, was proposed and fabricated. Sub-wavelength apertures on a Si substrate are successfully produced by means of standard photolithography techniques with properly selected masks. In particular, the anisotropic etching characteristics of Si substrate and the hardness of the Si 3 N 4 film are utilized. Probe-to-probe scanning of the fabricated near-field nano-probe slide shows sub-wavelength confinement of light and comparable throughput to the conventional optical fiber probe. We also show that the nano-probe slide can serve as a supporting base and a sub-wavelength aperture to obtain the near-field photoluminescence spectra of a limited number of CdSe nanocrystals.

Chemically etched fiber tips for near-field optical microscopy: a process for smoother tips.

Science.gov (United States)

Lambelet, P; Sayah, A; Pfeffer, M; Philipona, C; Marquis-Weible, F

1998-11-01

An improved method for producing fiber tips for scanning near-field optical microscopy is presented. The improvement consists of chemically etching quartz optical fibers through their acrylate jacket. This new method is compared with the previous one in which bare fibers were etched. With the new process the meniscus formed by the acid along the fiber does not move during etching, leading to a much smoother surface of the tip cone. Subsequent metallization is thus improved, resulting in better coverage of the tip with an aluminum opaque layer. Our results show that leakage can be avoided along the cone, and light transmission through the tip is spatially limited to an optical aperture of a 100-nm dimension.

Near-field scanning optical microscopy cross-sectional measurements of crystalline GaAs solar cells

International Nuclear Information System (INIS)

Herndon, M. K.; Bradford, W. C.; Collins, R. T.; Hawkins, B. E.; Kuech, T. F.; Friedman, D. J.; Kurtz, S. R.

2000-01-01

Near-field scanning optical microscopy (NSOM) was used to study cleaved edges of GaAs solar cell devices. Using visible light for excitation, the NSOM acquired spatially resolved traces of the photocurrent response across the various layers in the device. For excitation energies well above the band gap, carrier recombination at the cleaved surface had a strong influence on the photocurrent signal. Decreasing the excitation energy, which increased the optical penetration depth, allowed the effects of surface recombination to be separated from collection by the pn junction. Using this approach, the NSOM measurements directly observed the effects of a buried minority carrier reflector/passivation layer. (c) 2000 American Institute of Physics

Longitudinal correlation properties of an optical field with broad angular and frequency spectra and their manifestation in interference microscopy

International Nuclear Information System (INIS)

Lyakin, D V; Ryabukho, V P

2013-01-01

The results of theoretical and experimental studies of the longitudinal correlation properties of an optical field with broad angular and frequency spectra and manifestations of these properties in interference microscopy are presented. The joint and competitive influence of the angular and frequency spectra of the object-probing field on the longitudinal resolution and on the amplitude of the interference microscope signals from the interfaces between the media inside a multilayer object is demonstrated. The method of compensating the so-called defocusing effect that arises in the interference microscopy using objectives with a large numerical aperture is experimentally demonstrated, which consists in using as a light source in the interference microscope an illuminating interferometer with a frequency-broadband light source. This method of compensation may be used as the basis of simultaneous determination of geometric thickness and refractive index of media forming a multilayer object. (optical fields)

Aberrations and adaptive optics in super-resolution microscopy

Science.gov (United States)

Booth, Martin; Andrade, Débora; Burke, Daniel; Patton, Brian; Zurauskas, Mantas

2015-01-01

As one of the most powerful tools in the biological investigation of cellular structures and dynamic processes, fluorescence microscopy has undergone extraordinary developments in the past decades. The advent of super-resolution techniques has enabled fluorescence microscopy – or rather nanoscopy – to achieve nanoscale resolution in living specimens and unravelled the interior of cells with unprecedented detail. The methods employed in this expanding field of microscopy, however, are especially prone to the detrimental effects of optical aberrations. In this review, we discuss how super-resolution microscopy techniques based upon single-molecule switching, stimulated emission depletion and structured illumination each suffer from aberrations in different ways that are dependent upon intrinsic technical aspects. We discuss the use of adaptive optics as an effective means to overcome this problem. PMID:26124194

Investigation of optical nanostructures for photovoltaics with near-field scanning microscopy; Untersuchung optischer Nanostrukturen fuer die Photovoltaik mit Nahfeldmikroskopie

Energy Technology Data Exchange (ETDEWEB)

Beckers, Thomas

2011-09-26

Textured and rough surfaces are known to increase light trapping in solar cells significantly. The development and optimization of these nano-structures is essential to improve the energy conversion efficiency of thin-film solar cells. In the past, first research approaches covered classical and macroscopic investigations, e.g. determining the haze or angularly resolved scattering. These methods do not provide precise explanation for the optical improvement of the devices, because layer thicknesses and structure sizes in thin-film solar cells are smaller than the wavelength of visible light. The impact of local nano-structures and their contribution to the local absorption enhancement is not resolved by macroscopic measurements. In this thesis, near-field scanning optical microscopy is introduced as first near-field investigations of nano-structures for photovoltaics. This provides an insight into local optical effects for relevant surfaces of photovoltaic devices. Investigating the distribution of the electric fields in layer stacks is crucial to understand the absorption in solar cells. Evanescent fields, which occur due to total internal reflection at the interfaces, are measurable by near-field scanning optical microscopy and yield important information about local light trapping. Within the framework of this thesis, correlations between local surface structures and optical near-field effects are shown. In this case structure features of randomly textured surfaces, which optimize local light trapping, are identified. It paves the way to connect microscopic optical effects on the surface with the macroscopic performance of thin-film solar cells. Moreover, the measurement yields a 3D illustration of the electric field distribution over the sample surface. It is an important criterion to prove the results of rigorous diffraction theory. An excellent agreement between experiment and simulation is found. The simulations provide an insight into the material, which is

Single-spin stochastic optical reconstruction microscopy.

Science.gov (United States)

Pfender, Matthias; Aslam, Nabeel; Waldherr, Gerald; Neumann, Philipp; Wrachtrup, Jörg

2014-10-14

We experimentally demonstrate precision addressing of single-quantum emitters by combined optical microscopy and spin resonance techniques. To this end, we use nitrogen vacancy (NV) color centers in diamond confined within a few ten nanometers as individually resolvable quantum systems. By developing a stochastic optical reconstruction microscopy (STORM) technique for NV centers, we are able to simultaneously perform sub-diffraction-limit imaging and optically detected spin resonance (ODMR) measurements on NV spins. This allows the assignment of spin resonance spectra to individual NV center locations with nanometer-scale resolution and thus further improves spatial discrimination. For example, we resolved formerly indistinguishable emitters by their spectra. Furthermore, ODMR spectra contain metrology information allowing for sub-diffraction-limit sensing of, for instance, magnetic or electric fields with inherently parallel data acquisition. As an example, we have detected nuclear spins with nanometer-scale precision. Finally, we give prospects of how this technique can evolve into a fully parallel quantum sensor for nanometer resolution imaging of delocalized quantum correlations.

Rigorous numerical modeling of scattering-type scanning near-field optical microscopy and spectroscopy

Science.gov (United States)

Chen, Xinzhong; Lo, Chiu Fan Bowen; Zheng, William; Hu, Hai; Dai, Qing; Liu, Mengkun

2017-11-01

Over the last decade, scattering-type scanning near-field optical microscopy and spectroscopy have been widely used in nano-photonics and material research due to their fine spatial resolution and broad spectral range. A number of simplified analytical models have been proposed to quantitatively understand the tip-scattered near-field signal. However, a rigorous interpretation of the experimental results is still lacking at this stage. Numerical modelings, on the other hand, are mostly done by simulating the local electric field slightly above the sample surface, which only qualitatively represents the near-field signal rendered by the tip-sample interaction. In this work, we performed a more comprehensive numerical simulation which is based on realistic experimental parameters and signal extraction procedures. By directly comparing to the experiments as well as other simulation efforts, our methods offer a more accurate quantitative description of the near-field signal, paving the way for future studies of complex systems at the nanoscale.

A spatio-temporally compensated acousto-optic scanner for two-photon microscopy providing large field of view.

Science.gov (United States)

Kremer, Y; Léger, J-F; Lapole, R; Honnorat, N; Candela, Y; Dieudonné, S; Bourdieu, L

2008-07-07

Acousto-optic deflectors (AOD) are promising ultrafast scanners for non-linear microscopy. Their use has been limited until now by their small scanning range and by the spatial and temporal dispersions of the laser beam going through the deflectors. We show that the use of AOD of large aperture (13mm) compared to standard deflectors allows accessing much larger field of view while minimizing spatio-temporal distortions. An acousto-optic modulator (AOM) placed at distance of the AOD is used to compensate spatial and temporal dispersions. Fine tuning of the AOM-AOD setup using a frequency-resolved optical gating (GRENOUILLE) allows elimination of pulse front tilt whereas spatial chirp is minimized thanks to the large aperture AOD.

Feasibility of full-field optical coherence microscopy in ultra-structural imaging of human colon tissues

Energy Technology Data Exchange (ETDEWEB)

Choi, Eun Seo [Chosun University, Gwangju (Korea, Republic of); Choi, Woo June; Ryu, Seon Young; Lee, Byeong Ha [Gwangju Institute of Science and Technology, Gwangju (Korea, Republic of); Lee, Jae Hyuk; Bom, Hee Seung; Lee, Byeong Il [Chonnam National University Hospital, Gwangju (Korea, Republic of)

2010-06-15

We demonstrated the imaging feasibility of full-field optical coherence microscopy (FF-OCM) in pathological diagnosis of human colon tissues. FF-OCM images with high transverse resolution were obtained at different depths of the samples without any dye staining or physical slicing, and detailed microstructures of human colon tissues were visualized. Morphological differences in normal tissues, cancer tissues, and tissues under transition were observed and matched with results seen in conventional optical microscope images. The optical biopsy based on FF-OCM could overcome the limitations on the number of physical cuttings of tissues and could perform high-throughput mass diagnosis of diseased tissues. The proved utility of FF-OCM as a comprehensive and efficient imaging modality of human tissues showed it to be a good alternative to conventional biopsy.

Feasibility of full-field optical coherence microscopy in ultra-structural imaging of human colon tissues

International Nuclear Information System (INIS)

Choi, Eun Seo; Choi, Woo June; Ryu, Seon Young; Lee, Byeong Ha; Lee, Jae Hyuk; Bom, Hee Seung; Lee, Byeong Il

2010-01-01

We demonstrated the imaging feasibility of full-field optical coherence microscopy (FF-OCM) in pathological diagnosis of human colon tissues. FF-OCM images with high transverse resolution were obtained at different depths of the samples without any dye staining or physical slicing, and detailed microstructures of human colon tissues were visualized. Morphological differences in normal tissues, cancer tissues, and tissues under transition were observed and matched with results seen in conventional optical microscope images. The optical biopsy based on FF-OCM could overcome the limitations on the number of physical cuttings of tissues and could perform high-throughput mass diagnosis of diseased tissues. The proved utility of FF-OCM as a comprehensive and efficient imaging modality of human tissues showed it to be a good alternative to conventional biopsy.

Field-based dynamic light scattering microscopy: theory and numerical analysis.

Science.gov (United States)

Joo, Chulmin; de Boer, Johannes F

2013-11-01

We present a theoretical framework for field-based dynamic light scattering microscopy based on a spectral-domain optical coherence phase microscopy (SD-OCPM) platform. SD-OCPM is an interferometric microscope capable of quantitative measurement of amplitude and phase of scattered light with high phase stability. Field-based dynamic light scattering (F-DLS) analysis allows for direct evaluation of complex-valued field autocorrelation function and measurement of localized diffusive and directional dynamic properties of biological and material samples with high spatial resolution. In order to gain insight into the information provided by F-DLS microscopy, theoretical and numerical analyses are performed to evaluate the effect of numerical aperture of the imaging optics. We demonstrate that sharp focusing of fields affects the measured diffusive and transport velocity, which leads to smaller values for the dynamic properties in the sample. An approach for accurately determining the dynamic properties of the samples is discussed.

An Evanescent Field Optical Microscope. Scanning probe Microscopy

NARCIS (Netherlands)

van Hulst, N.F.; Segerink, Franciscus B.; Bölger, B.; Bölger, B.; Wickramasinghe, H. Kumar

1991-01-01

An Evanescent Field Optical Microscope (EFOM) is presented, which employs frustrated total internal reflection on a highly localized scale by means of a sharp dielectric tip. The coupling of the evanescent field to the sub-micrometer probe as a function of probe-sample distance, angle of incidence

Analysis of artificial opals by scanning near field optical microscopy

Science.gov (United States)

Barrio, J.; Lozano, G.; Lamela, J.; Lifante, G.; Dorado, L. A.; Depine, R. A.; Jaque, F.; Míguez, H.

2011-04-01

Herein we present a detailed analysis of the optical response of artificial opal films realized employing a near-field scanning optical microscope in collection and transmission modes. Near-field patterns measured at the rear surface when a plane wave impinges on the front face are presented with the finding that optical intensity maps present a clear correlation with the periodic arrangement of the outer surface. Calculations based on the vector Korringa-Kohn-Rostoker method reproduce the different profiles experimentally observed as well as the response to the polarization of the incident field. These observations constitute the first experimental confirmation of the collective lattice resonances that give rise to the optical response of these three dimensional periodic structures in the high-energy range.

Stochastic Optical Reconstruction Microscopy (STORM).

Science.gov (United States)

Xu, Jianquan; Ma, Hongqiang; Liu, Yang

2017-07-05

Super-resolution (SR) fluorescence microscopy, a class of optical microscopy techniques at a spatial resolution below the diffraction limit, has revolutionized the way we study biology, as recognized by the Nobel Prize in Chemistry in 2014. Stochastic optical reconstruction microscopy (STORM), a widely used SR technique, is based on the principle of single molecule localization. STORM routinely achieves a spatial resolution of 20 to 30 nm, a ten-fold improvement compared to conventional optical microscopy. Among all SR techniques, STORM offers a high spatial resolution with simple optical instrumentation and standard organic fluorescent dyes, but it is also prone to image artifacts and degraded image resolution due to improper sample preparation or imaging conditions. It requires careful optimization of all three aspects-sample preparation, image acquisition, and image reconstruction-to ensure a high-quality STORM image, which will be extensively discussed in this unit. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Near-field scanning optical microscopy based nanostructuring of glass

International Nuclear Information System (INIS)

Chimmalgi, A; Hwang, D J; Grigoropoulos, C P

2007-01-01

Nanofabrication, at lateral resolutions beyond the capability of conventional optical lithography techniques, is demonstrated here. Femtosecond laser was used in conjunction with Near-field Scanning Optical Microscopes (NSOMs) to nanostructure thin metal films. Also, the possibility of using these nanostructured metal films as masks to effectively transfer the pattern to the underlying substrate by wet etching process is shown. Two different optical nearfiled processing schemes were studied for near-field nanostructuring. In the first scheme, local field enhancement in the near-field of a scanning probe microscope (SPM) probe tip irradiated with femtosecond laser pulses was utilized (apertureless NSOM mode) and as a second approach, femtosecond laser beam was spatially confined by cantilevered NSOM fiber tip (apertured NOSM mode). The minimized heat- and shock-affected areas introduced during ultrafast laser based machining process, allows processing of even high conductivity thin metal films with minimized formation of any interfacial compounds between the metal films and the underlying substrate. Potential applications of this method may be in the fields of nanolithography, nanofluidics, nanoscale chemical and gas sensors, high-density data storage, nano-opto-electronics, as well as biotechnology related applications

Microsphere-aided optical microscopy and its applications for super-resolution imaging

Science.gov (United States)

Upputuri, Paul Kumar; Pramanik, Manojit

2017-12-01

The spatial resolution of a standard optical microscope (SOM) is limited by diffraction. In visible spectrum, SOM can provide ∼ 200 nm resolution. To break the diffraction limit several approaches were developed including scanning near field microscopy, metamaterial super-lenses, nanoscale solid immersion lenses, super-oscillatory lenses, confocal fluorescence microscopy, techniques that exploit non-linear response of fluorophores like stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, etc. Recently, photonic nanojet generated by a dielectric microsphere was used to break the diffraction limit. The microsphere-approach is simple, cost-effective and can be implemented under a standard microscope, hence it has gained enormous attention for super-resolution imaging. In this article, we briefly review the microsphere approach and its applications for super-resolution imaging in various optical imaging modalities.

Dark Field Microscopy for Analytical Laboratory Courses

Science.gov (United States)

Augspurger, Ashley E.; Stender, Anthony S.; Marchuk, Kyle; Greenbowe, Thomas J.; Fang, Ning

2014-01-01

An innovative and inexpensive optical microscopy experiment for a quantitative analysis or an instrumental analysis chemistry course is described. The students have hands-on experience with a dark field microscope and investigate the wavelength dependence of localized surface plasmon resonance in gold and silver nanoparticles. Students also…

In vivo calcium imaging from dentate granule cells with wide-field fluorescence microscopy.

Directory of Open Access Journals (Sweden)

Yuichiro Hayashi

Full Text Available A combination of genetically-encoded calcium indicators and micro-optics has enabled monitoring of large-scale dynamics of neuronal activity from behaving animals. In these studies, wide-field microscopy is often used to visualize neural activity. However, this method lacks optical sectioning capability, and therefore its axial resolution is generally poor. At present, it is unclear whether wide-field microscopy can visualize activity of densely packed small neurons at cellular resolution. To examine the applicability of wide-field microscopy for small-sized neurons, we recorded calcium activity of dentate granule cells having a small soma diameter of approximately 10 micrometers. Using a combination of high numerical aperture (0.8 objective lens and independent component analysis-based image segmentation technique, activity of putative single granule cell activity was separated from wide-field calcium imaging data. The result encourages wider application of wide-field microscopy in in vivo neurophysiology.

Holographic fluorescence microscopy with incoherent digital holographic adaptive optics.

Science.gov (United States)

Jang, Changwon; Kim, Jonghyun; Clark, David C; Lee, Seungjae; Lee, Byoungho; Kim, Myung K

2015-01-01

Introduction of adaptive optics technology into astronomy and ophthalmology has made great contributions in these fields, allowing one to recover images blurred by atmospheric turbulence or aberrations of the eye. Similar adaptive optics improvement in microscopic imaging is also of interest to researchers using various techniques. Current technology of adaptive optics typically contains three key elements: a wavefront sensor, wavefront corrector, and controller. These hardware elements tend to be bulky, expensive, and limited in resolution, involving, for example, lenslet arrays for sensing or multiactuator deformable mirrors for correcting. We have previously introduced an alternate approach based on unique capabilities of digital holography, namely direct access to the phase profile of an optical field and the ability to numerically manipulate the phase profile. We have also demonstrated that direct access and compensation of the phase profile are possible not only with conventional coherent digital holography, but also with a new type of digital holography using incoherent light: selfinterference incoherent digital holography (SIDH). The SIDH generates a complex—i.e., amplitude plus phase—hologram from one or several interferograms acquired with incoherent light, such as LEDs, lamps, sunlight, or fluorescence. The complex point spread function can be measured using guide star illumination and it allows deterministic deconvolution of the full-field image. We present experimental demonstration of aberration compensation in holographic fluorescence microscopy using SIDH. Adaptive optics by SIDH provides new tools for improved cellular fluorescence microscopy through intact tissue layers or other types of aberrant media.

Scanning near-field infrared microscopy on semiconductor structures

Energy Technology Data Exchange (ETDEWEB)

Jacob, Rainer

2011-01-15

Near-field optical microscopy has attracted remarkable attention, as it is the only technique that allows the investigation of local optical properties with a resolution far below the diffraction limit. Especially, the scattering-type near-field optical microscopy allows the nondestructive examination of surfaces without restrictions to the applicable wavelengths. However, its usability is limited by the availability of appropriate light sources. In the context of this work, this limit was overcome by the development of a scattering-type near-field microscope that uses a widely tunable free-electron laser as primary light source. In the theoretical part, it is shown that an optical near-field contrast can be expected when materials with different dielectric functions are combined. It is derived that these differences yield different scattering cross-sections for the coupled system of the probe and the sample. Those cross-sections define the strength of the near-field signal that can be measured for different materials. Hence, an optical contrast can be expected, when different scattering cross-sections are probed. This principle also applies to vertically stacked or even buried materials, as shown in this thesis experimentally for two sample systems. In the first example, the different dielectric functions were obtained by locally changing the carrier concentration in silicon by the implantation of boron. It is shown that the concentration of free charge-carriers can be deduced from the near-field contrast between implanted and pure silicon. For this purpose, two different experimental approaches were used, a non-interferometric one by using variable wavelengths and an interferometric one with a fixed wavelength. As those techniques yield complementary information, they can be used to quantitatively determine the effective carrier concentration. Both approaches yield consistent results for the carrier concentration, which excellently agrees with predictions from

Scanning near-field infrared microscopy on semiconductor structures

International Nuclear Information System (INIS)

Jacob, Rainer

2011-01-01

Near-field optical microscopy has attracted remarkable attention, as it is the only technique that allows the investigation of local optical properties with a resolution far below the diffraction limit. Especially, the scattering-type near-field optical microscopy allows the nondestructive examination of surfaces without restrictions to the applicable wavelengths. However, its usability is limited by the availability of appropriate light sources. In the context of this work, this limit was overcome by the development of a scattering-type near-field microscope that uses a widely tunable free-electron laser as primary light source. In the theoretical part, it is shown that an optical near-field contrast can be expected when materials with different dielectric functions are combined. It is derived that these differences yield different scattering cross-sections for the coupled system of the probe and the sample. Those cross-sections define the strength of the near-field signal that can be measured for different materials. Hence, an optical contrast can be expected, when different scattering cross-sections are probed. This principle also applies to vertically stacked or even buried materials, as shown in this thesis experimentally for two sample systems. In the first example, the different dielectric functions were obtained by locally changing the carrier concentration in silicon by the implantation of boron. It is shown that the concentration of free charge-carriers can be deduced from the near-field contrast between implanted and pure silicon. For this purpose, two different experimental approaches were used, a non-interferometric one by using variable wavelengths and an interferometric one with a fixed wavelength. As those techniques yield complementary information, they can be used to quantitatively determine the effective carrier concentration. Both approaches yield consistent results for the carrier concentration, which excellently agrees with predictions from

Visual-servoing optical microscopy

Science.gov (United States)

Callahan, Daniel E.; Parvin, Bahram

2009-06-09

The present invention provides methods and devices for the knowledge-based discovery and optimization of differences between cell types. In particular, the present invention provides visual servoing optical microscopy, as well as analysis methods. The present invention provides means for the close monitoring of hundreds of individual, living cells over time: quantification of dynamic physiological responses in multiple channels; real-time digital image segmentation and analysis; intelligent, repetitive computer-applied cell stress and cell stimulation; and the ability to return to the same field of cells for long-term studies and observation. The present invention further provides means to optimize culture conditions for specific subpopulations of cells.

Light depolarization induced by metallic tips in apertureless near-field optical microscopy and tip-enhanced Raman spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Gucciardi, P G [CNR-Istituto per i Processi Chimico-Fisici, sezione Messina, Salita Sperone, Contrada Papardo, I-98158 Faro Superiore, Messina (Italy); Lopes, M; Deturche, R; Julien, C; Barchiesi, D; Chapelle, M Lamy de la [Institut Charles Delaunay-CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de Technologie de Troyes, 12 rue Marie Curie, BP2060, 10010 Troyes (France)

2008-05-28

We have investigated the depolarization effects of light scattered by sharp tips used for apertureless near-field optical microscopy. Dielectric and metal coated tips have been investigated and depolarization factors between 5 and 30% have been measured, changing as a function of the incident light polarization and of the tip shape. The experimental results are in good agreement with theoretical calculations performed by the finite element method, giving a near-field depolarization factor close to 10%. The effect of depolarization has been investigated in polarized tip-enhanced Raman spectroscopy (TERS) experiments; the depolarization gives rise to forbidden Raman modes in Si crystals.

Near-field optical microscopy with a scanning tunneling microscope

International Nuclear Information System (INIS)

Barbara, A.; Lopez-Rios, T.; Quemerais, P.

2005-01-01

A homemade apertureless near-field optical microscope using a scanning tunneling microscope (STM) is described. The experimental set-up simultaneously provides optical and topographic images of the sample. Technical details and features of the set-up are presented, together with results demonstrating the sub-wavelength resolution achieved as well as its sensitivity to dielectric contrasts. We show that the use of a STM permits to precisely control very small distances between the tip and the sample which is a great advantage to excite localized optical resonances between the tip and the surface

Single spin stochastic optical reconstruction microscopy

OpenAIRE

Pfender, Matthias; Aslam, Nabeel; Waldherr, Gerald; Wrachtrup, Jörg

2014-01-01

We experimentally demonstrate precision addressing of single quantum emitters by combined optical microscopy and spin resonance techniques. To this end we utilize nitrogen-vacancy (NV) color centers in diamond confined within a few ten nanometers as individually resolvable quantum systems. By developing a stochastic optical reconstruction microscopy (STORM) technique for NV centers we are able to simultaneously perform sub diffraction-limit imaging and optically detected spin resonance (ODMR)...

Imaging slit-coupled surface plasmon polaritons using conventional optical microscopy.

Science.gov (United States)

Mehfuz, R; Chowdhury, F A; Chau, K J

2012-05-07

We develop a technique that now enables surface plasmon polaritons (SPPs) coupled by nano-patterned slits in a metal film to be detected using conventional optical microscopy with standard objective lenses. The crux of this method is an ultra-thin polymer layer on the metal surface, whose thickness can be varied over a nanoscale range to enable controllable tuning of the SPP momentum. At an optimal layer thickness for which the SPP momentum matches the momentum of light emerging from the slit, the SPP coupling efficiency is enhanced about six times relative to that without the layer. The enhanced efficiency results in distinctive and bright plasmonic signatures near the slit visible by naked eye under an optical microscope. We demonstrate how this capability can be used for parallel measurement through a simple experiment in which the SPP propagation distance is extracted from a single microscope image of an illuminated array of nano-patterned slits on a metal surface. We also use optical microscopy to image the focal region of a plasmonic lens and obtain results consistent with a previously-reported results using near-field optical microscopy. Measurement of SPPs near a nano-slit using conventional and widely-available optical microscopy is an important step towards making nano-plasmonic device technology highly accessible and easy-to-use.

Imaging of phase change materials below a capping layer using correlative infrared near-field microscopy and electron microscopy

Science.gov (United States)

Lewin, M.; Hauer, B.; Bornhöfft, M.; Jung, L.; Benke, J.; Michel, A.-K. U.; Mayer, J.; Wuttig, M.; Taubner, T.

2015-10-01

Phase Change Materials (PCM) show two stable states in the solid phase with significantly different optical and electronic properties. They can be switched reversibly between those two states and are promising candidates for future non-volatile memory applications. The development of phase change devices demands characterization tools, yielding information about the switching process at high spatial resolution. Scattering-type Scanning Near-field Optical Microscopy (s-SNOM) allows for spectroscopic analyses of the different optical properties of the PCMs on the nm-scale. By correlating the optical s-SNOM images with transmission electron microscopy images of the same sample, we unambiguously demonstrate the correlation of the infrared optical contrast with the structural state of the phase change material. The investigated sample consists of sandwiched amorphous and crystalline regions of Ag 4 In 3 Sb 67 Te 26 below a 100 nm thick ( ZnS ) 80 - ( SiO2 ) 20 capping layer. Our results demonstrate the sensitivity of s-SNOM to small dielectric near-field contrasts even below a comparably thick capping layer ( 100 nm ).

Fully time-resolved near-field scanning optical microscopy fluorescence imaging

International Nuclear Information System (INIS)

Kwak, Eun-Soo; Vanden Bout, David A.

2003-01-01

Time-correlated single photon counting has been coupled with near-field scanning optical microscopy (NSOM) to record complete fluorescence lifetime decays at each pixel in an NSOM image. The resulting three-dimensional data sets can be binned in the time dimension to create images of photons at particular time delays or images of the fluorescence lifetime. Alternatively, regions of interest identified in the topography and fluorescence images can be used to bin the data in the spatial dimensions resulting in high signal to noise fluorescence decays of particular regions of the sample. The technique has been demonstrated on films of poly(vinylalcohol), doped with the fluorescent dye, cascade blue (CB). The CB segregates into small circular regions of high concentration within the films during the drying process. The lifetime imaging shows that the spots have slightly faster excited state decays due to quenching of the luminescence as a result of the higher concentration. The technique is also used to image the fluorescence lifetime of an annealed film of poly(dihexylfluorene). The samples show high contrast in the total intensity fluorescence image, but the lifetime image reveals the sample to be extremely uniform

Polarization contrast in reflection near-field optical microscopy with uncoated fibre tips

DEFF Research Database (Denmark)

Bozhevolnyi, Sergey I.; Langbein, Wolfgang; Hvam, Jørn Märcher

1999-01-01

Using cross-hatched, patterned semiconductor surfaces and round 20-nm-thick gold pads on semiconductor wafers, we investigate the imaging characteristics of a reflection near-field optical microscope with an uncoated fibre tip for different polarization configurations and light wavelengths....... Is is shown that cross-polarized detection allows one to effectively suppress far-field components in the detected signal and to realise imaging of optical contrast on the sub-wavelength scale. The sensitivity window of our microscope, i.e. the scale on which near-field optical images represent mainly optical...

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy

Science.gov (United States)

Lim, Daryl; Ford, Tim N.; Chu, Kengyeh K.; Mertz, Jerome

2011-01-01

We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish.

Fast Calcium Imaging with Optical Sectioning via HiLo Microscopy.

Science.gov (United States)

Lauterbach, Marcel A; Ronzitti, Emiliano; Sternberg, Jenna R; Wyart, Claire; Emiliani, Valentina

2015-01-01

Imaging intracellular calcium concentration via reporters that change their fluorescence properties upon binding of calcium, referred to as calcium imaging, has revolutionized our way to probe neuronal activity non-invasively. To reach neurons densely located deep in the tissue, optical sectioning at high rate of acquisition is necessary but difficult to achieve in a cost effective manner. Here we implement an accessible solution relying on HiLo microscopy to provide robust optical sectioning with a high frame rate in vivo. We show that large calcium signals can be recorded from dense neuronal populations at high acquisition rates. We quantify the optical sectioning capabilities and demonstrate the benefits of HiLo microscopy compared to wide-field microscopy for calcium imaging and 3D reconstruction. We apply HiLo microscopy to functional calcium imaging at 100 frames per second deep in biological tissues. This approach enables us to discriminate neuronal activity of motor neurons from different depths in the spinal cord of zebrafish embryos. We observe distinct time courses of calcium signals in somata and axons. We show that our method enables to remove large fluctuations of the background fluorescence. All together our setup can be implemented to provide efficient optical sectioning in vivo at low cost on a wide range of existing microscopes.

Progress in the Correlative Atomic Force Microscopy and Optical Microscopy

Directory of Open Access Journals (Sweden)

Lulu Zhou

2017-04-01

Full Text Available Atomic force microscopy (AFM has evolved from the originally morphological imaging technique to a powerful and multifunctional technique for manipulating and detecting the interactions between molecules at nanometer resolution. However, AFM cannot provide the precise information of synchronized molecular groups and has many shortcomings in the aspects of determining the mechanism of the interactions and the elaborate structure due to the limitations of the technology, itself, such as non-specificity and low imaging speed. To overcome the technical limitations, it is necessary to combine AFM with other complementary techniques, such as fluorescence microscopy. The combination of several complementary techniques in one instrument has increasingly become a vital approach to investigate the details of the interactions among molecules and molecular dynamics. In this review, we reported the principles of AFM and optical microscopy, such as confocal microscopy and single-molecule localization microscopy, and focused on the development and use of correlative AFM and optical microscopy.

Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

Science.gov (United States)

Kim, Doory; Deerinck, Thomas J.; Sigal, Yaron M.; Babcock, Hazen P.; Ellisman, Mark H.; Zhuang, Xiaowei

2015-01-01

Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM) and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets. PMID:25874453

X-ray diffraction microscopy based on refractive optics

DEFF Research Database (Denmark)

Poulsen, Henning Friis; Jakobsen, A. C.; Simons, Hugh

2017-01-01

A formalism is presented for dark‐field X‐ray microscopy using refractive optics. The new technique can produce three‐dimensional maps of lattice orientation and axial strain within millimetre‐sized sampling volumes and is particularly suited to in situ studies of materials at hard X‐ray energies....... An objective lens in the diffracted beam magnifies the image and acts as a very efficient filter in reciprocal space, enabling the imaging of individual domains of interest with a resolution of 100 nm. Analytical expressions for optical parameters such as numerical aperture, vignetting, and the resolution...

Fluorescent nanoscale detection of biotin-streptavidin interaction using near-field scanning optical microscopy

International Nuclear Information System (INIS)

Park, Hyun Kyu; Chung, Bong Hyun; Gokarna, Anisha; Hulme, John P; Park, Hyun Gyu

2008-01-01

We describe a nanoscale strategy for detecting biotin-streptavidin binding using near-field scanning optical microscopy (NSOM) that exploits the fluorescence properties of single polydiacetylene (PDA) liposomes. NSOM is more useful to observe nanomaterials having optical properties with the help of topological information. We synthesized amine-terminated 10,12-pentacosadiynoic acid (PCDA) monomer (PCDA-NH 2 ) and used this derivatized monomer to prepare PCDA liposomes. PCDA-NH 2 liposomes were immobilized on an aldehyde-functionalized glass surface followed by photopolymerization by using a 254 nm light source. To measure the biotin-streptavidin binding, we conjugated photoactivatable biotin to immobilized PCDA-NH 2 liposomes by UV irradiation (365 nm) and subsequently allowed them to interact with streptavidin. We analyzed the fluorescence using a fluorescence scanner and observed single liposomes using NSOM. The average height and NSOM signal observed in a single liposome after binding were ∼31.3 to 8.5 ± 0.5 nm and 0.37 to 0.16 ± 0.6 kHz, respectively. This approach, which has the advantage of not requiring a fluorescent label, could prove highly beneficial for single molecule detection technology

Optical Imaging and Microscopy Techniques and Advanced Systems

CERN Document Server

Török, Peter

2007-01-01

This text on contemporary optical systems is intended for optical researchers and engineers, graduate students and optical microscopists in the biological and biomedical sciences. This second edition contains two completely new chapters. In addition most of the chapters from the first edition have been revised and updated. The book consists of three parts: The first discusses high-aperture optical systems, which form the backbone of optical microscopes. An example is a chapter new in the second edition on the emerging field of high numerical aperture diffractive lenses which seems to have particular promise in improving the correction of lenses. In this part particular attention is paid to optical data storage. The second part is on the use of non-linear optical techniques, including nonlinear optical excitation (total internal reflection fluorescence, second and third harmonic generation and two photon microscopy) and non-linear spectroscopy (CARS). The final part of the book presents miscellaneous technique...

Fiber optic probe of free electron evanescent fields in the optical frequency range

Energy Technology Data Exchange (ETDEWEB)

So, Jin-Kyu, E-mail: js1m10@orc.soton.ac.uk; MacDonald, Kevin F. [Optoelectronics Research Centre and Centre for Photonic Metamaterials, University of Southampton, Highfield, Southampton SO17 1BJ (United Kingdom); Zheludev, Nikolay I. [Optoelectronics Research Centre and Centre for Photonic Metamaterials, University of Southampton, Highfield, Southampton SO17 1BJ (United Kingdom); Centre for Disruptive Photonic Technologies, Nanyang Technological University, Singapore 637371 (Singapore)

2014-05-19

We introduce an optical fiber platform which can be used to interrogate proximity interactions between free-electron evanescent fields and photonic nanostructures at optical frequencies in a manner similar to that in which optical evanescent fields are sampled using nanoscale aperture probes in scanning near-field microscopy. Conically profiled optical fiber tips functionalized with nano-gratings are employed to couple electron evanescent fields to light via the Smith-Purcell effect. We demonstrate the interrogation of medium energy (30–50 keV) electron fields with a lateral resolution of a few micrometers via the generation and detection of visible/UV radiation in the 700–300 nm (free-space) wavelength range.

The development of optical microscopy techniques for the advancement of single-particle studies

Science.gov (United States)

Marchuk, Kyle

Single particle orientation and rotational tracking (SPORT) has recently become a powerful optical microscopy tool that can expose many molecular motions. Unfortunately, there is not yet a single microscopy technique that can decipher all particle motions in all environmental conditions, thus there are limitations to current technologies. Within, the two powerful microscopy tools of total internal reflection and interferometry are advanced to determine the position, orientation, and optical properties of metallic nanoparticles in a variety of environments. Total internal reflection is an optical phenomenon that has been applied to microscopy to produce either fluorescent or scattered light. The non-invasive far-field imaging technique is coupled with a near-field illumination scheme that allows for better axial resolution than confocal microscopy and epi-fluorescence microscopy. By controlling the incident illumination angle using total internal reflection fluorescence (TIRF) microscopy, a new type of imaging probe called "non-blinking" quantum dots (NBQDs) were super-localized in the axial direction to sub-10-nm precision. These particles were also used to study the rotational motion of microtubules being propelled by the motor protein kinesin across the substrate surface. The same instrument was modified to function under total internal reflection scattering (TIRS) microscopy to study metallic anisotropic nanoparticles and their dynamic interactions with synthetic lipid bilayers. Utilizing two illumination lasers with opposite polarization directions at wavelengths corresponding to the short and long axis surface plasmon resonance (SPR) of the nanoparticles, both the in-plane and out-of-plane movements of many particles could be tracked simultaneously. When combined with Gaussian point spread function (PSF) fitting for particle super-localization, the binding status and rotational movement could be resolved without degeneracy. TIRS microscopy was also used to

The development of optical microscopy techniques for the advancement of single-particle studies

Energy Technology Data Exchange (ETDEWEB)

Marchuk, Kyle [Iowa State Univ., Ames, IA (United States)

2013-05-15

Single particle orientation and rotational tracking (SPORT) has recently become a powerful optical microscopy tool that can expose many molecular motions. Unfortunately, there is not yet a single microscopy technique that can decipher all particle motions in all environmental conditions, thus there are limitations to current technologies. Within, the two powerful microscopy tools of total internal reflection and interferometry are advanced to determine the position, orientation, and optical properties of metallic nanoparticles in a variety of environments. Total internal reflection is an optical phenomenon that has been applied to microscopy to produce either fluorescent or scattered light. The non-invasive far-field imaging technique is coupled with a near-field illumination scheme that allows for better axial resolution than confocal microscopy and epi-fluorescence microscopy. By controlling the incident illumination angle using total internal reflection fluorescence (TIRF) microscopy, a new type of imaging probe called “non-blinking” quantum dots (NBQDs) were super-localized in the axial direction to sub-10-nm precision. These particles were also used to study the rotational motion of microtubules being propelled by the motor protein kinesin across the substrate surface. The same instrument was modified to function under total internal reflection scattering (TIRS) microscopy to study metallic anisotropic nanoparticles and their dynamic interactions with synthetic lipid bilayers. Utilizing two illumination lasers with opposite polarization directions at wavelengths corresponding to the short and long axis surface plasmon resonance (SPR) of the nanoparticles, both the in-plane and out-of-plane movements of many particles could be tracked simultaneously. When combined with Gaussian point spread function (PSF) fitting for particle super-localization, the binding status and rotational movement could be resolved without degeneracy. TIRS microscopy was also used to

Transfer functions in collection scanning near-field optical microscopy

DEFF Research Database (Denmark)

Bozhevolnyi, Sergey I.; Vohnsen, Brian; Bozhevolnaya, Elena A.

1999-01-01

are considered with respect to the relation between near-field optical images and the corresponding intensity distributions. Our conclusions are supported with numerical simulations and experimental results obtained by using a photon scanning tunneling microscope with an uncoated fiber tip....

Optimization of s-Polarization Sensitivity in Apertureless Near-Field Optical Microscopy

Directory of Open Access Journals (Sweden)

Yuika Saito

2012-01-01

Full Text Available It is a general belief in apertureless near-field microscopy that the so-called p-polarization configuration, where the incident light is polarized parallel to the axis of the probe, is advantageous to its counterpart, the s-polarization configuration, where the incident light is polarized perpendicular to the probe axis. While this is true for most samples under common near-field experimental conditions, there are samples which respond better to the s-polarization configuration due to their orientations. Indeed, there have been several reports that have discussed such samples. This leads us to an important requirement that the near-field experimental setup should be equipped with proper sensitivity for measurements with s-polarization configuration. This requires not only creation of effective s-polarized illumination at the near-field probe, but also proper enhancement of s-polarized light by the probe. In this paper, we have examined the s-polarization enhancement sensitivity of near-field probes by measuring and evaluating the near-field Rayleigh scattering images constructed by a variety of probes. We found that the s-polarization enhancement sensitivity strongly depends on the sharpness of the apex of near-field probes. We have discussed the efficient value of probe sharpness by considering a balance between the enhancement and the spatial resolution, both of which are essential requirements of apertureless near-field microscopy.

Subnanometric stabilization of plasmon-enhanced optical microscopy

International Nuclear Information System (INIS)

Yano, Taka-aki; Ichimura, Taro; Kuwahara, Shota; Verma, Prabhat; Kawata, Satoshi

2012-01-01

We have demonstrated subnanometric stabilization of tip-enhanced optical microscopy under ambient condition. Time-dependent thermal drift of a plasmonic metallic tip was optically sensed at subnanometer scale, and was compensated in real-time. In addition, mechanically induced displacement of the tip, which usually occurs when the amount of tip-applied force varies, was also compensated in situ. The stabilization of tip-enhanced optical microscopy enables us to perform long-time and robust measurement without any degradation of optical signal, resulting in true nanometric optical imaging with high reproducibility and high precision. The technique presented is applicable for AFM-based nanoindentation with subnanometric precision. (paper)

Waveguide evanescent field fluorescence microscopy & its application in cell biology

Science.gov (United States)

Hassanzadeh, Abdollah

There are many powerful microscopy technologies available for the investigation of bulk materials as well as for thin film samples. Nevertheless, for imaging an interface, especially live cells on a substrate and ultra thin-films, only Total Internal Reflection Fluorescence (TIRF) microscopy is available. This TIRF microscopy allows imaging without interference of the bulk. Various approaches are employed in fluorescence microscopy applications to restrict the excitation and detection of fluorophores to a thin region of the specimen. Elimination of background fluorescence from outside the focal plane can dramatically improve the signal-to-noise ratio, and consequently, the spatial resolution of the features or events of interest. TIRF microscopy is an evanescent field based microscopy. In this method, fluorescent dyes are only excited within an evanescent field: roughly within 100 nm above a glass coverslip. This will allow imaging surface and interfacial issues of the glass coverslip and an adjacent material. Waveguide evanescent field fluorescence (WEFF) microscopy is a new development for imaging cell-substrate interactions in real time and in vitro. It is an alternative to TIRF microscopy. In this method the light is coupled into a waveguide via an optical grating. The coupled light propagates as a waveguide mode and exhibits an evanescent field on top of the waveguide. This can be used as a surface-bound illumination source to excite fluorophores. This evanescent field serves as an extremely powerful tool for quality control of thin films, to study cell-substrate contacts, and investigating the effect of external agents and drugs on the cell-substrate interaction in real time and in vitro. This new method has been established and optimized to minimize non-uniformity, scattering and photo bleaching issues. Visualizing and quantifying of the cell-substrates and solid thin films have been carried out by WEFF microscopy. The images of the cell-substrate interface

Optical microscope illumination analysis using through-focus scanning optical microscopy.

Science.gov (United States)

Attota, Ravi Kiran; Park, Haesung

2017-06-15

Misalignment of the aperture diaphragm present in optical microscopes results in angular illumination asymmetry (ANILAS) at the sample plane. Here we show that through-focus propagation of ANILAS results in a lateral image shift with a focus position. This could lead to substantial errors in quantitative results for optical methods that use through-focus images such as three-dimensional nanoparticle tracking, confocal microscopy, and through-focus scanning optical microscopy (TSOM). A correlation exists between ANILAS and the slant in TSOM images. Hence, the slant in the TSOM image can be used to detect, analyze, and rectify the presence of ANILAS.

Optically sectioned imaging by oblique plane microscopy

Science.gov (United States)

Kumar, Sunil; Lin, Ziduo; Lyon, Alex R.; MacLeod, Ken T.; Dunsby, Chris

2011-03-01

Oblique Plane Microscopy (OPM) is a light sheet microscopy technique that combines oblique illumination with correction optics that tilt the focal plane of the collection system. OPM can be used to image conventionally mounted specimens on coverslips or tissue culture dishes and has low out-of-plane photobleaching and phototoxicity. No moving parts are required to achieve an optically sectioned image and so high speed optically sectioned imaging is possible. The first OPM results obtained using a high NA water immersion lens on a commercially available inverted microscope frame are presented, together with a measurement of the achievable optical resolution.

Correlated topographic and spectroscopic imaging by combined atomic force microscopy and optical microscopy

International Nuclear Information System (INIS)

Hu Dehong; Micic, Miodrag; Klymyshyn, Nicholas; Suh, Y.D.; Lu, H.P.

2004-01-01

Near-field scanning microscopy is a powerful approach to obtain topographic and spectroscopic characterization simultaneously for imaging biological and nanoscale systems. To achieve optical imaging at high spatial resolution beyond the diffraction limit, aperture-less metallic scanning tips have been utilized to enhance the laser illumination local electromagnetic field at the apex of the scanning tips. In this paper, we discuss and review our work on combined fluorescence imaging with AFM-metallic tip enhancement, finite element method simulation of the tip enhancement, and their applications on AFM-tip enhanced fluorescence lifetime imaging (AFM-FLIM) and correlated AFM and FLIM imaging of the living cells

Review of near-field optics and superlenses for sub-diffraction-limited nano-imaging

Directory of Open Access Journals (Sweden)

Wyatt Adams

2016-10-01

Full Text Available Near-field optics and superlenses for imaging beyond Abbe’s diffraction limit are reviewed. A comprehensive and contemporary background is given on scanning near-field microscopy and superlensing. Attention is brought to recent research leveraging scanning near-field optical microscopy with superlenses for new nano-imaging capabilities. Future research directions are explored for realizing the goal of low-cost and high-performance sub-diffraction-limited imaging systems.

Simultaneous differential spinning disk fluorescence optical sectioning microscopy and nanomechanical mapping atomic force microscopy

International Nuclear Information System (INIS)

Miranda, Adelaide; De Beule, Pieter A. A.; Martins, Marco

2015-01-01

Combined microscopy techniques offer the life science research community a powerful tool to investigate complex biological systems and their interactions. Here, we present a new combined microscopy platform based on fluorescence optical sectioning microscopy through aperture correlation microscopy with a Differential Spinning Disk (DSD) and nanomechanical mapping with an Atomic Force Microscope (AFM). The illumination scheme of the DSD microscope unit, contrary to standard single or multi-point confocal microscopes, provides a time-independent illumination of the AFM cantilever. This enables a distortion-free simultaneous operation of fluorescence optical sectioning microscopy and atomic force microscopy with standard probes. In this context, we discuss sample heating due to AFM cantilever illumination with fluorescence excitation light. Integration of a DSD fluorescence optical sectioning unit with an AFM platform requires mitigation of mechanical noise transfer of the spinning disk. We identify and present two solutions to almost annul this noise in the AFM measurement process. The new combined microscopy platform is applied to the characterization of a DOPC/DOPS (4:1) lipid structures labelled with a lipophilic cationic indocarbocyanine dye deposited on a mica substrate

Simultaneous differential spinning disk fluorescence optical sectioning microscopy and nanomechanical mapping atomic force microscopy

Energy Technology Data Exchange (ETDEWEB)

Miranda, Adelaide; De Beule, Pieter A. A., E-mail: pieter.de-beule@inl.int [Applied Nano-Optics Laboratory, International Iberian Nanotechnology Laboratory, Avenida Mestre José Veiga, s/n, 4715-330 Braga (Portugal); Martins, Marco [Nano-ICs Group, International Iberian Nanotechnology Laboratory, Avenida Mestre José Veiga, s/n, 4715-330 Braga (Portugal)

2015-09-15

Combined microscopy techniques offer the life science research community a powerful tool to investigate complex biological systems and their interactions. Here, we present a new combined microscopy platform based on fluorescence optical sectioning microscopy through aperture correlation microscopy with a Differential Spinning Disk (DSD) and nanomechanical mapping with an Atomic Force Microscope (AFM). The illumination scheme of the DSD microscope unit, contrary to standard single or multi-point confocal microscopes, provides a time-independent illumination of the AFM cantilever. This enables a distortion-free simultaneous operation of fluorescence optical sectioning microscopy and atomic force microscopy with standard probes. In this context, we discuss sample heating due to AFM cantilever illumination with fluorescence excitation light. Integration of a DSD fluorescence optical sectioning unit with an AFM platform requires mitigation of mechanical noise transfer of the spinning disk. We identify and present two solutions to almost annul this noise in the AFM measurement process. The new combined microscopy platform is applied to the characterization of a DOPC/DOPS (4:1) lipid structures labelled with a lipophilic cationic indocarbocyanine dye deposited on a mica substrate.

Coherent optical adaptive technique improves the spatial resolution of STED microscopy in thick samples

Science.gov (United States)

Yan, Wei; Yang, Yanlong; Tan, Yu; Chen, Xun; Li, Yang; Qu, Junle; Ye, Tong

2018-01-01

Stimulated emission depletion microscopy (STED) is one of far-field optical microscopy techniques that can provide sub-diffraction spatial resolution. The spatial resolution of the STED microscopy is determined by the specially engineered beam profile of the depletion beam and its power. However, the beam profile of the depletion beam may be distorted due to aberrations of optical systems and inhomogeneity of specimens’ optical properties, resulting in a compromised spatial resolution. The situation gets deteriorated when thick samples are imaged. In the worst case, the sever distortion of the depletion beam profile may cause complete loss of the super resolution effect no matter how much depletion power is applied to specimens. Previously several adaptive optics approaches have been explored to compensate aberrations of systems and specimens. However, it is hard to correct the complicated high-order optical aberrations of specimens. In this report, we demonstrate that the complicated distorted wavefront from a thick phantom sample can be measured by using the coherent optical adaptive technique (COAT). The full correction can effectively maintain and improve the spatial resolution in imaging thick samples. PMID:29400356

Nanometrology using a through-focus scanning optical microscopy method

International Nuclear Information System (INIS)

Attota, Ravikiran; Silver, Richard

2011-01-01

We present an initial review of a novel through-focus scanning optical microscopy (TSOM pronounced as 'tee-som') imaging method that produces nanometer-dimensional measurement sensitivity using a conventional bright-field optical microscope. In the TSOM method a target is scanned through the focus of an optical microscope, acquiring conventional optical images at different focal positions. The TSOM images are constructed using the through-focus optical images. A TSOM image is unique under given experimental conditions and is sensitive to changes in the dimensions of a target in a distinct way. We use this characteristic for nanoscale-dimensional metrology. This technique can be used to identify the dimension which is changing between two nanosized targets and to determine the dimensions using a library-matching method. This methodology has potential utility for a wide range of target geometries and application areas, including nanometrology, nanomanufacturing, defect analysis, inspection, process control and biotechnology

High-speed atomic force microscopy combined with inverted optical microscopy for studying cellular events.

OpenAIRE

Suzuki, Yuki; Sakai, Nobuaki; Yoshida, Aiko; Uekusa, Yoshitsugu; Yagi, Akira; Imaoka, Yuka; Ito, Shuichi; Karaki, Koichi; Takeyasu, Kunio

2013-01-01

A hybrid atomic force microscopy (AFM)-optical fluorescence microscopy is a powerful tool for investigating cellular morphologies and events. However, the slow data acquisition rates of the conventional AFM unit of the hybrid system limit the visualization of structural changes during cellular events. Therefore, high-speed AFM units equipped with an optical/fluorescence detection device have been a long-standing wish. Here we describe the implementation of high-speed AFM coupled with an optic...

Wave front engineering by means of diffractive optical elements for applications in microscopy

Science.gov (United States)

Cojoc, Dan; Ferrari, Enrico; Garbin, Valeria; Cabrini, Stefano; Carpentiero, Alessandro; Prasciolu, Mauro; Businaro, Luca; Kaulich, Burchard; Di Fabrizio, Enzo

2006-05-01

We present a unified view regarding the use of diffractive optical elements (DOEs) for microscopy applications a wide range of electromagnetic spectrum. The unified treatment is realized through the design and fabrication of DOE through which wave front beam shaping is obtained. In particular we show applications ranging from micromanipulation using optical tweezers to X-ray differential interference contrast (DIC) microscopy. We report some details on the design and physical implementation of diffractive elements that beside focusing perform also other optical functions: beam splitting, beam intensity and phase redistribution or mode conversion. Laser beam splitting is used for multiple trapping and independent manipulation of spherical micro beads and for direct trapping and manipulation of biological cells with non-spherical shapes. Another application is the Gauss to Laguerre-Gaussian mode conversion, which allows to trap and transfer orbital angular momentum of light to micro particles with high refractive index and to trap and manipulate low index particles. These experiments are performed in an inverted optical microscope coupled with an infrared laser beam and a spatial light modulator for DOEs implementation. High resolution optics, fabricated by means of e-beam lithography, are demonstrated to control the intensity and the phase of the sheared beams in X-ray DIC microscopy. DIC experiments with phase objects reveal a dramatic increase in image contrast compared to bright-field X-ray microscopy.

The 2015 super-resolution microscopy roadmap

International Nuclear Information System (INIS)

Hell, Stefan W; Sahl, Steffen J; Bates, Mark; Jakobs, Stefan; Zhuang, Xiaowei; Heintzmann, Rainer; Booth, Martin J; Bewersdorf, Joerg; Shtengel, Gleb; Hess, Harald; Tinnefeld, Philip; Honigmann, Alf; Testa, Ilaria; Cognet, Laurent; Lounis, Brahim; Ewers, Helge; Davis, Simon J; Eggeling, Christian; Klenerman, David; Willig, Katrin I

2015-01-01

Far-field optical microscopy using focused light is an important tool in a number of scientific disciplines including chemical, (bio)physical and biomedical research, particularly with respect to the study of living cells and organisms. Unfortunately, the applicability of the optical microscope is limited, since the diffraction of light imposes limitations on the spatial resolution of the image. Consequently the details of, for example, cellular protein distributions, can be visualized only to a certain extent. Fortunately, recent years have witnessed the development of ‘super-resolution’ far-field optical microscopy (nanoscopy) techniques such as stimulated emission depletion (STED), ground state depletion (GSD), reversible saturated optical (fluorescence) transitions (RESOLFT), photoactivation localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) or saturated structured illumination microscopy (SSIM), all in one way or another addressing the problem of the limited spatial resolution of far-field optical microscopy. While SIM achieves a two-fold improvement in spatial resolution compared to conventional optical microscopy, STED, RESOLFT, PALM/STORM, or SSIM have all gone beyond, pushing the limits of optical image resolution to the nanometer scale. Consequently, all super-resolution techniques open new avenues of biomedical research. Because the field is so young, the potential capabilities of different super-resolution microscopy approaches have yet to be fully explored, and uncertainties remain when considering the best choice of methodology. Thus, even for experts, the road to the future is sometimes shrouded in mist. The super-resolution optical microscopy roadmap of Journal of Physics D: Applied Physics addresses this need for clarity. It provides guidance to the outstanding questions through a collection of short review articles from experts in the field, giving a thorough

Extending Single-Molecule Microscopy Using Optical Fourier Processing

Science.gov (United States)

2015-01-01

This article surveys the recent application of optical Fourier processing to the long-established but still expanding field of single-molecule imaging and microscopy. A variety of single-molecule studies can benefit from the additional image information that can be obtained by modulating the Fourier, or pupil, plane of a widefield microscope. After briefly reviewing several current applications, we present a comprehensive and computationally efficient theoretical model for simulating single-molecule fluorescence as it propagates through an imaging system. Furthermore, we describe how phase/amplitude-modulating optics inserted in the imaging pathway may be modeled, especially at the Fourier plane. Finally, we discuss selected recent applications of Fourier processing methods to measure the orientation, depth, and rotational mobility of single fluorescent molecules. PMID:24745862

Near-field optical microscope using a silicon-nitride probe

NARCIS (Netherlands)

van Hulst, N.F.; Moers, M.H.P.; Moers, M.H.P.; Noordman, O.F.J.; Noordman, O.F.J.; Tack, R.G.; Segerink, Franciscus B.; Bölger, B.; Bölger, B.

1993-01-01

Operation of an alternative near-field optical microscope is presented. The microscope uses a microfabricated silicon- nitride probe with integrated cantilever, as originally developed for force microscopy. The cantilever allows routine close contact near-field imaging o­n arbitrary surfaces without

Full optical model of micro-endoscope with optical coherence microscopy, multiphoton microscopy and visible capabilities

Science.gov (United States)

Vega, David; Kiekens, Kelli C.; Syson, Nikolas C.; Romano, Gabriella; Baker, Tressa; Barton, Jennifer K.

2018-02-01

While Optical Coherence Microscopy (OCM), Multiphoton Microscopy (MPM), and narrowband imaging are powerful imaging techniques that can be used to detect cancer, each imaging technique has limitations when used by itself. Combining them into an endoscope to work in synergy can help achieve high sensitivity and specificity for diagnosis at the point of care. Such complex endoscopes have an elevated risk of failure, and performing proper modelling ensures functionality and minimizes risk. We present full 2D and 3D models of a multimodality optical micro-endoscope to provide real-time detection of carcinomas, called a salpingoscope. The models evaluate the endoscope illumination and light collection capabilities of various modalities. The design features two optical paths with different numerical apertures (NA) through a single lens system with a scanning optical fiber. The dual path is achieved using dichroic coatings embedded in a triplet. A high NA optical path is designed to perform OCM and MPM while a low NA optical path is designed for the visible spectrum to navigate the endoscope to areas of interest and narrowband imaging. Different tests such as the reflectance profile of homogeneous epithelial tissue were performed to adjust the models properly. Light collection models for the different modalities were created and tested for efficiency. While it is challenging to evaluate the efficiency of multimodality endoscopes, the models ensure that the system is design for the expected light collection levels to provide detectable signal to work for the intended imaging.

Novel concepts in near-field optics: from magnetic near-field to optical forces

Science.gov (United States)

Yang, Honghua

Driven by the progress in nanotechnology, imaging and spectroscopy tools with nanometer spatial resolution are needed for in situ material characterizations. Near-field optics provides a unique way to selectively excite and detect elementary electronic and vibrational interactions at the nanometer scale, through interactions of light with matter in the near-field region. This dissertation discusses the development and applications of near-field optical imaging techniques, including plasmonic material characterization, optical spectral nano-imaging and magnetic field detection using scattering-type scanning near-field optical microscopy (s-SNOM), and exploring new modalities of optical spectroscopy based on optical gradient force detection. Firstly, the optical dielectric functions of one of the most common plasmonic materials---silver is measured with ellipsometry, and analyzed with the Drude model over a broad spectral range from visible to mid-infrared. This work was motivated by the conflicting results of previous measurements, and the need for accurate values for a wide range of applications of silver in plasmonics, optical antennas, and metamaterials. This measurement provides a reference for dielectric functions of silver used in metamaterials, plasmonics, and nanophotonics. Secondly, I implemented an infrared s-SNOM instrument for spectroscopic nano-imaging at both room temperature and low temperature. As one of the first cryogenic s-SNOM instruments, the novel design concept and key specifications are discussed. Initial low-temperature and high-temperature performances of the instrument are examined by imaging of optical conductivity of vanadium oxides (VO2 and V2O 3) across their phase transitions. The spectroscopic imaging capability is demonstrated on chemical vibrational resonances of Poly(methyl methacrylate) (PMMA) and other samples. The third part of this dissertation explores imaging of optical magnetic fields. As a proof-of-principle, the magnetic

Quantitative Near-field Microscopy of Heterogeneous and Correlated Electron Oxides

Science.gov (United States)

McLeod, Alexander Swinton

Scanning near-field optical microscopy (SNOM) is a novel scanning probe microscopy technique capable of circumventing the conventional diffraction limit of light, affording unparalleled optical resolution (down to 10 nanometers) even for radiation in the infrared and terahertz energy regimes, with light wavelengths exceeding 10 micrometers. However, although this technique has been developed and employed for more than a decade to a qualitatively impressive effect, researchers have lacked a practically quantitative grasp of its capabilities, and its application scope has so far remained restricted by implementations limited to ambient atmospheric conditions. The two-fold objective of this dissertation work has been to address both these shortcomings. The first half of the dissertation presents a realistic, semi-analytic, and benchmarked theoretical description of probe-sample near-field interactions that form the basis of SNOM. Owing its name to the efficient nano-focusing of light at a sharp metallic apex, the "lightning rod model" of probe-sample near-field interactions is mathematically developed from a flexible and realistic scattering formalism. Powerful and practical applications are demonstrated through the accurate prediction of spectroscopic near-field optical contrasts, as well as the "inversion" of these spectroscopic contrasts into a quantitative description of material optical properties. Thus enabled, this thesis work proceeds to present quantitative applications of infrared near-field spectroscopy to investigate nano-resolved chemical compositions in a diverse host of samples, including technologically relevant lithium ion battery materials, astrophysical planetary materials, and invaluable returned extraterrestrial samples. The second half of the dissertation presents the design, construction, and demonstration of a sophisticated low-temperature scanning near-field infrared microscope. This instrument operates in an ultra-high vacuum environment

Wide-field two-photon microscopy with temporal focusing and HiLo background rejection

Science.gov (United States)

Yew, Elijah Y. S.; Choi, Heejin; Kim, Daekeun; So, Peter T. C.

2011-03-01

Scanningless depth-resolved microscopy is achieved through spatial-temporal focusing and has been demonstrated previously. The advantage of this method is that a large area may be imaged without scanning resulting in higher throughput of the imaging system. Because it is a widefield technique, the optical sectioning effect is considerably poorer than with conventional spatial focusing two-photon microscopy. Here we propose wide-field two-photon microscopy based on spatio-temporal focusing and employing background rejection based on the HiLo microscope principle. We demonstrate the effects of applying HiLo microscopy to widefield temporally focused two-photon microscopy.

Second-harmonic scanning optical microscopy of poled silica waveguides

DEFF Research Database (Denmark)

Pedersen, Kjeld; Bozhevolnyi, Sergey I.; Arentoft, Jesper

2000-01-01

Second-harmonic scanning optical microscopy (SHSOM) is performed on electric-field poled silica-based waveguides. Two operation modes of SHSOM are considered. Oblique transmission reflection and normal reflection modes are used to image the spatial distribution of nonlinear susceptibilities...... and limitations of the two operation modes when used for SHSOM studies of poled silica-based waveguides are discussed. The influence of surface defects on the resulting second-harmonic images is also considered. ©2000 American Institute of Physics....

Nonlinear microscopy of localized field enhancements in fractal shaped periodic metal nanostructures

DEFF Research Database (Denmark)

Beermann, I.; Evlyukhin, A.; Boltasseva, Alexandra

2008-01-01

Fractal shaped periodic nanostructures formed with a 100 nm period square lattice of gold nanoparticles placed on a gold film are characterized using far-field nonlinear scanning optical microscopy, in which two-photon photoluminescence (TPL) excited with a strongly focused femtosecond laser beam...

Biobeam—Multiplexed wave-optical simulations of light-sheet microscopy

Science.gov (United States)

Weigert, Martin; Bundschuh, Sebastian T.

2018-01-01

Sample-induced image-degradation remains an intricate wave-optical problem in light-sheet microscopy. Here we present biobeam, an open-source software package that enables simulation of operational light-sheet microscopes by combining data from 105–106 multiplexed and GPU-accelerated point-spread-function calculations. The wave-optical nature of these simulations leads to the faithful reproduction of spatially varying aberrations, diffraction artifacts, geometric image distortions, adaptive optics, and emergent wave-optical phenomena, and renders image-formation in light-sheet microscopy computationally tractable. PMID:29652879

Portable fiber-optic taper coupled optical microscopy platform

Science.gov (United States)

Wang, Weiming; Yu, Yan; Huang, Hui; Ou, Jinping

2017-04-01

The optical fiber taper coupled with CMOS has advantages of high sensitivity, compact structure and low distortion in the imaging platform. So it is widely used in low light, high speed and X-ray imaging systems. In the meanwhile, the peculiarity of the coupled structure can meet the needs of the demand in microscopy imaging. Toward this end, we developed a microscopic imaging platform based on the coupling of cellphone camera module and fiber optic taper for the measurement of the human blood samples and ascaris lumbricoides. The platform, weighing 70 grams, is based on the existing camera module of the smartphone and a fiber-optic array which providing a magnification factor of 6x.The top facet of the taper, on which samples are placed, serves as an irregular sampling grid for contact imaging. The magnified images of the sample, located on the bottom facet of the fiber, are then projected onto the CMOS sensor. This paper introduces the portable medical imaging system based on the optical fiber coupling with CMOS, and theoretically analyzes the feasibility of the system. The image data and process results either can be stored on the memory or transmitted to the remote medical institutions for the telemedicine. We validate the performance of this cell-phone based microscopy platform using human blood samples and test target, achieving comparable results to a standard bench-top microscope.

Deep Learning Microscopy

KAUST Repository

Rivenson, Yair

2017-05-12

We demonstrate that a deep neural network can significantly improve optical microscopy, enhancing its spatial resolution over a large field-of-view and depth-of-field. After its training, the only input to this network is an image acquired using a regular optical microscope, without any changes to its design. We blindly tested this deep learning approach using various tissue samples that are imaged with low-resolution and wide-field systems, where the network rapidly outputs an image with remarkably better resolution, matching the performance of higher numerical aperture lenses, also significantly surpassing their limited field-of-view and depth-of-field. These results are transformative for various fields that use microscopy tools, including e.g., life sciences, where optical microscopy is considered as one of the most widely used and deployed techniques. Beyond such applications, our presented approach is broadly applicable to other imaging modalities, also spanning different parts of the electromagnetic spectrum, and can be used to design computational imagers that get better and better as they continue to image specimen and establish new transformations among different modes of imaging.

Multimodal imaging of heterogeneous polymers at the nanoscale by AFM and scanning near-field ellipsometric microscopy

NARCIS (Netherlands)

Cumurcu, Aysegul; Duvigneau, Joost; Lindsay, I.D.; Schön, Peter Manfred; Vancso, Gyula J.

2013-01-01

Scanning near field ellipsometric microscopy (SNEM) was used to simultaneously obtain optical images and tapping mode topography images of the microphase separated morphology of PS-b-P2VP block copolymer thin films. Optical images revealed a spatial resolution well below the diffraction limit. The

Optical near-fields & nearfield optics

OpenAIRE

Meixner, Alfred J; Leiderer, Paul

2014-01-01

Optical methods provide exceedingly powerful tools in science and technology for measuring, analyzing and manipulating, from optical microscopy and spectroscopy to the characterization of ultrafast processes by femtosecond pulses and the modification of materials by intense laser radiation. However, when it comes to applications in the nanometer-regime, the conventional optical techniques suffer from the resolution limit – formulated by Ernst Abbe one and a half centuries ago – that light can...

Insights on proximity effect and multiphoton induced luminescence from gold nanospheres in far field optical microscopy

Energy Technology Data Exchange (ETDEWEB)

Borglin, Johan [Biomedical Photonics Group, Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Department of Physics, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Guldbrand, Stina [Department of Physics, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Evenbratt, Hanne [Pharmaceutical Technology, Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemigården 4, 412 96 Gothenburg (Sweden); Kirejev, Vladimir; Ericson, Marica B., E-mail: marica.ericson@chem.gu.se [Biomedical Photonics Group, Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Grönbeck, Henrik [Department of Applied Physics, Chalmers University of Technology, Kemivägen 9, 412 96 Gothenburg (Sweden)

2015-12-07

Gold nanoparticles can be visualized in far-field multiphoton laser-scanning microscopy (MPM) based on the phenomena of multiphoton induced luminescence (MIL). This is of interest for biomedical applications, e.g., for cancer diagnostics, as MPM allows for working in the near-infrared (NIR) optical window of tissue. It is well known that the aggregation of particles causes a redshift of the plasmon resonance, but its implications for MIL applying far-field MPM should be further exploited. Here, we explore MIL from 10 nm gold nanospheres that are chemically deposited on glass substrates in controlled coverage gradients using MPM operating in NIR range. The substrates enable studies of MIL as a function of inter-particle distance and clustering. It was shown that MIL was only detected from areas on the substrates where the particle spacing was less than one particle diameter, or where the particles have aggregated. The results are interpreted in the context that the underlying physical phenomenon of MIL is a sequential two-photon absorption process, where the first event is driven by the plasmon resonance. It is evident that gold nanospheres in this size range have to be closely spaced or clustered to exhibit detectable MIL using far-field MPM operating in the NIR region.

Insights on proximity effect and multiphoton induced luminescence from gold nanospheres in far field optical microscopy

International Nuclear Information System (INIS)

Borglin, Johan; Guldbrand, Stina; Evenbratt, Hanne; Kirejev, Vladimir; Ericson, Marica B.; Grönbeck, Henrik

2015-01-01

Gold nanoparticles can be visualized in far-field multiphoton laser-scanning microscopy (MPM) based on the phenomena of multiphoton induced luminescence (MIL). This is of interest for biomedical applications, e.g., for cancer diagnostics, as MPM allows for working in the near-infrared (NIR) optical window of tissue. It is well known that the aggregation of particles causes a redshift of the plasmon resonance, but its implications for MIL applying far-field MPM should be further exploited. Here, we explore MIL from 10 nm gold nanospheres that are chemically deposited on glass substrates in controlled coverage gradients using MPM operating in NIR range. The substrates enable studies of MIL as a function of inter-particle distance and clustering. It was shown that MIL was only detected from areas on the substrates where the particle spacing was less than one particle diameter, or where the particles have aggregated. The results are interpreted in the context that the underlying physical phenomenon of MIL is a sequential two-photon absorption process, where the first event is driven by the plasmon resonance. It is evident that gold nanospheres in this size range have to be closely spaced or clustered to exhibit detectable MIL using far-field MPM operating in the NIR region

DMD-based LED-illumination super-resolution and optical sectioning microscopy.

Science.gov (United States)

Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

2013-01-01

Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×10(7) pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

Nanometric locking of the tight focus for optical microscopy and tip-enhanced microscopy

International Nuclear Information System (INIS)

Hayazawa, N; Furusawa, K; Kawata, S

2012-01-01

We have successfully stabilized the tight focus onto the sample surface of an optical microscope within ±1.0 nm for a virtually unlimited time duration. The time-dependent thermal drift of the tight focus and the mechanical tilt of the sample surface were simultaneously sensed by a non-optical means based on a capacitive sensor and were compensated for in real-time. This non-optical scheme is promising for the suppression of background light sources for optical microscopy. The focus stabilization is crucial for microscopic measurement at an interface, particularly when scanning a large surface area, because there is always a certain amount of mechanical tilt of the sample substrate, which degrades the contrast of the image. When imaging nanoscopic materials such as carbon nanotubes or silicon nanowires, more stringent nanometric stabilization of the focus position relative to such samples is required, otherwise it is often difficult to interpret the results from the observations. Moreover, the smaller the sample volume is, the smaller the signal becomes, resulting in a long exposure time at each position. In this sense, long-term stability of the tight focus is essential for both microscopic large area scanning and nanosized sample scanning (high-resolution/large-area imaging). In addition, the recently developed tip-enhanced microscopy requires long-term stability of the relative position of the tip, sample and focus position. We were able to successfully demonstrate a stability improvement for tip-enhanced microscopy in the same manner. The stabilization of the tight focus enables us to perform long-term and robust measurements without any degradation of optical signal, resulting in the capability of true nanometric optical imaging with good reproducibility and high precision. The technique presented is a simple add-on for any kind of optical microscope. (paper)

True Tapping Mode Scanning Near-Field Optical Microscopy with Bent Glass Fiber Probes.

Science.gov (United States)

Smirnov, A; Yasinskii, V M; Filimonenko, D S; Rostova, E; Dietler, G; Sekatskii, S K

2018-01-01

In scanning near-field optical microscopy, the most popular probes are made of sharpened glass fiber attached to a quartz tuning fork (TF) and exploiting the shear force-based feedback. The use of tapping mode feedback could be preferable. Such an approach can be realized, for example, using bent fiber probes. Detailed analysis of fiber vibration modes shows that realization of truly tapping mode of the probe dithering requires an extreme caution. In case of using the second resonance mode, probes vibrate mostly in shear force mode unless the bending radius is rather small (ca. 0.3 mm) and the probe's tip is short. Otherwise, the shear force character of the dithering persists. Probes having these characteristics were prepared by irradiation of a tapered etched glass fiber with a CW CO 2 laser. These probes were attached to the TF in double resonance conditions which enables achieving significant quality factor (4000-6000) of the TF + probe system (Cherkun et al., 2006). We also show that, to achieve a truly tapping character, dithering, short, and not exceeding 3 mm lengths of a freestanding part of bent fiber probe beam should also be used in the case of nonresonant excitation.

True Tapping Mode Scanning Near-Field Optical Microscopy with Bent Glass Fiber Probes

Directory of Open Access Journals (Sweden)

A. Smirnov

2018-01-01

Full Text Available In scanning near-field optical microscopy, the most popular probes are made of sharpened glass fiber attached to a quartz tuning fork (TF and exploiting the shear force-based feedback. The use of tapping mode feedback could be preferable. Such an approach can be realized, for example, using bent fiber probes. Detailed analysis of fiber vibration modes shows that realization of truly tapping mode of the probe dithering requires an extreme caution. In case of using the second resonance mode, probes vibrate mostly in shear force mode unless the bending radius is rather small (ca. 0.3 mm and the probe’s tip is short. Otherwise, the shear force character of the dithering persists. Probes having these characteristics were prepared by irradiation of a tapered etched glass fiber with a CW CO2 laser. These probes were attached to the TF in double resonance conditions which enables achieving significant quality factor (4000–6000 of the TF + probe system (Cherkun et al., 2006. We also show that, to achieve a truly tapping character, dithering, short, and not exceeding 3 mm lengths of a freestanding part of bent fiber probe beam should also be used in the case of nonresonant excitation.

Characterization of power induced heating and damage in fiber optic probes for near-field scanning optical microscopy

Science.gov (United States)

Dickenson, Nicholas E.; Erickson, Elizabeth S.; Mooren, Olivia L.; Dunn, Robert C.

2007-05-01

Tip-induced sample heating in near-field scanning optical microscopy (NSOM) is studied for fiber optic probes fabricated using the chemical etching technique. To characterize sample heating from etched NSOM probes, the spectra of a thermochromic polymer sample are measured as a function of probe output power, as was previously reported for pulled NSOM probes. The results reveal that sample heating increases rapidly to ˜55-60°C as output powers reach ˜50nW. At higher output powers, the sample heating remains approximately constant up to the maximum power studied of ˜450nW. The sample heating profiles measured for etched NSOM probes are consistent with those previously measured for NSOM probes fabricated using the pulling method. At high powers, both pulled and etched NSOM probes fail as the aluminum coating is damaged. For probes fabricated in our laboratory we find failure occurring at input powers of 3.4±1.7 and 20.7±6.9mW for pulled and etched probes, respectively. The larger half-cone angle for etched probes (˜15° for etched and ˜6° for pulled probes) enables more light delivery and also apparently leads to a different failure mechanism. For pulled NSOM probes, high resolution images of NSOM probes as power is increased reveal the development of stress fractures in the coating at a taper diameter of ˜6μm. These stress fractures, arising from the differential heating expansion of the dielectric and the metal coating, eventually lead to coating removal and probe failure. For etched tips, the absence of clear stress fractures and the pooled morphology of the damaged aluminum coating following failure suggest that thermal damage may cause coating failure, although other mechanisms cannot be ruled out.

Poly(diacetylene) Monolayers Studied with a Fluorescence Scanning Near-Field Optical Microscope

NARCIS (Netherlands)

Moers, Marco H.P.; Moers, M.H.P.; Gaub, Hermann E.; van Hulst, N.F.

1994-01-01

A novel and powerful method to study the optical properties of thin lipid films which a resolution superior to confocal microscopy is presented. With a scanning near-field optical microscope, fluorescence images of a Langmuir-Blodgett film of diethylene glycol diamine pentacosadiynoic amide are

High-speed atomic force microscopy combined with inverted optical microscopy for studying cellular events.

Science.gov (United States)

Suzuki, Yuki; Sakai, Nobuaki; Yoshida, Aiko; Uekusa, Yoshitsugu; Yagi, Akira; Imaoka, Yuka; Ito, Shuichi; Karaki, Koichi; Takeyasu, Kunio

2013-01-01

A hybrid atomic force microscopy (AFM)-optical fluorescence microscopy is a powerful tool for investigating cellular morphologies and events. However, the slow data acquisition rates of the conventional AFM unit of the hybrid system limit the visualization of structural changes during cellular events. Therefore, high-speed AFM units equipped with an optical/fluorescence detection device have been a long-standing wish. Here we describe the implementation of high-speed AFM coupled with an optical fluorescence microscope. This was accomplished by developing a tip-scanning system, instead of a sample-scanning system, which operates on an inverted optical microscope. This novel device enabled the acquisition of high-speed AFM images of morphological changes in individual cells. Using this instrument, we conducted structural studies of living HeLa and 3T3 fibroblast cell surfaces. The improved time resolution allowed us to image dynamic cellular events.

Characterization and improvement of highly inclined optical sheet microscopy

Science.gov (United States)

Vignolini, T.; Curcio, V.; Gardini, L.; Capitanio, M.; Pavone, F. S.

2018-02-01

Highly Inclined and Laminated Optical sheet (HILO) microscopy is an optical technique that employs a highly inclined laser beam to illuminate the sample with a thin sheet of light that can be scanned through the sample volume1 . HILO is an efficient illumination technique when applied to fluorescence imaging of thick samples owing to the confined illumination volume that allows high contrast imaging while retaining deep scanning capability in a wide-field configuration. The restricted illumination volume is crucial to limit background fluorescence originating from portions of the sample far from the focal plane, especially in applications such as single molecule localization and super-resolution imaging2-4. Despite its widespread use, current literature lacks comprehensive reports of the actual advantages of HILO in these kinds of microscopies. Here, we thoroughly characterize the propagation of a highly inclined beam through fluorescently labeled samples and implement appropriate beam shaping for optimal application to single molecule and super-resolution imaging. We demonstrate that, by reducing the beam size along the refracted axis only, the excitation volume is consequently reduced while maintaining a field of view suitable for single cell imaging. We quantify the enhancement in signal-tobackground ratio with respect to the standard HILO technique and apply our illumination method to dSTORM superresolution imaging of the actin and vimentin cytoskeleton. We define the conditions to achieve localization precisions comparable to state-of-the-art reports, obtain a significant improvement in the image contrast, and enhanced plane selectivity within the sample volume due to the further confinement of the inclined beam.

Multiparallel Three-Dimensional Optical Microscopy

Science.gov (United States)

Nguyen, Lam K.; Price, Jeffrey H.; Kellner, Albert L.; Bravo-Zanoquera, Miguel

2010-01-01

Multiparallel three-dimensional optical microscopy is a method of forming an approximate three-dimensional image of a microscope sample as a collection of images from different depths through the sample. The imaging apparatus includes a single microscope plus an assembly of beam splitters and mirrors that divide the output of the microscope into multiple channels. An imaging array of photodetectors in each channel is located at a different distance along the optical path from the microscope, corresponding to a focal plane at a different depth within the sample. The optical path leading to each photodetector array also includes lenses to compensate for the variation of magnification with distance so that the images ultimately formed on all the photodetector arrays are of the same magnification. The use of optical components common to multiple channels in a simple geometry makes it possible to obtain high light-transmission efficiency with an optically and mechanically simple assembly. In addition, because images can be read out simultaneously from all the photodetector arrays, the apparatus can support three-dimensional imaging at a high scanning rate.

Proximal design for a multimodality endoscope with multiphoton microscopy, optical coherence microscopy and visual modalities

Science.gov (United States)

Kiekens, Kelli C.; Talarico, Olivia; Barton, Jennifer K.

2018-02-01

A multimodality endoscope system has been designed for early detection of ovarian cancer. Multiple illumination and detection systems must be integrated in a compact, stable, transportable configuration to meet the requirements of a clinical setting. The proximal configuration presented here supports visible light navigation with a large field of view and low resolution, high resolution multiphoton microscopy (MPM), and high resolution optical coherence microscopy (OCM). All modalities are integrated into a single optical system in the endoscope. The system requires two light sources: a green laser for visible light navigation and a compact fiber based femtosecond laser for MPM and OCM. Using an inline wavelength division multiplexer, the two sources are combined into a single mode fiber. To accomplish OCM, a fiber coupler is used to separate the femtosecond laser into a reference arm and signal arm. The reflected reference arm and the signal from the sample are interfered and wavelength separated by a reflection grating and detected using a linear array. The MPM signal is collimated and goes through a series of filters to separate the 2nd and 3rd harmonics as well as twophoton excitation florescence (2PEF) and 3PEF. Each signal is independently detected on a photo multiplier tube and amplified. The visible light is collected by multiple high numerical aperture fibers at the endoscope tip which are bundled into one SMA adapter at the proximal end and connected to a photodetector. This integrated system design is compact, efficient and meets both optical and mechanical requirements for clinical applications.

Advanced magneto-optical microscopy: Imaging from picoseconds to centimeters - imaging spin waves and temperature distributions (invited

Directory of Open Access Journals (Sweden)

Necdet Onur Urs

2016-05-01

Full Text Available Recent developments in the observation of magnetic domains and domain walls by wide-field optical microscopy based on the magneto-optical Kerr, Faraday, Voigt, and Gradient effect are reviewed. Emphasis is given to the existence of higher order magneto-optical effects for advanced magnetic imaging. Fundamental concepts and advances in methodology are discussed that allow for imaging of magnetic domains on various length and time scales. Time-resolved imaging of electric field induced domain wall rotation is shown. Visualization of magnetization dynamics down to picosecond temporal resolution for the imaging of spin-waves and magneto-optical multi-effect domain imaging techniques for obtaining vectorial information are demonstrated. Beyond conventional domain imaging, the use of a magneto-optical indicator technique for local temperature sensing is shown.

Nanospectrofluorometry inside single living cell by scanning near-field optical microscopy

Science.gov (United States)

Lei, F. H.; Shang, G. Y.; Troyon, M.; Spajer, M.; Morjani, H.; Angiboust, J. F.; Manfait, M.

2001-10-01

Near-field fluorescence spectra with subdiffraction limit spatial resolution have been taken in the proximity of mitochondrial membrane inside breast adenocarcinoma cells (MCF7) treated with the fluorescent dye (JC-1) by using a scanning near-field optical microscope coupled with a confocal laser microspectrofluorometer. The probe-sample distance control is based on a piezoelectric bimorph shear force sensor having a static spring constant k=5 μN/nm and a quality factor Q=40 in a physiological medium of viscosity η=1.0 cp. The sensitivity of the force sensor has been tested by imaging a MCF7 cell surface.

An optical super-microscope for far-field, real-time imaging beyond the diffraction limit.

Science.gov (United States)

Wong, Alex M H; Eleftheriades, George V

2013-01-01

Optical microscopy suffers from a fundamental resolution limitation arising from the diffractive nature of light. While current solutions to sub-diffraction optical microscopy involve combinations of near-field, non-linear and fine scanning operations, we hereby propose and demonstrate the optical super-microscope (OSM) - a superoscillation-based linear imaging system with far-field working and observation distances - which can image an object in real-time and with sub-diffraction resolution. With our proof-of-principle prototype we report a point spread function with a spot size clearly reduced from the diffraction limit, and demonstrate corresponding improvements in two-point resolution experiments. Harnessing a new understanding of superoscillations, based on antenna array theory, our OSM achieves far-field, sub-diffraction optical imaging of an object without the need for fine scanning, data post-processing or object pre-treatment. Hence the OSM can be used in a wide variety of imaging applications beyond the diffraction limit, including real-time imaging of moving objects.

Adaptive optics stochastic optical reconstruction microscopy (AO-STORM) by particle swarm optimization.

Science.gov (United States)

Tehrani, Kayvan F; Zhang, Yiwen; Shen, Ping; Kner, Peter

2017-11-01

Stochastic optical reconstruction microscopy (STORM) can achieve resolutions of better than 20nm imaging single fluorescently labeled cells. However, when optical aberrations induced by larger biological samples degrade the point spread function (PSF), the localization accuracy and number of localizations are both reduced, destroying the resolution of STORM. Adaptive optics (AO) can be used to correct the wavefront, restoring the high resolution of STORM. A challenge for AO-STORM microscopy is the development of robust optimization algorithms which can efficiently correct the wavefront from stochastic raw STORM images. Here we present the implementation of a particle swarm optimization (PSO) approach with a Fourier metric for real-time correction of wavefront aberrations during STORM acquisition. We apply our approach to imaging boutons 100 μm deep inside the central nervous system (CNS) of Drosophila melanogaster larvae achieving a resolution of 146 nm.

A correlative optical microscopy and scanning electron microscopy approach to locating nanoparticles in brain tumors.

Science.gov (United States)

Kempen, Paul J; Kircher, Moritz F; de la Zerda, Adam; Zavaleta, Cristina L; Jokerst, Jesse V; Mellinghoff, Ingo K; Gambhir, Sanjiv S; Sinclair, Robert

2015-01-01

The growing use of nanoparticles in biomedical applications, including cancer diagnosis and treatment, demands the capability to exactly locate them within complex biological systems. In this work a correlative optical and scanning electron microscopy technique was developed to locate and observe multi-modal gold core nanoparticle accumulation in brain tumor models. Entire brain sections from mice containing orthotopic brain tumors injected intravenously with nanoparticles were imaged using both optical microscopy to identify the brain tumor, and scanning electron microscopy to identify the individual nanoparticles. Gold-based nanoparticles were readily identified in the scanning electron microscope using backscattered electron imaging as bright spots against a darker background. This information was then correlated to determine the exact location of the nanoparticles within the brain tissue. The nanoparticles were located only in areas that contained tumor cells, and not in the surrounding healthy brain tissue. This correlative technique provides a powerful method to relate the macro- and micro-scale features visible in light microscopy with the nanoscale features resolvable in scanning electron microscopy. Copyright © 2014 Elsevier Ltd. All rights reserved.

Stray-field-induced Faraday contributions in wide-field Kerr microscopy and -magnetometry

International Nuclear Information System (INIS)

Markó, D.; Soldatov, I.; Tekielak, M.; Schäfer, R.

2015-01-01

The magnetic domain contrast in wide-field Kerr microscopy on bulk specimens can be substantially distorted by non-linear, field-dependent Faraday rotations in the objective lens that are caused by stray-field components emerging from the specimen. These Faraday contributions, which were detected by Kerr-magnetometry on grain-oriented iron–silicon steel samples, are thoroughly elaborated and characterized. They express themselves as a field-dependent gray-scale offset to the domain contrast and in highly distorted surface magnetization curves if optically measured in a wide field Kerr microscope. An experimental method to avoid such distortions is suggested. In the course of these studies, a low-permeability part in the surface magnetization loop of slightly misoriented (110)-surfaces in iron–silicon sheets was discovered that is attributed to demagnetization effects in direction perpendicular to the sheet surface. - Highlights: • Magnetizing a finite sample in a Kerr microscope leads to sample-generated stray-fields. • They cause non-linear, field- and position-dependent Faraday rotations in the objective. • This leads to a modulation of the Kerr contrast and to distorted MOKE loops. • A method to compensate these Faraday rotations is presented

Chain end distribution of block copolymer in two-dimensional microphase-separated structure studied by scanning near-field optical microscopy.

Science.gov (United States)

Sekine, Ryojun; Aoki, Hiroyuki; Ito, Shinzaburo

2009-10-01

The chain end distribution of a block copolymer in a two-dimensional microphase-separated structure was studied by scanning near-field optical microscopy (SNOM). In the monolayer of poly(octadecyl methacrylate)-block-poly(isobutyl methacrylate) (PODMA-b-PiBMA), the free end of the PiBMA subchain was directly observed by SNOM, and the spatial distributions of the whole block and the chain end are examined and compared with the convolution of the point spread function of the microscope and distribution function of the model structures. It was found that the chain end distribution of the block copolymer confined in two dimensions has a peak near the domain center, being concentrated in the narrower region, as compared with three-dimensional systems.

Conference Paper NFO-7:7^th International Conference on Near-Field Optics and Related Technologies

Energy Technology Data Exchange (ETDEWEB)

Novotny, Lukas [Univ. of Rochester, NY (United States)

2004-10-18

The seventh conference in the NFO conference series, held here in Rochester, provided to be the principal forum for advances in sub-wavelength optics, near-field optical microscopy, local field enhancement, instrumental developments and the ever-increasing range of applications. This conference brought together the diverse scientific communities working on the theory and application of near-field optics (NFO) and related techniques.

Microscopy of biological sample through advanced diffractive optics from visible to X-ray wavelength regime.

Science.gov (United States)

Di Fabrizio, Enzo; Cojoc, Dan; Emiliani, Valentina; Cabrini, Stefano; Coppey-Moisan, Maite; Ferrari, Enrico; Garbin, Valeria; Altissimo, Matteo

2004-11-01

The aim of this report is to demonstrate a unified version of microscopy through the use of advanced diffractive optics. The unified scheme derives from the technical possibility of realizing front wave engineering in a wide range of electromagnetic spectrum. The unified treatment is realized through the design and nanofabrication of phase diffractive elements (PDE) through which wave front beam shaping is obtained. In particular, we will show applications, by using biological samples, ranging from micromanipulation using optical tweezers to X-ray differential interference contrast (DIC) microscopy combined with X-ray fluorescence. We report some details on the design and physical implementation of diffractive elements that besides focusing also perform other optical functions: beam splitting, beam intensity, and phase redistribution or mode conversion. Laser beam splitting is used for multiple trapping and independent manipulation of micro-beads surrounding a cell as an array of tweezers and for arraying and sorting microscopic size biological samples. Another application is the Gauss to Laguerre-Gauss mode conversion, which allows for trapping and transfering orbital angular momentum of light to micro-particles immersed in a fluid. These experiments are performed in an inverted optical microscope coupled with an infrared laser beam and a spatial light modulator for diffractive optics implementation. High-resolution optics, fabricated by means of e-beam lithography, are demonstrated to control the intensity and the phase of the sheared beams in x-ray DIC microscopy. DIC experiments with phase objects reveal a dramatic increase in image contrast compared to bright-field x-ray microscopy. Besides the topographic information, fluorescence allows detection of certain chemical elements (Cl, P, Sc, K) in the same setup, by changing the photon energy of the x-ray beam. (c) 2005 Wiley-Liss, Inc.

Enhanced Emission from Single Isolated Gold Quantum Dots Investigated Using Two-Photon-Excited Fluorescence Near-Field Scanning Optical Microscopy.

Science.gov (United States)

Abeyasinghe, Neranga; Kumar, Santosh; Sun, Kai; Mansfield, John F; Jin, Rongchao; Goodson, Theodore

2016-12-21

New approaches in molecular nanoscopy are greatly desired for interrogation of biological, organic, and inorganic objects with sizes below the diffraction limit. Our current work investigates emergent monolayer-protected gold quantum dots (nanoclusters, NCs) composed of 25 Au atoms by utilizing two-photon-excited fluorescence (TPEF) near-field scanning optical microscopy (NSOM) at single NC concentrations. Here, we demonstrate an approach to synthesize and isolate single NCs on solid glass substrates. Subsequent investigation of the NCs using TPEF NSOM reveals that, even when they are separated by distances of several tens of nanometers, we can excite and interrogate single NCs individually. Interestingly, we observe an enhanced two-photon absorption (TPA) cross section for single Au 25 NCs that can be attributed to few-atom local field effects and to local field-induced microscopic cascading, indicating their potential for use in ultrasensitive sensing, disease diagnostics, cancer cell therapy, and molecular computers. Finally, we report room-temperature aperture-based TPEF NSOM imaging of these NCs for the first time at 30 nm point resolution, which is a ∼5-fold improvement compared to the previous best result for the same technique. This report unveils the unique combination of an unusually large TPA cross section and the high photostability of Au NCs to (non-destructively) investigate stable isolated single NCs using TPEF NSOM. This is the first reported optical study of monolayer-protected single quantum clusters, opening some very promising opportunities in spectroscopy of nanosized objects, bioimaging, ultrasensitive sensing, molecular computers, and high-density data storage.

Leakage radiation interference microscopy.

Science.gov (United States)

Descrovi, Emiliano; Barakat, Elsie; Angelini, Angelo; Munzert, Peter; De Leo, Natascia; Boarino, Luca; Giorgis, Fabrizio; Herzig, Hans Peter

2013-09-01

We present a proof of principle for a new imaging technique combining leakage radiation microscopy with high-resolution interference microscopy. By using oil immersion optics it is demonstrated that amplitude and phase can be retrieved from optical fields, which are evanescent in air. This technique is illustratively applied for mapping a surface mode propagating onto a planar dielectric multilayer on a thin glass substrate. The surface mode propagation constant estimated after Fourier transformation of the measured complex field is well matched with an independent measurement based on back focal plane imaging.

Differential Polarization Nonlinear Optical Microscopy with Adaptive Optics Controlled Multiplexed Beams

Directory of Open Access Journals (Sweden)

Virginijus Barzda

2013-09-01

Full Text Available Differential polarization nonlinear optical microscopy has the potential to become an indispensable tool for structural investigations of ordered biological assemblies and microcrystalline aggregates. Their microscopic organization can be probed through fast and sensitive measurements of nonlinear optical signal anisotropy, which can be achieved with microscopic spatial resolution by using time-multiplexed pulsed laser beams with perpendicular polarization orientations and photon-counting detection electronics for signal demultiplexing. In addition, deformable membrane mirrors can be used to correct for optical aberrations in the microscope and simultaneously optimize beam overlap using a genetic algorithm. The beam overlap can be achieved with better accuracy than diffraction limited point-spread function, which allows to perform polarization-resolved measurements on the pixel-by-pixel basis. We describe a newly developed differential polarization microscope and present applications of the differential microscopy technique for structural studies of collagen and cellulose. Both, second harmonic generation, and fluorescence-detected nonlinear absorption anisotropy are used in these investigations. It is shown that the orientation and structural properties of the fibers in biological tissue can be deduced and that the orientation of fluorescent molecules (Congo Red, which label the fibers, can be determined. Differential polarization microscopy sidesteps common issues such as photobleaching and sample movement. Due to tens of megahertz alternating polarization of excitation pulses fast data acquisition can be conveniently applied to measure changes in the nonlinear signal anisotropy in dynamically changing in vivo structures.

Evaluations of carbon nanotube field emitters for electron microscopy

Science.gov (United States)

Nakahara, Hitoshi; Kusano, Yoshikazu; Kono, Takumi; Saito, Yahachi

2009-11-01

Brightness of carbon nanotube (CNT) emitters was already reported elsewhere. However, brightness of electron emitter is affected by a virtual source size of the emitter, which strongly depends on electron optical configuration around the emitter. In this work, I- V characteristics and brightness of a CNT emitter are measured under a practical field emission electron gun (e-gun) configuration to investigate availability of CNT for electron microscopy. As a result, it is obtained that an emission area of MWNT is smaller than its tip surface area, and the emission area corresponds to a five-membered-ring with 2nd nearest six-membered-rings on the MWNT cap surface. Reduced brightness of MWNT is measured as at least 2.6×109 A/m 2 sr V. It is concluded that even a thick MWNT has enough brightness under a practical e-gun electrode configuration and suitable for electron microscopy.

Fluorescence microscopy.

Science.gov (United States)

Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

2014-10-01

Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

Assessment of nerve ultrastructure by fibre-optic confocal microscopy.

Science.gov (United States)

Cushway, T R; Lanzetta, M; Cox, G; Trickett, R; Owen, E R

1996-01-01

Fibre-optic technology combined with confocality produces a microscope capable of optical thin sectioning. In this original study, tibial nerves have been stained in a rat model with a vital dye, 4-(4-diethylaminostyryl)-N-methylpyridinium iodide, and analysed by fibre-optic confocal microscopy to produce detailed images of nerve ultrastructure. Schwann cells, nodes of Ranvier and longitudinal myelinated sheaths enclosing axons were clearly visible. Single axons appeared as brightly staining longitudinal structures. This allowed easy tracing of multiple signal axons within the nerve tissue. An accurate measurement of internodal lengths was easily accomplished. This technique is comparable to current histological techniques, but does not require biopsy, thin sectioning or tissue fixing. This study offers a standard for further in vivo microscopy, including the possibility of monitoring the progression of nerve regeneration following microsurgical neurorraphy.

Through-focus scanning optical microscopy (TSOM) with adaptive optics

Science.gov (United States)

Lee, Jun Ho; Park, Gyunam; Jeong, Junhee; Park, Chris

2018-03-01

Through-focus optical microscopy (TSOM) with nanometer-scale lateral and vertical sensitivity levels matching those of scanning electron microscopy has been demonstrated to be useful both for 3D inspections and metrology assessments. In 2014, funded by two private companies (Nextin/Samsung Electronics) and the Korea Evaluation Institute of Industrial Technology (KEIT), a research team from four universities in South Korea set out to investigate core technologies for developing in-line TSOM inspection and metrology tools, with the respective teams focusing on optics implementation, defect inspection, computer simulation and high-speed metrology matching. We initially confirmed the reported validity of the TSOM operation through a computer simulation, after which we implemented the TSOM operation by throughfocus scanning of existing UV (355nm) and IR (800nm) inspection tools. These tools have an identical sampling distance of 150 nm but have different resolving distances (310 and 810 nm, respectively). We initially experienced some improvement in the defect inspection sensitivity level over TSV (through-silicon via) samples with 6.6 μm diameters. However, during the experiment, we noted sensitivity and instability issues when attempting to acquire TSOM images. As TSOM 3D information is indirectly extracted by differentiating a target TSOM image from reference TSOM images, any instability or mismatch in imaging conditions can result in measurement errors. As a remedy to such a situation, we proposed the application of adaptive optics to the TSOM operation and developed a closed-loop system with a tip/tilt mirror and a Shack-Hartmann sensor on an optical bench. We were able to keep the plane position within in RMS 0.4 pixel by actively compensating for any position instability which arose during the TSOM scanning process along the optical axis. Currently, we are also developing another TSOM tool with a deformable mirror instead of a tip/tilt mirror, in which case we

3D wide field-of-view Gabor-domain optical coherence microscopy advancing real-time in-vivo imaging and metrology

Science.gov (United States)

Canavesi, Cristina; Cogliati, Andrea; Hayes, Adam; Tankam, Patrice; Santhanam, Anand; Rolland, Jannick P.

2017-02-01

Real-time volumetric high-definition wide-field-of-view in-vivo cellular imaging requires micron-scale resolution in 3D. Compactness of the handheld device and distortion-free images with cellular resolution are also critically required for onsite use in clinical applications. By integrating a custom liquid lens-based microscope and a dual-axis MEMS scanner in a compact handheld probe, Gabor-domain optical coherence microscopy (GD-OCM) breaks the lateral resolution limit of optical coherence tomography through depth, overcoming the tradeoff between numerical aperture and depth of focus, enabling advances in biotechnology. Furthermore, distortion-free imaging with no post-processing is achieved with a compact, lightweight handheld MEMS scanner that obtained a 12-fold reduction in volume and 17-fold reduction in weight over a previous dual-mirror galvanometer-based scanner. Approaching the holy grail of medical imaging - noninvasive real-time imaging with histologic resolution - GD-OCM demonstrates invariant resolution of 2 μm throughout a volume of 1 x 1 x 0.6 mm3, acquired and visualized in less than 2 minutes with parallel processing on graphics processing units. Results on the metrology of manufactured materials and imaging of human tissue with GD-OCM are presented.

Near-field microscopy with a microfabricated solid immersion lens

Science.gov (United States)

Fletcher, Daniel Alden

2001-07-01

Diffraction of focused light prevents optical microscopes from resolving features in air smaller than half the wavelength, λ Spatial resolution can be improved by passing light through a sub-wavelength metal aperture scanned close to a sample, but aperture-based probes suffer from low optical throughput, typically below 10-4. An alternate and more efficient technique is solid immersion microscopy in which light is focused through a high refractive index Solid Immersion Lens (SIL). This work describes the fabrication, modeling, and use of a microfabricated SIL to obtain spatial resolution better than the diffraction limit in air with high optical throughput for infrared applications. SILs on the order of 10 μm in diameter are fabricated from single-crystal silicon and integrated onto silicon cantilevers with tips for scanning. We measure a focused spot size of λ/5 with optical throughput better than 10-1 at a wavelength of λ = 9.3 μm. Spatial resolution is improved to λ/10 with metal apertures fabricated directly on the tip of the silicon SIL. Microlenses have reduced spherical aberration and better transparency than large lenses but cannot be made arbitrarily small and still focus. We model the advantages and limitations of focusing in lenses close to the wavelength in diameter using an extension of Mie theory. We also investigate a new contrast mechanism unique to microlenses resulting from the decrease in field-of-view with lens diameter. This technique is shown to achieve λ/4 spatial resolution. We explore applications of the microfabricated silicon SIL for high spatial resolution thermal microscopy and biological spectroscopy. Thermal radiation is collected through the SIL from a heated surface with spatial resolution four times better than that of a diffraction- limited infrared microscope. Using a Fourier-transform infrared spectrometer, we observe absorption peaks in bacteria cells positioned at the focus of the silicon SIL.

Fractal vector optical fields.

Science.gov (United States)

Pan, Yue; Gao, Xu-Zhen; Cai, Meng-Qiang; Zhang, Guan-Lin; Li, Yongnan; Tu, Chenghou; Wang, Hui-Tian

2016-07-15

We introduce the concept of a fractal, which provides an alternative approach for flexibly engineering the optical fields and their focal fields. We propose, design, and create a new family of optical fields-fractal vector optical fields, which build a bridge between the fractal and vector optical fields. The fractal vector optical fields have polarization states exhibiting fractal geometry, and may also involve the phase and/or amplitude simultaneously. The results reveal that the focal fields exhibit self-similarity, and the hierarchy of the fractal has the "weeding" role. The fractal can be used to engineer the focal field.

Evaluations of carbon nanotube field emitters for electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Nakahara, Hitoshi, E-mail: nakahara@nagoya-u.jp [Department of Quantum Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan); Kusano, Yoshikazu; Kono, Takumi; Saito, Yahachi [Department of Quantum Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan)

2009-11-30

Brightness of carbon nanotube (CNT) emitters was already reported elsewhere. However, brightness of electron emitter is affected by a virtual source size of the emitter, which strongly depends on electron optical configuration around the emitter. In this work, I-V characteristics and brightness of a CNT emitter are measured under a practical field emission electron gun (e-gun) configuration to investigate availability of CNT for electron microscopy. As a result, it is obtained that an emission area of MWNT is smaller than its tip surface area, and the emission area corresponds to a five-membered-ring with 2nd nearest six-membered-rings on the MWNT cap surface. Reduced brightness of MWNT is measured as at least 2.6x10{sup 9} A/m{sup 2} sr V. It is concluded that even a thick MWNT has enough brightness under a practical e-gun electrode configuration and suitable for electron microscopy.

Single molecule detection on the cell membrane with Near-field Scanning Optical Microscopy

NARCIS (Netherlands)

de Bakker, B.I.

2004-01-01

In this research we have developed a dedicated near- field scanning optical microscope (NSOM) for molecular biology and applied it to study the spatial organization of (fluorescently labeled) proteins at the cell surface. For the first time, protein clusters and individual molecules are resolved at

Plasmonic optical antenna design for performing tip-enhanced Raman spectroscopy and microscopy

International Nuclear Information System (INIS)

Kharintsev, S S; Fishman, A I; Salakhov, M Kh; Hoffmann, G G

2013-01-01

This paper highlights optical plasmonic antennas designed with dc-pulsed low-voltage electrochemical etching of a gold wire for implementing tip-enhanced Raman scattering (TERS) measurements. We demonstrate a versatile electrochemical system that allows one to engineer TERS-active metallic gold tips with diverse shapes and sizes in a highly reproducible fashion. The underlying etching mechanism at a voltage-driven meniscus around a gold wire immersed into an electrolyte is discussed in detail. We show that the developed method is suitable to produce not only the simplest geometries such as cones and spheroids, but more complex designs. Attempts have been made to design plasmonic tapered antennas with quasi-uniformly spaced nano-sized bumps on the mesoscopic zone for the extra surface plasmon-light coupling. The capability of the patterned antenna to enhance and localize optical fields is demonstrated with near-field Raman microscopy and spectroscopy of single-walled carbon nanotubes bundles. (paper)

Brain plasticity and functionality explored by nonlinear optical microscopy

Science.gov (United States)

Sacconi, L.; Allegra, L.; Buffelli, M.; Cesare, P.; D'Angelo, E.; Gandolfi, D.; Grasselli, G.; Lotti, J.; Mapelli, J.; Strata, P.; Pavone, F. S.

2010-02-01

In combination with fluorescent protein (XFP) expression techniques, two-photon microscopy has become an indispensable tool to image cortical plasticity in living mice. In parallel to its application in imaging, multi-photon absorption has also been used as a tool for the dissection of single neurites with submicrometric precision without causing any visible collateral damage to the surrounding neuronal structures. In this work, multi-photon nanosurgery is applied to dissect single climbing fibers expressing GFP in the cerebellar cortex. The morphological consequences are then characterized with time lapse 3-dimensional two-photon imaging over a period of minutes to days after the procedure. Preliminary investigations show that the laser induced fiber dissection recalls a regenerative process in the fiber itself over a period of days. These results show the possibility of this innovative technique to investigate regenerative processes in adult brain. In parallel with imaging and manipulation technique, non-linear microscopy offers the opportunity to optically record electrical activity in intact neuronal networks. In this work, we combined the advantages of second-harmonic generation (SHG) with a random access (RA) excitation scheme to realize a new microscope (RASH) capable of optically recording fast membrane potential events occurring in a wide-field of view. The RASH microscope, in combination with bulk loading of tissue with FM4-64 dye, was used to simultaneously record electrical activity from clusters of Purkinje cells in acute cerebellar slices. Complex spikes, both synchronous and asynchronous, were optically recorded simultaneously across a given population of neurons. Spontaneous electrical activity was also monitored simultaneously in pairs of neurons, where action potentials were recorded without averaging across trials. These results show the strength of this technique in describing the temporal dynamics of neuronal assemblies, opening promising

The effect of the ferroelectric domain walls in the scanning near field optical microscopy response of periodically poled Ba2NaNb5O15 and LiNbO3 crystals

International Nuclear Information System (INIS)

Han, T P J; Jaque, F; Lamela, J; Jaque, D; Lifante, G; Cusso, F; Kamiskii, A A

2009-01-01

A study of Ba 2 NaNb 5 O 15 and LiNbO 3 crystals with periodic ferroelectric domain structures using the scanning near field optical microscopy technique is reported. Optical contrast is observed in the regions of ferroelectric domain boundaries and it is analysed using beam propagation method modelling. This reveals that the optical contrast, a consequence of changes in the refractive index, is not due to variation of the waveguide-coupling efficiency, and supports the hypothesis that it is associated with the domain array, which is related to the size of the domain. (fast track communication)

Microsphere imaging with confocal microscopy and two photon microscopy

International Nuclear Information System (INIS)

Chun, Hyung Su; An, Kyung Won; Lee, Jai Hyung

2002-01-01

We have acquired images of polystyrene and fused-silica microsphere by using conventional optical microscopy, confocal microscopy and two-photon microscopy, and performed comparative analysis of these images. Different from conventional optical microscopy, confocal and two-photon microscopy had good optical sectioning capability. In addition, confocal microscopy and two-photon microscopy had better lateral resolution than conventional optical microscopy. These results are attributed to confocality and nonlinearity of confocal microscopy and two photon microscopy, respectively.

Computers in field ion microscopy

International Nuclear Information System (INIS)

Suvorov, A.L.; Razinkova, T.L.; Sokolov, A.G.

1980-01-01

A review is presented of computer applications in field ion microscopy (FIM). The following topics are discussed in detail: (1) modeling field ion images in perfect crystals, (2) a general scheme of modeling, (3) modeling of the process of field evaporation, (4) crystal structure defects, (5) alloys, and (6) automation of FIM experiments and computer-assisted processing of real images. 146 references are given

FDTD simulated observation of a gold nanorod by scanning near-field optical microscopy

International Nuclear Information System (INIS)

Sawada, Keiji; Maruoka, Teruto; Nakamura, Hiroaki; Tamura, Yuichi; Imura, Kohei; Saiki, Toshiharu; Okamoto, Hiromi

2010-01-01

The optical properties of a gold nanorod were investigated by Imura et. al. using an apertured-type scanning near-field optical microscope (SNOM). The observed transmission image showed an oscillating pattern along the long axis of the nanorod. We obtain the image using the finite-difference time-domain (FDTD) method. Our model includes a nanorod on a glass substrate, a SNOM, and current as a light source. We develop a simple method for including the Drude-Lorentz dispersion relation of Vial et. al. for gold in the FDTD. The oscillating pattern is explained by the total current in the nanorod, tip of the SNOM, and light source. (author)

Confocal scanning microscopy with multiple optical probes for high speed measurements and better imaging

Science.gov (United States)

Chun, Wanhee; Lee, SeungWoo; Gweon, Dae-Gab

2008-02-01

Confocal scanning microscopy (CSM) needs a scanning mechanism because only one point information of specimen can be obtained. Therefore the speed of the confocal scanning microscopy is limited by the speed of the scanning tool. To overcome this limitation from scanning tool we propose another scanning mechanism. We make three optical probes in the specimen under confocal condition of each point. Three optical probes are moved by beam scanning mechanism with shared resonant scanning mirror (RM) and galvanometer driven mirror (GM). As each optical probe scan allocated region of the specimen, information from three points is obtained simultaneously and image acquisition time is reduced. Therefore confocal scanning microscopy with multiple optical probes is expected to have three times faster speed of the image acquisition than conventional one. And as another use, multiple optical probes to which different light wavelength is applied can scan whole same region respectively. It helps to obtain better contrast image in case of specimens having different optical characteristics for specific light wavelength. In conclusion confocal scanning microscopy with multiple optical probes is useful technique for views of image acquisition speed and image quality.

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis.

Science.gov (United States)

Agarwal, Nitin; Biancardi, Alberto M; Patten, Florence W; Reeves, Anthony P; Seibel, Eric J

2014-04-01

Aneuploidy is typically assessed by flow cytometry (FCM) and image cytometry (ICM). We used optical projection tomographic microscopy (OPTM) for assessing cellular DNA content using absorption and fluorescence stains. OPTM combines some of the attributes of both FCM and ICM and generates isometric high-resolution three-dimensional (3-D) images of single cells. Although the depth of field of the microscope objective was in the submicron range, it was extended by scanning the objective's focal plane. The extended depth of field image is similar to a projection in a conventional x-ray computed tomography. These projections were later reconstructed using computed tomography methods to form a 3-D image. We also present an automated method for 3-D nuclear segmentation. Nuclei of chicken, trout, and triploid trout erythrocyte were used to calibrate OPTM. Ratios of integrated optical densities extracted from 50 images of each standard were compared to ratios of DNA indices from FCM. A comparison of mean square errors with thionin, hematoxylin, Feulgen, and SYTOX green was done. Feulgen technique was preferred as it showed highest stoichiometry, least variance, and preserved nuclear morphology in 3-D. The addition of this quantitative biomarker could further strengthen existing classifiers and improve early diagnosis of cancer using 3-D microscopy.

Direct imaging of phase objects enables conventional deconvolution in bright field light microscopy.

Directory of Open Access Journals (Sweden)

Carmen Noemí Hernández Candia

Full Text Available In transmitted optical microscopy, absorption structure and phase structure of the specimen determine the three-dimensional intensity distribution of the image. The elementary impulse responses of the bright field microscope therefore consist of separate absorptive and phase components, precluding general application of linear, conventional deconvolution processing methods to improve image contrast and resolution. However, conventional deconvolution can be applied in the case of pure phase (or pure absorptive objects if the corresponding phase (or absorptive impulse responses of the microscope are known. In this work, we present direct measurements of the phase point- and line-spread functions of a high-aperture microscope operating in transmitted bright field. Polystyrene nanoparticles and microtubules (biological polymer filaments serve as the pure phase point and line objects, respectively, that are imaged with high contrast and low noise using standard microscopy plus digital image processing. Our experimental results agree with a proposed model for the response functions, and confirm previous theoretical predictions. Finally, we use the measured phase point-spread function to apply conventional deconvolution on the bright field images of living, unstained bacteria, resulting in improved definition of cell boundaries and sub-cellular features. These developments demonstrate practical application of standard restoration methods to improve imaging of phase objects such as cells in transmitted light microscopy.

All-optical optoacoustic microscopy based on probe beam deflection technique

Directory of Open Access Journals (Sweden)

Saher M. Maswadi

2016-09-01

Full Text Available Optoacoustic (OA microscopy using an all-optical system based on the probe beam deflection technique (PBDT for detection of laser-induced acoustic signals was investigated as an alternative to conventional piezoelectric transducers. PBDT provides a number of advantages for OA microscopy including (i efficient coupling of laser excitation energy to the samples being imaged through the probing laser beam, (ii undistorted coupling of acoustic waves to the detector without the need for separation of the optical and acoustic paths, (iii high sensitivity and (iv ultrawide bandwidth. Because of the unimpeded optical path in PBDT, diffraction-limited lateral resolution can be readily achieved. The sensitivity of the current PBDT sensor of 22 μV/Pa and its noise equivalent pressure (NEP of 11.4 Pa are comparable with these parameters of the optical micro-ring resonator and commercial piezoelectric ultrasonic transducers. Benefits of the present prototype OA microscope were demonstrated by successfully resolving micron-size details in histological sections of cardiac muscle.

All-optical optoacoustic microscopy based on probe beam deflection technique.

Science.gov (United States)

Maswadi, Saher M; Ibey, Bennett L; Roth, Caleb C; Tsyboulski, Dmitri A; Beier, Hope T; Glickman, Randolph D; Oraevsky, Alexander A

2016-09-01

Optoacoustic (OA) microscopy using an all-optical system based on the probe beam deflection technique (PBDT) for detection of laser-induced acoustic signals was investigated as an alternative to conventional piezoelectric transducers. PBDT provides a number of advantages for OA microscopy including (i) efficient coupling of laser excitation energy to the samples being imaged through the probing laser beam, (ii) undistorted coupling of acoustic waves to the detector without the need for separation of the optical and acoustic paths, (iii) high sensitivity and (iv) ultrawide bandwidth. Because of the unimpeded optical path in PBDT, diffraction-limited lateral resolution can be readily achieved. The sensitivity of the current PBDT sensor of 22 μV/Pa and its noise equivalent pressure (NEP) of 11.4 Pa are comparable with these parameters of the optical micro-ring resonator and commercial piezoelectric ultrasonic transducers. Benefits of the present prototype OA microscope were demonstrated by successfully resolving micron-size details in histological sections of cardiac muscle.

Electric-field induced surface instabilities of soft dielectrics and their effects on optical transmittance and scattering

Science.gov (United States)

Shian, Samuel; Kjeer, Peter; Clarke, David R.

2018-03-01

When a voltage is applied to a percolative, mechanically compliant mat of carbon nanotubes (CNTs) on a smooth elastomer bilayer attached to an ITO coated glass substrate, the in-line optical transmittance decreases with increasing voltage. Two regimes of behavior have been identified based on optical scattering, bright field optical microscopy, and confocal optical microscopy. In the low field regime, the electric field produces a spatially inhomogeneous surface deformation of the elastomer that causes local variations in optical refraction and modulates the light transmittance. The spatial variation is associated with the distribution of the CNTs over the surface. At higher fields, above a threshold voltage, an array of pits in the surface form by a nucleation and growth mechanism and these also scatter light. The formation of pits, and creases, in the thickness of the elastomer, is due to a previously identified electro-mechanical surface instability. When the applied voltage is decreased from its maximum, the transmittance returns to its original value although there is a transmittance hysteresis and a complicated time response. When the applied voltage exceeds the threshold voltage, there can be remnant optical contrast associated with creasing of the elastomer and the recovery time appears to be dependent on local jamming of CNTs in areas where the pits formed. A potential application of this work as an electrically tunable privacy window or camouflaging devices is demonstrated.

Practical guidelines for implementing adaptive optics in fluorescence microscopy

Science.gov (United States)

Wilding, Dean; Pozzi, Paolo; Soloviev, Oleg; Vdovin, Gleb; Verhaegen, Michel

2018-02-01

In life sciences, interest in the microscopic imaging of increasingly complex three dimensional samples, such as cell spheroids, zebrafish embryos, and in vivo applications in small animals, is growing quickly. Due to the increasing complexity of samples, more and more life scientists are considering the implementation of adaptive optics in their experimental setups. While several approaches to adaptive optics in microscopy have been reported, it is often difficult and confusing for the microscopist to choose from the array of techniques and equipment. In this poster presentation we offer a small guide to adaptive optics providing general guidelines for successful adaptive optics implementation.

Dimensional metrology of lab-on-a-chip internal structures: a comparison of optical coherence tomography with confocal fluorescence microscopy.

Science.gov (United States)

Reyes, D R; Halter, M; Hwang, J

2015-07-01

The characterization of internal structures in a polymeric microfluidic device, especially of a final product, will require a different set of optical metrology tools than those traditionally used for microelectronic devices. We demonstrate that optical coherence tomography (OCT) imaging is a promising technique to characterize the internal structures of poly(methyl methacrylate) devices where the subsurface structures often cannot be imaged by conventional wide field optical microscopy. The structural details of channels in the devices were imaged with OCT and analyzed with an in-house written ImageJ macro in an effort to identify the structural details of the channel. The dimensional values obtained with OCT were compared with laser-scanning confocal microscopy images of channels filled with a fluorophore solution. Attempts were also made using confocal reflectance and interferometry microscopy to measure the channel dimensions, but artefacts present in the images precluded quantitative analysis. OCT provided the most accurate estimates for the channel height based on an analysis of optical micrographs obtained after destructively slicing the channel with a microtome. OCT may be a promising technique for the future of three-dimensional metrology of critical internal structures in lab-on-a-chip devices because scans can be performed rapidly and noninvasively prior to their use. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Hybrid Imaging for Extended Depth of Field Microscopy

Science.gov (United States)

Zahreddine, Ramzi Nicholas

An inverse relationship exists in optical systems between the depth of field (DOF) and the minimum resolvable feature size. This trade-off is especially detrimental in high numerical aperture microscopy systems where resolution is pushed to the diffraction limit resulting in a DOF on the order of 500 nm. Many biological structures and processes of interest span over micron scales resulting in significant blurring during imaging. This thesis explores a two-step computational imaging technique known as hybrid imaging to create extended DOF (EDF) microscopy systems with minimal sacrifice in resolution. In the first step a mask is inserted at the pupil plane of the microscope to create a focus invariant system over 10 times the traditional DOF, albeit with reduced contrast. In the second step the contrast is restored via deconvolution. Several EDF pupil masks from the literature are quantitatively compared in the context of biological microscopy. From this analysis a new mask is proposed, the incoherently partitioned pupil with binary phase modulation (IPP-BPM), that combines the most advantageous properties from the literature. Total variation regularized deconvolution models are derived for the various noise conditions and detectors commonly used in biological microscopy. State of the art algorithms for efficiently solving the deconvolution problem are analyzed for speed, accuracy, and ease of use. The IPP-BPM mask is compared with the literature and shown to have the highest signal-to-noise ratio and lowest mean square error post-processing. A prototype of the IPP-BPM mask is fabricated using a combination of 3D femtosecond glass etching and standard lithography techniques. The mask is compared against theory and demonstrated in biological imaging applications.

Coherent light microscopy

CERN Document Server

Ferraro, Pietro; Zalevsky, Zeev

2011-01-01

This book deals with the latest achievements in the field of optical coherent microscopy. While many other books exist on microscopy and imaging, this book provides a unique resource dedicated solely to this subject. Similarly, many books describe applications of holography, interferometry and speckle to metrology but do not focus on their use for microscopy. The coherent light microscopy reference provided here does not focus on the experimental mechanics of such techniques but instead is meant to provide a users manual to illustrate the strengths and capabilities of developing techniques. Th

Interference electron microscopy of one-dimensional electron-optical phase objects

International Nuclear Information System (INIS)

Fazzini, P.F.; Ortolani, L.; Pozzi, G.; Ubaldi, F.

2006-01-01

The application of interference electron microscopy to the investigation of electron optical one-dimensional phase objects like reverse biased p-n junctions and ferromagnetic domain walls is considered. In particular the influence of diffraction from the biprism edges on the interference images is analyzed and the range of applicability of the geometric optical equation for the interpretation of the interference fringe shifts assessed by comparing geometric optical images with full wave-optical simulations. Finally, the inclusion of partial spatial coherence effects are discussed

Portable optical-resolution photoacoustic microscopy for volumetric imaging of multiscale organisms.

Science.gov (United States)

Jin, Tian; Guo, Heng; Yao, Lei; Xie, Huikai; Jiang, Huabei; Xi, Lei

2018-04-01

Photoacoustic microscopy (PAM) provides a fundamentally new tool for a broad range of studies of biological structures and functions. However, the use of PAM has been largely limited to small vertebrates due to the large size/weight and the inconvenience of the equipment. Here, we describe a portable optical-resolution photoacoustic microscopy (pORPAM) system for 3-dimensional (3D) imaging of small-to-large rodents and humans with a high spatiotemporal resolution and a large field of view. We show extensive applications of pORPAM to multiscale animals including mice and rabbits. In addition, we image the 3D vascular networks of human lips, and demonstrate the feasibility of pORPAM to observe the recovery process of oral ulcer and cancer-associated capillary loops in human oral cavities. This technology is promising for broad biomedical studies from fundamental biology to clinical diseases. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Applications of multiphoton microscopy in the field of colorectal cancer

Science.gov (United States)

Wang, Shu; Li, Lianhuang; Zhu, Xiaoqin; Zheng, Liqin; Zhuo, Shuangmu; Chen, Jianxin

2018-06-01

Multiphoton microscopy (MPM) is a powerful tool for visualizing cellular and subcellular details within living tissue by its unique advantages of being label-free, its intrinsic optical sectioning ability, near-infrared excitation for deep penetration depth into tissue, reduced photobleaching and phototoxicity in the out-of-focus regions, and being capable of providing quantitative information. In this review, we focus on applications of MPM in the field of colorectal cancer, including monitoring cancer progression, detecting tumor metastasis and microenvironment, evaluating the cancer therapy response, and visualizing and ablating pre-invasive cancer cells. We also present one of the major challenges and the future research direction to exploit a colorectal multiphoton endoscope.

Hybrid Microscopy: Enabling Inexpensive High-Performance Imaging through Combined Physical and Optical Magnifications.

Science.gov (United States)

Zhang, Yu Shrike; Chang, Jae-Byum; Alvarez, Mario Moisés; Trujillo-de Santiago, Grissel; Aleman, Julio; Batzaya, Byambaa; Krishnadoss, Vaishali; Ramanujam, Aishwarya Aravamudhan; Kazemzadeh-Narbat, Mehdi; Chen, Fei; Tillberg, Paul W; Dokmeci, Mehmet Remzi; Boyden, Edward S; Khademhosseini, Ali

2016-03-15

To date, much effort has been expended on making high-performance microscopes through better instrumentation. Recently, it was discovered that physical magnification of specimens was possible, through a technique called expansion microscopy (ExM), raising the question of whether physical magnification, coupled to inexpensive optics, could together match the performance of high-end optical equipment, at a tiny fraction of the price. Here we show that such "hybrid microscopy" methods--combining physical and optical magnifications--can indeed achieve high performance at low cost. By physically magnifying objects, then imaging them on cheap miniature fluorescence microscopes ("mini-microscopes"), it is possible to image at a resolution comparable to that previously attainable only with benchtop microscopes that present costs orders of magnitude higher. We believe that this unprecedented hybrid technology that combines expansion microscopy, based on physical magnification, and mini-microscopy, relying on conventional optics--a process we refer to as Expansion Mini-Microscopy (ExMM)--is a highly promising alternative method for performing cost-effective, high-resolution imaging of biological samples. With further advancement of the technology, we believe that ExMM will find widespread applications for high-resolution imaging particularly in research and healthcare scenarios in undeveloped countries or remote places.

Conformation of single block copolymer chain in two-dimensional microphase-separated structure studied by scanning near-field optical microscopy.

Science.gov (United States)

Sekine, Ryojun; Aoki, Hiroyuki; Ito, Shinzaburo

2009-05-21

The localization and orientation of the symmetric diblock copolymer chain in a quasi-two-dimensional microphase-separated structure were studied by scanning near-field optical microscopy (SNOM). In the monolayer of poly(isobutyl methacrylate)-block-poly(octadecyl methacrylate) (PiBMA-b-PODMA), the individual PiBMA subchains were directly observed by SNOM, and the center of mass (CM) and orientational angle relative to the phase interface were examined at the single chain level. It was found that the position of the CM and the orientation of the PiBMA subchain in the lamellar structure were dependent on the curvature of the PiBMA/PODMA interface. As the interface was bent toward the objective chain, the block chain preferred the CM position closer to the domain center, and the conformation was strongly oriented perpendicularly to the domain interface. With increase of the curvature, the steric hindrance among the block chain increases, resulting in the stretched conformation.

Atom probe field ion microscopy and related topics: A bibliography 1992

Energy Technology Data Exchange (ETDEWEB)

Russell, K.F.; Godfrey, R.D.; Miller, M.K.

1993-12-01

This bibliography contains citations of books, conference proceedings, journals, and patents published in 1992 on the following types of microscopy: atom probe field ion microscopy (108 items); field emission microscopy (101 items); and field ion microscopy (48 items). An addendum of 34 items missed in previous bibliographies is included.

Advanced Wide-Field Interferometric Microscopy for Nanoparticle Sensing and Characterization

Science.gov (United States)

Avci, Oguzhan

Nanoparticles have a key role in today's biotechnological research owing to the rapid advancement of nanotechnology. While metallic, polymer, and semiconductor based artificial nanoparticles are widely used as labels or targeted drug delivery agents, labeled and label-free detection of natural nanoparticles promise new ways for viral diagnostics and therapeutic applications. The increasing impact of nanoparticles in bio- and nano-technology necessitates the development of advanced tools for their accurate detection and characterization. Optical microscopy techniques have been an essential part of research for visualizing micron-scale particles. However, when it comes to the visualization of individual nano-scale particles, they have shown inadequate success due to the resolution and visibility limitations. Interferometric microscopy techniques have gained significant attention for providing means to overcome the nanoparticle visibility issue that is often the limiting factor in the imaging techniques based solely on the scattered light. In this dissertation, we develop a rigorous physical model to simulate the single nanoparticle optical response in a common-path wide-field interferometric microscopy (WIM) system. While the fundamental elements of the model can be used to analyze nanoparticle response in any generic wide-field imaging systems, we focus on imaging with a layered substrate (common-path interferometer) where specular reflection of illumination provides the reference light for interferometry. A robust physical model is quintessential in realizing the full potential of an optical system, and throughout this dissertation, we make use of it to benchmark our experimental findings, investigate the utility of various optical configurations, reconstruct weakly scattering nanoparticle images, as well as to characterize and discriminate interferometric nanoparticle responses. This study investigates the integration of advanced optical schemes in WIM with two

Super-resolution fluorescence microscopy by stepwise optical saturation

Science.gov (United States)

Zhang, Yide; Nallathamby, Prakash D.; Vigil, Genevieve D.; Khan, Aamir A.; Mason, Devon E.; Boerckel, Joel D.; Roeder, Ryan K.; Howard, Scott S.

2018-01-01

Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the super-resolution microscopy is not feasible in many applications. In this paper, we propose and demonstrate a saturation-based super-resolution fluorescence microscopy technique that can be easily implemented and requires neither additional hardware nor complex post-processing. The method is based on the principle of stepwise optical saturation (SOS), where M steps of raw fluorescence images are linearly combined to generate an image with a M-fold increase in resolution compared with conventional diffraction-limited images. For example, linearly combining (scaling and subtracting) two images obtained at regular powers extends the resolution by a factor of 1.4 beyond the diffraction limit. The resolution improvement in SOS microscopy is theoretically infinite but practically is limited by the signal-to-noise ratio. We perform simulations and experimentally demonstrate super-resolution microscopy with both one-photon (confocal) and multiphoton excitation fluorescence. We show that with the multiphoton modality, the SOS microscopy can provide super-resolution imaging deep in scattering samples. PMID:29675306

Quantitative optical microscopy: measurement of cellular biophysical features with a standard optical microscope.

Science.gov (United States)

Phillips, Kevin G; Baker-Groberg, Sandra M; McCarty, Owen J T

2014-04-07

We describe the use of a standard optical microscope to perform quantitative measurements of mass, volume, and density on cellular specimens through a combination of bright field and differential interference contrast imagery. Two primary approaches are presented: noninterferometric quantitative phase microscopy (NIQPM), to perform measurements of total cell mass and subcellular density distribution, and Hilbert transform differential interference contrast microscopy (HTDIC) to determine volume. NIQPM is based on a simplified model of wave propagation, termed the paraxial approximation, with three underlying assumptions: low numerical aperture (NA) illumination, weak scattering, and weak absorption of light by the specimen. Fortunately, unstained cellular specimens satisfy these assumptions and low NA illumination is easily achieved on commercial microscopes. HTDIC is used to obtain volumetric information from through-focus DIC imagery under high NA illumination conditions. High NA illumination enables enhanced sectioning of the specimen along the optical axis. Hilbert transform processing on the DIC image stacks greatly enhances edge detection algorithms for localization of the specimen borders in three dimensions by separating the gray values of the specimen intensity from those of the background. The primary advantages of NIQPM and HTDIC lay in their technological accessibility using "off-the-shelf" microscopes. There are two basic limitations of these methods: slow z-stack acquisition time on commercial scopes currently abrogates the investigation of phenomena faster than 1 frame/minute, and secondly, diffraction effects restrict the utility of NIQPM and HTDIC to objects from 0.2 up to 10 (NIQPM) and 20 (HTDIC) μm in diameter, respectively. Hence, the specimen and its associated time dynamics of interest must meet certain size and temporal constraints to enable the use of these methods. Excitingly, most fixed cellular specimens are readily investigated with

X-ray optics for scanning fluorescence microscopy and other applications

International Nuclear Information System (INIS)

Ryon, R.W.; Warburton, W.K.

1992-05-01

Scanning x-ray fluorescence microscopy is analogous to scanning electron microscopy. Maps of chemical element distribution are produced by scanning with a very small x-ray beam. Goal is to perform such scanning microscopy with resolution in the range of <1 to 10 μm, using standard laboratory x-ray tubes. We are investigating mirror optics in the Kirkpatrick-Baez (K-B) configuration. K-B optics uses two curved mirrors mounted orthogonally along the optical axis. The first mirror provides vertical focus, the second mirror provides horizontal focus. We have used two types of mirrors: synthetic multilayers and crystals. Multilayer mirrors are used with lower energy radiation such as Cu Kα. At higher energies such as Ag Kα, silicon wafers are used in order to increase the incidence angles and thereby the photon collection efficiency. In order to increase the surface area of multilayers which reflects x-rays at the Bragg angle, we have designed mirrors with the spacing between layers graded along the optic axis in order to compensate for the changing angle of incidence. Likewise, to achieve a large reflecting surface with silicon, the wafers are placed on a specially designed lever arm which is bent into a log spiral by applying force at one end. In this way, the same diffracting angle is maintained over the entire surface of the wafer, providing a large solid angle for photon collection

Polarization Control with Plasmonic Antenna Tips: A Universal Approach to Optical Nanocrystallography and Vector-Field Imaging

Science.gov (United States)

Park, Kyoung-Duck; Raschke, Markus B.

2018-05-01

Controlling the propagation and polarization vectors in linear and nonlinear optical spectroscopy enables to probe the anisotropy of optical responses providing structural symmetry selective contrast in optical imaging. Here we present a novel tilted antenna-tip approach to control the optical vector-field by breaking the axial symmetry of the nano-probe in tip-enhanced near-field microscopy. This gives rise to a localized plasmonic antenna effect with significantly enhanced optical field vectors with control of both \\textit{in-plane} and \\textit{out-of-plane} components. We use the resulting vector-field specificity in the symmetry selective nonlinear optical response of second-harmonic generation (SHG) for a generalized approach to optical nano-crystallography and -imaging. In tip-enhanced SHG imaging of monolayer MoS$_2$ films and single-crystalline ferroelectric YMnO$_3$, we reveal nano-crystallographic details of domain boundaries and domain topology with enhanced sensitivity and nanoscale spatial resolution. The approach is applicable to any anisotropic linear and nonlinear optical response, and provides for optical nano-crystallographic imaging of molecular or quantum materials.

An introduction to optical super-resolution microscopy for the adventurous biologist

Science.gov (United States)

Vangindertael, J.; Camacho, R.; Sempels, W.; Mizuno, H.; Dedecker, P.; Janssen, K. P. F.

2018-04-01

Ever since the inception of light microscopy, the laws of physics have seemingly thwarted every attempt to visualize the processes of life at its most fundamental, sub-cellular, level. The diffraction limit has restricted our view to length scales well above 250 nm and in doing so, severely compromised our ability to gain true insights into many biological systems. Fortunately, continuous advancements in optics, electronics and mathematics have since provided the means to once again make physics work to our advantage. Even though some of the fundamental concepts enabling super-resolution light microscopy have been known for quite some time, practically feasible implementations have long remained elusive. It should therefore not come as a surprise that the 2014 Nobel Prize in Chemistry was awarded to the scientists who, each in their own way, contributed to transforming super-resolution microscopy from a technological tour de force to a staple of the biologist’s toolkit. By overcoming the diffraction barrier, light microscopy could once again be established as an indispensable tool in an age where the importance of understanding life at the molecular level cannot be overstated. This review strives to provide the aspiring life science researcher with an introduction to optical microscopy, starting from the fundamental concepts governing compound and fluorescent confocal microscopy to the current state-of-the-art of super-resolution microscopy techniques and their applications.

Atom probe field ion microscopy and related topics: A bibliography 1991

International Nuclear Information System (INIS)

Russell, K.F.; Miller, M.K.

1993-01-01

This report contains a bibliography for 1991 on the following topics: Atom probe field ion microscopy; field desorption mass spectrometry; field emission; field ion microscopy; and field emission theory

Tissue imaging using full field optical coherence microscopy with short multimode fiber probe

Science.gov (United States)

Sato, Manabu; Eto, Kai; Goto, Tetsuhiro; Kurotani, Reiko; Abe, Hiroyuki; Nishidate, Izumi

2018-03-01

In achieving minimally invasive accessibility to deeply located regions the size of the imaging probes is important. We demonstrated full-field optical coherence tomography (FF-OCM) using an ultrathin forward-imaging short multimode fiber (SMMF) probe of 50 μm core diameter, 125 μm diameter, and 7.4 mm length for optical communications. The axial resolution was measured to be 2.14 μm and the lateral resolution was also evaluated to be below 4.38 μm using a test pattern (TP). The spatial mode and polarization characteristics of SMMF were evaluated. Inserting SMMF to in vivo rat brain, 3D images were measured and 2D information of nerve fibers was obtained. The feasibility of an SMMF as an ultrathin forward-imaging probe in FF-OCM has been demonstrated.

Characterization and fabrication of fully metal-coated scanning near-field optical microscopy SiO2 tips.

Science.gov (United States)

Aeschimann, L; Akiyama, T; Staufer, U; De Rooij, N F; Thiery, L; Eckert, R; Heinzelmann, H

2003-03-01

The fabrication of silicon cantilever-based scanning near-field optical microscope probes with fully aluminium-coated quartz tips was optimized to increase production yield. Different cantilever designs for dynamic- and contact-mode force feedback were implemented. Light transmission through the tips was investigated experimentally in terms of the metal coating and the tip cone-angle. We found that transmittance varies with the skin depth of the metal coating and is inverse to the cone angle, meaning that slender tips showed higher transmission. Near-field optical images of individual fluorescing molecules showed a resolution thermocouple showed no evidence of mechanical defect or orifice formation by thermal effects.

Wide-field imaging of birefringent synovial fluid crystals using lens-free polarized microscopy for gout diagnosis

Science.gov (United States)

Zhang, Yibo; Lee, Seung Yoon Celine; Zhang, Yun; Furst, Daniel; Fitzgerald, John; Ozcan, Aydogan

2016-06-01

Gout is a form of crystal arthropathy where monosodium urate (MSU) crystals deposit and elicit inflammation in a joint. Diagnosis of gout relies on identification of MSU crystals under a compensated polarized light microscope (CPLM) in synovial fluid aspirated from the patient’s joint. The detection of MSU crystals by optical microscopy is enhanced by their birefringent properties. However, CPLM partially suffers from the high-cost and bulkiness of conventional lens-based microscopy, and its relatively small field-of-view (FOV) limits the efficiency and accuracy of gout diagnosis. Here we present a lens-free polarized microscope which adopts a novel differential and angle-mismatched polarizing optical design achieving wide-field and high-resolution holographic imaging of birefringent objects with a color contrast similar to that of a standard CPLM. The performance of this computational polarization microscope is validated by imaging MSU crystals made from a gout patient’s tophus and steroid crystals used as negative control. This lens-free polarized microscope, with its wide FOV (>20 mm2), cost-effectiveness and field-portability, can significantly improve the efficiency and accuracy of gout diagnosis, reduce costs, and can be deployed even at the point-of-care and in resource-limited clinical settings.

Probing graphene defects and estimating graphene quality with optical microscopy

International Nuclear Information System (INIS)

Lai, Shen; Kyu Jang, Sung; Jae Song, Young; Lee, Sungjoo

2014-01-01

We report a simple and accurate method for detecting graphene defects that utilizes the mild, dry annealing of graphene/Cu films in air. In contrast to previously reported techniques, our simple approach with optical microscopy can determine the density and degree of dislocation of defects in a graphene film without inducing water-related damage or functionalization. Scanning electron microscopy, confocal Raman and atomic force microscopy, and X-ray photoelectron spectroscopy analysis were performed to demonstrate that our nondestructive approach to characterizing graphene defects with optimized thermal annealing provides rapid and comprehensive determinations of graphene quality

Isometric multimodal photoacoustic microscopy based on optically transparent micro-ring ultrasonic detection.

Science.gov (United States)

Dong, Biqin; Li, Hao; Zhang, Zhen; Zhang, Kevin; Chen, Siyu; Sun, Cheng; Zhang, Hao F

2015-01-01

Photoacoustic microscopy (PAM) is an attractive imaging tool complementary to established optical microscopic modalities by providing additional molecular specificities through imaging optical absorption contrast. While the development of optical resolution photoacoustic microscopy (ORPAM) offers high lateral resolution, the acoustically-determined axial resolution is limited due to the constraint in ultrasonic detection bandwidth. ORPAM with isometric spatial resolution along both axial and lateral direction is yet to be developed. Although recently developed sophisticated optical illumination and reconstruction methods offer improved axial resolution in ORPAM, the image acquisition procedures are rather complicated, limiting their capabilities for high-speed imaging and being easily integrated with established optical microscopic modalities. Here we report an isometric ORPAM based on an optically transparent micro-ring resonator ultrasonic detector and a commercial inverted microscope platform. Owing to the superior spatial resolution and the ease of integrating our ORPAM with established microscopic modalities, single cell imaging with extrinsic fluorescence staining, intrinsic autofluorescence, and optical absorption can be achieved simultaneously. This technique holds promise to greatly improve the accessibility of PAM to the broader biomedical researchers.

Synthetic optical holography for rapid nanoimaging.

Science.gov (United States)

Schnell, M; Carney, P S; Hillenbrand, R

2014-03-20

Holography has paved the way for phase imaging in a variety of wide-field techniques, including electron, X-ray and optical microscopy. In scanning optical microscopy, however, the serial fashion of image acquisition seems to challenge a direct implementation of traditional holography. Here we introduce synthetic optical holography (SOH) for quantitative phase-resolved imaging in scanning optical microscopy. It uniquely combines fast phase imaging, technical simplicity and simultaneous operation at visible and infrared frequencies with a single reference arm. We demonstrate SOH with a scattering-type scanning near-field optical microscope (s-SNOM) where it enables reliable quantitative phase-resolved near-field imaging with unprecedented speed. We apply these capabilities to nanoscale, non-invasive and rapid screening of grain boundaries in CVD-grown graphene, by recording 65 kilopixel near-field images in 26 s and 2.3 megapixel images in 13 min. Beyond s-SNOM, the SOH concept could boost the implementation of holography in other scanning imaging applications such as confocal microscopy.

Combined multi-plane phase retrieval and super-resolution optical fluctuation imaging for 4D cell microscopy

Science.gov (United States)

Descloux, A.; Grußmayer, K. S.; Bostan, E.; Lukes, T.; Bouwens, A.; Sharipov, A.; Geissbuehler, S.; Mahul-Mellier, A.-L.; Lashuel, H. A.; Leutenegger, M.; Lasser, T.

2018-03-01

Super-resolution fluorescence microscopy provides unprecedented insight into cellular and subcellular structures. However, going `beyond the diffraction barrier' comes at a price, since most far-field super-resolution imaging techniques trade temporal for spatial super-resolution. We propose the combination of a novel label-free white light quantitative phase imaging with fluorescence to provide high-speed imaging and spatial super-resolution. The non-iterative phase retrieval relies on the acquisition of single images at each z-location and thus enables straightforward 3D phase imaging using a classical microscope. We realized multi-plane imaging using a customized prism for the simultaneous acquisition of eight planes. This allowed us to not only image live cells in 3D at up to 200 Hz, but also to integrate fluorescence super-resolution optical fluctuation imaging within the same optical instrument. The 4D microscope platform unifies the sensitivity and high temporal resolution of phase imaging with the specificity and high spatial resolution of fluorescence microscopy.

Electron microscopy of primary cell cultures in solution and correlative optical microscopy using ASEM

International Nuclear Information System (INIS)

Hirano, Kazumi; Kinoshita, Takaaki; Uemura, Takeshi; Motohashi, Hozumi; Watanabe, Yohei; Ebihara, Tatsuhiko; Nishiyama, Hidetoshi; Sato, Mari; Suga, Mitsuo; Maruyama, Yuusuke; Tsuji, Noriko M.; Yamamoto, Masayuki; Nishihara, Shoko; Sato, Chikara

2014-01-01

Correlative light-electron microscopy of cells in a natural environment of aqueous liquid facilitates high-throughput observation of protein complex formation. ASEM allows the inverted SEM to observe the wet sample from below, while an optical microscope observes it from above quasi-simultaneously. The disposable ASEM dish with a silicon nitride (SiN) film window can be coated variously to realize the primary-culture of substrate-sensitive cells in a few milliliters of culture medium in a stable incubator environment. Neuron differentiation, neural networking, proplatelet-formation and phagocytosis were captured by optical or fluorescence microscopy, and imaged at high resolution by gold-labeled immuno-ASEM with/without metal staining. Fas expression on the cell surface was visualized, correlated to the spatial distribution of F-actin. Axonal partitioning was studied using primary-culture neurons, and presynaptic induction by GluRδ2-N-terminus-linked fluorescent magnetic beads was correlated to the presynaptic-marker Bassoon. Further, megakaryocytes secreting proplatelets were captured, and P-selectins with adherence activity were localized to some of the granules present by immuno-ASEM. The phagocytosis of lactic acid bacteria by dendritic cells was also imaged. Based on these studies, ASEM correlative microscopy promises to allow the study of various mesoscopic-scale dynamics in the near future. - Highlights: • In situ correlative light electron microscopy of samples in open solution by ASEM. • Primary cultures for in-solution CLEM by developing SiN-film coating methods • First visualization of fluorescent magnetic beads in aqueous solution by CLEM. • Presynaptic induction of neurons by GluRδ2-N-terminus-coated beads studied by CLEM. • Axonal partitioning, bacterial phagocytosis, platelet formation imaged by CLEM

Electron microscopy of primary cell cultures in solution and correlative optical microscopy using ASEM

Energy Technology Data Exchange (ETDEWEB)

Hirano, Kazumi; Kinoshita, Takaaki [Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577 (Japan); Uemura, Takeshi [Department of Molecular Neurobiology and Pharmacology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Department of Molecular and Cellular Physiology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621 (Japan); Motohashi, Hozumi [Department of Gene Expression Regulation, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryo-cho, Aoba-ku, Sendai 980-8575 (Japan); Watanabe, Yohei; Ebihara, Tatsuhiko [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Nishiyama, Hidetoshi [JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558 (Japan); Sato, Mari [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Suga, Mitsuo [JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558 (Japan); Maruyama, Yuusuke; Tsuji, Noriko M. [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Yamamoto, Masayuki [Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai 980-8575 (Japan); Nishihara, Shoko, E-mail: shoko@soka.ac.jp [Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577 (Japan); Sato, Chikara, E-mail: ti-sato@aist.go.jp [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan)

2014-08-01

Correlative light-electron microscopy of cells in a natural environment of aqueous liquid facilitates high-throughput observation of protein complex formation. ASEM allows the inverted SEM to observe the wet sample from below, while an optical microscope observes it from above quasi-simultaneously. The disposable ASEM dish with a silicon nitride (SiN) film window can be coated variously to realize the primary-culture of substrate-sensitive cells in a few milliliters of culture medium in a stable incubator environment. Neuron differentiation, neural networking, proplatelet-formation and phagocytosis were captured by optical or fluorescence microscopy, and imaged at high resolution by gold-labeled immuno-ASEM with/without metal staining. Fas expression on the cell surface was visualized, correlated to the spatial distribution of F-actin. Axonal partitioning was studied using primary-culture neurons, and presynaptic induction by GluRδ2-N-terminus-linked fluorescent magnetic beads was correlated to the presynaptic-marker Bassoon. Further, megakaryocytes secreting proplatelets were captured, and P-selectins with adherence activity were localized to some of the granules present by immuno-ASEM. The phagocytosis of lactic acid bacteria by dendritic cells was also imaged. Based on these studies, ASEM correlative microscopy promises to allow the study of various mesoscopic-scale dynamics in the near future. - Highlights: • In situ correlative light electron microscopy of samples in open solution by ASEM. • Primary cultures for in-solution CLEM by developing SiN-film coating methods • First visualization of fluorescent magnetic beads in aqueous solution by CLEM. • Presynaptic induction of neurons by GluRδ2-N-terminus-coated beads studied by CLEM. • Axonal partitioning, bacterial phagocytosis, platelet formation imaged by CLEM.

Investigation of porous asphalt microstructure using optical and electron microscopy.

Science.gov (United States)

Poulikakos, L D; Partl, M N

2010-11-01

Direct observations of porous asphalt concrete samples in their natural state using optical and electron microscopy techniques led to useful information regarding the microstructure of two mixes and indicated a relationship between microstructure and in situ performance. This paper presents evidence that suboptimal microstructure can lead to premature failure thus making a first step in defining well or suboptimal performing pavements with a bottom-up approach (microstructure). Laboratory and field compaction produce different samples in terms of the microstructure. Laboratory compaction using the gyratory method has produced more microcracks in mineral aggregates after the binder had cooled. Well-performing mixes used polymer-modified binders, had a more homogeneous void structure with fewer elongated voids and better interlocking of the aggregates. Furthermore, well-performing mixes showed better distribution of the mastic and better coverage of the aggregates with bitumen. Low vacuum scanning electron microscopy showed that styrene butadiene styrene polymer modification in binder exists in the form of discontinuous globules and not continuous networks. A reduction in the polymer phase was observed as a result of aging and in-service use. © 2010 The Authors Journal compilation © 2010 The Royal Microscopical Society.

Direct characterization of ultraviolet-light-induced refractive index structures by scanning near-field optical microscopy

DEFF Research Database (Denmark)

Svalgaard, Mikael; Madsen, S.; Hvam, Jørn Märcher

1998-01-01

We have applied a reflection scanning near-field optical microscope to directly probe ultraviolet (UV)-light-induced refractive index structures in planar glass samples. This technique permits direct comparison between topography and refractive index changes (10(-5)-10(-3)) with submicrometer...

Dictionary of Microscopy

Science.gov (United States)

Heath, Julian

2005-10-01

The past decade has seen huge advances in the application of microscopy in all areas of science. This welcome development in microscopy has been paralleled by an expansion of the vocabulary of technical terms used in microscopy: terms have been coined for new instruments and techniques and, as microscopes reach even higher resolution, the use of terms that relate to the optical and physical principles underpinning microscopy is now commonplace. The Dictionary of Microscopy was compiled to meet this challenge and provides concise definitions of over 2,500 terms used in the fields of light microscopy, electron microscopy, scanning probe microscopy, x-ray microscopy and related techniques. Written by Dr Julian P. Heath, Editor of Microscopy and Analysis, the dictionary is intended to provide easy navigation through the microscopy terminology and to be a first point of reference for definitions of new and established terms. The Dictionary of Microscopy is an essential, accessible resource for: students who are new to the field and are learning about microscopes equipment purchasers who want an explanation of the terms used in manufacturers' literature scientists who are considering using a new microscopical technique experienced microscopists as an aide mémoire or quick source of reference librarians, the press and marketing personnel who require definitions for technical reports.

Field guide to nonlinear optics

CERN Document Server

Powers, Peter E

2013-01-01

Optomechanics is a field of mechanics that addresses the specific design challenges associated with optical systems. This [i]Field Guide [/i]describes how to mount optical components, as well as how to analyze a given design. It is intended for practicing optical and mechanical engineers whose work requires knowledge in both optics and mechanics. This Field Guide is designed for those looking for a condensed and concise source of key concepts, equations, and techniques for nonlinear optics. Topics covered include technologically important effects, recent developments in nonlinear optics

Incorporating Basic Optical Microscopy in the Instrumental Analysis Laboratory

Science.gov (United States)

Flowers, Paul A.

2011-01-01

A simple and versatile approach to incorporating basic optical microscopy in the undergraduate instrumental analysis laboratory is described. Attaching a miniature CCD spectrometer to the video port of a standard compound microscope yields a visible microspectrophotometer suitable for student investigations of fundamental spectrometry concepts,…

Doppler optical coherence microscopy and tomography applied to inner ear mechanics

Energy Technology Data Exchange (ETDEWEB)

Page, Scott; Freeman, Dennis M. [Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Ghaffari, Roozbeh [Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States)

2015-12-31

While it is clear that cochlear traveling waves underlie the extraordinary sensitivity, frequency selectivity, and dynamic range of mammalian hearing, the underlying micromechanical mechanisms remain unresolved. Recent advances in low coherence measurement techniques show promise over traditional laser Doppler vibrometry and video microscopy, which are limited by low reflectivities of cochlear structures and restricted optical access. Doppler optical coherence tomography (DOCT) and Doppler optical coherence microscopy (DOCM) both utilize a broadband source to limit constructive interference of scattered light to a small axial depth called a coherence gate. The coherence gate can be swept axially to image and measure sub-nanometer motions of cochlear structures throughout the cochlear partition. The coherence gate of DOCT is generally narrower than the confocal gate of the focusing optics, enabling increased axial resolution (typically 15 μm) within optical sections of the cochlear partition. DOCM, frequently implemented in the time domain, centers the coherence gate on the focal plane, achieving enhanced lateral and axial resolution when the confocal gate is narrower than the coherence gate. We compare these two complementary systems and demonstrate their utility in studying cellular and micromechanical mechanisms involved in mammalian hearing.

Growth and decay dynamics of a stable microbubble produced at the end of a near-field scanning optical microscopy fiber probe

International Nuclear Information System (INIS)

Taylor, R.S.; Hnatovsky, C.

2004-01-01

Low power cw laser radiation coupled into a near-field scanning optical microscopy fiber probe has been used to generate a stable microbubble in water. A probe tip which was selectively chemically etched and metallized served as a microheater for the generation of the stable bubble. Bubble diameters in the range of 40-400 μm and lifetimes of over an hour have been obtained. The microbubble exhibited a linear growth phase over a period of a few seconds before reaching a maximum diameter which depended on the laser power. When the laser beam was blocked the microbubble decayed with a rate which was inversely proportional to the bubble diameter. The bubble lifetime depended on the square of the initial bubble diameter. Instabilities which transform a large stable bubble into a microjet stream of micron sized bubbles as the laser power was increased is also described

All-optical optoacoustic microscopy based on probe beam deflection technique

OpenAIRE

Maswadi, Saher M.; Ibey, Bennett L.; Roth, Caleb C.; Tsyboulski, Dmitri A.; Beier, Hope T.; Glickman, Randolph D.; Oraevsky, Alexander A.

2016-01-01

Optoacoustic (OA) microscopy using an all-optical system based on the probe beam deflection technique (PBDT) for detection of laser-induced acoustic signals was investigated as an alternative to conventional piezoelectric transducers. PBDT provides a number of advantages for OA microscopy including (i) efficient coupling of laser excitation energy to the samples being imaged through the probing laser beam, (ii) undistorted coupling of acoustic waves to the detector without the need for separa...

Second-harmonic scanning optical microscopy of semiconductor quantum dots

DEFF Research Database (Denmark)

Vohnsen, B.; Bozhevolnyi, S.I.; Pedersen, K.

2001-01-01

Second-harmonic (SH) optical imaging of self-assembled InAlGaAs quantum dots (QD's) grown on a GaAs(0 0 1) substrate has been accomplished at room temperature by use of respectively a scanning far-field optical microscope in reflection mode and a scanning near-field optical microscope...... in transmission mode. In both cases the SH signal peaks at a pump wavelength of similar to 885 nm in correspondence to the maximum in the photoluminescence spectrum of the QD sample. SH near-field optical images exhibit spatial signal variations on a subwavelength scale that depend on the pump wavelength. We...

Contrast Induced by a Static Magnetic Field for Improved Detection in Nanodiamond Fluorescence Microscopy

Science.gov (United States)

Singam, Shashi K. R.; Motylewski, Jaroslaw; Monaco, Antonina; Gjorgievska, Elena; Bourgeois, Emilie; Nesládek, Milos; Giugliano, Michele; Goovaerts, Etienne

2016-12-01

Diamond nanoparticles with negatively charged nitrogen-vacancy (NV) centers are highly efficient nonblinking emitters that exhibit spin-dependent intensity. An attractive application of these emitters is background-free fluorescence microscopy exploiting the fluorescence quenching induced either by resonant microwaves (RMWs) or by an applied static magnetic field (SMF). Here, we compare RMW- and SMF-induced contrast measurements over a wide range of optical excitation rates for fluorescent nanodiamonds (FNDs) and for NV centers shallowly buried under the (100)-oriented surface of a diamond single crystal (SC). Contrast levels are found to be systematically lower in the FNDs than in the SC. At low excitation rates, the RMW contrast initially rises to a maximum (up to 7% in FNDs and 13% in the SC) but then decreases steadily at higher intensities. Conversely, the SMF contrast increases from approximately 12% at low excitation rates to high values of 20% and 38% for the FNDs and SC, respectively. These observations are well described in a rate-equations model for the charged NV defect using parameters in good agreement with the literature. The SMF approach yields higher induced contrast in image collection under commonly applied optical excitation. Unlike the RMW method, there is no thermal load exerted on the aqueous media in biological samples in the SMF approach. We demonstrate imaging by SMF-induced contrast in neuronal cultures incorporating FNDs (i) in a setup for patch-clamp experiments in parallel with differential-interference-contrast microscopy, (ii) after a commonly used staining procedure as an illustration of the high selectivity against background fluorescence, and (iii) in a confocal fluorescence microscope in combination with bright-field microscopy.

Quantitative photoacoustic microscopy of optical absorption coefficients from acoustic spectra in the optical diffusive regime.

Science.gov (United States)

Guo, Zijian; Favazza, Christopher; Garcia-Uribe, Alejandro; Wang, Lihong V

2012-06-01

Photoacoustic (PA) microscopy (PAM) can image optical absorption contrast with ultrasonic spatial resolution in the optical diffusive regime. Conventionally, accurate quantification in PAM requires knowledge of the optical fluence attenuation, acoustic pressure attenuation, and detection bandwidth. We circumvent this requirement by quantifying the optical absorption coefficients from the acoustic spectra of PA signals acquired at multiple optical wavelengths. With the acoustic spectral method, the absorption coefficients of an oxygenated bovine blood phantom at 560, 565, 570, and 575 nm were quantified with errors of <3%. We also quantified the total hemoglobin concentration and hemoglobin oxygen saturation in a live mouse. Compared with the conventional amplitude method, the acoustic spectral method provides greater quantification accuracy in the optical diffusive regime. The limitations of the acoustic spectral method was also discussed.

Near field plasmon and force microscopy

NARCIS (Netherlands)

de Hollander, R.B.G.; van Hulst, N.F.; Kooyman, R.P.H.

1995-01-01

A scanning plasmon near field optical microscope (SPNM) is presented which combines a conventional far field surface plasmon microscope with a stand-alone atomic force microscope (AFM). Near field plasmon and force images are recorded simultaneously both with a lateral resolution limited by the

Dual-polarization interference microscopy for advanced quantification of phase associated with the image field.

Science.gov (United States)

Bouchal, Petr; Chmelík, Radim; Bouchal, Zdeněk

2018-02-01

A new concept of dual-polarization spatial light interference microscopy (DPSLIM) is proposed and demonstrated experimentally. The method works with two orthogonally polarized modes in which signal and reference waves are combined to realize the polarization-sensitive phase-shifting, thus allowing advanced reconstruction of the phase associated with the image field. The image phase is reconstructed directly from four polarization encoded interference records by a single step processing. This is a progress compared with common methods, in which the phase of the image field is reconstructed using the optical path difference and the amplitudes of interfering waves, which are calculated in multiple-step processing of the records. The DPSLIM is implemented in a common-path configuration using a spatial light modulator, which is connected to a commercial microscope Nikon E200. The optical performance of the method is demonstrated in experiments using both polystyrene microspheres and live LW13K2 cells.

Bright field electron microscopy of biological specimens

International Nuclear Information System (INIS)

Johansen, B.V.

1976-01-01

A preirradiation procedure is described which preserves negatively stained morphological features in bright field electron micrographs to a resolution of about 1.2 nm. Prior to microscopy the pre-irradiation dose (1.6 x 10 -3 C cm -2 ) is given at low electron optical magnification at five different areas on the grid (the centre plus four 'corners'). This pre-irradiation can be measured either with a Faraday cage or through controlled exposure-developing conditions. Uranyl formate stained T2 bacteriophages and stacked disk aggregates of Tobacco Mosaic Virus (TMV) protein served as test objects. A comparative study was performed on specimens using either the pre-irradiation procedure or direct irradiation by the 'minimum beam exposure' technique. Changes in the electron diffraction pattern of the stain-protein complex and the disappearance of certain morphological features in the specimens were both used in order to compare the pre-irradiation method with the direct exposure technique. After identical electron exposures the pre-irradiation approach gave a far better preservation of specimen morphology. Consequently this procedure gives the microscopist more time to select and focus appropriate areas for imaging before deteriorations take place. The investigation also suggested that microscopy should be carried out between 60,000 and 100,000 times magnification. Within this magnification range, it is possible to take advantage of the phase contrast transfer characteristics of the objective lens while the electron load on the object is kept at a moderate level. Using the pre-irradiation procedure special features of the T2 bacteriophage morphology could be established. (author)

Elliptic-symmetry vector optical fields.

Science.gov (United States)

Pan, Yue; Li, Yongnan; Li, Si-Min; Ren, Zhi-Cheng; Kong, Ling-Jun; Tu, Chenghou; Wang, Hui-Tian

2014-08-11

We present in principle and demonstrate experimentally a new kind of vector fields: elliptic-symmetry vector optical fields. This is a significant development in vector fields, as this breaks the cylindrical symmetry and enriches the family of vector fields. Due to the presence of an additional degrees of freedom, which is the interval between the foci in the elliptic coordinate system, the elliptic-symmetry vector fields are more flexible than the cylindrical vector fields for controlling the spatial structure of polarization and for engineering the focusing fields. The elliptic-symmetry vector fields can find many specific applications from optical trapping to optical machining and so on.

Atom probe field ion microscopy and related topics: A bibliography 1989

International Nuclear Information System (INIS)

Miller, M.K.; Hawkins, A.R.; Russell, K.F.

1990-12-01

This bibliography includes references related to the following topics: atom probe field ion microscopy (APFIM), field ion spectroscopy (FIM), field emission microscopy (FEM), liquid metal ion sources (LMIS), scanning tunneling microscopy (STM), and theory. Technique-orientated studies and applications are included. This bibliography covers the period 1989. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications

Exploring neural cell dynamics with digital holographic microscopy

KAUST Repository

Marquet, Pierre; Depeursinge, Christian D.; Magistretti, Pierre J.

2013-01-01

In this review, we summarize how the new concept of digital optics applied to the field of holographic microscopy has allowed the development of a reliable and flexible digital holographic quantitative phase microscopy (DH-QPM) technique at the nanoscale particularly suitable for cell imaging. Particular emphasis is placed on the original biological ormation provided by the quantitative phase signal. We present the most relevant DH-QPM applications in the field of cell biology, including automated cell counts, recognition, classification, three-dimensional tracking, discrimination between physiological and pathophysiological states, and the study of cell membrane fluctuations at the nanoscale. In the last part, original results show how DH-QPM can address two important issues in the field of neurobiology, namely, multiple-site optical recording of neuronal activity and noninvasive visualization of dendritic spine dynamics resulting from a full digital holographic microscopy tomographic approach. Copyright © 2013 by Annual Reviews.

Exploring neural cell dynamics with digital holographic microscopy

KAUST Repository

Marquet, Pierre

2013-07-11

In this review, we summarize how the new concept of digital optics applied to the field of holographic microscopy has allowed the development of a reliable and flexible digital holographic quantitative phase microscopy (DH-QPM) technique at the nanoscale particularly suitable for cell imaging. Particular emphasis is placed on the original biological ormation provided by the quantitative phase signal. We present the most relevant DH-QPM applications in the field of cell biology, including automated cell counts, recognition, classification, three-dimensional tracking, discrimination between physiological and pathophysiological states, and the study of cell membrane fluctuations at the nanoscale. In the last part, original results show how DH-QPM can address two important issues in the field of neurobiology, namely, multiple-site optical recording of neuronal activity and noninvasive visualization of dendritic spine dynamics resulting from a full digital holographic microscopy tomographic approach. Copyright © 2013 by Annual Reviews.

Comparison between optical techniques and confocal microscopy for defect detection on thin wires

International Nuclear Information System (INIS)

Siegmann, Philip; Sanchez-Brea, Luis Miguel; Martinez-Anton, Juan Carlos; Bernabeu, Eusebio

2004-01-01

Conventional microscopy techniques, such as atomic force microscopy (AFM), scanning electron microscopy (SEM), and confocal microscopy (CM) are not suitable for on-line surface inspection of fine metallic wires. In the recent years, some optical techniques have been developed to be used for those tasks. However, they need a rigorous validation. In this work, we have used confocal microscopy to obtain the topography z(x,y) of wires with longitudinal defects, such as dielines. The topography has been used to predict the light scattered by the wire. These simulations have been compared with experimental results, showing a good agreement

Waveguide analysis of heat-drawn and chemically etched probe tips for scanning near-field optical microscopy.

Science.gov (United States)

Moar, Peter N; Love, John D; Ladouceur, François; Cahill, Laurence W

2006-09-01

We analyze two basic aspects of a scanning near-field optical microscope (SNOM) probe's operation: (i) spot-size evolution of the electric field along the probe with and without a metal layer, and (ii) a modal analysis of the SNOM probe, particularly in close proximity to the aperture. A slab waveguide model is utilized to minimize the analytical complexity, yet provides useful quantitative results--including losses associated with the metal coating--which can then be used as design rules.

Vectorial optical fields fundamentals and applications

CERN Document Server

2014-01-01

Polarization is a vector nature of light that plays an important role in optical science and engineering. While existing textbook treatments of light assume beams with spatially homogeneous polarization, there is an increasing interest in vectorial optical fields with spatially engineered states of polarization. New effects and phenomena have been predicted and observed for light beams with these unconventional polarization states. This edited review volume aims to provide a comprehensive overview and summarize the latest developments in this important emerging field of optics. This book will cover the fundamentals including mathematical and physical descriptions, experimental generation, manipulation, focusing, propagation, and the applications of the engineered vectorial optical fields in focal field engineering, plasmonic focusing and optical antenna, single molecular imaging, optical tweezers/trapping, as well as optical measurements and instrumentations. Readership: Students, professionals, post-graduat...

Dark field X-ray microscopy for studies of recrystallization

DEFF Research Database (Denmark)

Ahl, Sonja Rosenlund; Simons, Hugh; Jakobsen, Anders Clemen

2015-01-01

We present the recently developed technique of Dark Field X-Ray Microscopy that utilizes the diffraction of hard X-rays from individual grains or subgrains at the (sub)micrometre- scale embedded within mm-sized samples. By magnifying the diffracted signal, 3D mapping of orientations and strains...... external influences. The capabilities of Dark Field X- Ray Microscopy are illustrated by examples from an ongoing study of recrystallization of 50% cold-rolled Al1050 specimens....

Progress in nano-electro optics characterization of nano-optical materials and optical near-field interactions

CERN Document Server

Ohtsu, Motoichi

2005-01-01

This volume focuses on the characterization of nano-optical materials and optical-near field interactions. It begins with the techniques for characterizing the magneto-optical Kerr effect and continues with methods to determine structural and optical properties in high-quality quantum wires with high spatial uniformity. Further topics include: near-field luminescence mapping in InGaN/GaN single quantum well structures in order to interpret the recombination mechanism in InGaN-based nano-structures; and theoretical treatment of the optical near field and optical near-field interactions, providing the basis for investigating the signal transport and associated dissipation in nano-optical devices. Taken as a whole, this overview will be a valuable resource for engineers and scientists working in the field of nano-electro-optics.

Magnetoelectric force microscopy based on magnetic force microscopy with modulated electric field.

Science.gov (United States)

Geng, Yanan; Wu, Weida

2014-05-01

We present the realization of a mesoscopic imaging technique, namely, the Magnetoelectric Force Microscopy (MeFM), for visualization of local magnetoelectric effect. The basic principle of MeFM is the lock-in detection of local magnetoelectric response, i.e., the electric field-induced magnetization, using magnetic force microscopy. We demonstrate MeFM capability by visualizing magnetoelectric domains on single crystals of multiferroic hexagonal manganites. Results of several control experiments exclude artifacts or extrinsic origins of the MeFM signal. The parameters are tuned to optimize the signal to noise ratio.

Mapping the Landscape of Domain-Wall Pinning in Ferromagnetic Films Using Differential Magneto-Optical Microscopy

Science.gov (United States)

Badea, Robert; Berezovsky, Jesse

2016-06-01

The propagation of domain walls in a ferromagnetic film is largely determined by domain-wall pinning at defects in the material. In this article, we map the effective potential landscape for domain-wall pinning in permalloy films by raster scanning a single ferromagnetic vortex and monitoring the hysteretic vortex displacement vs applied magnetic field. The measurement is carried out using a differential magneto-optical microscopy technique which yields spatial sensitivity of approximately 10 nm. We present a simple algorithm for extracting an effective pinning potential from the measurement of vortex displacement vs applied field. The resulting maps of the pinning potential reveal distinct types of pinning sites, which we attribute to quasi-zero-, one-, and two-dimensional defects in the permalloy film.

Image correction in magneto-optical microscopy

DEFF Research Database (Denmark)

Paturi, P.; Larsen, B.H.; Jacobsen, B.A.

2003-01-01

An image-processing procedure that assures correct determination of the magnetic field distribution of magneto-optical images is presented. The method remedies image faults resulting from sources that are proportional to the incident light intensity, such as different types of defects...

SEM/EDS and optical microscopy analyses of microplastics in ocean trawl and fish guts.

Science.gov (United States)

Wang, Zhong-Min; Wagner, Jeff; Ghosal, Sutapa; Bedi, Gagandeep; Wall, Stephen

2017-12-15

Microplastic particles from Atlantic and Pacific Ocean trawls, lab-fed fish guts and ocean fish guts have been characterized using optical microscopy and SEM/EDS in terms of size, morphology, and chemistry. We assessed whether these measurements could serve as a rapid screening process for subsequent identification of the likely microplastic candidates by micro-spectroscopy. Optical microscopy enabled morphological classification of the types of particles or fibers present in the sample, as well as the quantification of particle size ranges and fiber lengths. SEM/EDS analysis was used to rule out non-plastic particles and screen the prepared samples for potential microplastic, based on their element signatures and surface characteristics. Chlorinated plastics such as polyvinyl chloride (PVC) could be easily identified with SEM/EDS due to their unique elemental signatures including chlorine, as could mineral species that are falsely identified as plastics by optical microscopy. Particle morphology determined by optical microscopy and SEM suggests the fish ingested particles contained both degradation fragments from larger plastic pieces and also manufactured microplastics. SEM images of microplastic particle surfaces revealed characteristic cracks consistent with environmental exposure, as well as pigment particles consistent with manufactured materials. Most of the microplastic surfaces in the fish guts and ocean trawls were covered with biofilms, radiolarians, and crustaceans. Many of the fish stomachs contained micro-shell pieces which visually resembled microplastics. Copyright © 2017 Elsevier B.V. All rights reserved.

Scanning probe microscopy in material science and biology

International Nuclear Information System (INIS)

Cricenti, A; Colonna, S; Girasole, M; Gori, P; Ronci, F; Longo, G; Dinarelli, S; Luce, M; Rinaldi, M; Ortenzi, M

2011-01-01

A review of the activity of scanning probe microscopy at our Institute is presented, going from instrumentation to software development of scanning tunnelling microscopy, atomic force microscopy and scanning near-field optical microscopy (SNOM). Some of the most important experiments in material science and biology performed by our group through the years with these SPM techniques will be presented. Finally, infrared applications by coupling a SNOM with a free electron laser will also be presented.

Asymmetric-detection time-stretch optical microscopy (ATOM) for ultrafast high-contrast cellular imaging in flow

Science.gov (United States)

Wong, Terence T. W.; Lau, Andy K. S.; Ho, Kenneth K. Y.; Tang, Matthew Y. H.; Robles, Joseph D. F.; Wei, Xiaoming; Chan, Antony C. S.; Tang, Anson H. L.; Lam, Edmund Y.; Wong, Kenneth K. Y.; Chan, Godfrey C. F.; Shum, Ho Cheung; Tsia, Kevin K.

2014-01-01

Accelerating imaging speed in optical microscopy is often realized at the expense of image contrast, image resolution, and detection sensitivity – a common predicament for advancing high-speed and high-throughput cellular imaging. We here demonstrate a new imaging approach, called asymmetric-detection time-stretch optical microscopy (ATOM), which can deliver ultrafast label-free high-contrast flow imaging with well delineated cellular morphological resolution and in-line optical image amplification to overcome the compromised imaging sensitivity at high speed. We show that ATOM can separately reveal the enhanced phase-gradient and absorption contrast in microfluidic live-cell imaging at a flow speed as high as ~10 m/s, corresponding to an imaging throughput of ~100,000 cells/sec. ATOM could thus be the enabling platform to meet the pressing need for intercalating optical microscopy in cellular assay, e.g. imaging flow cytometry – permitting high-throughput access to the morphological information of the individual cells simultaneously with a multitude of parameters obtained in the standard assay. PMID:24413677

Fluorescence microscopy for the characterization of structural integrity

Science.gov (United States)

Street, Kenneth W.; Leonhardt, Todd A.

1991-01-01

The absorption characteristics of light and the optical technique of fluorescence microscopy for enhancing metallographic interpretation are presented. Characterization of thermally sprayed coatings by optical microscopy suffers because of the tendency for misidentification of the microstructure produced by metallographic preparation. Gray scale, in bright field microscopy, is frequently the only means of differentiating the actual structural details of porosity, cracking, and debonding of coatings. Fluorescence microscopy is a technique that helps to distinguish the artifacts of metallographic preparation (pullout, cracking, debonding) from the microstructure of the specimen by color contrasting structural differences. Alternative instrumentation and the use of other dye systems are also discussed. The combination of epoxy vacuum infiltration with fluorescence microscopy to verify microstructural defects is an effective means to characterize advanced materials and to assess structural integrity.

Optical characterication of probes for photon scanning tunnelling microscopy

DEFF Research Database (Denmark)

Vohnsen, Brian; Bozhevolnyi, Sergey I.

1999-01-01

The photon scanning tunnelling microscope is a well-established member of the family of scanning near-field optical microscopes used for optical imaging at the sub-wavelength scale. The quality of the probes, typically pointed uncoated optical fibres, used is however difficult to evaluate...

Brain refractive index measured in vivo with high-NA defocus-corrected full-field OCT and consequences for two-photon microscopy.

Science.gov (United States)

Binding, Jonas; Ben Arous, Juliette; Léger, Jean-François; Gigan, Sylvain; Boccara, Claude; Bourdieu, Laurent

2011-03-14

Two-photon laser scanning microscopy (2PLSM) is an important tool for in vivo tissue imaging with sub-cellular resolution, but the penetration depth of current systems is potentially limited by sample-induced optical aberrations. To quantify these, we measured the refractive index n' in the somatosensory cortex of 7 rats in vivo using defocus optimization in full-field optical coherence tomography (ff-OCT). We found n' to be independent of imaging depth or rat age. From these measurements, we calculated that two-photon imaging beyond 200 µm into the cortex is limited by spherical aberration, indicating that adaptive optics will improve imaging depth.

Scanning Tunneling Optical Resonance Microscopy

Science.gov (United States)

Bailey, Sheila; Wilt, Dave; Raffaelle, Ryne; Gennett, Tom; Tin, Padetha; Lau, Janice; Castro, Stephanie; Jenkins, Philip; Scheiman, Dave

2003-01-01

Scanning tunneling optical resonance microscopy (STORM) is a method, now undergoing development, for measuring optoelectronic properties of materials and devices on the nanoscale by means of a combination of (1) traditional scanning tunneling microscopy (STM) with (2) tunable laser spectroscopy. In STORM, an STM tip probing a semiconductor is illuminated with modulated light at a wavelength in the visible-to-near-infrared range and the resulting photoenhancement of the tunneling current is measured as a function of the illuminating wavelength. The photoenhancement of tunneling current occurs when the laser photon energy is sufficient to excite charge carriers into the conduction band of the semiconductor. Figure 1 schematically depicts a proposed STORM apparatus. The light for illuminating the semiconductor specimen at the STM would be generated by a ring laser that would be tunable across the wavelength range of interest. The laser beam would be chopped by an achromatic liquid-crystal modulator. A polarization-maintaining optical fiber would couple the light to the tip/sample junction of a commercial STM. An STM can be operated in one of two modes: constant height or constant current. A STORM apparatus would be operated in the constant-current mode, in which the height of the tip relative to the specimen would be varied in order to keep the tunneling current constant. In this mode, a feedback control circuit adjusts the voltage applied to a piezoelectric actuator in the STM that adjusts the height of the STM tip to keep the tunneling current constant. The exponential relationship between the tunneling current and tip-to-sample distance makes it relatively easy to implement this mode of operation. The choice of method by which the photoenhanced portion of the tunneling current would be measured depends on choice of the frequency at which the input illumination would be modulated (chopped). If the frequency of modulation were low enough (typically tunneling current

A new microscope optics for laser dark-field illumination applied to high precision two dimensional measurement of specimen displacement.

Science.gov (United States)

Noda, Naoki; Kamimura, Shinji

2008-02-01

With conventional light microscopy, precision in the measurement of the displacement of a specimen depends on the signal-to-noise ratio when we measure the light intensity of magnified images. This implies that, for the improvement of precision, getting brighter images and reducing background light noise are both inevitably required. For this purpose, we developed a new optics for laser dark-field illumination. For the microscopy, we used a laser beam and a pair of axicons (conical lenses) to get an optimal condition for dark-field observations. The optics was applied to measuring two dimensional microbead displacements with subnanometer precision. The bandwidth of our detection system overall was 10 kHz. Over most of this bandwidth, the observed noise level was as small as 0.1 nm/radicalHz.

Multifocal multiphoton microscopy with adaptive optical correction

Science.gov (United States)

Coelho, Simao; Poland, Simon; Krstajic, Nikola; Li, David; Monypenny, James; Walker, Richard; Tyndall, David; Ng, Tony; Henderson, Robert; Ameer-Beg, Simon

2013-02-01

Fluorescence lifetime imaging microscopy (FLIM) is a well established approach for measuring dynamic signalling events inside living cells, including detection of protein-protein interactions. The improvement in optical penetration of infrared light compared with linear excitation due to Rayleigh scattering and low absorption have provided imaging depths of up to 1mm in brain tissue but significant image degradation occurs as samples distort (aberrate) the infrared excitation beam. Multiphoton time-correlated single photon counting (TCSPC) FLIM is a method for obtaining functional, high resolution images of biological structures. In order to achieve good statistical accuracy TCSPC typically requires long acquisition times. We report the development of a multifocal multiphoton microscope (MMM), titled MegaFLI. Beam parallelization performed via a 3D Gerchberg-Saxton (GS) algorithm using a Spatial Light Modulator (SLM), increases TCSPC count rate proportional to the number of beamlets produced. A weighted 3D GS algorithm is employed to improve homogeneity. An added benefit is the implementation of flexible and adaptive optical correction. Adaptive optics performed by means of Zernike polynomials are used to correct for system induced aberrations. Here we present results with significant improvement in throughput obtained using a novel complementary metal-oxide-semiconductor (CMOS) 1024 pixel single-photon avalanche diode (SPAD) array, opening the way to truly high-throughput FLIM.

Fluorescent dyes with large Stokes shifts for super-resolution optical microscopy of biological objects: a review

International Nuclear Information System (INIS)

Sednev, Maksim V; Belov, Vladimir N; Hell, Stefan W

2015-01-01

The review deals with commercially available organic dyes possessing large Stokes shifts and their applications as fluorescent labels in optical microscopy based on stimulated emission depletion (STED). STED microscopy breaks Abbe’s diffraction barrier and provides optical resolution beyond the diffraction limit. STED microscopy is non-invasive and requires photostable fluorescent markers attached to biomolecules or other objects of interest. Up to now, in most biology-related STED experiments, bright and photoresistant dyes with small Stokes shifts of 20–40 nm were used. The rapid progress in STED microscopy showed that organic fluorophores possessing large Stokes shifts are indispensable in multi-color super-resolution techniques. The ultimate result of the imaging relies on the optimal combination of a dye, the bio-conjugation procedure and the performance of the optical microscope. Modern bioconjugation methods, basics of STED microscopy, as well as structures and spectral properties of the presently available fluorescent markers are reviewed and discussed. In particular, the spectral properties of the commercial dyes are tabulated and correlated with the available depletion wavelengths found in STED microscopes produced by LEICA Microsytems, Abberior Instruments and Picoquant GmbH. (topical review)

Identification of nodal tissue in the living heart using rapid scanning fiber-optics confocal microscopy and extracellular fluorophores.

Science.gov (United States)

Huang, Chao; Kaza, Aditya K; Hitchcock, Robert W; Sachse, Frank B

2013-09-01

Risks associated with pediatric reconstructive heart surgery include injury of the sinoatrial node (SAN) and atrioventricular node (AVN), requiring cardiac rhythm management using implantable pacemakers. These injuries are the result of difficulties in identifying nodal tissues intraoperatively. Here we describe an approach based on confocal microscopy and extracellular fluorophores to quantify tissue microstructure and identify nodal tissue. Using conventional 3-dimensional confocal microscopy we investigated the microstructural arrangement of SAN, AVN, and atrial working myocardium (AWM) in fixed rat heart. AWM exhibited a regular striated arrangement of the extracellular space. In contrast, SAN and AVN had an irregular, reticulated arrangement. AWM, SAN, and AVN tissues were beneath a thin surface layer of tissue that did not obstruct confocal microscopic imaging. Subsequently, we imaged tissues in living rat hearts with real-time fiber-optics confocal microscopy. Fiber-optics confocal microscopy images resembled images acquired with conventional confocal microscopy. We investigated spatial regularity of tissue microstructure from Fourier analysis and second-order image moments. Fourier analysis of fiber-optics confocal microscopy images showed that the spatial regularity of AWM was greater than that of nodal tissues (37.5 ± 5.0% versus 24.3 ± 3.9% for SAN and 23.8 ± 3.7% for AVN; Pfiber-optics confocal microscopy. Application of the approach in pediatric reconstructive heart surgery may reduce risks of injuring nodal tissues.

Near field plasmon and force microscopy

OpenAIRE

de Hollander, R.B.G.; van Hulst, N.F.; Kooyman, R.P.H.

1995-01-01

A scanning plasmon near field optical microscope (SPNM) is presented which combines a conventional far field surface plasmon microscope with a stand-alone atomic force microscope (AFM). Near field plasmon and force images are recorded simultaneously both with a lateral resolution limited by the probe size to about 20 nm. At variance to previous work, utilizing a scanning tunneling microscope (STM) with a metallic tip, a dielectric silicon-nitride tip is used in contact mode. This arrangement ...

Simultaneous topographical, electrical and optical microscopy of optoelectronic devices at the nanoscale

KAUST Repository

Kumar, Naresh; Zoladek-Lemanczyk, Alina; Guilbert, Anne A. Y.; Su, Weitao; Tuladhar, Sachetan M.; Kirchartz, Thomas; Schroeder, Bob C.; McCulloch, Iain; Nelson, Jenny; Roy, Debdulal; Castro, Fernando A.

2017-01-01

resolution by combining plasmonic optical signal enhancement with electrical-mode scanning probe microscopy. We demonstrate that this combined approach offers subsurface sensitivity that can be exploited to provide molecular information with a nanoscale

Characterization of near-field optical probes

DEFF Research Database (Denmark)

Vohnsen, Brian; Bozhevolnyi, Sergey I.

1999-01-01

Radiation and collection characteristics of four different near-field optical-fiber probes, namely, three uncoated probes and an aluminium-coated small-aperture probe, are investigated and compared. Their radiation properties are characterized by observation of light-induced topography changes...... in a photo-sensitive film illuminated with the probes, and it is confirmed that the radiated optical field is unambigiously confined only for the coated probe. Near-field optical imaging of a standing evanescent-wave pattern is used to compare the detection characteristics of the probes, and it is concluded...... that, for the imaging of optical-field intensity distributions containing predominantly evanescent-wave components, a sharp uncoated tip is the probe of choice. Complementary results obtained with optical phase-conjugation experiments with he uncoated probes are discussed in relation to the probe...

Near Field Magneto-Optical Microscope

Science.gov (United States)

Vlasko-Vlasov, Vitalii K.; Welp, Ulrich; Crabtree, George W.

2005-12-06

A device and method for mapping magnetic fields of a sample at a resolution less than the wavelength of light without altering the magnetic field of the sample is disclosed. A device having a tapered end portion with a magneto-optically active particle positioned at the distal end thereof in communication with a fiber optic for transferring incoming linearly polarized light from a source thereof to the particle and for transferring reflected light from the particle is provided. The fiber optic has a reflective material trapping light within the fiber optic and in communication with a light detector for determining the polarization of light reflected from the particle as a function of the strength and direction of the magnetic field of the sample. Linearly polarized light from the source thereof transferred to the particle positioned proximate the sample is affected by the magnetic field of the sample sensed by the particle such that the difference in polarization of light entering and leaving the particle is due to the magnetic field of the sample. Relative movement between the particle and sample enables mapping.

Near-Field Magneto-Optical Microscope

Science.gov (United States)

Vlasko-Vlasov, Vitalii; Welp, Ulrich; and Crabtree, George W.

2005-12-06

A device and method for mapping magnetic fields of a sample at a resolution less than the wavelength of light without altering the magnetic field of the sample is disclosed. A device having a tapered end portion with a magneto-optically active particle positioned at the distal end thereof in communication with a fiber optic for transferring incoming linearly polarized light from a source thereof to the particle and for transferring reflected light from the particle is provided. The fiber optic has a reflective material trapping light within the fiber optic and in communication with a light detector for determining the polarization of light reflected from the particle as a function of the strength and direction of the magnetic field of the sample. Linearly polarized light from the source thereof transferred to the particle positioned proximate the sample is affected by the magnetic field of the sample sensed by the particle such that the difference in polarization of light entering and leaving the particle is due to the magnetic field of the sample. Relative movement between the particle and sample enables mapping.

Tunable thin-film optical filters for hyperspectral microscopy

Science.gov (United States)

Favreau, Peter F.; Rich, Thomas C.; Prabhat, Prashant; Leavesley, Silas J.

2013-02-01

Hyperspectral imaging was originally developed for use in remote sensing applications. More recently, it has been applied to biological imaging systems, such as fluorescence microscopes. The ability to distinguish molecules based on spectral differences has been especially advantageous for identifying fluorophores in highly autofluorescent tissues. A key component of hyperspectral imaging systems is wavelength filtering. Each filtering technology used for hyperspectral imaging has corresponding advantages and disadvantages. Recently, a new optical filtering technology has been developed that uses multi-layered thin-film optical filters that can be rotated, with respect to incident light, to control the center wavelength of the pass-band. Compared to the majority of tunable filter technologies, these filters have superior optical performance including greater than 90% transmission, steep spectral edges and high out-of-band blocking. Hence, tunable thin-film optical filters present optical characteristics that may make them well-suited for many biological spectral imaging applications. An array of tunable thin-film filters was implemented on an inverted fluorescence microscope (TE 2000, Nikon Instruments) to cover the full visible wavelength range. Images of a previously published model, GFP-expressing endothelial cells in the lung, were acquired using a charge-coupled device camera (Rolera EM-C2, Q-Imaging). This model sample presents fluorescently-labeled cells in a highly autofluorescent environment. Linear unmixing of hyperspectral images indicates that thin-film tunable filters provide equivalent spectral discrimination to our previous acousto-optic tunable filter-based approach, with increased signal-to-noise characteristics. Hence, tunable multi-layered thin film optical filters may provide greatly improved spectral filtering characteristics and therefore enable wider acceptance of hyperspectral widefield microscopy.

Structured light optical microscopy for three-dimensional reconstruction of technical surfaces

Science.gov (United States)

Kettel, Johannes; Reinecke, Holger; Müller, Claas

2016-04-01

In microsystems technology quality control of micro structured surfaces with different surface properties is playing an ever more important role. The process of quality control incorporates three-dimensional (3D) reconstruction of specularand diffusive reflecting technical surfaces. Due to the demand on high measurement accuracy and data acquisition rates, structured light optical microscopy has become a valuable solution to solve this problem providing high vertical and lateral resolution. However, 3D reconstruction of specular reflecting technical surfaces still remains a challenge to optical measurement principles. In this paper we present a measurement principle based on structured light optical microscopy which enables 3D reconstruction of specular- and diffusive reflecting technical surfaces. It is realized using two light paths of a stereo microscope equipped with different magnification levels. The right optical path of the stereo microscope is used to project structured light onto the object surface. The left optical path is used to capture the structured illuminated object surface with a camera. Structured light patterns are generated by a Digital Light Processing (DLP) device in combination with a high power Light Emitting Diode (LED). Structured light patterns are realized as a matrix of discrete light spots to illuminate defined areas on the object surface. The introduced measurement principle is based on multiple and parallel processed point measurements. Analysis of the measured Point Spread Function (PSF) by pattern recognition and model fitting algorithms enables the precise calculation of 3D coordinates. Using exemplary technical surfaces we demonstrate the successful application of our measurement principle.

Multiscale 3D characterization with dark-field x-ray microscopy

DEFF Research Database (Denmark)

Simons, Hugh; Jakobsen, Anders Clemen; Ahl, Sonja Rosenlund

2016-01-01

Dark-field x-ray microscopy is a new way to three-dimensionally map lattice strain and orientation in crystalline matter. It is analogous to dark-field electron microscopy in that an objective lens magnifies diffracting features of the sample; however, the use of high-energy synchrotron x-rays me......, multiscale phenomena in situ is a key step toward formulating and validating multiscale models that account for the entire heterogeneity of materials....

U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

Energy Technology Data Exchange (ETDEWEB)

Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

2016-10-01

The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

Energy Technology Data Exchange (ETDEWEB)

Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

2016-03-30

The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

Live cell refractometry using Hilbert phase microscopy and confocal reflectance microscopy.

Science.gov (United States)

Lue, Niyom; Choi, Wonshik; Popescu, Gabriel; Yaqoob, Zahid; Badizadegan, Kamran; Dasari, Ramachandra R; Feld, Michael S

2009-11-26

Quantitative chemical analysis has served as a useful tool for understanding cellular metabolisms in biology. Among many physical properties used in chemical analysis, refractive index in particular has provided molecular concentration that is an important indicator for biological activities. In this report, we present a method of extracting full-field refractive index maps of live cells in their native states. We first record full-field optical thickness maps of living cells by Hilbert phase microscopy and then acquire physical thickness maps of the same cells using a custom-built confocal reflectance microscope. Full-field and axially averaged refractive index maps are acquired from the ratio of optical thickness to physical thickness. The accuracy of the axially averaged index measurement is 0.002. This approach can provide novel biological assays of label-free living cells in situ.

Discontinuous precipitation in a Ni-In alloy studied by analytical field ion microscopy

International Nuclear Information System (INIS)

Geber, G.P.; Kirchheim, R.

1997-01-01

Discontinuous precipitation (DP) in a Ni-7.5 at.% In alloy has been studied by means of atom probe field ion microscopy (APFIM). The morphology of the lamellar microstructure and the growth kinetics of the decomposition reaction have been investigated by optical and scanning electron microscopy (LM, SEM). A back polishing method has been developed to prepare APFIM specimens in a systematic manner for transmission electron microscopy (TEM), in order to obtain a definite inclination of different interfaces in the tip apex for subsequent analysis of their chemical composition. APFIM results of the Ni- and In-concentration across the interfaces are presented. Based on the APFIM results it is possible to distinguish between the theoretical description of the concentration profiles given by the models of Cahn and Hillert. It is shown by both FIM images and concentration depth profiles that the α/β-interface has a thickness of about 0.5 nm. In addition it is shown for the first time that the concentration across a β-lamella is not constant. This experimental finding was used to develop a generalization of Cahn's model on discontinuous precipitation including the solute flux within the interface between β and the parent phase α 0

Comprehensive study of unexpected microscope condensers formed in sample arrangements commonly used in optical microscopy.

Science.gov (United States)

Desai, Darshan B; Aldawsari, Mabkhoot Mudith S; Alharbi, Bandar Mohammed H; Sen, Sanchari; Grave de Peralta, Luis

2015-09-01

We show that various setups for optical microscopy which are commonly used in biomedical laboratories behave like efficient microscope condensers that are responsible for observed subwavelength resolution. We present a series of experiments and simulations that reveal how inclined illumination from such unexpected condensers occurs when the sample is perpendicularly illuminated by a microscope's built-in white-light source. In addition, we demonstrate an inexpensive add-on optical module that serves as an efficient and lightweight microscope condenser. Using such add-on optical module in combination with a low-numerical-aperture objective lens and Fourier plane imaging microscopy technique, we demonstrate detection of photonic crystals with a period nearly eight times smaller than the Rayleigh resolution limit.

Photoionization microscopy of Rydberg hydrogen atom in a non-uniform electrical field

International Nuclear Information System (INIS)

Cheng Shao-Hao; Wang De-Hua; Chen Zhao-Hang; Chen Qiang

2016-01-01

In this paper, we investigate the photoionization microscopy of the Rydberg hydrogen atom in a gradient electric field for the first time. The observed oscillatory patterns in the photoionization microscopy are explained within the framework of the semiclassical theory, which can be considered as a manifestation of interference between various electron trajectories arriving at a given point on the detector plane. In contrast with the photoionization microscopy in the uniform electric field, the trajectories of the ionized electron in the gradient electric field will become chaotic. An infinite set of different electron trajectories can arrive at a given point on the detector plane, which makes the interference pattern of the electron probability density distribution extremely complicated. Our calculation results suggest that the oscillatory pattern in the electron probability density distribution depends sensitively on the electric field gradient, the scaled energy and the position of the detector plane. Through our research, we predict that the interference pattern in the electron probability density distribution can be observed in an actual photoionization microscopy experiment once the external electric field strength and the position of the electron detector plane are reasonable. This study provides some references for the future experimental research on the photoionization microscopy of the Rydberg atom in the non-uniform external fields. (paper)

Automated seeding-based nuclei segmentation in nonlinear optical microscopy.

Science.gov (United States)

Medyukhina, Anna; Meyer, Tobias; Heuke, Sandro; Vogler, Nadine; Dietzek, Benjamin; Popp, Jürgen

2013-10-01

Nonlinear optical (NLO) microscopy based, e.g., on coherent anti-Stokes Raman scattering (CARS) or two-photon-excited fluorescence (TPEF) is a fast label-free imaging technique, with a great potential for biomedical applications. However, NLO microscopy as a diagnostic tool is still in its infancy; there is a lack of robust and durable nuclei segmentation methods capable of accurate image processing in cases of variable image contrast, nuclear density, and type of investigated tissue. Nonetheless, such algorithms specifically adapted to NLO microscopy present one prerequisite for the technology to be routinely used, e.g., in pathology or intraoperatively for surgical guidance. In this paper, we compare the applicability of different seeding and boundary detection methods to NLO microscopic images in order to develop an optimal seeding-based approach capable of accurate segmentation of both TPEF and CARS images. Among different methods, the Laplacian of Gaussian filter showed the best accuracy for the seeding of the image, while a modified seeded watershed segmentation was the most accurate in the task of boundary detection. The resulting combination of these methods followed by the verification of the detected nuclei performs high average sensitivity and specificity when applied to various types of NLO microscopy images.

Adaptive optics in spinning disk microscopy: improved contrast and brightness by a simple and fast method.

Science.gov (United States)

Fraisier, V; Clouvel, G; Jasaitis, A; Dimitrov, A; Piolot, T; Salamero, J

2015-09-01

Multiconfocal microscopy gives a good compromise between fast imaging and reasonable resolution. However, the low intensity of live fluorescent emitters is a major limitation to this technique. Aberrations induced by the optical setup, especially the mismatch of the refractive index and the biological sample itself, distort the point spread function and further reduce the amount of detected photons. Altogether, this leads to impaired image quality, preventing accurate analysis of molecular processes in biological samples and imaging deep in the sample. The amount of detected fluorescence can be improved with adaptive optics. Here, we used a compact adaptive optics module (adaptive optics box for sectioning optical microscopy), which was specifically designed for spinning disk confocal microscopy. The module overcomes undesired anomalies by correcting for most of the aberrations in confocal imaging. Existing aberration detection methods require prior illumination, which bleaches the sample. To avoid multiple exposures of the sample, we established an experimental model describing the depth dependence of major aberrations. This model allows us to correct for those aberrations when performing a z-stack, gradually increasing the amplitude of the correction with depth. It does not require illumination of the sample for aberration detection, thus minimizing photobleaching and phototoxicity. With this model, we improved both signal-to-background ratio and image contrast. Here, we present comparative studies on a variety of biological samples. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Super-resolution optical microscopy for studying membrane structure and dynamics.

Science.gov (United States)

Sezgin, Erdinc

2017-07-12

Investigation of cell membrane structure and dynamics requires high spatial and temporal resolution. The spatial resolution of conventional light microscopy is limited due to the diffraction of light. However, recent developments in microscopy enabled us to access the nano-scale regime spatially, thus to elucidate the nanoscopic structures in the cellular membranes. In this review, we will explain the resolution limit, address the working principles of the most commonly used super-resolution microscopy techniques and summarise their recent applications in the biomembrane field.

Fabrication and characterization of a nanometer-sized optical fiber electrode based on selective chemical etching for scanning electrochemical/optical microscopy.

Science.gov (United States)

Maruyama, Kenichi; Ohkawa, Hiroyuki; Ogawa, Sho; Ueda, Akio; Niwa, Osamu; Suzuki, Koji

2006-03-15

We have already reported a method for fabricating ultramicroelectrodes (Suzuki, K. JP Patent, 2004-45394, 2004). This method is based on the selective chemical etching of optical fibers. In this work, we undertake a detailed investigation involving a combination of etched optical fibers with various types of tapered tip (protruding-shape, double- (or pencil-) shape and triple-tapered electrode) and insulation with electrophoretic paint. Our goal is to establish a method for fabricating nanometer-sized optical fiber electrodes with high reproducibility. As a result, we realized pencil-shaped and triple-tapered electrodes that had radii in the nanometer range with high reproducibility. These nanometer-sized electrodes showed well-defined sigmoidal curves and stable diffusion-limited responses with cyclic voltammetry. The pencil-shaped optical fiber, which has a conical tip with a cone angle of 20 degrees , was effective for controlling the electrode radius. The pencil-shaped electrodes had higher reproducibility and smaller electrode radii (r(app) etched optical fiber electrodes. By using a pencil-shaped electrode with a 105-nm radius as a probe, we obtained simultaneous electrochemical and optical images of an implantable interdigitated array electrode. We achieved nanometer-scale resolution with a combination of scanning electrochemical microscopy SECM and optical microscopy. The resolution of the electrochemical and optical images indicated sizes of 300 and 930 nm, respectively. The neurites of living PC12 cells were also successfully imaged on a 1.6-microm scale by using the negative feedback mode of an SECM.

Field programmable gate array based reconfigurable scanning probe/optical microscope.

Science.gov (United States)

Nowak, Derek B; Lawrence, A J; Dzegede, Zechariah K; Hiester, Justin C; Kim, Cliff; Sánchez, Erik J

2011-10-01

The increasing popularity of nanometrology and nanospectroscopy has pushed researchers to develop complex new analytical systems. This paper describes the development of a platform on which to build a microscopy tool that will allow for flexibility of customization to suit research needs. The novelty of the described system lies in its versatility of capabilities. So far, one version of this microscope has allowed for successful near-field and far-field fluorescence imaging with single molecule detection sensitivity. This system is easily adapted for reflection, polarization (Kerr magneto-optical (MO)), Raman, super-resolution techniques, and other novel scanning probe imaging and spectroscopic designs. While collecting a variety of forms of optical images, the system can simultaneously monitor topographic information of a sample with an integrated tuning fork based shear force system. The instrument has the ability to image at room temperature and atmospheric pressure or under liquid. The core of the design is a field programmable gate array (FPGA) data acquisition card and a single, low cost computer to control the microscope with analog control circuitry using off-the-shelf available components. A detailed description of electronics, mechanical requirements, and software algorithms as well as examples of some different forms of the microscope developed so far are discussed.

Full-field parallel interferometry coherence probe microscope for high-speed optical metrology.

Science.gov (United States)

Safrani, A; Abdulhalim, I

2015-06-01

Parallel detection of several achromatic phase-shifted images is used to obtain a high-speed, high-resolution, full-field, optical coherence probe tomography system based on polarization interferometry. The high enface imaging speed, short coherence gate, and high lateral resolution provided by the system are exploited to determine microbump height uniformity in an integrated semiconductor chip at 50 frames per second. The technique is demonstrated using the Linnik microscope, although it can be implemented on any polarization-based interference microscopy system.

Design of angle-resolved illumination optics using nonimaging bi-telecentricity for 193 nm scatterfield microscopy.

Science.gov (United States)

Sohn, Martin Y; Barnes, Bryan M; Silver, Richard M

2018-03-01

Accurate optics-based dimensional measurements of features sized well-below the diffraction limit require a thorough understanding of the illumination within the optical column and of the three-dimensional scattered fields that contain the information required for quantitative metrology. Scatterfield microscopy can pair simulations with angle-resolved tool characterization to improve agreement between the experiment and calculated libraries, yielding sub-nanometer parametric uncertainties. Optimized angle-resolved illumination requires bi-telecentric optics in which a telecentric sample plane defined by a Köhler illumination configuration and a telecentric conjugate back focal plane (CBFP) of the objective lens; scanning an aperture or an aperture source at the CBFP allows control of the illumination beam angle at the sample plane with minimal distortion. A bi-telecentric illumination optics have been designed enabling angle-resolved illumination for both aperture and source scanning modes while yielding low distortion and chief ray parallelism. The optimized design features a maximum chief ray angle at the CBFP of 0.002° and maximum wavefront deviations of less than 0.06 λ for angle-resolved illumination beams at the sample plane, holding promise for high quality angle-resolved illumination for improved measurements of deep-subwavelength structures using deep-ultraviolet light.

New fluorinated rhodamines for optical microscopy and nanoscopy.

Science.gov (United States)

Mitronova, Gyuzel Yu; Belov, Vladimir N; Bossi, Mariano L; Wurm, Christian A; Meyer, Lars; Medda, Rebecca; Moneron, Gael; Bretschneider, Stefan; Eggeling, Christian; Jakobs, Stefan; Hell, Stefan W

2010-04-19

New photostable rhodamine dyes represented by the compounds 1 a-r and 3-5 are proposed as efficient fluorescent markers with unique combination of structural features. Unlike rhodamines with monoalkylated nitrogen atoms, N',N-bis(2,2,2-trifluoroethyl) derivatives 1 e, 1 i, 1 j, 3-H and 5 were found to undergo sulfonation of the xanthene fragment at the positions 4' and 5'. Two fluorine atoms were introduced into the positions 2' and 7' of the 3',6'-diaminoxanthene fragment in compounds 1 a-d, 1 i-l and 1 m-r. The new rhodamine dyes may be excited with λ=488 or 514 nm light; most of them emit light at λ=512-554 nm (compounds 1 q and 1r at λ=576 and 589 nm in methanol, respectively) and have high fluorescence quantum yields in solution (up to 98 %), relatively long excited-state lifetimes (>3 ns) and are resistant against photobleaching, especially at high laser intensities, as is usually applied in confocal microscopy. Sulfonation of the xanthene fragment with 30 % SO3 in H2SO4 is compatible with the secondary amide bond (rhodamine-CON(Me)CH2CH2COOH) formed with MeNHCH2CH2COOCH3 to providing the sterically unhindered carboxylic group required for further (bio)conjugation reactions. After creating the amino reactive sites, the modified derivatives may be used as fluorescent markers and labels for (bio)molecules in optical microscopy and nanoscopy with very-high light intensities. Further, the new rhodamine dyes are able to pass the plasma membrane of living cells, introducing them as potential labels for recent live-cell-tag approaches. We exemplify the excellent performance of the fluorinated rhodamines in optical microscopy by fluorescence correlation spectroscopy (FCS) and stimulated emission depletion (STED) nanoscopy experiments. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Workshop on the coupling of synchrotron radiation IR and X-rays with tip based scanning probe microscopies X-TIP

Energy Technology Data Exchange (ETDEWEB)

Comin, F.; Martinez-Criado, G.; Mundboth, K.; Susini, J. [European Synchrotron Radiation Facility (ESRF), 38 - Grenoble (France); Purans, J.; Sammelselg, V. [Tartu Univ. (Estonia); Chevrier, J.; Huant, S. [Universite Joseph-Fourier, Grenoble I, LEPES, 38 (France); Hamilton, B. [School of Electrical Engineering and Electronics, Manchester (United Kingdom); Saito, A. [Osaka Univ., RIKEN/SPring8 (Japan); Dhez, O. [OGG, INFM/CNR, 38 - Grenoble (France); Brocklesby, W.S. [Southampton Univ., Optoelectronics Research Centre (United Kingdom); Alvarez-Prado, L.M. [Ovieado, Dept. de Fisica (Spain); Kuzmin, A. [Institute of Solid State Physics - Riga (Latvia); Pailharey, D. [CRMC-N - CNRS, 13 - Marseille (France); Tonneau, D. [CRMCN - Faculte des sciences de Luminy, 13 - Marseille (France); Chretien, P. [Laboratoire de Genie Electrique de Paris, 75 - Paris (France); Cricenti, A. [ISM-CNR, Rome (Italy); DeWilde, Y. [ESPCI, 75 - Paris (France)

2005-07-01

The coupling of scanning probe microscopy (SPM) with synchrotron radiation is attracting increasing attention from nano-science community. By combining these 2 tools one can visualize, for example, the sample nano-structure prior to any X-ray characterization. Coupled with focusing devices or independently, SPM can provide spatial resolution below the optical limits. Furthermore, the possibility of employing SPM to manipulate nano-objects under X-ray beams is another exciting perspective. This document gathers the transparencies of 6 of the presentations made at the workshop: 1) the combination of atomic force microscopy and X-ray beam - experimental set-up and objectives; 2) the combination of scanning probe microscope and X-rays for detection of electrons; 3) towards soft X-ray scanning microscopy using tapered capillaries and laser-based high harmonic sources; 4) near-field magneto-optical microscopy; 5) near-field scanning optical microscopy - a brief overview -; and 6) from aperture-less near-field optical microscopy to infra-red near-field night vision. 4 posters entitled: 1) development of laboratory setup for X-ray/AFM experiments, 2) towards X-ray diffraction on single islands, 3) nano-XEOL using near-field detection, and 4) local collection with a STM tip of photoelectrons emitted by a surface irradiated by visible of UV laser beam, are included in the document.

Workshop on the coupling of synchrotron radiation IR and X-rays with tip based scanning probe microscopies X-TIP

International Nuclear Information System (INIS)

Comin, F.; Martinez-Criado, G.; Mundboth, K.; Susini, J.; Purans, J.; Sammelselg, V.; Chevrier, J.; Huant, S.; Hamilton, B.; Saito, A.; Dhez, O.; Brocklesby, W.S.; Alvarez-Prado, L.M.; Kuzmin, A.; Pailharey, D.; Tonneau, D.; Chretien, P.; Cricenti, A.; DeWilde, Y.

2005-01-01

The coupling of scanning probe microscopy (SPM) with synchrotron radiation is attracting increasing attention from nano-science community. By combining these 2 tools one can visualize, for example, the sample nano-structure prior to any X-ray characterization. Coupled with focusing devices or independently, SPM can provide spatial resolution below the optical limits. Furthermore, the possibility of employing SPM to manipulate nano-objects under X-ray beams is another exciting perspective. This document gathers the transparencies of 6 of the presentations made at the workshop: 1) the combination of atomic force microscopy and X-ray beam - experimental set-up and objectives; 2) the combination of scanning probe microscope and X-rays for detection of electrons; 3) towards soft X-ray scanning microscopy using tapered capillaries and laser-based high harmonic sources; 4) near-field magneto-optical microscopy; 5) near-field scanning optical microscopy - a brief overview -; and 6) from aperture-less near-field optical microscopy to infra-red near-field night vision. 4 posters entitled: 1) development of laboratory setup for X-ray/AFM experiments, 2) towards X-ray diffraction on single islands, 3) nano-XEOL using near-field detection, and 4) local collection with a STM tip of photoelectrons emitted by a surface irradiated by visible of UV laser beam, are included in the document

Polarized Light Microscopy

Science.gov (United States)

Frandsen, Athela F.

2016-01-01

Polarized light microscopy (PLM) is a technique which employs the use of polarizing filters to obtain substantial optical property information about the material which is being observed. This information can be combined with other microscopy techniques to confirm or elucidate the identity of an unknown material, determine whether a particular contaminant is present (as with asbestos analysis), or to provide important information that can be used to refine a manufacturing or chemical process. PLM was the major microscopy technique in use for identification of materials for nearly a century since its introduction in 1834 by William Fox Talbot, as other techniques such as SEM (Scanning Electron Microscopy), FTIR (Fourier Transform Infrared spectroscopy), XPD (X-ray Powder Diffraction), and TEM (Transmission Electron Microscopy) had not yet been developed. Today, it is still the only technique approved by the Environmental Protection Agency (EPA) for asbestos analysis, and is often the technique first applied for identification of unknown materials. PLM uses different configurations in order to determine different material properties. With each configuration additional clues can be gathered, leading to a conclusion of material identity. With no polarizing filter, the microscope can be used just as a stereo optical microscope, and view qualities such as morphology, size, and number of phases. With a single polarizing filter (single polars), additional properties can be established, such as pleochroism, individual refractive indices, and dispersion staining. With two polarizing filters (crossed polars), even more can be deduced: isotropy vs. anisotropy, extinction angle, birefringence/degree of birefringence, sign of elongation, and anomalous polarization colors, among others. With the use of PLM many of these properties can be determined in a matter of seconds, even for those who are not highly trained. McCrone, a leader in the field of polarized light microscopy, often

Quantitative readout of optically encoded gold nanorods using an ordinary dark-field microscope.

Science.gov (United States)

Mercatelli, Raffaella; Ratto, Fulvio; Centi, Sonia; Soria, Silvia; Romano, Giovanni; Matteini, Paolo; Quercioli, Franco; Pini, Roberto; Fusi, Franco

2013-10-21

In this paper we report on a new use for dark-field microscopy in order to retrieve two-dimensional maps of optical parameters of a thin sample such as a cryptograph, a histological section, or a cell monolayer. In particular, we discuss the construction of quantitative charts of light absorbance and scattering coefficients of a polyvinyl alcohol film that was embedded with gold nanorods and then etched using a focused mode-locked Ti:Sapphire oscillator. Individual pulses from this laser excite plasmonic oscillations of the gold nanorods, thus triggering plastic deformations of the particles and their environment, which are confined within a few hundred nm of the light focus. In turn, these deformations modify the light absorbance and scattering landscape, which can be measured with optical resolution in a dark-field microscope equipped with an objective of tuneable numerical aperture. This technique may prove to be valuable for various applications, such as the fast readout of optically encoded data or to model functional interactions between light and biological tissue at the level of cellular organelles, including the photothermolysis of cancer.

A robotized six degree of freedom stage for optical microscopy

Science.gov (United States)

Avramov, M. Z.; Ivanov, I.; Pavlov, V.; Zaharieva, K.

2013-04-01

This work represents an investigation of the possibility to use a hexapod system for optical microscopy investigation and measurements. An appropriate hexapod stage has been developed. The stage has been calibrated and used for several different optical microscopy applications. The construction of the stage is based on the classic Stewart platform and thus represents a parallel robot with 6 degree of freedom. Appropriate software is controlling the transformation of the 3 position coordinates of the moving plate and the 3 Euler angles in position velocities and accelerations of the plate motion. An embedded microcontroller is implementing the motion plan and the PID controller regulating the kinematics. By difference to the available in the market hexapods the proposed solution is with lower precision but is significantly cheaper and simple to maintain. The repeatability obtained with current implementation is 0,05 mm and 0,001 rad. A specialized DSP based video processing engine is used for both feedback computation and application specific image processing in real-time. To verify the concept some applications has been developed for specific tasks and has been used for specific measurements.

An integrated instrumental setup for the combination of atomic force microscopy with optical spectroscopy.

Science.gov (United States)

Owen, R J; Heyes, C D; Knebel, D; Röcker, C; Nienhaus, G U

2006-07-01

In recent years, the study of single biomolecules using fluorescence microscopy and atomic force microscopy (AFM) techniques has resulted in a plethora of new information regarding the physics underlying these complex biological systems. It is especially advantageous to be able to measure the optical, topographical, and mechanical properties of single molecules simultaneously. Here an AFM is used that is especially designed for integration with an inverted optical microscope and that has a near-infrared light source (850 nm) to eliminate interference between the optical experiment and the AFM operation. The Tip Assisted Optics (TAO) system consists of an additional 100 x 100-microm(2) X-Y scanner for the sample, which can be independently and simultaneously used with the AFM scanner. This allows the offset to be removed between the confocal optical image obtained with the sample scanner and the simultaneously acquired AFM topography image. The tip can be positioned exactly into the optical focus while the user can still navigate within the AFM image for imaging or manipulation of the sample. Thus the tip-enhancement effect can be maximized and it becomes possible to perform single molecule manipulation experiments within the focus of a confocal optical image. Here this is applied to simultaneous measurement of single quantum dot fluorescence and topography with high spatial resolution. (c) 2006 Wiley Periodicals, Inc.

Laser Light-field Fusion for Wide-field Lensfree On-chip Phase Contrast Microscopy of Nanoparticles

Science.gov (United States)

Kazemzadeh, Farnoud; Wong, Alexander

2016-12-01

Wide-field lensfree on-chip microscopy, which leverages holography principles to capture interferometric light-field encodings without lenses, is an emerging imaging modality with widespread interest given the large field-of-view compared to lens-based techniques. In this study, we introduce the idea of laser light-field fusion for lensfree on-chip phase contrast microscopy for detecting nanoparticles, where interferometric laser light-field encodings acquired using a lensfree, on-chip setup with laser pulsations at different wavelengths are fused to produce marker-free phase contrast images of particles at the nanometer scale. As a proof of concept, we demonstrate, for the first time, a wide-field lensfree on-chip instrument successfully detecting 300 nm particles across a large field-of-view of ~30 mm2 without any specialized or intricate sample preparation, or the use of synthetic aperture- or shift-based techniques.

Cytology 3D structure formation based on optical microscopy images

Science.gov (United States)

Pronichev, A. N.; Polyakov, E. V.; Shabalova, I. P.; Djangirova, T. V.; Zaitsev, S. M.

2017-01-01

The article the article is devoted to optimization of the parameters of imaging of biological preparations in optical microscopy using a multispectral camera in visible range of electromagnetic radiation. A model for the image forming of virtual preparations was proposed. The optimum number of layers was determined for the object scan in depth and holistic perception of its switching according to the results of the experiment.

Cytology 3D structure formation based on optical microscopy images

International Nuclear Information System (INIS)

Pronichev, A N; Polyakov, E V; Zaitsev, S M; Shabalova, I P; Djangirova, T V

2017-01-01

The article the article is devoted to optimization of the parameters of imaging of biological preparations in optical microscopy using a multispectral camera in visible range of electromagnetic radiation. A model for the image forming of virtual preparations was proposed. The optimum number of layers was determined for the object scan in depth and holistic perception of its switching according to the results of the experiment. (paper)

Multifocus microscopy with precise color multi-phase diffractive optics applied in functional neuronal imaging.

Science.gov (United States)

Abrahamsson, Sara; Ilic, Rob; Wisniewski, Jan; Mehl, Brian; Yu, Liya; Chen, Lei; Davanco, Marcelo; Oudjedi, Laura; Fiche, Jean-Bernard; Hajj, Bassam; Jin, Xin; Pulupa, Joan; Cho, Christine; Mir, Mustafa; El Beheiry, Mohamed; Darzacq, Xavier; Nollmann, Marcelo; Dahan, Maxime; Wu, Carl; Lionnet, Timothée; Liddle, J Alexander; Bargmann, Cornelia I

2016-03-01

Multifocus microscopy (MFM) allows high-resolution instantaneous three-dimensional (3D) imaging and has been applied to study biological specimens ranging from single molecules inside cells nuclei to entire embryos. We here describe pattern designs and nanofabrication methods for diffractive optics that optimize the light-efficiency of the central optical component of MFM: the diffractive multifocus grating (MFG). We also implement a "precise color" MFM layout with MFGs tailored to individual fluorophores in separate optical arms. The reported advancements enable faster and brighter volumetric time-lapse imaging of biological samples. In live microscopy applications, photon budget is a critical parameter and light-efficiency must be optimized to obtain the fastest possible frame rate while minimizing photodamage. We provide comprehensive descriptions and code for designing diffractive optical devices, and a detailed methods description for nanofabrication of devices. Theoretical efficiencies of reported designs is ≈90% and we have obtained efficiencies of > 80% in MFGs of our own manufacture. We demonstrate the performance of a multi-phase MFG in 3D functional neuronal imaging in living C. elegans.

Optically Sectioned Imaging of Microvasculature of In-Vivo and Ex-Vivo Thick Tissue Models with Speckle-illumination HiLo Microscopy and HiLo Image Processing Implementation in MATLAB Architecture

Science.gov (United States)

Suen, Ricky Wai

The work described in this thesis covers the conversion of HiLo image processing into MATLAB architecture and the use of speckle-illumination HiLo microscopy for use of ex-vivo and in-vivo imaging of thick tissue models. HiLo microscopy is a wide-field fluorescence imaging technique and has been demonstrated to produce optically sectioned images comparable to confocal in thin samples. The imaging technique was developed by Jerome Mertz and the Boston University Biomicroscopy Lab and has been implemented in our lab as a stand-alone optical setup and a modification to a conventional fluorescence microscope. Speckle-illumination HiLo microscopy combines two images taken under speckle-illumination and standard uniform-illumination to generate an optically sectioned image that reject out-of-focus fluorescence. The evaluated speckle contrast in the images is used as a weighting function where elements that move out-of-focus have a speckle contrast that decays to zero. The experiments shown here demonstrate the capability of our HiLo microscopes to produce optically-sectioned images of the microvasculature of ex-vivo and in-vivo thick tissue models. The HiLo microscope were used to image the microvasculature of ex-vivo mouse heart sections prepared for optical histology and the microvasculature of in-vivo rodent dorsal window chamber models. Studies in label-free surface profiling with HiLo microscopy is also presented.

Laser terahertz emission microscopy with near-field probes

DEFF Research Database (Denmark)

Pedersen, Pernille Klarskov; Mittleman, Daniel M.

2016-01-01

Using an AFM, an optical near-field image at 800 nm of a dipole antenna for THz emission is measured, and by simultaneously collecting the emitted THz radiation, the laser light confined under the AFM probe gives a THz emission resolution of less than 50 nm.......Using an AFM, an optical near-field image at 800 nm of a dipole antenna for THz emission is measured, and by simultaneously collecting the emitted THz radiation, the laser light confined under the AFM probe gives a THz emission resolution of less than 50 nm....

Nonlinear optical spectroscopy and microscopy of model random and biological media

Science.gov (United States)

Guo, Yici

Nonlinear optical (NLO) spectroscopy and microscopy applied to biomedical science are emerging as new and rapidly growing areas which offer important insight into basic phenomena. Ultrafast NLO processes provide temporal, spectral and spatial sensitivities complementary or superior to those achieved through conventional linear optical approaches. The goal of this thesis is to explore the potential of two fundamental NLO processes to produce noninvasive histological maps of biological tissues. Within the goal of the thesis, steady state intensity, polarization and angular measurements of second- and third-harmonic generations (SHG, THG) have been performed on model random scattering and animal tissue samples. The nonlinear optical effects have been evaluated using models. Conversion efficiencies of SHG and THG from animal tissue interfaces have been determined, ranging from 10-7 to 10-10. The changes in the multiharmonic signals were found to depend on both local and overall histological structures of biological samples. The spectral signatures of two photon excitation induced fluorescence from intrinsic fluorophores have been acquired and used to characterize the physical state and types of tissues. Two dimensional scanning SHG and TPF tomographic images have been obtained from in vitro animal tissues, normal and diseased human breast tissues, and resolved subsurface layers and histo-chemical distributions. By combining consecutive 2D maps, a 3D image can be produced. The structure and morphology dependence of the SH signal has been utilized to image and evaluate subsurface tumor progression depth. Second harmonic microscopy in model random and biological cells has been studied using a CCD camera to obtain direct images from subcellular structures. Finally, near infrared (NIR) NLO spectroscopy and microscopy based on SHG and TPF have demonstrated high spatial resolution, deeper penetration depth, low level photo-damaging and enhanced morphological sensitivity for

Atom probe field ion microscopy and related topics: A bibliography 1993

Energy Technology Data Exchange (ETDEWEB)

Godfrey, R.D.; Miller, M.K.; Russell, K.F.

1994-10-01

This bibliography, covering the period 1993, includes references related to the following topics: atom probe field ion microscopy (APFIM), field emission (FE), and field ion microscopy (FIM). Technique-oriented studies and applications are included. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications. To reduce the length of this document, the references have been reduced to the minimum necessary to locate the articles. The references are listed alphabetically by authors, an Addendum of references missed in previous bibliographies is included.

Atom probe field ion microscopy and related topics: A bibliography 1993

International Nuclear Information System (INIS)

Godfrey, R.D.; Miller, M.K.; Russell, K.F.

1994-10-01

This bibliography, covering the period 1993, includes references related to the following topics: atom probe field ion microscopy (APFIM), field emission (FE), and field ion microscopy (FIM). Technique-oriented studies and applications are included. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications. To reduce the length of this document, the references have been reduced to the minimum necessary to locate the articles. The references are listed alphabetically by authors, an Addendum of references missed in previous bibliographies is included

Photoinduced force microscopy: A technique for hyperspectral nanochemical mapping

Science.gov (United States)

Murdick, Ryan A.; Morrison, William; Nowak, Derek; Albrecht, Thomas R.; Jahng, Junghoon; Park, Sung

2017-08-01

Advances in nanotechnology have intensified the need for tools that can characterize newly synthesized nanomaterials. A variety of techniques has recently been shown which combines atomic force microscopy (AFM) with optical illumination including tip-enhanced Raman spectroscopy (TERS), scattering-type scanning near-field optical microscopy (sSNOM), and photothermal induced resonance microscopy (PTIR). To varying degrees, these existing techniques enable optical spectroscopy with the nanoscale spatial resolution inherent to AFM, thereby providing nanochemical interrogation of a specimen. Here we discuss photoinduced force microscopy (PiFM), a recently developed technique for nanoscale optical spectroscopy that exploits image forces acting between an AFM tip and sample to detect wavelength-dependent polarization within the sample to generate absorption spectra. This approach enables ∼10 nm spatial resolution with spectra that show correlation with macroscopic optical absorption spectra. Unlike other techniques, PiFM achieves this high resolution with virtually no constraints on sample or substrate properties. The applicability of PiFM to a variety of archetypal systems is reported here, highlighting the potential of PiFM as a useful tool for a wide variety of industrial and academic investigations, including semiconducting nanoparticles, nanocellulose, block copolymers, and low dimensional systems, as well as chemical and morphological mixing at interfaces.

Light propagation studies on laser modified waveguides using scanning near-field optical microscopy

DEFF Research Database (Denmark)

Borrise, X.; Berini, Abadal Gabriel; Jimenez, D.

2001-01-01

By means of direct laser writing on Al, a new method to locally modify optical waveguides is proposed. This technique has been applied to silicon nitride waveguides, allowing modifications of the optical propagation along the guide. To study the formed structures, a scanning near-held optical mic...

Three-dimensional scanning near field optical microscopy (3D-SNOM) imaging of random arrays of copper nanoparticles: implications for plasmonic solar cell enhancement.

Science.gov (United States)

Ezugwu, Sabastine; Ye, Hanyang; Fanchini, Giovanni

2015-01-07

In order to investigate the suitability of random arrays of nanoparticles for plasmonic enhancement in the visible-near infrared range, we introduced three-dimensional scanning near-field optical microscopy (3D-SNOM) imaging as a useful technique to probe the intensity of near-field radiation scattered by random systems of nanoparticles at heights up to several hundred nm from their surface. We demonstrated our technique using random arrays of copper nanoparticles (Cu-NPs) at different particle diameter and concentration. Bright regions in the 3D-SNOM images, corresponding to constructive interference of forward-scattered plasmonic waves, were obtained at heights Δz ≥ 220 nm from the surface for random arrays of Cu-NPs of ∼ 60-100 nm in diameter. These heights are too large to use Cu-NPs in contact of the active layer for light harvesting in thin organic solar cells, which are typically no thicker than 200 nm. Using a 200 nm transparent spacer between the system of Cu-NPs and the solar cell active layer, we demonstrate that forward-scattered light can be conveyed in 200 nm thin film solar cells. This architecture increases the solar cell photoconversion efficiency by a factor of 3. Our 3D-SNOM technique is general enough to be suitable for a large number of other applications in nanoplasmonics.

Simultaneous characterization of rotational and translational diffusion of optically anisotropic particles by optical microscopy

International Nuclear Information System (INIS)

Giavazzi, Fabio; Cerbino, Roberto; Haro-Pérez, Catalina

2016-01-01

We probe the roto-translational Brownian motion of optically anisotropic particles suspended in water with a simple and straightforward optical microscopy experiment that does not require positional or rotational particle tracking. We acquire a movie of the suspension placed between two polarizing elements and we extract the translational diffusion coefficient D T and the rotational diffusion coefficient D R from the analysis of the temporal correlation properties of the spatial Fourier modes of the intensity fluctuations in the movie. Our method is successfully tested with a dilute suspension of birefringent spherical colloidal particles obtained by polymerizing an emulsion of droplets of liquid crystal in a nematic phase, whose roto-translational dynamics is found to be well described by theory. The simplicity of our approach makes our method a viable alternative to particle tracking and depolarized dynamic light scattering. (paper)

The mechanism of borax crystallization using in situ optical microscopy and AFM

International Nuclear Information System (INIS)

Suharso, G.; Parkinson, M.; Ogden, M.

2002-01-01

Full text: The quality of high-purity borax depends both on the concentrations of the impurities and the product appearance, which are mainly determined by the size and morphology of the crystals. Thus, knowledge about crystallization of borax is of direct relevance to the industrial production of borax. In addition, fundamental studies of borax crystallization will provide results of relevance to the crystallization of other economically important materials. An investigation into the fundamental mechanism of crystal growth of borax from aqueous solution was carried out, as a model system. The investigation focussed on the growth mechanism, and the influence of factors such as solution supersaturation, temperature, crystal size and solution flow on the rate of crystal growth. In situ optical microscopy was used to determine growth rates of three different faces of borax crystals at 20, 25, 30, and 35 deg C, at various concentrations. It was found that the growth rate increases with increasing temperature and supersaturation. At low concentration , growth on the (010), (001), and (111) faces occurs via a spiral growth mechanism and at high concentration birth and spread is the principal mechanism operating. The activation energy for the different mechanisms was determined. Examination by ex situ Atomic Force Microscopy (AFM) showed features suggesting that the (100), (010), (001) faces of borax crystals grow by spiral mechanism at low concentration and two dimensional nucleation at high concentration. These experiments support the data obtained from in situ optical microscopy. Copyright (2002) Australian Society for Electron Microscopy Inc

Scanning tunneling microscopy II further applications and related scanning techniques

CERN Document Server

Güntherodt, Hans-Joachim

1992-01-01

Scanning Tunneling Microscopy II, like its predecessor, presents detailed and comprehensive accounts of the basic principles and broad range of applications of STM and related scanning probe techniques. The applications discussed in this volume come predominantly from the fields of electrochemistry and biology. In contrast to those described in Vol. I, these sudies may be performed in air and in liquids. The extensions of the basic technique to map other interactions are described inchapters on scanning force microscopy, magnetic force microscopy, scanning near-field optical microscopy, together with a survey of other related techniques. Also described here is the use of a scanning proximal probe for surface modification. Togehter, the two volumes give a comprehensive account of experimental aspcets of STM. They provide essentialreading and reference material for all students and researchers involvedin this field.

Structured caustic vector vortex optical field: manipulating optical angular momentum flux and polarization rotation.

Science.gov (United States)

Chen, Rui-Pin; Chen, Zhaozhong; Chew, Khian-Hooi; Li, Pei-Gang; Yu, Zhongliang; Ding, Jianping; He, Sailing

2015-05-29

A caustic vector vortex optical field is experimentally generated and demonstrated by a caustic-based approach. The desired caustic with arbitrary acceleration trajectories, as well as the structured states of polarization (SoP) and vortex orders located in different positions in the field cross-section, is generated by imposing the corresponding spatial phase function in a vector vortex optical field. Our study reveals that different spin and orbital angular momentum flux distributions (including opposite directions) in different positions in the cross-section of a caustic vector vortex optical field can be dynamically managed during propagation by intentionally choosing the initial polarization and vortex topological charges, as a result of the modulation of the caustic phase. We find that the SoP in the field cross-section rotates during propagation due to the existence of the vortex. The unique structured feature of the caustic vector vortex optical field opens the possibility of multi-manipulation of optical angular momentum fluxes and SoP, leading to more complex manipulation of the optical field scenarios. Thus this approach further expands the functionality of an optical system.

Imaging subsurface damage of grinded fused silica optics by confocal fluorescence microscopy

International Nuclear Information System (INIS)

Neauport, J.; Cormont, P.; Destribats, J.; Legros, P.; Ambard, C.

2009-01-01

We report an experimental investigation of fluorescence confocal microscopy as a tool to measure subsurface damage on grinded fused silica optics. Confocal fluorescence microscopy was performed with an excitation at the wavelength of 405 nm on fixed abrasive diamond grinded fused silica samples. We detail the measured fluorescence spectrums and compare them to those of oil based coolants and grinding slurries. We evidence that oil based coolant used in diamond grinding induces a fluorescence that marks the subsurface damages and eases its observation. Such residual traces might also be involved in the laser damage process. (authors)

Light distribution analysis of optical fibre probe-based near-field optical tweezers using FDTD

Energy Technology Data Exchange (ETDEWEB)

Liu, B H; Yang, L J; Wang, Y [School of Mechanical and Electrical Engineering, Harbin Institute of Technology, Heilongjiang, Harbin, 150001 (China)], E-mail: richelaw@163.com

2009-09-01

Optical fibre probe-based near-field optical tweezers overcomes the diffraction limit of conventional optical tweezers, utilizing strong mechanical forces and torque associated with highly enhanced electric fields to trap and manipulate nano-scale particles. Near-field evanescent wave generated at optical fibre probe decays rapidly with the distance that results a significant reduced trapping volume, thus it is necessary to analyze the near-field distribution of optical fibre probe. The finite difference time domain (FDTD) method is applied to characterize the near-field distribution of optical fibre probe. In terms of the distribution patterns, depolarization and polarization, the near-field distributions in longitudinal sections and cross-sections of tapered metal-coated optical fibre probe are calculated. The calculation results reveal that the incident polarized wave becomes depolarized after exiting from the nano-scale aperture of probe. The near-field distribution of the probe is unsymmetrical, and the near-field distribution in the cross-section vertical to the incident polarized wave is different from that in the cross-section parallel to the incident polarized wave. Moreover, the polarization of incident wave has a great impact on the light intensity distribution.

Optimising electron microscopy experiment through electron optics simulation

Energy Technology Data Exchange (ETDEWEB)

Kubo, Y. [CEMES-CNRS, 29 Rue Jeanne Marvig, 31055 Toulouse France (France); Hitachi High-Technologies Corporation, 882, Ichige, Hitachinaka, Ibaraki 312-8504 (Japan); Gatel, C.; Snoeck, E. [CEMES-CNRS, 29 Rue Jeanne Marvig, 31055 Toulouse France (France); Houdellier, F., E-mail: florent.houdellier@cemes.fr [CEMES-CNRS, 29 Rue Jeanne Marvig, 31055 Toulouse France (France)

2017-04-15

We developed a new type of electron trajectories simulation inside a complete model of a modern transmission electron microscope (TEM). Our model incorporates the precise and real design of each element constituting a TEM, i.e. the field emission (FE) cathode, the extraction optic and acceleration stages of a 300 kV cold field emission gun, the illumination lenses, the objective lens, the intermediate and projection lenses. Full trajectories can be computed using magnetically saturated or non-saturated round lenses, magnetic deflectors and even non-cylindrical symmetry elements like electrostatic biprism. This multi-scale model gathers nanometer size components (FE tip) with parts of meter length (illumination and projection systems). We demonstrate that non-trivial TEM experiments requiring specific and complex optical configurations can be simulated and optimized prior to any experiment using such model. We show that all the currents set in all optical elements of the simulated column can be implemented in the real column (I2TEM in CEMES) and used as starting alignment for the requested experiment. We argue that the combination of such complete electron trajectory simulations in the whole TEM column with automatic optimization of the microscope parameters for optimal experimental data (images, diffraction, spectra) allows drastically simplifying the implementation of complex experiments in TEM and will facilitate the development of advanced use of the electron microscope in the near future. - Highlights: • Using dedicated electron optics software, we calculate full electrons trajectories inside a modern transmission electron microscope. • We have determined how to deal with multi-scale electron optics elements like high voltage cold field emission source. • W • e have succeed to model both weak and strong magnetic lenses whether in saturated or unsaturated conditions as well as electrostatic biprism and magnetic deflectors. • We have applied this model

Optimising electron microscopy experiment through electron optics simulation

International Nuclear Information System (INIS)

Kubo, Y.; Gatel, C.; Snoeck, E.; Houdellier, F.

2017-01-01

We developed a new type of electron trajectories simulation inside a complete model of a modern transmission electron microscope (TEM). Our model incorporates the precise and real design of each element constituting a TEM, i.e. the field emission (FE) cathode, the extraction optic and acceleration stages of a 300 kV cold field emission gun, the illumination lenses, the objective lens, the intermediate and projection lenses. Full trajectories can be computed using magnetically saturated or non-saturated round lenses, magnetic deflectors and even non-cylindrical symmetry elements like electrostatic biprism. This multi-scale model gathers nanometer size components (FE tip) with parts of meter length (illumination and projection systems). We demonstrate that non-trivial TEM experiments requiring specific and complex optical configurations can be simulated and optimized prior to any experiment using such model. We show that all the currents set in all optical elements of the simulated column can be implemented in the real column (I2TEM in CEMES) and used as starting alignment for the requested experiment. We argue that the combination of such complete electron trajectory simulations in the whole TEM column with automatic optimization of the microscope parameters for optimal experimental data (images, diffraction, spectra) allows drastically simplifying the implementation of complex experiments in TEM and will facilitate the development of advanced use of the electron microscope in the near future. - Highlights: • Using dedicated electron optics software, we calculate full electrons trajectories inside a modern transmission electron microscope. • We have determined how to deal with multi-scale electron optics elements like high voltage cold field emission source. • W • e have succeed to model both weak and strong magnetic lenses whether in saturated or unsaturated conditions as well as electrostatic biprism and magnetic deflectors. • We have applied this model

CARS microscopy for imaging

International Nuclear Information System (INIS)

Arzumanyan Grigory; Voskanyan Karine

2013-01-01

Optical microscopy grows in its importance with the development of modern nanotechnology, biotechnology, methods of diagnostics and treatment of most dangerous diseases for mankind. There are several important goals of optical microscopy for biomedical studies among which the next three may be distinguished: fast imaging with high lateral spatial resolution, 3-D sectioning capability and high contrast for chemical selectivity. To meet these specific requirements, various types of both linear and nonlinear optical microscopy were elaborated. (authors)

Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy

Science.gov (United States)

Neuman, Keir C.; Nagy, Attila

2012-01-01

Single-molecule force spectroscopy has emerged as a powerful tool to investigate the forces and motions associated with biological molecules and enzymatic activity. The most common force spectroscopy techniques are optical tweezers, magnetic tweezers and atomic force microscopy. These techniques are described and illustrated with examples highlighting current capabilities and limitations. PMID:18511917

Field-induced optically isotropic state in bent core nematic liquid crystals: unambiguous proof of field-induced optical biaxiality

International Nuclear Information System (INIS)

Elamain, Omaima; Komitov, Lachezar; Hegde, Gurumurthy; Fodor-Csorba, Katalin

2013-01-01

The behaviour of bent core (BC) nematic liquid crystals was investigated under dc applied electric field. The optically isotropic state of a sample containing BC nematic was observed under application of low dc electric fields. The quality of the dark state when the sample was inserted between two crossed polarizers was found to be superb and it did not change when rotating the sample between the polarizers. The coupling between the net molecular dipole moment and the applied dc electric field was considered as the origin of the out-of-plane switching of the BC molecules resulting in switching from the field-off bright state to the field-on dark state. The field-induced optically isotropic state is an unambiguous proof of the field-induced biaxiality in the BC nematic liquid crystal. A simple model explaining the appearance of the isotropic optical state in BC nematics and the switching of the sample slow axis between three mutually orthogonal directions under dc applied electric field is proposed. (paper)

U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy. Rev. 1

International Nuclear Information System (INIS)

Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.; Schemer-Kohrn, Alan L.; Guzman, Anthony D.; Lavender, Curt A.

2016-01-01

The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

Noninvasive label-free monitoring of cosmetics and pharmaceuticals in human skin using nonlinear optical microscopy (Conference Presentation)

Science.gov (United States)

Osseiran, Sam; Wang, Hequn; Evans, Conor L.

2017-02-01

Over the past decade, nonlinear optical microscopy has seen a dramatic rise in its use in research settings due to its noninvasiveness, enhanced penetration depth, intrinsic optical sectioning, and the ability to probe chemical compounds with molecular specificity without exogenous contrast agents. Nonlinear optical techniques including two-photon excitation fluorescence (2PEF), fluorescence lifetime imaging microscopy (FLIM), second harmonic generation (SHG), coherent anti-Stokes and stimulated Raman scattering (CARS and SRS, respectively), as well as transient and sum frequency absorption (TA and SFA, respectively), have been widely used to explore the physiology and microanatomy of skin. Recently, these modalities have shed light on dermal processes that could not have otherwise been observed, including the spatiotemporal monitoring of cosmetics and pharmaceuticals. However, a challenge quickly arises when studying such chemicals in a dermatological context: many exogenous compounds have optical signatures that can interfere with the signals that would otherwise be acquired from intact skin. For example, oily solvents exhibit strong signals when probing CH2 vibrations with CARS/SRS; chemical sun filters appear bright in 2PEF microscopy; and darkly colored compounds readily absorb light across a broad spectrum, producing strong TA/SFA signals. Thus, this discussion will first focus on the molecular contrast in skin that can be probed using the aforementioned nonlinear optical techniques. This will be followed by an overview of strategies that take advantage of the exogenous compounds' optical signatures to probe spatiotemporal dynamics while preserving endogenous information from skin.

Magneto-Optic Field Coupling in Optical Fiber Bragg Gratings

Science.gov (United States)

Carman, Gregory P. (Inventor); Mohanchandra, Panduranga K. (Inventor); Emmons, Michael C. (Inventor); Richards, William Lance (Inventor)

2016-01-01

The invention is a magneto-optic coupled magnetic sensor that comprises a standard optical fiber Bragg grating system. The system includes an optical fiber with at least one Bragg grating therein. The optical fiber has at least an inner core and a cladding that surrounds the inner core. The optical fiber is part of an optical system that includes an interrogation device that provides a light wave through the optical fiber and a system to determine the change in the index of refraction of the optical fiber. The cladding of the optical fiber comprises at least a portion of which is made up of ferromagnetic particles so that the ferromagnetic particles are subject to the light wave provided by the interrogation system. When a magnetic field is present, the ferromagnetic particles change the optical properties of the sensor directly.

Label-free imaging of acanthamoeba using multimodal nonlinear optical microscopy

Science.gov (United States)

Kobayashi, Tsubasa; Cha, Yu-Rok; Kaji, Yuichi; Oshika, Tetsuro; Leproux, Philippe; Couderc, Vincent; Kano, Hideaki

2018-02-01

Acanthamoeba keratitis is a disease in which amoebae named Acanthamoeba invade the cornea of an eye. To diagnose this disease before it becomes serious, it is important to detect the cyst state of Acanthamoeba in the early stage of infection. In the present study, we explored spectroscopic signitures of the cyst state of Acanthamoeba using multimodal nonlinear optical microscopy with the channels of multiplex coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), and third harmonic generation (THG). A sharp band at around 1603 cm-1 in the CARS (Im[χ(3)]) spectrum was found at the cyst state of Acanthamoeba, which possibly originates from ergosterol and/or 7-dehydrostigmasterol. It can be used as a maker band of Acanthamoeba for medical treatment. Keyword: Acanthamoeba keratitis, coherent anti-Stokes Raman scattering, CARS, second harmonic generation, SHG, microspectroscopy, multiphoton microscopy

Field guide to geometrical optics

CERN Document Server

Greivenkamp, John E

2004-01-01

This Field Guide derives from the treatment of geometrical optics that has evolved from both the undergraduate and graduate programs at the Optical Sciences Center at the University of Arizona. The development is both rigorous and complete, and it features a consistent notation and sign convention. This volume covers Gaussian imagery, paraxial optics, first-order optical system design, system examples, illumination, chromatic effects, and an introduction to aberrations. The appendices provide supplemental material on radiometry and photometry, the human eye, and several other topics.

Cycloid scanning for wide field optical coherence tomography endomicroscopy and angiography in vivo

Science.gov (United States)

Liang, Kaicheng; Wang, Zhao; Ahsen, Osman O.; Lee, Hsiang-Chieh; Potsaid, Benjamin M.; Jayaraman, Vijaysekhar; Cable, Alex; Mashimo, Hiroshi; Li, Xingde; Fujimoto, James G.

2018-01-01

Devices that perform wide field-of-view (FOV) precision optical scanning are important for endoscopic assessment and diagnosis of luminal organ disease such as in gastroenterology. Optical scanning for in vivo endoscopic imaging has traditionally relied on one or more proximal mechanical actuators, limiting scan accuracy and imaging speed. There is a need for rapid and precise two-dimensional (2D) microscanning technologies to enable the translation of benchtop scanning microscopies to in vivo endoscopic imaging. We demonstrate a new cycloid scanner in a tethered capsule for ultrahigh speed, side-viewing optical coherence tomography (OCT) endomicroscopy in vivo. The cycloid capsule incorporates two scanners: a piezoelectrically actuated resonant fiber scanner to perform a precision, small FOV, fast scan and a micromotor scanner to perform a wide FOV, slow scan. Together these scanners distally scan the beam circumferentially in a 2D cycloid pattern, generating an unwrapped 1 mm × 38 mm strip FOV. Sequential strip volumes can be acquired with proximal pullback to image centimeter-long regions. Using ultrahigh speed 1.3 μm wavelength swept-source OCT at a 1.17 MHz axial scan rate, we imaged the human rectum at 3 volumes/s. Each OCT strip volume had 166 × 2322 axial scans with 8.5 μm axial and 30 μm transverse resolution. We further demonstrate OCT angiography at 0.5 volumes/s, producing volumetric images of vasculature. In addition to OCT applications, cycloid scanning promises to enable precision 2D optical scanning for other imaging modalities, including fluorescence confocal and nonlinear microscopy. PMID:29682598

Polarization contrast in photon scanning tunnelling microscopy combined with atomic force microscopy

NARCIS (Netherlands)

Propstra, K.; Propstra, K.; van Hulst, N.F.

1995-01-01

Photon scanning tunnelling microscopy combined with atomic force microscopy allows simultaneous acquisition and direct comparison of optical and topographical images, both with a lateral resolution of about 30 nm, far beyond the optical diffraction limit. The probe consists of a modified

Optimal model-based sensorless adaptive optics for epifluorescence microscopy.

Science.gov (United States)

Pozzi, Paolo; Soloviev, Oleg; Wilding, Dean; Vdovin, Gleb; Verhaegen, Michel

2018-01-01

We report on a universal sample-independent sensorless adaptive optics method, based on modal optimization of the second moment of the fluorescence emission from a point-like excitation. Our method employs a sample-independent precalibration, performed only once for the particular system, to establish the direct relation between the image quality and the aberration. The method is potentially applicable to any form of microscopy with epifluorescence detection, including the practically important case of incoherent fluorescence emission from a three dimensional object, through minor hardware modifications. We have applied the technique successfully to a widefield epifluorescence microscope and to a multiaperture confocal microscope.

Detection, mapping, and quantification of single walled carbon nanotubes in histological specimens with photoacoustic microscopy.

Science.gov (United States)

Avti, Pramod K; Hu, Song; Favazza, Christopher; Mikos, Antonios G; Jansen, John A; Shroyer, Kenneth R; Wang, Lihong V; Sitharaman, Balaji

2012-01-01

In the present study, the efficacy of multi-scale photoacoustic microscopy (PAM) was investigated to detect, map, and quantify trace amounts [nanograms (ng) to micrograms (µg)] of SWCNTs in a variety of histological tissue specimens consisting of cancer and benign tissue biopsies (histological specimens from implanted tissue engineering scaffolds). Optical-resolution (OR) and acoustic-resolution (AR)--Photoacoustic microscopy (PAM) was employed to detect, map and quantify the SWCNTs in a variety of tissue histological specimens and compared with other optical techniques (bright-field optical microscopy, Raman microscopy, near infrared (NIR) fluorescence microscopy). Both optical-resolution and acoustic-resolution PAM, allow the detection and quantification of SWCNTs in histological specimens with scalable spatial resolution and depth penetration. The noise-equivalent detection sensitivity to SWCNTs in the specimens was calculated to be as low as ∼7 pg. Image processing analysis further allowed the mapping, distribution, and quantification of the SWCNTs in the histological sections. The results demonstrate the potential of PAM as a promising imaging technique to detect, map, and quantify SWCNTs in histological specimens, and could complement the capabilities of current optical and electron microscopy techniques in the analysis of histological specimens containing SWCNTs.

Detection, mapping, and quantification of single walled carbon nanotubes in histological specimens with photoacoustic microscopy.

Directory of Open Access Journals (Sweden)

Pramod K Avti

Full Text Available In the present study, the efficacy of multi-scale photoacoustic microscopy (PAM was investigated to detect, map, and quantify trace amounts [nanograms (ng to micrograms (µg] of SWCNTs in a variety of histological tissue specimens consisting of cancer and benign tissue biopsies (histological specimens from implanted tissue engineering scaffolds.Optical-resolution (OR and acoustic-resolution (AR--Photoacoustic microscopy (PAM was employed to detect, map and quantify the SWCNTs in a variety of tissue histological specimens and compared with other optical techniques (bright-field optical microscopy, Raman microscopy, near infrared (NIR fluorescence microscopy.Both optical-resolution and acoustic-resolution PAM, allow the detection and quantification of SWCNTs in histological specimens with scalable spatial resolution and depth penetration. The noise-equivalent detection sensitivity to SWCNTs in the specimens was calculated to be as low as ∼7 pg. Image processing analysis further allowed the mapping, distribution, and quantification of the SWCNTs in the histological sections.The results demonstrate the potential of PAM as a promising imaging technique to detect, map, and quantify SWCNTs in histological specimens, and could complement the capabilities of current optical and electron microscopy techniques in the analysis of histological specimens containing SWCNTs.

Generalized spectral method for near-field optical microscopy

Energy Technology Data Exchange (ETDEWEB)

Jiang, B.-Y.; Zhang, L. M.; Basov, D. N.; Fogler, M. M. [Department of Physics, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093 (United States); Castro Neto, A. H. [Department of Physics, Boston University, 590 Commonwealth Avenue, Boston, Massachusetts 02215 (United States); Centre for Advanced 2D Materials and Graphene Research Centre, National University of Singapore, Singapore, Singapore 117542 (Singapore)

2016-02-07

Electromagnetic interaction between a sub-wavelength particle (the “probe”) and a material surface (the “sample”) is studied theoretically. The interaction is shown to be governed by a series of resonances corresponding to surface polariton modes localized near the probe. The resonance parameters depend on the dielectric function and geometry of the probe as well as on the surface reflectivity of the material. Calculation of such resonances is carried out for several types of axisymmetric probes: spherical, spheroidal, and pear-shaped. For spheroids, an efficient numerical method is developed, capable of handling cases of large or strongly momentum-dependent surface reflectivity. Application of the method to highly resonant materials, such as aluminum oxide (by itself or covered with graphene), reveals a rich structure of multi-peak spectra and nonmonotonic approach curves, i.e., the probe-sample distance dependence. These features also strongly depend on the probe shape and optical constants of the model. For less resonant materials such as silicon oxide, the dependence is weak, so that the spheroidal model is reliable. The calculations are done within the quasistatic approximation with radiative damping included perturbatively.

Exploring lipids with nonlinear optical microscopy in multiple biological systems

Science.gov (United States)

Alfonso-Garcia, Alba

Lipids are crucial biomolecules for the well being of humans. Altered lipid metabolism may give rise to a variety of diseases that affect organs from the cardiovascular to the central nervous system. A deeper understanding of lipid metabolic processes would spur medical research towards developing precise diagnostic tools, treatment methods, and preventive strategies for reducing the impact of lipid diseases. Lipid visualization remains a complex task because of the perturbative effect exerted by traditional biochemical assays and most fluorescence markers. Coherent Raman scattering (CRS) microscopy enables interrogation of biological samples with minimum disturbance, and is particularly well suited for label-free visualization of lipids, providing chemical specificity without compromising on spatial resolution. Hyperspectral imaging yields large datasets that benefit from tailored multivariate analysis. In this thesis, CRS microscopy was combined with Raman spectroscopy and other label-free nonlinear optical techniques to analyze lipid metabolism in multiple biological systems. We used nonlinear Raman techniques to characterize Meibum secretions in the progression of dry eye disease, where the lipid and protein contributions change in ratio and phase segregation. We employed similar tools to examine lipid droplets in mice livers aboard a spaceflight mission, which lose their retinol content contributing to the onset of nonalcoholic fatty-liver disease. We also focused on atherosclerosis, a disease that revolves around lipid-rich plaques in arterial walls. We examined the lipid content of macrophages, whose variable phenotype gives rise to contrasting healing and inflammatory activities. We also proposed new label-free markers, based on lifetime imaging, for macrophage phenotype, and to detect products of lipid oxidation. Cholesterol was also detected in hepatitis C virus infected cells, and in specific strains of age-related macular degeneration diseased cells by

Near field optics and nanoscopy

CERN Document Server

Fillard, J P

1996-01-01

This book contains the most recent information on optical nanoscopy. Far-Field and Near-Field properties on e.m. waves are presented which illustrate how optical images can be obtained from sub-micron objects. Scanning Probe techniques and computer processing are covered here. An explanation is given on how propagating photons or evanescent waves can behave over distances shorter than the wavelength, taking into account the presence of small objects. Quantum tunneling of photons is explained comparatively with the electron mechanism. Technical details are given on photon tunneling microscopes.

Expansions of general stationary stochastic optical fields: general formalism

International Nuclear Information System (INIS)

Martinez-Herrero, R.; Mejias, P.M.

1985-01-01

A new expansion of a general stationary stochastic optical field is derived. Each term of the series is seen to represent a recently defined new class of optical fields, the so-called spectrally quasi-factorizable fields. Alternative expansion in terms of nonstationary fields that obey the wave equation is also shown. A relationship between temporal and spatial features of stationary free optical fields is discussed

Mapping exciton quenching in photovoltaic-applicable polymer blends using time-resolved scanning near-field optical microscopy

Science.gov (United States)

Cadby, A.; Khalil, G.; Fox, A. M.; Lidzey, D. G.

2008-05-01

We have used time-resolved scanning near-field microscopy to image the fluorescence decay lifetime across a phase-separated blend of the photovoltaic-applicable polymers poly(9,9'-dioctylfluorene-alt-benzothiadiazole) (F8BT) and poly(9,9'-dioctylfluorene-alt-bis- N ,N'-(4-butylphenyl)-bis-N ,N'-phenyl-1,4-phenylenediamine) (PFB). We show that the efficiency of local fluorescence quenching is composition dependent, with excitons on F8BT molecules being more effectively quenched when F8BT is trapped at a low concentration in a PFB-rich phase. Despite such presumed differences in charge-carrier generation efficiency, our results demonstrate that charge extraction from F8BT:PFB devices is the most dominant mechanism limiting their operational efficiency.

Fiber-optic annular detector array for large depth of field photoacoustic macroscopy

Directory of Open Access Journals (Sweden)

Johannes Bauer-Marschallinger

2017-03-01

Full Text Available We report on a novel imaging system for large depth of field photoacoustic scanning macroscopy. Instead of commonly used piezoelectric transducers, fiber-optic based ultrasound detection is applied. The optical fibers are shaped into rings and mainly receive ultrasonic signals stemming from the ring symmetry axes. Four concentric fiber-optic rings with varying diameters are used in order to increase the image quality. Imaging artifacts, originating from the off-axis sensitivity of the rings, are reduced by coherence weighting. We discuss the working principle of the system and present experimental results on tissue mimicking phantoms. The lateral resolution is estimated to be below 200 μm at a depth of 1.5 cm and below 230 μm at a depth of 4.5 cm. The minimum detectable pressure is in the order of 3 Pa. The introduced method has the potential to provide larger imaging depths than acoustic resolution photoacoustic microscopy and an imaging resolution similar to that of photoacoustic computed tomography.

Fiber-optic annular detector array for large depth of field photoacoustic macroscopy.

Science.gov (United States)

Bauer-Marschallinger, Johannes; Höllinger, Astrid; Jakoby, Bernhard; Burgholzer, Peter; Berer, Thomas

2017-03-01

We report on a novel imaging system for large depth of field photoacoustic scanning macroscopy. Instead of commonly used piezoelectric transducers, fiber-optic based ultrasound detection is applied. The optical fibers are shaped into rings and mainly receive ultrasonic signals stemming from the ring symmetry axes. Four concentric fiber-optic rings with varying diameters are used in order to increase the image quality. Imaging artifacts, originating from the off-axis sensitivity of the rings, are reduced by coherence weighting. We discuss the working principle of the system and present experimental results on tissue mimicking phantoms. The lateral resolution is estimated to be below 200 μm at a depth of 1.5 cm and below 230 μm at a depth of 4.5 cm. The minimum detectable pressure is in the order of 3 Pa. The introduced method has the potential to provide larger imaging depths than acoustic resolution photoacoustic microscopy and an imaging resolution similar to that of photoacoustic computed tomography.

Imaging arterial cells, atherosclerosis, and restenosis by multimodal nonlinear optical microscopy

Science.gov (United States)

Wang, Han-Wei; Simianu, Vlad; Locker, Matthew J.; Sturek, Michael; Cheng, Ji-Xin

2008-02-01

By integrating sum-frequency generation (SFG), and two-photon excitation fluorescence (TPEF) on a coherent anti-Stokes Raman scattering (CARS) microscope platform, multimodal nonlinear optical (NLO) imaging of arteries and atherosclerotic lesions was demonstrated. CARS signals arising from CH II-rich membranes allowed visualization of endothelial cells and smooth muscle cells in a carotid artery. Additionally, CARS microscopy allowed vibrational imaging of elastin and collagen fibrils which are rich in CH II bonds in their cross-linking residues. The extracellular matrix organization was further confirmed by TPEF signals arising from elastin's autofluorescence and SFG signals arising from collagen fibrils' non-centrosymmetric structure. The system is capable of identifying different atherosclerotic lesion stages with sub-cellular resolution. The stages of atherosclerosis, such as macrophage infiltration, lipid-laden foam cell accumulation, extracellular lipid distribution, fibrous tissue deposition, plaque establishment, and formation of other complicated lesions could be viewed by our multimodal CARS microscope. Collagen percentages in the region adjacent to coronary artery stents were resolved. High correlation between NLO and histology imaging evidenced the validity of the NLO imaging. The capability of imaging significant components of an arterial wall and distinctive stages of atherosclerosis in a label-free manner suggests the potential application of multimodal nonlinear optical microscopy to monitor the onset and progression of arterial diseases.

Vector optical fields with bipolar symmetry of linear polarization.

Science.gov (United States)

Pan, Yue; Li, Yongnan; Li, Si-Min; Ren, Zhi-Cheng; Si, Yu; Tu, Chenghou; Wang, Hui-Tian

2013-09-15

We focus on a new kind of vector optical field with bipolar symmetry of linear polarization instead of cylindrical and elliptical symmetries, enriching members of family of vector optical fields. We design theoretically and generate experimentally the demanded vector optical fields and then explore some novel tightly focusing properties. The geometric configurations of states of polarization provide additional degrees of freedom assisting in engineering the field distribution at the focus to the specific applications such as lithography, optical trapping, and material processing.

3D imaging of optically cleared tissue using a simplified CLARITY method and on-chip microscopy

KAUST Repository

Zhang, Yibo; Shin, Yoonjung; Sung, Kevin; Yang, Sam; Chen, Harrison; Wang, Hongda; Teng, Da; Rivenson, Yair; Kulkarni, Rajan P.; Ozcan, Aydogan

2017-01-01

High-throughput sectioning and optical imaging of tissue samples using traditional immunohistochemical techniques can be costly and inaccessible in resource-limited areas. We demonstrate three-dimensional (3D) imaging and phenotyping in optically transparent tissue using lens-free holographic on-chip microscopy as a low-cost, simple, and high-throughput alternative to conventional approaches. The tissue sample is passively cleared using a simplified CLARITY method and stained using 3,3′-diaminobenzidine to target cells of interest, enabling bright-field optical imaging and 3D sectioning of thick samples. The lens-free computational microscope uses pixel super-resolution and multi-height phase recovery algorithms to digitally refocus throughout the cleared tissue and obtain a 3D stack of complex-valued images of the sample, containing both phase and amplitude information. We optimized the tissue-clearing and imaging system by finding the optimal illumination wavelength, tissue thickness, sample preparation parameters, and the number of heights of the lens-free image acquisition and implemented a sparsity-based denoising algorithm to maximize the imaging volume and minimize the amount of the acquired data while also preserving the contrast-to-noise ratio of the reconstructed images. As a proof of concept, we achieved 3D imaging of neurons in a 200-μm-thick cleared mouse brain tissue over a wide field of view of 20.5 mm2. The lens-free microscope also achieved more than an order-of-magnitude reduction in raw data compared to a conventional scanning optical microscope imaging the same sample volume. Being low cost, simple, high-throughput, and data-efficient, we believe that this CLARITY-enabled computational tissue imaging technique could find numerous applications in biomedical diagnosis and research in low-resource settings.

3D imaging of optically cleared tissue using a simplified CLARITY method and on-chip microscopy

KAUST Repository

Zhang, Yibo

2017-08-12

High-throughput sectioning and optical imaging of tissue samples using traditional immunohistochemical techniques can be costly and inaccessible in resource-limited areas. We demonstrate three-dimensional (3D) imaging and phenotyping in optically transparent tissue using lens-free holographic on-chip microscopy as a low-cost, simple, and high-throughput alternative to conventional approaches. The tissue sample is passively cleared using a simplified CLARITY method and stained using 3,3′-diaminobenzidine to target cells of interest, enabling bright-field optical imaging and 3D sectioning of thick samples. The lens-free computational microscope uses pixel super-resolution and multi-height phase recovery algorithms to digitally refocus throughout the cleared tissue and obtain a 3D stack of complex-valued images of the sample, containing both phase and amplitude information. We optimized the tissue-clearing and imaging system by finding the optimal illumination wavelength, tissue thickness, sample preparation parameters, and the number of heights of the lens-free image acquisition and implemented a sparsity-based denoising algorithm to maximize the imaging volume and minimize the amount of the acquired data while also preserving the contrast-to-noise ratio of the reconstructed images. As a proof of concept, we achieved 3D imaging of neurons in a 200-μm-thick cleared mouse brain tissue over a wide field of view of 20.5 mm2. The lens-free microscope also achieved more than an order-of-magnitude reduction in raw data compared to a conventional scanning optical microscope imaging the same sample volume. Being low cost, simple, high-throughput, and data-efficient, we believe that this CLARITY-enabled computational tissue imaging technique could find numerous applications in biomedical diagnosis and research in low-resource settings.

Near-field reflection backscattering apertureless optical microscopy: Application to spectroscopy experiments on opaque samples, comparison between lock-in and digital photon counting detection techniques

International Nuclear Information System (INIS)

Diziain, S.; Bijeon, J.-L.; Adam, P.-M.; Lamy de la Chapelle, M.; Thomas, B.; Deturche, R.; Royer, P.

2007-01-01

An apertureless scanning near-field optical microscope (ASNOM) in reflection backscattering configuration is designed to conduct spectroscopic experiments on opaque samples constituted of latex beads. The ASNOM proposed takes advantage of the depth-discrimination properties of confocal microscopes to efficiently extract the near-field optical signal. Given their importance in a spectroscopic experiment, we systematically compare the lock-in and synchronous photon counting detection methods. Some results of Rayleigh's scattering in the near field of the test samples are used to illustrate the possibilities of this technique for reflection backscattering spectroscopy

Near-field reflection backscattering apertureless optical microscopy: Application to spectroscopy experiments on opaque samples, comparison between lock-in and digital photon counting detection techniques

Energy Technology Data Exchange (ETDEWEB)

Diziain, S. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Bijeon, J.-L. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France)]. E-mail: bijeon@utt.fr; Adam, P.-M. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Lamy de la Chapelle, M. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Thomas, B. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Deturche, R. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Royer, P. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France)

2007-01-15

An apertureless scanning near-field optical microscope (ASNOM) in reflection backscattering configuration is designed to conduct spectroscopic experiments on opaque samples constituted of latex beads. The ASNOM proposed takes advantage of the depth-discrimination properties of confocal microscopes to efficiently extract the near-field optical signal. Given their importance in a spectroscopic experiment, we systematically compare the lock-in and synchronous photon counting detection methods. Some results of Rayleigh's scattering in the near field of the test samples are used to illustrate the possibilities of this technique for reflection backscattering spectroscopy.

Photonic Torque Microscopy of the Nonconservative Force Field for Optically Trapped Silicon Nanowires

Czech Academy of Sciences Publication Activity Database

Irrera, A.; Maggazu, A.; Artoni, P.; Simpson, Stephen Hugh; Hanna, S.; Jones, P.H.; Priolo, F.; Gucciardi, P. G.; Marago, O.M.

2016-01-01

Roč. 16, č. 7 (2016), s. 4181-4188 ISSN 1530-6984 R&D Projects: GA ČR GB14-36681G Institutional support: RVO:68081731 Keywords : optical tweezers * silicon nanowires * nonequilibrium dynamics * Brownian motion Subject RIV: BH - Optics, Masers, Lasers Impact factor: 12.712, year: 2016

Unconventional methods of imaging: computational microscopy and compact implementations

Science.gov (United States)

McLeod, Euan; Ozcan, Aydogan

2016-07-01

In the past two decades or so, there has been a renaissance of optical microscopy research and development. Much work has been done in an effort to improve the resolution and sensitivity of microscopes, while at the same time to introduce new imaging modalities, and make existing imaging systems more efficient and more accessible. In this review, we look at two particular aspects of this renaissance: computational imaging techniques and compact imaging platforms. In many cases, these aspects go hand-in-hand because the use of computational techniques can simplify the demands placed on optical hardware in obtaining a desired imaging performance. In the first main section, we cover lens-based computational imaging, in particular, light-field microscopy, structured illumination, synthetic aperture, Fourier ptychography, and compressive imaging. In the second main section, we review lensfree holographic on-chip imaging, including how images are reconstructed, phase recovery techniques, and integration with smart substrates for more advanced imaging tasks. In the third main section we describe how these and other microscopy modalities have been implemented in compact and field-portable devices, often based around smartphones. Finally, we conclude with some comments about opportunities and demand for better results, and where we believe the field is heading.

Heavy-ion microscopy

International Nuclear Information System (INIS)

Kraft, G.; Yang, T.C.H.; Richards, T.; Tobias, C.A.

1980-01-01

This chapter briefly describes the techniques of optical microscopy, scanning and transmission electron microscopy, soft x-ray microscopy and compares these latter techniques with heavy-ion microscopy. The resolution obtained with these various types of microscopy are compared and the influence of the etching procedure on total resolution is discussed. Several micrographs of mammalian cells are included

Synthesis, field emission properties and optical properties of ZnSe nanoflowers

Energy Technology Data Exchange (ETDEWEB)

Xue, S.L., E-mail: slxue@dhu.edu.cn [Department of Applied Physics, College of Science, Donghua University, Shanghai 201620 (China); Wu, S.X.; Zeng, Q.Z.; Xie, P.; Gan, K.X.; Wei, J.; Bu, S.Y.; Ye, X.N.; Xie, L. [Department of Applied Physics, College of Science, Donghua University, Shanghai 201620 (China); Zou, R.J. [State Key Laboratory for Modification and Chemical Fibers and Polymer Materials, Donghua University, Shanghai 201620 (China); Zhang, C.M.; Zhu, P.F. [Department of Physics, School of Fundamental Studies, Shanghai University of Engineering Science, Shanghai 201620 (China)

2016-03-01

Graphical abstract: Unique ZnSe nanoflowers have been successfully synthesized by reaction of Se powder with Zn substrates. They are characterized by XRD, SEM, TEM, XPS, EDS and Raman spectroscopy and were single crystals with cubic zinc blende (ZB) structure. They also have excellent field emission properties and optical properties. - Highlights: • Novel ZnSe nanoflowers are grown on Zn foils. • ZnSe nanoflowers are characterized by XRD, SEM, TEM, XPS and Raman spectra. • ZnSe nanoflowers on Zn foils as cathodes possess good FE properties. - Abstract: ZnSe nanoflowers have been synthesized by reaction of Se powder with Zn substrates at low temperature. The as-prepared ZnSe nanoflowers were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), X-ray photoelectron spectroscopy (XPS), X-ray energy dispersive spectroscope (EDS) and Raman spectroscopy measurements. It was found that the morphologies of the as-prepared samples highly depended on reaction time. ZnSe nanoclusters and nanoflowers formed at 573 K when the reaction time was 20 and 60 min, respectively. The as-prepared ZnSe nanoflowers were composed of radically aligned ZnSe nanorods with smooth surfaces. The results of XRD, XPS, EDS, TEM and Raman showed that the as-prepared ZnSe nanocrystals were single crystals with cubic zinc blende (ZB) structure. The formation mechanism of the as-prepared ZnSe nanoflowers was also discussed. In addition, the as-prepared ZnSe nanoflowers had excellent electron emission properties. The turn-on field of the as-prepared ZnSe nanoflowers was 3.5 V/μm and the enhancement factor was 3499. The optical properties of the as-prepared ZnSe nanoflowers were also investigated. The results demonstrated that the as-prepared ZnSe nanoflowers were potential candidates for optoelectronic devices.

Fluorescence detection of single molecules using pulsed near-field optical excitation and time correlated photon counting

International Nuclear Information System (INIS)

Ambrose, W.P.; Goodwin, P.M.; Martin, J.C.; Keller, R.A.

1994-01-01

Pulsed excitation, time correlated single photon counting and time gated detection are used in near-field optical microscopy to enhance fluorescence images and measure the fluorescence lifetimes of single molecules of Rhodamine 6G on silica surfaces. Time gated detection is used to reject prompt scattered background and to improve the image signal to noise ratio. The excited state lifetime of a single Rhodamine 6G molecule is found to depend on the position of the near-field probe. We attribute the lifetime variations to spontaneous emission rate alterations by the fluorescence reflected from and quenching by the aluminum coated probe

Statistical behaviour of optical vortex fields

CSIR Research Space (South Africa)

Roux, FS

2009-09-01

Full Text Available ) Density limitation→ effective profile for point vortex (remove evanescent field) . – p.10/37 Scintillated optical beams Optical beam in a turbulent atmosphere: → index variations cause random phase modulations → leads to distortion of the optical beam.... Weak scintillation→ continuous phase distortions that can be corrected by an adaptive optical system: Wavefront sensor Beam splitter Scintillated beam Corrected beam Deformable mirror Control signal . – p.11/37 Strong scintillation Strong scintillation...

Correlation of ''twins'' observed by optical microscopy and transmission electron microscopy in YBa2Cu3O7/sub -//sub x/ superconductors

International Nuclear Information System (INIS)

Hoff, H.A.; Singh, A.K.; Pande, C.S.

1988-01-01

By using transmission electron microscopy and optical microscopy on the same specimens, the patterns of light- and dark-contrast lines seen in reflected polarized light were shown to be an interference pattern due to the variable spacing of suboptical microtwins. These microtwins are mostly [110] reflection twins. The [110] twinning was observed to be cyclic and occasionally pseudotetragonal because of the progressive cycling of contact twin domains. Within a domain, and occasionally in a whole grain, the [110] reflection twins occurred as polysynthetic lamellae. The morphology of the domain structure can be explained from the theory of martensitic transformation

In situ 3D characterization of historical coatings and wood using multimodal nonlinear optical microscopy

OpenAIRE

Latour , Gaël; Echard , Jean-Philippe; Didier , Marie; Schanne-Klein , Marie-Claire

2012-01-01

International audience; We demonstrate multimodal nonlinear optical imaging of historical artifacts by combining Second Harmonic Generation (SHG) and Two-Photon Excited Fluorescence (2PEF) microscopies. We first identify the nonlinear optical response of materials commonly encountered in coatings of cultural heritage artifacts by analyzing one- and multi-layered model samples. We observe 2PEF signals from cochineal lake and sandarac and show that pigments and varnish films can be discriminate...

Assessment of fibrotic liver disease with multimodal nonlinear optical microscopy

Science.gov (United States)

Lu, Fake; Zheng, Wei; Tai, Dean C. S.; Lin, Jian; Yu, Hanry; Huang, Zhiwei

2010-02-01

Liver fibrosis is the excessive accumulation of extracellular matrix proteins such as collagens, which may result in cirrhosis, liver failure, and portal hypertension. In this study, we apply a multimodal nonlinear optical microscopy platform developed to investigate the fibrotic liver diseases in rat models established by performing bile duct ligation (BDL) surgery. The three nonlinear microscopy imaging modalities are implemented on the same sectioned tissues of diseased model sequentially: i.e., second harmonic generation (SHG) imaging quantifies the contents of the collagens, the two-photon excitation fluorescence (TPEF) imaging reveals the morphology of hepatic cells, while coherent anti-Stokes Raman scattering (CARS) imaging maps the distributions of fats or lipids quantitatively across the tissue. Our imaging results show that during the development of liver fibrosis (collagens) in BDL model, fatty liver disease also occurs. The aggregated concentrations of collagen and fat constituents in liver fibrosis model show a certain correlationship between each other.

Emerging optical nanoscopy techniques

Directory of Open Access Journals (Sweden)

Montgomery PC

2015-09-01

Full Text Available Paul C Montgomery, Audrey Leong-Hoi Laboratoire des Sciences de l'Ingénieur, de l'Informatique et de l'Imagerie (ICube, Unistra-CNRS, Strasbourg, France Abstract: To face the challenges of modern health care, new imaging techniques with subcellular resolution or detection over wide fields are required. Far field optical nanoscopy presents many new solutions, providing high resolution or detection at high speed. We present a new classification scheme to help appreciate the growing number of optical nanoscopy techniques. We underline an important distinction between superresolution techniques that provide improved resolving power and nanodetection techniques for characterizing unresolved nanostructures. Some of the emerging techniques within these two categories are highlighted with applications in biophysics and medicine. Recent techniques employing wider angle imaging by digital holography and scattering lens microscopy allow superresolution to be achieved for subcellular and even in vivo, imaging without labeling. Nanodetection techniques are divided into four subcategories using contrast, phase, deconvolution, and nanomarkers. Contrast enhancement is illustrated by means of a polarized light-based technique and with strobed phase-contrast microscopy to reveal nanostructures. Very high sensitivity phase measurement using interference microscopy is shown to provide nanometric surface roughness measurement or to reveal internal nanometric structures. Finally, the use of nanomarkers is illustrated with stochastic fluorescence microscopy for mapping intracellular structures. We also present some of the future perspectives of optical nanoscopy. Keywords: microscopy, imaging, superresolution, nanodetection, biophysics, medical imaging

Robustness of plasmonic angular momentum confinement in cross resonant optical antennas

Energy Technology Data Exchange (ETDEWEB)

Klaer, Peter; Lehr, Martin; Krewer, Keno; Schertz, Florian; Schönhense, Gerd; Elmers, Hans Joachim, E-mail: elmers@uni-mainz.de [Institut für Physik, Johannes Gutenberg-Universität, Staudingerweg 7, D-55099 Mainz (Germany); Razinskas, Gary; Wu, Xiao-Fei; Hecht, Bert [Institut für Physik, Julius-Maximilians-Universität, Am Hubland, 97074 Würzburg (Germany)

2015-06-29

Using a combination of photoemission electron microscopy and numerical simulations, we investigated the angular moment transfer in strongly enhanced optical near-fields of artificially fabricated optical antennas. The polarization dependence of the optical near-field enhancement has been measured in a maximum symmetric geometry, i.e., excitation by a normal incident planar wave. Finite-difference time-domain simulations for the realistic antenna geometries as determined by high-resolution electron microscopy reveal a very good agreement with experimental data. The agreement confirms that the geometrical asymmetries and inhomogeneities due to the nanoscale fabrication process preserve the circular polarization in the gap regions with strong near-field enhancement.

Single myelin fiber imaging in living rodents without labeling by deep optical coherence microscopy

Science.gov (United States)

Ben Arous, Juliette; Binding, Jonas; Léger, Jean-François; Casado, Mariano; Topilko, Piotr; Gigan, Sylvain; Claude Boccara, A.; Bourdieu, Laurent

2011-11-01

Myelin sheath disruption is responsible for multiple neuropathies in the central and peripheral nervous system. Myelin imaging has thus become an important diagnosis tool. However, in vivo imaging has been limited to either low-resolution techniques unable to resolve individual fibers or to low-penetration imaging of single fibers, which cannot provide quantitative information about large volumes of tissue, as required for diagnostic purposes. Here, we perform myelin imaging without labeling and at micron-scale resolution with >300-μm penetration depth on living rodents. This was achieved with a prototype [termed deep optical coherence microscopy (deep-OCM)] of a high-numerical aperture infrared full-field optical coherence microscope, which includes aberration correction for the compensation of refractive index mismatch and high-frame-rate interferometric measurements. We were able to measure the density of individual myelinated fibers in the rat cortex over a large volume of gray matter. In the peripheral nervous system, deep-OCM allows, after minor surgery, in situ imaging of single myelinated fibers over a large fraction of the sciatic nerve. This allows quantitative comparison of normal and Krox20 mutant mice, in which myelination in the peripheral nervous system is impaired. This opens promising perspectives for myelin chronic imaging in demyelinating diseases and for minimally invasive medical diagnosis.

Single myelin fiber imaging in living rodents without labeling by deep optical coherence microscopy.

Science.gov (United States)

Ben Arous, Juliette; Binding, Jonas; Léger, Jean-François; Casado, Mariano; Topilko, Piotr; Gigan, Sylvain; Boccara, A Claude; Bourdieu, Laurent

2011-11-01

Myelin sheath disruption is responsible for multiple neuropathies in the central and peripheral nervous system. Myelin imaging has thus become an important diagnosis tool. However, in vivo imaging has been limited to either low-resolution techniques unable to resolve individual fibers or to low-penetration imaging of single fibers, which cannot provide quantitative information about large volumes of tissue, as required for diagnostic purposes. Here, we perform myelin imaging without labeling and at micron-scale resolution with >300-μm penetration depth on living rodents. This was achieved with a prototype [termed deep optical coherence microscopy (deep-OCM)] of a high-numerical aperture infrared full-field optical coherence microscope, which includes aberration correction for the compensation of refractive index mismatch and high-frame-rate interferometric measurements. We were able to measure the density of individual myelinated fibers in the rat cortex over a large volume of gray matter. In the peripheral nervous system, deep-OCM allows, after minor surgery, in situ imaging of single myelinated fibers over a large fraction of the sciatic nerve. This allows quantitative comparison of normal and Krox20 mutant mice, in which myelination in the peripheral nervous system is impaired. This opens promising perspectives for myelin chronic imaging in demyelinating diseases and for minimally invasive medical diagnosis.

160 Gb/s all-optical packet switching field experiment

DEFF Research Database (Denmark)

Dorren, H.J.S.; Herrera, J.; Raz, O.

2007-01-01

We discus an all-optical packet switching experiment over 110 km of field installed optical fiber. The switching node is controlled by solely photonic control circuits.......We discus an all-optical packet switching experiment over 110 km of field installed optical fiber. The switching node is controlled by solely photonic control circuits....

Optical imaging beyond the diffraction limit by SNEM: Effects of AFM tip modifications with thiol monolayers on imaging quality

NARCIS (Netherlands)

Cumurcu, Aysegul; Diaz, J.; Lindsay, I.D.; de Beer, Sissi; Duvigneau, Joost; Schön, Peter Manfred; Vancso, Gyula J.

2015-01-01

Tip-enhanced nanoscale optical imaging techniques such as apertureless scanning near-field optical microscopy (a-SNOM) and scanning near-field ellipsometric microscopy (SNEM) applications can suffer from a steady degradation in performance due to adhesion of atmospheric contaminants to the metal

Scanning tunneling microscopy II further applications and related scanning techniques

CERN Document Server

Güntherodt, Hans-Joachim

1995-01-01

Scanning Tunneling Microscopy II, like its predecessor, presents detailed and comprehensive accounts of the basic principles and broad range of applications of STM and related scanning probe techniques. The applications discussed in this volume come predominantly from the fields of electrochemistry and biology. In contrast to those described in STM I, these studies may be performed in air and in liquids. The extensions of the basic technique to map other interactions are described in chapters on scanning force microscopy, magnetic force microscopy, and scanning near-field optical microscopy, together with a survey of other related techniques. Also described here is the use of a scanning proximal probe for surface modification. Together, the two volumes give a comprehensive account of experimental aspects of STM. They provide essential reading and reference material for all students and researchers involved in this field. In this second edition the text has been updated and new methods are discussed.

Dark-field X-ray microscopy for multiscale structural characterization

DEFF Research Database (Denmark)

Simons, Hugh; King, A.; Ludwig, W.

2015-01-01

of the interactions between crystalline elements is a key step towards the formulation and validation of multiscale models that account for the entire heterogeneity of a material. Furthermore, dark-field X-ray microscopy is well suited to applied topics, where the structural evolution of internal nanoscale elements...

Two-Photon Fluorescence Microscopy Developed for Microgravity Fluid Physics

Science.gov (United States)

Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

2004-01-01

Recent research efforts within the Microgravity Fluid Physics Branch of the NASA Glenn Research Center have necessitated the development of a microscope capable of high-resolution, three-dimensional imaging of intracellular structure and tissue morphology. Standard optical microscopy works well for thin samples, but it does not allow the imaging of thick samples because of severe degradation caused by out-of-focus object structure. Confocal microscopy, which is a laser-based scanning microscopy, provides improved three-dimensional imaging and true optical sectioning by excluding the out-of-focus light. However, in confocal microscopy, out-of-focus object structure is still illuminated by the incoming beam, which can lead to substantial photo-bleaching. In addition, confocal microscopy is plagued by limited penetration depth, signal loss due to the presence of a confocal pinhole, and the possibility of live-cell damage. Two-photon microscopy is a novel form of laser-based scanning microscopy that allows three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon microscopy, it utilizes the nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption because of the nonlinear (i.e., quadratic) electric field dependence, so an ultrafast pulsed laser source must typically be employed. On the other hand, this stringent energy density requirement effectively localizes fluorophore excitation to the focal volume. Consequently, two-photon microscopy provides optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction in photo-damage because of the longer excitation wavelength, a reduction in background fluorescence, and a 4 increase in penetration depth over confocal methods because of the reduction in Rayleigh scattering.

Vector optical fields with polarization distributions similar to electric and magnetic field lines.

Science.gov (United States)

Pan, Yue; Li, Si-Min; Mao, Lei; Kong, Ling-Jun; Li, Yongnan; Tu, Chenghou; Wang, Pei; Wang, Hui-Tian

2013-07-01

We present, design and generate a new kind of vector optical fields with linear polarization distributions modeling to electric and magnetic field lines. The geometric configurations of "electric charges" and "magnetic charges" can engineer the spatial structure and symmetry of polarizations of vector optical field, providing additional degrees of freedom assisting in controlling the field symmetry at the focus and allowing engineering of the field distribution at the focus to the specific applications.

Establishing the suitability of quantitative optical CT microscopy of PRESAGE® radiochromic dosimeters for the verification of synchrotron microbeam therapy

Science.gov (United States)

Doran, Simon J.; Rahman, A. T. Abdul; Bräuer-Krisch, Elke; Brochard, Thierry; Adamovics, John; Nisbet, Andrew; Bradley, David

2013-09-01

Previous research on optical computed tomography (CT) microscopy in the context of the synchrotron microbeam has shown the potential of the technique and demonstrated high quality images, but has left two questions unanswered: (i) are the images suitably quantitative for 3D dosimetry? and (ii) what is the impact on the spatial resolution of the system of the limited depth-of-field of the microscope optics? Cuvette and imaging studies are reported here that address these issues. Two sets of cuvettes containing the radiochromic plastic PRESAGE® were irradiated at the ID17 biomedical beamline of the European Synchrotron Radiation facility over the ranges 0-20 and 0-35 Gy and a third set of cuvettes was irradiated over the range 0-20 Gy using a standard medical linac. In parallel, three cylindrical PRESAGE® samples of diameter 9.7 mm were irradiated with test patterns that allowed the quantitative capabilities of the optical CT microscope to be verified, and independent measurements of the imaging modulation transfer function (MTF) to be made via two different methods. Both spectrophotometric analysis and imaging gave a linear dose response, with gradients ranging from 0.036-0.041 cm-1 Gy-1 in the three sets of cuvettes and 0.037 (optical CT units) Gy-1 for the imaging. High-quality, quantitative imaging results were obtained throughout the 3D volume, as illustrated by depth-dose profiles. These profiles are shown to be monoexponential, and the linear attention coefficient of PRESAGE® for the synchrotron-generated x-ray beam is measured to be (0.185 ± 0.02) cm-1 in excellent agreement with expectations. Low-level (<5%) residual image artefacts are discussed in detail. It was possible to resolve easily slit patterns of width 37 µm (which are smaller than many of the microbeams used on ID-17), but some uncertainty remains as to whether the low values of MTF for the higher spatial frequencies are scanner related or a result of genuine (but non-ideal) dose

Laboratory source based full-field x-ray microscopy at 9 keV

Energy Technology Data Exchange (ETDEWEB)

Fella, C.; Balles, A.; Wiest, W. [Lehrstuhl für Röntgenmikroskopie, Julius-Maximilians-Universität, 97074 Würzburg (Germany); Zabler, S.; Hanke, R. [Lehrstuhl für Röntgenmikroskopie, Julius-Maximilians-Universität, 97074 Würzburg (Germany); Fraunhofer Development Center X-Ray Technology (EZRT), Flugplatzstrasse 75, 90768 Fürth (Germany)

2016-01-28

In the past decade, hard x-ray transmission microscopy experienced tremendous developments. With the avail-ability of efficient Fresnel zone plates, even set-ups utilizing laboratory sources were developed [1]. In order to improve the performance of these x-ray microscopes, novel approaches to fabricate optical elements [2] and brighter x-ray tubes [3] are promising candidates. We are currently building a laboratory transmission x-ray microscope for 9.25 keV, using an electron impact liquid-metal-jet anode source. Up to now, the further elements of our setup are: a polycapillary condenser, a tungsten zone plate, and a scintillator which is optically coupled to a CMOS camera. However, further variations in terms of optical elements are intended. Here we present the current status of our work, as well as first experimental results.

Optical design of an athermalised dual field of view step zoom optical system in MWIR

Science.gov (United States)

Kucukcelebi, Doruk

2017-08-01

In this paper, the optical design of an athermalised dual field of view step zoom optical system in MWIR (3.7μm - 4.8μm) is described. The dual field of view infrared optical system is designed based on the principle of passive athermalization method not only to achieve athermal optical system but also to keep the high image quality within the working temperature between -40°C and +60°C. The infrared optical system used in this study had a 320 pixel x 256 pixel resolution, 20μm pixel pitch size cooled MWIR focal plane array detector. In this study, the step zoom mechanism, which has the axial motion due to consisting of a lens group, is considered to simplify mechanical structure. The optical design was based on moving a single lens along the optical axis for changing the optical system's field of view not only to reduce the number of moving parts but also to athermalize for the optical system. The optical design began with an optimization process using paraxial optics when first-order optics parameters are determined. During the optimization process, in order to reduce aberrations, such as coma, astigmatism, spherical and chromatic aberrations, aspherical surfaces were used. As a result, athermalised dual field of view step zoom optical design is proposed and the performance of the design using proposed method was verified by providing the focus shifts, spot diagrams and MTF analyzes' plots.

Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope

Energy Technology Data Exchange (ETDEWEB)

Wang, Peng; Behan, Gavin; Kirkland, Angus I. [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Nellist, Peter D., E-mail: peter.nellist@materials.ox.ac.uk [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Cosgriff, Eireann C.; D' Alfonso, Adrian J.; Morgan, Andrew J.; Allen, Leslie J. [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Hashimoto, Ayako [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Takeguchi, Masaki [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Mitsuishi, Kazutaka [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Quantum Dot Research Center, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Shimojo, Masayuki [High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Advanced Science Research Laboratory, Saitama Institute of Technology, 1690 Fusaiji, Fukaya 369-0293 (Japan)

2011-06-15

Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored. -- Research Highlights: {yields} The confocal probe image in a scanning confocal electron microscopy image reveals information about the thickness and height of the crystalline layer. {yields} The form of the contrast in a three-dimensional bright-field scanning confocal electron microscopy image can be explained in terms of the confocal probe image. {yields} Despite the complicated form of the contrast in bright-field scanning confocal electron microscopy, we see that depth information is transferred on a 10 nm scale.

Dark field differential dynamic microscopy enables accurate characterization of the roto-translational dynamics of bacteria and colloidal clusters

Science.gov (United States)

Cerbino, Roberto; Piotti, Davide; Buscaglia, Marco; Giavazzi, Fabio

2018-01-01

Micro- and nanoscale objects with anisotropic shape are key components of a variety of biological systems and inert complex materials, and represent fundamental building blocks of novel self-assembly strategies. The time scale of their thermal motion is set by their translational and rotational diffusion coefficients, whose measurement may become difficult for relatively large particles with small optical contrast. Here we show that dark field differential dynamic microscopy is the ideal tool for probing the roto-translational Brownian motion of anisotropic shaped particles. We demonstrate our approach by successful application to aqueous dispersions of non-motile bacteria and of colloidal aggregates of spherical particles.

The optical analogy for vector fields

Science.gov (United States)

Parker, E. N. (Editor)

1991-01-01

This paper develops the optical analogy for a general vector field. The optical analogy allows the examination of certain aspects of a vector field that are not otherwise readily accessible. In particular, in the cases of a stationary Eulerian flow v of an ideal fluid and a magnetostatic field B, the vectors v and B have surface loci in common with their curls. The intrinsic discontinuities around local maxima in absolute values of v and B take the form of vortex sheets and current sheets, respectively, the former playing a fundamental role in the development of hydrodyamic turbulence and the latter playing a major role in heating the X-ray coronas of stars and galaxies.

Quantitative optical microscopy and micromanipulation studies on the lipid bilayer membranes of giant unilamellar vesicles

DEFF Research Database (Denmark)

Bagatolli, Luis; Needham, David

2014-01-01

to study composition-structure-property materials relationships of free-standing lipid bilayer membranes. Because their size (~5 to 100 m diameter) that is well above the resolution limit of regular light microscopes, GUVs are suitable membrane models for optical microscopy and micromanipulation...

Polarization resolved imaging with a reflection near-field optical microscope

DEFF Research Database (Denmark)

Bozhevolnyi, Sergey I.; Xiao, Mufei; Hvam, Jørn Märcher

1999-01-01

Using a rigorous microscopic point-dipole description of probe-sample interactions, we study imaging with a reflection scanning near-field optical microscope. Optical content, topographical artifacts, sensitivity window-i.e., the scale on which near-field optical images represent mainly optical...... configuration is preferable to the cross-linear one, since it ensures more isotropic (in the surface plane) near-field imaging of surface features. The numerical results are supported with experimental near-field images obtained by using a reflection microscope with an uncoated fiber tip....

Accurate Rapid Lifetime Determination on Time-Gated FLIM Microscopy with Optical Sectioning.

Science.gov (United States)

Silva, Susana F; Domingues, José Paulo; Morgado, António Miguel

2018-01-01

Time-gated fluorescence lifetime imaging microscopy (FLIM) is a powerful technique to assess the biochemistry of cells and tissues. When applied to living thick samples, it is hampered by the lack of optical sectioning and the need of acquiring many images for an accurate measurement of fluorescence lifetimes. Here, we report on the use of processing techniques to overcome these limitations, minimizing the acquisition time, while providing optical sectioning. We evaluated the application of the HiLo and the rapid lifetime determination (RLD) techniques for accurate measurement of fluorescence lifetimes with optical sectioning. HiLo provides optical sectioning by combining the high-frequency content from a standard image, obtained with uniform illumination, with the low-frequency content of a second image, acquired using structured illumination. Our results show that HiLo produces optical sectioning on thick samples without degrading the accuracy of the measured lifetimes. We also show that instrument response function (IRF) deconvolution can be applied with the RLD technique on HiLo images, improving greatly the accuracy of the measured lifetimes. These results open the possibility of using the RLD technique with pulsed diode laser sources to determine accurately fluorescence lifetimes in the subnanosecond range on thick multilayer samples, providing that offline processing is allowed.

High-Density Near-Field Optical Disc Recording

Science.gov (United States)

Shinoda, Masataka; Saito, Kimihiro; Ishimoto, Tsutomu; Kondo, Takao; Nakaoki, Ariyoshi; Ide, Naoki; Furuki, Motohiro; Takeda, Minoru; Akiyama, Yuji; Shimouma, Takashi; Yamamoto, Masanobu

2005-05-01

We developed a high-density near-field optical recording disc system using a solid immersion lens. The near-field optical pick-up consists of a solid immersion lens with a numerical aperture of 1.84. The laser wavelength for recording is 405 nm. In order to realize the near-field optical recording disc, we used a phase-change recording media and a molded polycarbonate substrate. A clear eye pattern of 112 GB capacity with 160 nm track pitch and 50 nm bit length was observed. The equivalent areal density is 80.6 Gbit/in2. The bottom bit error rate of 3 tracks-write was 4.5× 10-5. The readout power margin and the recording power margin were ± 30.4% and ± 11.2%, respectively.

Direct detection of the optical field beyond single polarization mode.

Science.gov (United States)

Che, Di; Sun, Chuanbowen; Shieh, William

2018-02-05

Direct detection is traditionally regarded as a detection method that recovers only the optical intensity. Compared with coherent detection, it owns a natural advantage-the simplicity-but lacks a crucial capability of field recovery that enables not only the multi-dimensional modulation, but also the digital compensation of the fiber impairments linear with the optical field. Full-field detection is crucial to increase the capacity-distance product of optical transmission systems. A variety of methods have been investigated to directly detect the optical field of the single polarization mode, which normally sends a carrier traveling with the signal for self-coherent detection. The crux, however, is that any optical transmission medium supports at least two propagating modes (e.g. single mode fiber supports two polarization modes), and until now there is no direct detection that can recover the complete set of optical fields beyond one polarization, due to the well-known carrier fading issue after mode demultiplexing induced by the random mode coupling. To avoid the fading, direct detection receivers should recover the signal in an intensity space isomorphic to the optical field without loss of any degrees of freedom, and a bridge should be built between the field and its isomorphic space for the multi-mode field recovery. Based on this thinking, we propose, for the first time, the direct detection of dual polarization modes by a novel receiver concept, the Stokes-space field receiver (SSFR) and its extension, the generalized SSFR for multiple spatial modes. The idea is verified by a dual-polarization field recovery of a polarization-multiplexed complex signal over an 80-km single mode fiber transmission. SSFR can be applied to a much wider range of fields beyond optical communications such as coherent sensing and imaging, where simple field recovery without an extra local laser is desired for enhanced system performance.

Building Practical Apertureless Scanning Near-Field Microscopy

Science.gov (United States)

Gungordu, M. Zeki

The fundamental objective of this study is to establish a functional, practical apertureless type scanning near-field optical microscope, and to figure out the working mechanism behind it. Whereas a far-field microscope can measure the propagating field's components, this gives us little information about the features of the sample. The resolution is limited to about half of the wavelength of the illuminating light. On the other hand, the a-SNOM system enables achieving non-propagating components of the field, which provides more details about the sample's features. It is really difficult to measure because the amplitude of this field decays exponentially when the tip is moved away from the sample. The sharpness of the tip is the only limitation for resolution of the a-SNOM system. Consequently, the sharp tips are achieved by using electrochemical etching, and these tips are used to detect near-field signal. Separating the weak a-SNOM system signals from the undesired background signal, the higher demodulation background suppression is utilized by lock-in detection.

Digital Holographic Microscopy Principles, Techniques, and Applications

CERN Document Server

Kim, Myung K

2011-01-01

Digital holography is an emerging field of new paradigm in general imaging applications. By replacing the photochemical procedures with electronic imaging and having a direct numerical access to the complex optical field, a wide range of new imaging capabilities become available, many of them difficult or infeasible in conventional holography. An increasing number of researchers—not only in optical physics and optical engineering, but also in diverse applications areas such as microbiology, medicine, marine science, particle analysis, microelectromechanics, and metrology—are realizing and exploiting the new capabilities of digital holography. Digital Holographic Microscopy: Principles, Techniques, and Applications, by Dr. Myung K. Kim, is intended to provide a brief but consistent introduction to the principles of digital holography as well as to give an organized overview of the large number of techniques and applications being developed. This will also shed some light on the range of possibilities for f...

Sub-40 fs, 1060-nm Yb-fiber laser enhances penetration depth in nonlinear optical microscopy of human skin

Science.gov (United States)

Balu, Mihaela; Saytashev, Ilyas; Hou, Jue; Dantus, Marcos; Tromberg, Bruce J.

2015-12-01

Advancing the practical utility of nonlinear optical microscopy requires continued improvement in imaging depth and contrast. We evaluated second-harmonic generation (SHG) and third-harmonic generation images from ex vivo human skin and showed that a sub-40 fs, 1060-nm Yb-fiber laser can enhance SHG penetration depth by up to 80% compared to a >100 fs, 800 nm Ti:sapphire source. These results demonstrate the potential of fiber-based laser systems to address a key performance limitation related to nonlinear optical microscopy (NLOM) technology while providing a low-barrier-to-access alternative to Ti:sapphire sources that could help accelerate the movement of NLOM into clinical practice.

Transmission electron and optical microscopy of the domain structure of Ni3B7O13Br ferroic boracite

International Nuclear Information System (INIS)

Castellanos-Guzman, A.G.; Trujillo-Torrez, M.; Czank, M.

2005-01-01

The study investigated the domain structure of nickel bromine boracite single crystals, by means of polarised-light in conjunction with transmission electron microscopy. Single crystals of Ni 3 B 7 O 13 Br were grown by chemical transport reactions in closed quartz ampoules, in the temperature range of 1130 K and were examined by polarising optical microscopy (PLM), and transmission electron microscopy (TEM). PLM was also used in order to study the behaviour of birefringence as a function of temperature. For TEM the single crystals were crushed and mounted on holey carbon films. Comparative electron microscope images were useful for revealing the domain structure of this fully ferroelectric/fully ferroelastic material previously observed between the crossed polars of an optical microscope. X-ray diffraction analysis of the crystal under study was performed at room temperature

Imaging optical fields below metal films and metal-dielectric waveguides by a scanning microscope

Science.gov (United States)

Zhu, Liangfu; Wang, Yong; Zhang, Douguo; Wang, Ruxue; Qiu, Dong; Wang, Pei; Ming, Hai; Badugu, Ramachandram; Rosenfeld, Mary; Lakowicz, Joseph R.

2017-09-01

Laser scanning confocal fluorescence microscopy (LSCM) is now an important method for tissue and cell imaging when the samples are located on the surfaces of glass slides. In the past decade, there has been extensive development of nano-optical structures that display unique effects on incident and transmitted light, which will be used with novel configurations for medical and consumer products. For these applications, it is necessary to characterize the light distribution within short distances from the structures for efficient detection and elimination of bulky optical components. These devices will minimize or possibly eliminate the need for free-space light propagation outside of the device itself. We describe the use of the scanning function of a LSCM to obtain 3D images of the light intensities below the surface of nano-optical structures. More specifically, we image the spatial distributions inside the substrate of fluorescence emission coupled to waveguide modes after it leaks through thin metal films or dielectric-coated metal films. The observed spatial distribution were in general agreement with far-field calculations, but the scanning images also revealed light intensities at angles not observed with classical back focal plane imaging. Knowledge of the subsurface optical intensities will be crucial in the combination of nano-optical structures with rapidly evolving imaging detectors.

Wide-field optical mapping of neural activity and brain haemodynamics: considerations and novel approaches

Science.gov (United States)

Ma, Ying; Shaik, Mohammed A.; Kozberg, Mariel G.; Thibodeaux, David N.; Zhao, Hanzhi T.; Yu, Hang

2016-01-01

Although modern techniques such as two-photon microscopy can now provide cellular-level three-dimensional imaging of the intact living brain, the speed and fields of view of these techniques remain limited. Conversely, two-dimensional wide-field optical mapping (WFOM), a simpler technique that uses a camera to observe large areas of the exposed cortex under visible light, can detect changes in both neural activity and haemodynamics at very high speeds. Although WFOM may not provide single-neuron or capillary-level resolution, it is an attractive and accessible approach to imaging large areas of the brain in awake, behaving mammals at speeds fast enough to observe widespread neural firing events, as well as their dynamic coupling to haemodynamics. Although such wide-field optical imaging techniques have a long history, the advent of genetically encoded fluorophores that can report neural activity with high sensitivity, as well as modern technologies such as light emitting diodes and sensitive and high-speed digital cameras have driven renewed interest in WFOM. To facilitate the wider adoption and standardization of WFOM approaches for neuroscience and neurovascular coupling research, we provide here an overview of the basic principles of WFOM, considerations for implementation of wide-field fluorescence imaging of neural activity, spectroscopic analysis and interpretation of results. This article is part of the themed issue ‘Interpreting BOLD: a dialogue between cognitive and cellular neuroscience’. PMID:27574312

Wide-field optical mapping of neural activity and brain haemodynamics: considerations and novel approaches.

Science.gov (United States)

Ma, Ying; Shaik, Mohammed A; Kim, Sharon H; Kozberg, Mariel G; Thibodeaux, David N; Zhao, Hanzhi T; Yu, Hang; Hillman, Elizabeth M C

2016-10-05

Although modern techniques such as two-photon microscopy can now provide cellular-level three-dimensional imaging of the intact living brain, the speed and fields of view of these techniques remain limited. Conversely, two-dimensional wide-field optical mapping (WFOM), a simpler technique that uses a camera to observe large areas of the exposed cortex under visible light, can detect changes in both neural activity and haemodynamics at very high speeds. Although WFOM may not provide single-neuron or capillary-level resolution, it is an attractive and accessible approach to imaging large areas of the brain in awake, behaving mammals at speeds fast enough to observe widespread neural firing events, as well as their dynamic coupling to haemodynamics. Although such wide-field optical imaging techniques have a long history, the advent of genetically encoded fluorophores that can report neural activity with high sensitivity, as well as modern technologies such as light emitting diodes and sensitive and high-speed digital cameras have driven renewed interest in WFOM. To facilitate the wider adoption and standardization of WFOM approaches for neuroscience and neurovascular coupling research, we provide here an overview of the basic principles of WFOM, considerations for implementation of wide-field fluorescence imaging of neural activity, spectroscopic analysis and interpretation of results.This article is part of the themed issue 'Interpreting BOLD: a dialogue between cognitive and cellular neuroscience'. © 2016 The Authors.

Compact diffraction phase microscopy for quantitative visualization of cells in biomedical applications

International Nuclear Information System (INIS)

Talaikova, N A; Ryabukho, V P

2016-01-01

We consider a simplified and compact scheme of interference phase microscopy using a diffraction grating and spatial filtering of the diffracted field, i.e., diffraction phase microscopy. The scheme and the parameters of the device with the possibility of using the optical system of a smartphone and its software are analysed. The results of experimental determination of the spatial structure parameters of erythrocytes are presented. (paper)

Structured Illumination-Based Super-Resolution Optical Microscopy for Hemato- and Cyto-Pathology Applications

Directory of Open Access Journals (Sweden)

Tieqiao Zhang

2013-01-01

Full Text Available Structured illumination fluorescence microscopy utilizes interfering light and the moiré effect to enhance spatial resolution to about a half of that of conventional light microscopy, i.e. approximately 90 nm. In addition to the enhancement in the x and y directions, it also allows enhancement of resolution in the z- direction by the same factor of two (to approximately 220 nm, making it a powerful tool for 3-D morphology studies of fluorescently labeled cells or thin tissue sections. In this report, we applied this technique to several types of blood cells that are commonly seen in hematopathology. Compared with standard brightfield and ordinary fluorescence microscopy images, the 3-D morphology results clearly reveal the morphological features of different types of normal blood cells. We have also used this technique to evaluate morphologies of abnormal erythrocytes and compare them with those recorded on normal cells. The results give a very intuitive presentation of morphological structures of erythrocytes with great details. This research illustrates the potential of this technique to be used in hematology and cyto-pathology studies aimed at identifying nanometer-sized features that cannot be distinguished otherwise with conventional optical microscopy.

Sensitivity and Specificity of Cardiac Tissue Discrimination Using Fiber-Optics Confocal Microscopy.

Science.gov (United States)

Huang, Chao; Sachse, Frank B; Hitchcock, Robert W; Kaza, Aditya K

2016-01-01

Disturbances of the cardiac conduction system constitute a major risk after surgical repair of complex cases of congenital heart disease. Intraoperative identification of the conduction system may reduce the incidence of these disturbances. We previously developed an approach to identify cardiac tissue types using fiber-optics confocal microscopy and extracellular fluorophores. Here, we applied this approach to investigate sensitivity and specificity of human and automated classification in discriminating images of atrial working myocardium and specialized tissue of the conduction system. Two-dimensional image sequences from atrial working myocardium and nodal tissue of isolated perfused rodent hearts were acquired using a fiber-optics confocal microscope (Leica FCM1000). We compared two methods for local application of extracellular fluorophores: topical via pipette and with a dye carrier. Eight blinded examiners evaluated 162 randomly selected images of atrial working myocardium (n = 81) and nodal tissue (n = 81). In addition, we evaluated the images using automated classification. Blinded examiners achieved a sensitivity and specificity of 99.2 ± 0.3% and 98.0 ± 0.7%, respectively, with the dye carrier method of dye application. Sensitivity and specificity was similar for dye application via a pipette (99.2 ± 0.3% and 94.0 ± 2.4%, respectively). Sensitivity and specificity for automated methods of tissue discrimination were similarly high. Human and automated classification achieved high sensitivity and specificity in discriminating atrial working myocardium and nodal tissue. We suggest that our findings facilitate clinical translation of fiber-optics confocal microscopy as an intraoperative imaging modality to reduce the incidence of conduction disturbances during surgical correction of congenital heart disease.

[Does dark field microscopy according to Enderlein allow for cancer diagnosis? A prospective study].

Science.gov (United States)

El-Safadi, Samer; Tinneberg, Hans-Rudolf; von Georgi, Richard; Münstedt, Karsten; Brück, Friede

2005-06-01

Dark field microscopy according to Enderlin claims to be able to detect forthcoming or beginning cancer at an early stage through minute abnormalities in the blood. In Germany and the USA, this method is used by an increasing number of physicians and health practitioners (non-medically qualified complementary practitioners), because this easy test seems to give important information about patients' health status. Can dark field microscopy reliably detect cancer? In the course of a prospective study on iridology, blood samples were drawn for dark field microscopy in 110 patients. A health practitioner with several years of training in the field carried out the examination without prior information about the patients. Out of 12 patients with present tumor metastasis as confirmed by radiological methods (CT, MRI or ultra-sound) 3 were correctly identified. Analysis of sensitivity (0.25), specificity (0.64), positive (0.09) and negative (0.85) predictive values revealed unsatisfactory results. Dark field micoroscopy does not seem to reliably detect the presence of cancer. Clinical use of the method can therefore not be recommended until future studies are conducted.

Tailoring optical complex field with spiral blade plasmonic vortex lens

Science.gov (United States)

Rui, Guanghao; Zhan, Qiwen; Cui, Yiping

2015-01-01

Optical complex fields have attracted increasing interests because of the novel effects and phenomena arising from the spatially inhomogeneous state of polarizations and optical singularities of the light beam. In this work, we propose a spiral blade plasmonic vortex lens (SBPVL) that offers unique opportunities to manipulate these novel fields. The strong interaction between the SBPVL and the optical complex fields enable the synthesis of highly tunable plasmonic vortex. Through theoretical derivations and numerical simulations we demonstrated that the characteristics of the plasmonic vortex are determined by the angular momentum (AM) of the light, and the geometrical topological charge of the SBPVL, which is govern by the nonlinear superposition of the pitch and the number of blade element. In addition, it is also shown that by adjusting the geometric parameters, SBPVL can be utilized to focus and manipulate optical complex field with fractional AM. This miniature plasmonic device may find potential applications in optical trapping, optical data storage and many other related fields. PMID:26335894

Photocurrent mapping of near-field optical antenna resonances

KAUST Repository

Barnard, Edward S.; Pala, Ragip A.; Brongersma, Mark L.

2011-01-01

An increasing number of photonics applications make use of nanoscale optical antennas that exhibit a strong, resonant interaction with photons of a specific frequency. The resonant properties of such antennas are conventionally characterized by far-field light-scattering techniques. However, many applications require quantitative knowledge of the near-field behaviour, and existing local field measurement techniques provide only relative, rather than absolute, data. Here, we demonstrate a photodetector platform that uses a silicon-on-insulator substrate to spectrally and spatially map the absolute values of enhanced fields near any type of optical antenna by transducing local electric fields into photocurrent. We are able to quantify the resonant optical and materials properties of nanoscale (∼50nm) and wavelength-scale (∼1μm) metallic antennas as well as high-refractive-index semiconductor antennas. The data agree well with light-scattering measurements, full-field simulations and intuitive resonator models. © 2011 Macmillan Publishers Limited. All rights reserved.

Photocurrent mapping of near-field optical antenna resonances

KAUST Repository

Barnard, Edward S.

2011-08-21

An increasing number of photonics applications make use of nanoscale optical antennas that exhibit a strong, resonant interaction with photons of a specific frequency. The resonant properties of such antennas are conventionally characterized by far-field light-scattering techniques. However, many applications require quantitative knowledge of the near-field behaviour, and existing local field measurement techniques provide only relative, rather than absolute, data. Here, we demonstrate a photodetector platform that uses a silicon-on-insulator substrate to spectrally and spatially map the absolute values of enhanced fields near any type of optical antenna by transducing local electric fields into photocurrent. We are able to quantify the resonant optical and materials properties of nanoscale (∼50nm) and wavelength-scale (∼1μm) metallic antennas as well as high-refractive-index semiconductor antennas. The data agree well with light-scattering measurements, full-field simulations and intuitive resonator models. © 2011 Macmillan Publishers Limited. All rights reserved.

Contributed review: Review of integrated correlative light and electron microscopy.

Science.gov (United States)

Timmermans, F J; Otto, C

2015-01-01

New developments in the field of microscopy enable to acquire increasing amounts of information from large sample areas and at an increased resolution. Depending on the nature of the technique, the information may reveal morphological, structural, chemical, and still other sample characteristics. In research fields, such as cell biology and materials science, there is an increasing demand to correlate these individual levels of information and in this way to obtain a better understanding of sample preparation and specific sample properties. To address this need, integrated systems were developed that combine nanometer resolution electron microscopes with optical microscopes, which produce chemically or label specific information through spectroscopy. The complementary information from electron microscopy and light microscopy presents an opportunity to investigate a broad range of sample properties in a correlated fashion. An important part of correlating the differences in information lies in bridging the different resolution and image contrast features. The trend to analyse samples using multiple correlated microscopes has resulted in a new research field. Current research is focused, for instance, on (a) the investigation of samples with nanometer scale distribution of inorganic and organic materials, (b) live cell analysis combined with electron microscopy, and (c) in situ spectroscopic and electron microscopy analysis of catalytic materials, but more areas will benefit from integrated correlative microscopy.

Contributed Review: Review of integrated correlative light and electron microscopy

International Nuclear Information System (INIS)

Timmermans, F. J.; Otto, C.

2015-01-01

New developments in the field of microscopy enable to acquire increasing amounts of information from large sample areas and at an increased resolution. Depending on the nature of the technique, the information may reveal morphological, structural, chemical, and still other sample characteristics. In research fields, such as cell biology and materials science, there is an increasing demand to correlate these individual levels of information and in this way to obtain a better understanding of sample preparation and specific sample properties. To address this need, integrated systems were developed that combine nanometer resolution electron microscopes with optical microscopes, which produce chemically or label specific information through spectroscopy. The complementary information from electron microscopy and light microscopy presents an opportunity to investigate a broad range of sample properties in a correlated fashion. An important part of correlating the differences in information lies in bridging the different resolution and image contrast features. The trend to analyse samples using multiple correlated microscopes has resulted in a new research field. Current research is focused, for instance, on (a) the investigation of samples with nanometer scale distribution of inorganic and organic materials, (b) live cell analysis combined with electron microscopy, and (c) in situ spectroscopic and electron microscopy analysis of catalytic materials, but more areas will benefit from integrated correlative microscopy

En face speckle reduction in optical coherence microscopy by frequency compounding.

Science.gov (United States)

Magnain, Caroline; Wang, Hui; Sakadžić, Sava; Fischl, Bruce; Boas, David A

2016-05-01

We report the use of frequency compounding to significantly reduce speckle noise in optical coherence microscopy, more specifically on the en face images. This method relies on the fact that the speckle patterns recorded from different wavelengths simultaneously are independent; hence their summation yields significant reduction in noise, with only a single acquisition. The results of our experiments with microbeads show that the narrow confocal parameter, due to a high numerical aperture objective, restricts the axial resolution loss that would otherwise theoretically broaden linearly with the number of optical frequency bands used. This speckle reduction scheme preserves the lateral resolution since it is performed on individual A-scans. Finally, we apply this technique to images of fixed human brain tissue, showing significant improvements in contrast-to-noise ratio with only moderate loss of axial resolution, in an effort to improve automatic three-dimensional detection of cells and fibers in the cortex.

Fault localization and analysis in semiconductor devices with optical-feedback infrared confocal microscopy

International Nuclear Information System (INIS)

Sarmiento, Raymund; Cemine, Vernon Julius; Tagaca, Imee Rose; Salvador, Arnel; Mar Blanca, Carlo; Saloma, Caesar

2007-01-01

We report on a cost-effective optical setup for characterizing light-emitting semiconductor devices with optical-feedback confocal infrared microscopy and optical beam-induced resistance change.We utilize the focused beam from an infrared laser diode to induce local thermal resistance changes across the surface of a biased integrated circuit (IC) sample. Variations in the multiple current paths are mapped by scanning the IC across the focused beam. The high-contrast current maps allow accurate differentiation of the functional and defective sites, or the isolation of the surface-emittingp-i-n devices in the IC. Optical beam-induced current (OBIC) is not generated since the incident beam energy is lower than the bandgap energy of the p-i-n device. Inhomogeneous current distributions in the IC become apparent without the strong OBIC background. They are located at a diffraction-limited resolution by referencing the current maps against the confocal reflectance image that is simultaneously acquired via optical-feedback detection. Our technique permits the accurate identification of metal and semiconductor sites as well as the classification of different metallic structures according to thickness, composition, or spatial inhomogeneity

Oxidation study by Moessbauer and optic microscopy of steels from boiler tubes used in sugar industry

International Nuclear Information System (INIS)

Fajardo, M.; Perez Alcazar, G.A.; Aguilar, Y.

1998-01-01

Optic microscopy and Moessbauer spectroscopy were used to study the fail and the inner rusted surface of two boiler tubes used in the sugar industry, respectively. The studied tubes, of the type ASTM A 192, were found to have cracks. By optic microscopy it was observed that the failure begins in the inner surface with circumferential cracking. Also, inside and around the surface close to the cracks a rusted layer was detected. Powder from these layers was collected for Moessbauer spectroscopy analysis. By this method the presence of two or three types of Fe oxides such as wuestite, magnetite and hematite, was proved. These results permit to conclude that the failure mechanism was the thermal fatigue due to a hot work in an O 2 -rich vapor atmosphere. The rusted products are stable at high temperatures

Visible light optical coherence microscopy imaging of the mouse cortex with femtoliter volume resolution

Science.gov (United States)

Merkle, Conrad W.; Chong, Shau Poh; Kho, Aaron M.; Zhu, Jun; Kholiqov, Oybek; Dubra, Alfredo; Srinivasan, Vivek J.

2018-02-01

Most flying-spot Optical Coherence Tomography (OCT) and Optical Coherence Microscopy (OCM) systems use a symmetric confocal geometry, where the detection path retraces the illumination path starting from and ending with the spatial mode of a single mode optical fiber. Here, we describe a visible light OCM instrument that breaks this symmetry to improve transverse resolution without sacrificing collection efficiency in scattering tissue. This was achieved by overfilling a 0.3 numerical aperture (NA) water immersion objective on the illumination path, while maintaining a conventional Gaussian mode detection path (1/e2 intensity diameter 0.82 Airy disks), enabling 1.1 μm full-width at half-maximum (FWHM) transverse resolution. At the same time, a 0.9 μm FWHM axial resolution in tissue, achieved by a broadband visible light source, enabled femtoliter volume resolution. We characterized this instrument according to paraxial coherent microscopy theory, and then used it to image the meningeal layers, intravascular red blood cell-free layer, and myelinated axons in the mouse neocortex in vivo through the thinned skull. Finally, by introducing a 0.8 NA water immersion objective, we improved the lateral resolution to 0.44 μm FWHM, which provided a volumetric resolution of 0.2 fL, revealing cell bodies in cortical layer I of the mouse brain with OCM for the first time.

Optical currents in vector fields

DEFF Research Database (Denmark)

Angelsky, O. V.; Gorsky, M. P.; Maksimyak, P. P.

2011-01-01

The influence of phase relations and the degree of mutual coherence of superimposing waves in the arrangements of twowave superposition on the characteristics of the microparticle's motion has been analyzed. The prospects of studying temporal coherence using the proposed approach are made. For th....... For the first time, we have shown experimentally the possibility of diagnostics the optical currents in liquids caused by polarization characteristics of an optical field alone, using test metallic particles of nanoscale....

Theory of aberration fields for general optical systems with freeform surfaces.

Science.gov (United States)

Fuerschbach, Kyle; Rolland, Jannick P; Thompson, Kevin P

2014-11-03

This paper utilizes the framework of nodal aberration theory to describe the aberration field behavior that emerges in optical systems with freeform optical surfaces, particularly φ-polynomial surfaces, including Zernike polynomial surfaces, that lie anywhere in the optical system. If the freeform surface is located at the stop or pupil, the net aberration contribution of the freeform surface is field constant. As the freeform optical surface is displaced longitudinally away from the stop or pupil of the optical system, the net aberration contribution becomes field dependent. It is demonstrated that there are no new aberration types when describing the aberration fields that arise with the introduction of freeform optical surfaces. Significantly it is shown that the aberration fields that emerge with the inclusion of freeform surfaces in an optical system are exactly those that have been described by nodal aberration theory for tilted and decentered optical systems. The key contribution here lies in establishing the field dependence and nodal behavior of each freeform term that is essential knowledge for effective application to optical system design. With this development, the nodes that are distributed throughout the field of view for each aberration type can be anticipated and targeted during optimization for the correction or control of the aberrations in an optical system with freeform surfaces. This work does not place any symmetry constraints on the optical system, which could be packaged in a fully three dimensional geometry, without fold mirrors.

Experimental Investigation of Integrated Optical Intensive Impulse Electric Field Sensors

International Nuclear Information System (INIS)

Bao, Sun; Fu-Shen, Chen

2009-01-01

We design and fabricate an integrated optical electric field sensor with segmented electrode for intensive impulse electric field measurement. The integrated optical sensor is based on a Mach–Zehnder interferometer with segmented electrodes. The output/input character of the sensing system is analysed and measured. The maximal detectable electric field range (−75 kV/m to 245 kV/m) is obtained by analysing the results. As a result, the integrated optics electric field sensing system is suitable for transient intensive electric field measurement investigation

Development of hard X-ray dark-field microscope using full-field optics

International Nuclear Information System (INIS)

Takano, Hidekazu; Azuma, Hiroaki; Shimomura, Sho; Tsuji, Takuya; Tsusaka, Yoshiyuki; Kagoshima, Yasushi

2016-01-01

We develop a dark-field X-ray microscope using full-field optics based on a synchrotron beamline. Our setup consists of a condenser system and a microscope objective with an angular acceptance larger than that of the condenser. The condenser system is moved downstream from its regular position such that the focus of the condenser is behind the objective. The dark-field microscope optics are configured by excluding the converging beam from the condenser at the focal point. The image properties of the system are evaluated by observing and calculating a Siemens star test chart with 10 keV X-rays. Our setup allows easy switching to bright-field imaging. (author)

Low-cost mobile phone microscopy with a reversed mobile phone camera lens.

Directory of Open Access Journals (Sweden)

Neil A Switz

Full Text Available The increasing capabilities and ubiquity of mobile phones and their associated digital cameras offer the possibility of extending low-cost, portable diagnostic microscopy to underserved and low-resource areas. However, mobile phone microscopes created by adding magnifying optics to the phone's camera module have been unable to make use of the full image sensor due to the specialized design of the embedded camera lens, exacerbating the tradeoff between resolution and field of view inherent to optical systems. This tradeoff is acutely felt for diagnostic applications, where the speed and cost of image-based diagnosis is related to the area of the sample that can be viewed at sufficient resolution. Here we present a simple and low-cost approach to mobile phone microscopy that uses a reversed mobile phone camera lens added to an intact mobile phone to enable high quality imaging over a significantly larger field of view than standard microscopy. We demonstrate use of the reversed lens mobile phone microscope to identify red and white blood cells in blood smears and soil-transmitted helminth eggs in stool samples.

Low-cost mobile phone microscopy with a reversed mobile phone camera lens.

Science.gov (United States)

Switz, Neil A; D'Ambrosio, Michael V; Fletcher, Daniel A

2014-01-01

The increasing capabilities and ubiquity of mobile phones and their associated digital cameras offer the possibility of extending low-cost, portable diagnostic microscopy to underserved and low-resource areas. However, mobile phone microscopes created by adding magnifying optics to the phone's camera module have been unable to make use of the full image sensor due to the specialized design of the embedded camera lens, exacerbating the tradeoff between resolution and field of view inherent to optical systems. This tradeoff is acutely felt for diagnostic applications, where the speed and cost of image-based diagnosis is related to the area of the sample that can be viewed at sufficient resolution. Here we present a simple and low-cost approach to mobile phone microscopy that uses a reversed mobile phone camera lens added to an intact mobile phone to enable high quality imaging over a significantly larger field of view than standard microscopy. We demonstrate use of the reversed lens mobile phone microscope to identify red and white blood cells in blood smears and soil-transmitted helminth eggs in stool samples.

Observer Performance in the Use of Digital and Optical Microscopy for the Interpretation of Tissue-Based Biomarkers

Directory of Open Access Journals (Sweden)

Marios A. Gavrielides

2014-01-01

Full Text Available Background. We conducted a validation study of digital pathology for the quantitative assessment of tissue-based biomarkers with immunohistochemistry. Objective.\tTo examine observer agreement as a function of viewing modality (digital versus optical microscopy, whole slide versus tissue microarray (TMA review, biomarker type (HER2 incorporating membranous staining and Ki-67 with nuclear staining, and data type (continuous and categorical. Methods.\tEight pathologists reviewed 50 breast cancer whole slides (25 stained with HER2 and 25 with Ki-67 and 2 TMAs (1 stained with HER2, 1 with Ki-67, each containing 97 cores, using digital and optical microscopy. Results. Results showed relatively high overall interobserver and intermodality agreement, with different patterns specific to biomarker type. For HER2, there was better interobserver agreement for optical compared to digital microscopy for whole slides as well as better interobserver and intermodality agreement for TMAs. For Ki-67, those patterns were not observed. Conclusions. The differences in agreement patterns when examining different biomarkers and different scoring methods and reviewing whole slides compared to TMA stress the need for validation studies focused on specific pathology tasks to eliminate sources of variability that might dilute findings. The statistical uncertainty observed in our analyses calls for adequate sampling for each individual task rather than pooling cases.

Advanced Microscopy of Microbial Cells

DEFF Research Database (Denmark)

Haagensen, Janus Anders Juul; Regenberg, Birgitte; Sternberg, Claus

2011-01-01

microscopy, super-resolution optical microscopy (STED, SIM, PALM) as well as atomic force microscopy and Raman spectroscopy. Using examples of bistability in microbial populations as well as biofilm development and differentiation in bacterial and yeast consortia, we demonstrate the importance of microscopy...

A minimal optical trapping and imaging microscopy system.

Directory of Open Access Journals (Sweden)

Carmen Noemí Hernández Candia

Full Text Available We report the construction and testing of a simple and versatile optical trapping apparatus, suitable for visualizing individual microtubules (∼25 nm in diameter and performing single-molecule studies, using a minimal set of components. This design is based on a conventional, inverted microscope, operating under plain bright field illumination. A single laser beam enables standard optical trapping and the measurement of molecular displacements and forces, whereas digital image processing affords real-time sample visualization with reduced noise and enhanced contrast. We have tested our trapping and imaging instrument by measuring the persistence length of individual double-stranded DNA molecules, and by following the stepping of single kinesin motor proteins along clearly imaged microtubules. The approach presented here provides a straightforward alternative for studies of biomaterials and individual biomolecules.

High-resolution electron microscopy

CERN Document Server

Spence, John C H

2013-01-01

This new fourth edition of the standard text on atomic-resolution transmission electron microscopy (TEM) retains previous material on the fundamentals of electron optics and aberration correction, linear imaging theory (including wave aberrations to fifth order) with partial coherence, and multiple-scattering theory. Also preserved are updated earlier sections on practical methods, with detailed step-by-step accounts of the procedures needed to obtain the highest quality images of atoms and molecules using a modern TEM or STEM electron microscope. Applications sections have been updated - these include the semiconductor industry, superconductor research, solid state chemistry and nanoscience, and metallurgy, mineralogy, condensed matter physics, materials science and material on cryo-electron microscopy for structural biology. New or expanded sections have been added on electron holography, aberration correction, field-emission guns, imaging filters, super-resolution methods, Ptychography, Ronchigrams, tomogr...

A hard X-ray nanoprobe beamline for nanoscale microscopy

Energy Technology Data Exchange (ETDEWEB)

Winarski, Robert P., E-mail: winarski@anl.gov; Holt, Martin V. [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States); Rose, Volker [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States); Fuesz, Peter; Carbaugh, Dean; Benson, Christa; Shu, Deming; Kline, David; Stephenson, G. Brian; McNulty, Ian [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States); Maser, Jörg [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States)

2012-11-01

The Hard X-ray Nanoprobe Beamline is a precision platform for scanning probe and full-field microscopy with 3–30 keV X-rays. A combination of high-stability X-ray optics and precision motion sensing and control enables detailed studies of the internal features of samples with resolutions approaching 30 nm. The Hard X-ray Nanoprobe Beamline (or Nanoprobe Beamline) is an X-ray microscopy facility incorporating diffraction, fluorescence and full-field imaging capabilities designed and operated by the Center for Nanoscale Materials and the Advanced Photon Source at Sector 26 of the Advanced Photon Source at Argonne National Laboratory. This facility was constructed to probe the nanoscale structure of biological, environmental and material sciences samples. The beamline provides intense focused X-rays to the Hard X-ray Nanoprobe (or Nanoprobe) which incorporates Fresnel zone plate optics and a precision laser sensing and control system. The beamline operates over X-ray energies from 3 to 30 keV, enabling studies of most elements in the periodic table, with a particular emphasis on imaging transition metals.

Optical spectroscopy and microscopy of radiation-induced light-emitting point defects in lithium fluoride crystals and films

Science.gov (United States)

Montereali, R. M.; Bonfigli, F.; Menchini, F.; Vincenti, M. A.

2012-08-01

Broad-band light-emitting radiation-induced F2 and F3+ electronic point defects, which are stable and laser-active at room temperature in lithium fluoride crystals and films, are used in dosimeters, tuneable color-center lasers, broad-band miniaturized light sources and novel radiation imaging detectors. A brief review of their photoemission properties is presented, and their behavior at liquid nitrogen temperatures is discussed. Some experimental data from optical spectroscopy and fluorescence microscopy of these radiation-induced point defects in LiF crystals and thin films are used to obtain information about the coloration curves, the efficiency of point defect formation, the effects of photo-bleaching processes, etc. Control of the local formation, stabilization, and transformation of radiation-induced light-emitting defect centers is crucial for the development of optically active micro-components and nanostructures. Some of the advantages of low temperature measurements for novel confocal laser scanning fluorescence microscopy techniques, widely used for spatial mapping of these point defects through the optical reading of their visible photoluminescence, are highlighted.

Super-resolution imaging of subcortical white matter using stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI)

Science.gov (United States)

Hainsworth, A. H.; Lee, S.; Patel, A.; Poon, W. W.; Knight, A. E.

2018-01-01

Aims The spatial resolution of light microscopy is limited by the wavelength of visible light (the ‘diffraction limit’, approximately 250 nm). Resolution of sub-cellular structures, smaller than this limit, is possible with super resolution methods such as stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI). We aimed to resolve subcellular structures (axons, myelin sheaths and astrocytic processes) within intact white matter, using STORM and SOFI. Methods Standard cryostat-cut sections of subcortical white matter from donated human brain tissue and from adult rat and mouse brain were labelled, using standard immunohistochemical markers (neurofilament-H, myelin-associated glycoprotein, glial fibrillary acidic protein, GFAP). Image sequences were processed for STORM (effective pixel size 8–32 nm) and for SOFI (effective pixel size 80 nm). Results In human, rat and mouse, subcortical white matter high-quality images for axonal neurofilaments, myelin sheaths and filamentous astrocytic processes were obtained. In quantitative measurements, STORM consistently underestimated width of axons and astrocyte processes (compared with electron microscopy measurements). SOFI provided more accurate width measurements, though with somewhat lower spatial resolution than STORM. Conclusions Super resolution imaging of intact cryo-cut human brain tissue is feasible. For quantitation, STORM can under-estimate diameters of thin fluorescent objects. SOFI is more robust. The greatest limitation for super-resolution imaging in brain sections is imposed by sample preparation. We anticipate that improved strategies to reduce autofluorescence and to enhance fluorophore performance will enable rapid expansion of this approach. PMID:28696566

Super-resolution imaging of subcortical white matter using stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI).

Science.gov (United States)

Hainsworth, A H; Lee, S; Foot, P; Patel, A; Poon, W W; Knight, A E

2017-07-11

The spatial resolution of light microscopy is limited by the wavelength of visible light (the 'diffraction limit', approximately 250 nm). Resolution of sub-cellular structures, smaller than this limit, is possible with super resolution methods such as stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI). We aimed to resolve subcellular structures (axons, myelin sheaths and astrocytic processes) within intact white matter, using STORM and SOFI. Standard cryostat-cut sections of subcortical white matter from donated human brain tissue and from adult rat and mouse brain were labelled, using standard immunohistochemical markers (neurofilament-H, myelin-associated glycoprotein, glial fibrillary acidic protein, GFAP). Image sequences were processed for STORM (effective pixel size 8-32 nm) and for SOFI (effective pixel size 80 nm). In human, rat and mouse, subcortical white matter high-quality images for axonal neurofilaments, myelin sheaths and filamentous astrocytic processes were obtained. In quantitative measurements, STORM consistently underestimated width of axons and astrocyte processes (compared with electron microscopy measurements). SOFI provided more accurate width measurements, though with somewhat lower spatial resolution than STORM. Super resolution imaging of intact cryo-cut human brain tissue is feasible. For quantitation, STORM can under-estimate diameters of thin fluorescent objects. SOFI is more robust. The greatest limitation for super-resolution imaging in brain sections is imposed by sample preparation. We anticipate that improved strategies to reduce autofluorescence and to enhance fluorophore performance will enable rapid expansion of this approach. © 2017 British Neuropathological Society.

Dual-wavelength optical-resolution photoacoustic microscopy for cells with gold nanoparticle bioconjugates in three-dimensional cultures

Science.gov (United States)

Lee, Po-Yi; Liu, Wei-Wen; Chen, Shu-Ching; Li, Pai-Chi

2016-03-01

Three-dimensional (3D) in vitro models bridge the gap between typical two-dimensional cultures and in vivo conditions. However, conventional optical imaging methods such as confocal microscopy and two-photon microscopy cannot accurately depict cellular processing in 3D models due to limited penetration of photons. We developed a dualwavelength optical-resolution photoacoustic microscopy (OR-PAM), which provides sufficient penetration depth and spatial resolution, for studying CD8+ cytotoxic T lymphocytes (CTLs) trafficking in an in vitro 3D tumor microenvironment. CTLs play a cardinal role in host defense against tumor. Efficient trafficking of CTLs to the tumor microenvironment is a critical step for cancer immunotherapy. For the proposed system, gold nanospheres and indocyanine green (ICG) have been remarkable choices for contrast agents for photoacoustic signals due to their excellent biocompatibility and high optical absorption. With distinct absorption spectrums, targeted cells with gold nanospheres and ICG respectively can be identified by switching 523-nm and 800-nm laser irradiation. Moreover, we use an x-y galvanometer scanner to obtain high scanning rate. In the developed system, lateral and axial resolutions were designed at 1.6 μm and 5 μm, respectively. We successfully showed that dual-spectral OR-PAM can map either the distribution of CTLs with gold nanospheres at a visible wavelength of 523 nm or the 3D structure of tumor spheres with ICG in an in vitro 3D microenvironment. Our OR-PAM can provide better biological relevant information in cellular interaction and is potential for preclinical screening of anti-cancer drugs.

Segmentation of Drosophila Heart in Optical Coherence Microscopy Images Using Convolutional Neural Networks

OpenAIRE

Duan, Lian; Qin, Xi; He, Yuanhao; Sang, Xialin; Pan, Jinda; Xu, Tao; Men, Jing; Tanzi, Rudolph E.; Li, Airong; Ma, Yutao; Zhou, Chao

2018-01-01

Convolutional neural networks are powerful tools for image segmentation and classification. Here, we use this method to identify and mark the heart region of Drosophila at different developmental stages in the cross-sectional images acquired by a custom optical coherence microscopy (OCM) system. With our well-trained convolutional neural network model, the heart regions through multiple heartbeat cycles can be marked with an intersection over union (IOU) of ~86%. Various morphological and dyn...

Spatiotemporal polarization modulation microscopy with a microretarder array

Science.gov (United States)

Ding, Changqin; Ulcickas, James R. W.; Simpson, Garth J.

2018-02-01

A patterned microretarder array positioned in the rear conjugate plane of a microscope enables rapid polarizationdependent nonlinear optical microscopy. The pattern introduced to the array results in periodic modulation of the polarization-state of the incident light as a function of position within the field of view with no moving parts or active control. Introduction of a single stationary optical element and a fixed polarizer into the beam of a nonlinear optical microscope enabled nonlinear optical tensor recovery, which informs on local structure and orientation. Excellent agreement was observed between the measured and predicted second harmonic generation (SHG) of z-cut quartz, selected as a test system with well-established nonlinear optical properties. Subsequent studies of spatially varying samples further support the general applicability of this relatively simple strategy for detailed polarization analysis in both conventional and nonlinear optical imaging of structurally diverse samples.

Dynamic contrast enhancement in widefield microscopy using projector-generated illumination patterns

International Nuclear Information System (INIS)

Samson, Edward Carlo; Blanca, Carlo Mar

2007-01-01

We present a simple and cost-effective optical protocol to realize contrast-enhancement imaging (such as dark-field, optical-staining and oblique illumination microscopy) of transparent samples on a conventional widefield microscope using commercial multimedia projectors. The projector functions as both light source and mask generator implemented by creating slideshows of the filters projected along the illumination planes of the microscope. The projected optical masks spatially modulate the distribution of the incident light to selectively enhance structures within the sample according to spatial frequency thereby increasing the image contrast of translucent biological specimens. Any amplitude filter can be customized and dynamically controlled so that switching from one imaging modality to another involves a simple slide transition and can be executed at a keystroke with no physical filters and no moving optical parts. The method yields an image contrast of 89-96% comparable with standard enhancement techniques. The polarization properties of the projector are then utilized to discriminate birefringent and non-birefringent sites on the sample using single-shot, simultaneous polarization and optical-staining microscopy. In addition to dynamic pattern generation and polarization, the projector also provides high illumination power and spectral excitation selectivity through its red-green-blue (RGB) channels. We exploit this last property to explore the feasibility of using video projectors to selectively excite stained samples and perform fluorescence imaging in tandem with reflectance and polarization reflectance microscopy

Selective sensitivity in Kerr microscopy.

Science.gov (United States)

Soldatov, I V; Schäfer, R

2017-07-01

A new technique for contrast separation in wide-field magneto-optical Kerr microscopy is introduced. Utilizing the light from eight light emitting diodes, guided to the microscope by glass fibers and being switched synchronously with the camera exposure, domain images with orthogonal in-plane sensitivity can be displayed simultaneously at real-time, and images with pure in-plane or polar contrast can be obtained. The benefit of this new method of contrast separation is demonstrated for Permalloy films, a NdFeB sinter magnet, and a cobalt crystal. Moreover, the new technique is shown to strongly enhance the sensitivity of Kerr microscopy by eliminating parasitic contrast contributions occurring in conventional setups. A doubling of the in-plane domain contrast and a sensitivity to Kerr rotations as low as 0.6 mdeg is demonstrated.

Selective sensitivity in Kerr microscopy

Science.gov (United States)

Soldatov, I. V.; Schäfer, R.

2017-07-01

A new technique for contrast separation in wide-field magneto-optical Kerr microscopy is introduced. Utilizing the light from eight light emitting diodes, guided to the microscope by glass fibers and being switched synchronously with the camera exposure, domain images with orthogonal in-plane sensitivity can be displayed simultaneously at real-time, and images with pure in-plane or polar contrast can be obtained. The benefit of this new method of contrast separation is demonstrated for Permalloy films, a NdFeB sinter magnet, and a cobalt crystal. Moreover, the new technique is shown to strongly enhance the sensitivity of Kerr microscopy by eliminating parasitic contrast contributions occurring in conventional setups. A doubling of the in-plane domain contrast and a sensitivity to Kerr rotations as low as 0.6 mdeg is demonstrated.

Investigation of autofocus algorithms for brightfield microscopy of unstained cells

Science.gov (United States)

Wu, Shu Yu; Dugan, Nazim; Hennelly, Bryan M.

2014-05-01

In the past decade there has been significant interest in image processing for brightfield cell microscopy. Much of the previous research on image processing for microscopy has focused on fluorescence microscopy, including cell counting, cell tracking, cell segmentation and autofocusing. Fluorescence microscopy provides functional image information that involves the use of labels in the form of chemical stains or dyes. For some applications, where the biochemical integrity of the cell is required to remain unchanged so that sensitive chemical testing can later be applied, it is necessary to avoid staining. For this reason the challenge of processing images of unstained cells has become a topic of increasing attention. These cells are often effectively transparent and appear to have a homogenous intensity profile when they are in focus. Bright field microscopy is the most universally available and most widely used form of optical microscopy and for this reason we are interested in investigating image processing of unstained cells recorded using a standard bright field microscope. In this paper we investigate the application of a range of different autofocus metrics applied to unstained bladder cancer cell lines using a standard inverted bright field microscope with microscope objectives that have high magnification and numerical aperture. We present a number of conclusions on the optimum metrics and the manner in which they should be applied for this application.

Analytic Optimization of Near-Field Optical Chirality Enhancement

Science.gov (United States)

2017-01-01

We present an analytic derivation for the enhancement of local optical chirality in the near field of plasmonic nanostructures by tuning the far-field polarization of external light. We illustrate the results by means of simulations with an achiral and a chiral nanostructure assembly and demonstrate that local optical chirality is significantly enhanced with respect to circular polarization in free space. The optimal external far-field polarizations are different from both circular and linear. Symmetry properties of the nanostructure can be exploited to determine whether the optimal far-field polarization is circular. Furthermore, the optimal far-field polarization depends on the frequency, which results in complex-shaped laser pulses for broadband optimization. PMID:28239617

Magnetic imaging with full-field soft X-ray microscopies

International Nuclear Information System (INIS)

Fischer, Peter; Im, Mi-Young; Baldasseroni, Chloe; Bordel, Catherine; Hellman, Frances; Lee, Jong-Soo; Fadley, Charles S.

2013-01-01

Progress toward a fundamental understanding of magnetism continues to be of great scientific interest and high technological relevance. To control magnetization on the nanoscale, external magnetic fields and spin polarized currents are commonly used. In addition, novel concepts based on spin manipulation by electric fields or photons are emerging which benefit from advances in tailoring complex magnetic materials. Although the nanoscale is at the very origin of magnetic behavior, there is a new trend toward investigating mesoscale magnetic phenomena, thus adding complexity and functionality, both of which will become crucial for future magnetic devices. Advanced analytical tools are thus needed for the characterization of magnetic properties spanning the nano- to the meso-scale. Imaging magnetic structures with high spatial and temporal resolution over a large field of view and in three dimensions is therefore a key challenge. A variety of spectromicroscopic techniques address this challenge by taking advantage of variable-polarization soft X-rays, thus enabling X-ray dichroism effects provide magnetic contrast. These techniques are also capable of quantifying in an element-, valence- and site-sensitive way the basic properties of ferro(i)- and antiferro-magnetic systems, such as spin and orbital moments, spin configurations from the nano- to the meso-scale and spin dynamics with sub-ns time resolution. This paper reviews current achievements and outlines future trends with one of these spectromicroscopies, magnetic full field transmission soft X-ray microscopy (MTXM) using a few selected examples of recent research on nano- and meso-scale magnetic phenomena. The complementarity of MTXM to X-ray photoemission electron microscopy (X-PEEM) is also emphasized

Magnetic imaging with full-field soft X-ray microscopies

Energy Technology Data Exchange (ETDEWEB)

Fischer, Peter, E-mail: PJFischer@lbl.gov [Center for X-ray Optics, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States); Im, Mi-Young [Center for X-ray Optics, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States); Baldasseroni, Chloe [Department of Materials Science and Engineering, University of California Berkeley, Berkeley, CA 94720 (United States); Bordel, Catherine; Hellman, Frances [Department of Physics, University of California Berkeley, Berkeley, CA 94720 (United States); Material Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94270 (United States); Lee, Jong-Soo [Department of Energy Systems Engineering, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 711-873 (Korea, Republic of); Fadley, Charles S. [Department of Physics, University of California Davis, Davis, CA 95616 (United States); Material Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94270 (United States)

2013-08-15

Progress toward a fundamental understanding of magnetism continues to be of great scientific interest and high technological relevance. To control magnetization on the nanoscale, external magnetic fields and spin polarized currents are commonly used. In addition, novel concepts based on spin manipulation by electric fields or photons are emerging which benefit from advances in tailoring complex magnetic materials. Although the nanoscale is at the very origin of magnetic behavior, there is a new trend toward investigating mesoscale magnetic phenomena, thus adding complexity and functionality, both of which will become crucial for future magnetic devices. Advanced analytical tools are thus needed for the characterization of magnetic properties spanning the nano- to the meso-scale. Imaging magnetic structures with high spatial and temporal resolution over a large field of view and in three dimensions is therefore a key challenge. A variety of spectromicroscopic techniques address this challenge by taking advantage of variable-polarization soft X-rays, thus enabling X-ray dichroism effects provide magnetic contrast. These techniques are also capable of quantifying in an element-, valence- and site-sensitive way the basic properties of ferro(i)- and antiferro-magnetic systems, such as spin and orbital moments, spin configurations from the nano- to the meso-scale and spin dynamics with sub-ns time resolution. This paper reviews current achievements and outlines future trends with one of these spectromicroscopies, magnetic full field transmission soft X-ray microscopy (MTXM) using a few selected examples of recent research on nano- and meso-scale magnetic phenomena. The complementarity of MTXM to X-ray photoemission electron microscopy (X-PEEM) is also emphasized.

Massively parallel data processing for quantitative total flow imaging with optical coherence microscopy and tomography

Science.gov (United States)

Sylwestrzak, Marcin; Szlag, Daniel; Marchand, Paul J.; Kumar, Ashwin S.; Lasser, Theo

2017-08-01

We present an application of massively parallel processing of quantitative flow measurements data acquired using spectral optical coherence microscopy (SOCM). The need for massive signal processing of these particular datasets has been a major hurdle for many applications based on SOCM. In view of this difficulty, we implemented and adapted quantitative total flow estimation algorithms on graphics processing units (GPU) and achieved a 150 fold reduction in processing time when compared to a former CPU implementation. As SOCM constitutes the microscopy counterpart to spectral optical coherence tomography (SOCT), the developed processing procedure can be applied to both imaging modalities. We present the developed DLL library integrated in MATLAB (with an example) and have included the source code for adaptations and future improvements. Catalogue identifier: AFBT_v1_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/AFBT_v1_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: GNU GPLv3 No. of lines in distributed program, including test data, etc.: 913552 No. of bytes in distributed program, including test data, etc.: 270876249 Distribution format: tar.gz Programming language: CUDA/C, MATLAB. Computer: Intel x64 CPU, GPU supporting CUDA technology. Operating system: 64-bit Windows 7 Professional. Has the code been vectorized or parallelized?: Yes, CPU code has been vectorized in MATLAB, CUDA code has been parallelized. RAM: Dependent on users parameters, typically between several gigabytes and several tens of gigabytes Classification: 6.5, 18. Nature of problem: Speed up of data processing in optical coherence microscopy Solution method: Utilization of GPU for massively parallel data processing Additional comments: Compiled DLL library with source code and documentation, example of utilization (MATLAB script with raw data) Running time: 1,8 s for one B-scan (150 × faster in comparison to the CPU

Optimizing Nanoscale Quantitative Optical Imaging of Subfield Scattering Targets

Science.gov (United States)

Henn, Mark-Alexander; Barnes, Bryan M.; Zhou, Hui; Sohn, Martin; Silver, Richard M.

2016-01-01

The full 3-D scattered field above finite sets of features has been shown to contain a continuum of spatial frequency information, and with novel optical microscopy techniques and electromagnetic modeling, deep-subwavelength geometrical parameters can be determined. Similarly, by using simulations, scattering geometries and experimental conditions can be established to tailor scattered fields that yield lower parametric uncertainties while decreasing the number of measurements and the area of such finite sets of features. Such optimized conditions are reported through quantitative optical imaging in 193 nm scatterfield microscopy using feature sets up to four times smaller in area than state-of-the-art critical dimension targets. PMID:27805660

Oxidation study by Mössbauer and optic microscopy of steels from boiler tubes used in sugar industry

Science.gov (United States)

Fajardo, M.; Pérez Alcázar, G. A.; Aguilar, Y.

1998-08-01

Optic microscopy and Mössbauer spectroscopy were used to study the fail and the inner rusted surface of two boiler tubes used in the sugar industry, respectively. The studied tubes, of the type ASTM A 192, were found to have cracks. By optic microscopy it was observed that the failure begins in the inner surface with circumferential cracking. Also, inside and around the surface close to the cracks a rusted layer was detected. Powder from these layers was collected for Mössbauer spectroscopy analysis. By this method the presence of two or three types of Fe oxides such as wüstite, magnetite and hematite, was proved. These results permit to conclude that the failure mechanism was the thermal fatigue due to a hot work in an O2 -rich vapor atmosphere. The rusted products are stable at high temperatures.

Advanced MOKE magnetometry in wide-field Kerr-microscopy

Science.gov (United States)

Soldatov, I. V.; Schäfer, R.

2017-10-01

The measurement of MOKE (Magneto-Optical Kerr Effect) magnetization loops in a wide-field Kerr microscope offers the advantage that the relevant domain images along the loop can be readily recorded. As the microscope's objective lens is exposed to the magnetic field, the loops are usually strongly distorted by non-linear Faraday rotations of the polarized light that occur in the objective lens and that are superimposed to the MOKE signal. In this paper, an experimental method, based on a motorized analyzer, is introduced which allows to compensate the Faraday contributions, thus leading to pure MOKE loops. A wide field Kerr microscope, equipped with this technology, works well as a laser-based MOKE magnetometer, additionally offering domain images and thus providing the basis for loop interpretation.

Investigation of shape memory of red blood cells using optical tweezers and quantitative phase microscopy

Science.gov (United States)

Cardenas, Nelson; Mohanty, Samarendra K.

2012-03-01

RBC has been shown to possess shape memory subsequent to shear-induced shape transformation. However, this property of RBC may not be generalized to all kinds of stresses. Here, we report our observation on the action of radiation pressure forces on RBC's shape memory using optical manipulation and quantitative phase microscopy (OMQPM). QPM, based on Mach-Zehnder interferrometry, allowed measurement of dynamic changes of shape of RBC in optical tweezers at different trapping laser powers. In high power near-infrared optical tweezers (>200mW), the RBC was found to deform significantly due to optical forces. Upon removal of the tweezers, hysteresis in recovering its original resting shape was observed. In very high power tweezers or long-term stretching events, shape memory was almost erased. This irreversibility of the deformation may be due to temperature rise or stress-induced phase transformation of lipids in RBC membrane.

Nonlinear Polarimetric Microscopy for Biomedical Imaging

Science.gov (United States)

Samim, Masood

A framework for the nonlinear optical polarimetry and polarimetric microscopy is developed. Mathematical equations are derived in terms of linear and nonlinear Stokes Mueller formalism, which comprehensively characterize the polarization properties of the incoming and outgoing radiations, and provide structural information about the organization of the investigated materials. The algebraic formalism developed in this thesis simplifies many predictions for a nonlinear polarimetry study and provides an intuitive understanding of various polarization properties for radiations and the intervening medium. For polarimetric microscopy experiments, a custom fast-scanning differential polarization microscope is developed, which is also capable of real-time three-dimensional imaging. The setup is equipped with a pair of high-speed resonant and galvanometric scanning mirrors, and supplemented by advanced adaptive optics and data acquisition modules. The scanning mirrors when combined with the adaptive optics deformable mirror enable fast 3D imaging. Deformable membrane mirrors and genetic algorithm optimization routines are employed to improve the imaging conditions including correcting the optical aberrations, maximizing signal intensities, and minimizing point-spread-functions of the focal volume. A field-programmable-gate array (FPGA) chip is exploited to rapidly acquire and process the multidimensional data. Using the nonlinear optical polarimetry framework and the home-built polarization microscope, a few biologically important tissues are measured and analyzed to gain insight as to their structure and dynamics. The structure and distribution of muscle sarcomere myosins, connective tissue collagen, carbohydrate-rich starch, and fruit fly eye retinal molecules are characterized with revealing polarization studies. In each case, using the theoretical framework, polarization sensitive data are analyzed to decipher the molecular orientations and nonlinear optical

Comparison of rigorous modelling of different structure profiles on photomasks for quantitative linewidth measurements by means of UV- or DUV-optical microscopy

Science.gov (United States)

Ehret, Gerd; Bodermann, Bernd; Woehler, Martin

2007-06-01

The optical microscopy is an important instrument for dimensional characterisation or calibration of micro- and nanostructures, e.g. chrome structures on photomasks. In comparison to scanning electron microscopy (possible contamination of the sample) and atomic force microscopy (slow, risk of damage) optical microscopy is a fast and non destructive metrology method. The precise quantitative determination of the linewidth from the microscope image is, however, only possible by knowledge of the geometry of the structures and their consideration in the optical modelling. We compared two different rigorous model approaches, the Rigorous Coupled Wave Analysis (RCWA) and the Finite Elements Method (FEM) for modelling of structures with different edge angles, linewidths, line to space ratios and polarisations. The RCWA method can adapt inclined edges profiles only by a staircase approximation leading to increased modelling errors of the RCWA method. Even today's sophisticated rigorous methods still show problems with TM-polarisation. Therefore both rigorous methods are compared in terms of their convergence for TE and TM- polarisation. Beyond that also the influence of typical illumination wavelengths (365 nm, 248 nm and 193 nm) on the microscope images and their contribution to the measuring uncertainty budget will be discussed.

Design and Development of Nonlinear Optical Microscope System: Simple Implementation with epi-Illumination Platform

Directory of Open Access Journals (Sweden)

Ryu Jiheun

2015-01-01

Full Text Available During the research using fluorescence-tagged or auto-fluorescence molecules, meaningful information is often buried deep inside the tissue, not its surface. Therefore, especially in the field of biomedical imaging, acquiring optically sectioned images from deep inside the tissue is very important. As well know already, confocal laser scanning microscopy (the most well-known optical sectioning microscopy gives axially-resolved fluorescence information using the physical background blocking component called pinhole. However, the axial range of imaging is practically limited due to such optical phenomena as the light scattered and absorbed in the tissue. However, nonlinear optical microscopy (e.g. Multiphoton microscopy, harmonic generation microscopy, coherent anti-Stokes Raman spectroscopy realized by the development of ultrafast light sources has been used for visualizing various tissues, especially in vivo, because of their low sensitivity to the limitation caused by the scattering and the absorption of light. Although nonlinear optical microscopy gives deep tissue image, it is not easy for many researcher to build customized nonlinear system. Here, we introduce an easy and simple way designing and developing such nonlinear optical microscope with upright or inverted epi-illumination platform using commercial optical components only.

Manipulation of local optical properties and structures in molybdenum-disulfide monolayers using electric field-assisted near-field techniques.

Science.gov (United States)

Nozaki, Junji; Fukumura, Musashi; Aoki, Takaaki; Maniwa, Yutaka; Yomogida, Yohei; Yanagi, Kazuhiro

2017-04-05

Remarkable optical properties, such as quantum light emission and large optical nonlinearity, have been observed in peculiar local sites of transition metal dichalcogenide monolayers, and the ability to tune such properties is of great importance for their optoelectronic applications. For that purpose, it is crucial to elucidate and tune their local optical properties simultaneously. Here, we develop an electric field-assisted near-field technique. Using this technique we can clarify and tune the local optical properties simultaneously with a spatial resolution of approximately 100 nm due to the electric field from the cantilever. The photoluminescence at local sites in molybdenum-disulfide (MoS 2 ) monolayers is reversibly modulated, and the inhomogeneity of the charge neutral points and quantum yields is suggested. We successfully etch MoS 2 crystals and fabricate nanoribbons using near-field techniques in combination with an electric field. This study creates a way to tune the local optical properties and to freely design the structural shapes of atomic monolayers using near-field optics.

Nonlinear optical microscopy for histology of fresh normal and cancerous pancreatic tissues.

Directory of Open Access Journals (Sweden)

Wenyan Hu

Full Text Available BACKGROUND: Pancreatic cancer is a lethal disease with a 5-year survival rate of only 1-5%. The acceleration of intraoperative histological examination would be beneficial for better management of pancreatic cancer, suggesting an improved survival. Nonlinear optical methods based on two-photon excited fluorescence (TPEF and second harmonic generation (SHG of intrinsic optical biomarkers show the ability to visualize the morphology of fresh tissues associated with histology, which is promising for real-time intraoperative evaluation of pancreatic cancer. METHODOLOGY/PRINCIPAL FINDINGS: In order to investigate whether the nonlinear optical imaging methods have the ability to characterize pancreatic histology at cellular resolution, we studied different types of pancreatic tissues by using label-free TPEF and SHG. Compared with other routine methods for the preparation of specimens, fresh tissues without processing were found to be most suitable for nonlinear optical imaging of pancreatic tissues. The detailed morphology of the normal rat pancreas was observed and related with the standard histological images. Comparatively speaking, the preliminary images of a small number of chemical-induced pancreatic cancer tissues showed visible neoplastic differences in the morphology of cells and extracellular matrix. The subcutaneous pancreatic tumor xenografts were further observed using the nonlinear optical microscopy, showing that most cells are leucocytes at 5 days after implantation, the tumor cells begin to proliferate at 10 days after implantation, and the extracellular collagen fibers become disordered as the xenografts grow. CONCLUSIONS/SIGNIFICANCE: In this study, nonlinear optical imaging was used to characterize the morphological details of fresh pancreatic tissues for the first time. We demonstrate that it is possible to provide real-time histological evaluation of pancreatic cancer by the nonlinear optical methods, which present an

Rod-like plasmonic nanoparticles as optical building blocks: how differences in particle shape and structural geometry influence optical signal

Energy Technology Data Exchange (ETDEWEB)

Stender, Anthony [Iowa State Univ., Ames, IA (United States)

2013-01-01

Gold nanoparticles, particularly those with an anisotropic shape, have become a popular optical probe for experiments involving work on the nanoscale. However, to carry out such delicate and intricate experiments, it is first necessary to understand the detailed behavior of individual nanoparticles. In this series of experiments, optical and electron microscopy were utilized for the characterization of individual nanoparticles and small assemblies of nanoparticles. In the first experiment, gold nanorods were investigated. Single, isolated nanorods exhibit two maxima of localized surface plasmon resonance (LSPR), which are associated with the two nanorod axes. Upon the physical rotation of a nanorod at one of its LSPR wavelengths under polarized illumination, the optical behavior varies in a sinusoidal fashion. A dimer of nanorods exhibits optical behavior quite similar to a nanorod, except the LSPR maxima are shifted and broader. Under differential interference contrast (DIC) microscopy, a pair of nanorods separated by a distance below the diffraction limit can be distinguished from a single nanorod due to its optical behavior upon rotation. Dark field microscopy is unable to distinguish the two geometries. For the second set of experiments, the optical behavior of single gold nanorods at non-plasmonic wavelengths was investigated. The same nanorod was rotated with respect to a polarized light source under DIC, dark field, and polarized light microscopy. DIC microscopy was found to produce diffraction pattern peaks at non-plasmonic wavelengths, which could be altered by adjusting the setting of the polarizer. In the third set of experiments, the optical behavior of a single gold dumbbell and several simple dumbbell geometries were investigated with microscopy and simulations. The single dumbbell displayed behavior quite similar to that of a nanorod, but dumbbells exhibit a shift in both LSPR wavebands. Moreover, the shape of dumbbell particles allows them to

Miniature fiber-optic multiphoton microscopy system using frequency-doubled femtosecond Er-doped fiber laser.

Science.gov (United States)

Huang, Lin; Mills, Arthur K; Zhao, Yuan; Jones, David J; Tang, Shuo

2016-05-01

We report on a miniature fiber-optic multiphoton microscopy (MPM) system based on a frequency-doubled femtosecond Er-doped fiber laser. The femtosecond pulses from the laser source are delivered to the miniature fiber-optic probe at 1.58 µm wavelength, where a standard single mode fiber is used for delivery without the need of free-space dispersion compensation components. The beam is frequency-doubled inside the probe by a periodically poled MgO:LiNbO3 crystal. Frequency-doubled pulses at 786 nm with a maximum power of 80 mW and a pulsewidth of 150 fs are obtained and applied to excite intrinsic signals from tissues. A MEMS scanner, a miniature objective, and a multimode collection fiber are further used to make the probe compact. The miniature fiber-optic MPM system is highly portable and robust. Ex vivo multiphoton imaging of mammalian skins demonstrates the capability of the system in imaging biological tissues. The results show that the miniature fiber-optic MPM system using frequency-doubled femtosecond fiber laser can potentially bring the MPM imaging for clinical applications.

French Society of Microscopies, 11. Colloquium. SFM Paris 2009. Compilation of summaries

International Nuclear Information System (INIS)

2009-06-01

The 11. conference of the SFM (French Society of Microscopies), held in Paris in 2009, was divided into 14 symposiums, 4 GN-MEBA symposiums, and 10 workshops. The titles of the symposiums are: homage to Nicolas Boisset, advanced microscopies, alternative microscopies, new optical and plasmonic imaging microscopies, dynamic and quantitative microscopy of the living matter, photonic and correlative electronic microscopy, near field microscopy, molecular and cellular electronic cryo-microscopy, cellular compartmentation and dynamics (CFPU), microscopy and materials, dynamical microscopy in materials science, minerals/bio-minerals and environment, structure and properties of nano-materials, sub-eV and sub-nm chemical bonds imaging. The titles of the GN-MEBA symposiums are: microscopy and metals, microscopy and minerals, microscopy and living beings, microscopy and new materials. The titles of the workshops are: Correlative Light and Electron Microscopy (CLEM), Cryo and electronic tomography in cellular biology, Cryo electronic microscopy of vitreous sections (CEMOVIS), Atomic Force Microscopy (AFM), ULTRASTEM, Digital Micrograph programming, Cryo-Microscopy and molecular tomography, Cryo-ultra-microtomy and immuno-marking, FIB, ASTAR(EBSD-MET) - rapid mapping of crystalline orientations and phases

Shaded-Mask Filtering for Extended Depth-of-Field Microscopy

OpenAIRE

Escobar García, Isabel María; Saavedra Tortosa, Genaro; Martínez Corral, Manuel; Calatayud, Arnau; Doblas Expósito, Ana Isabel

2013-01-01

This paper proposes a new spatial filtering approach for increasing the depth-of-field (DOF) of imaging systems, which is very useful for obtaining sharp images for a wide range of axial positions of the object. Many different techniques have been reported to increase the depth of field. However the main advantage in our method is its simplicity, since we propose the use of purely absorbing beam-shaping elements, which allows a high focal depth with a minimum modification of the optical archi...

Resolution enhancement techniques in microscopy

Science.gov (United States)

Cremer, Christoph; Masters, Barry R.

2013-05-01

We survey the history of resolution enhancement techniques in microscopy and their impact on current research in biomedicine. Often these techniques are labeled superresolution, or enhanced resolution microscopy, or light-optical nanoscopy. First, we introduce the development of diffraction theory in its relation to enhanced resolution; then we explore the foundations of resolution as expounded by the astronomers and the physicists and describe the conditions for which they apply. Then we elucidate Ernst Abbe's theory of optical formation in the microscope, and its experimental verification and dissemination to the world wide microscope communities. Second, we describe and compare the early techniques that can enhance the resolution of the microscope. Third, we present the historical development of various techniques that substantially enhance the optical resolution of the light microscope. These enhanced resolution techniques in their modern form constitute an active area of research with seminal applications in biology and medicine. Our historical survey of the field of resolution enhancement uncovers many examples of reinvention, rediscovery, and independent invention and development of similar proposals, concepts, techniques, and instruments. Attribution of credit is therefore confounded by the fact that for understandable reasons authors stress the achievements from their own research groups and sometimes obfuscate their contributions and the prior art of others. In some cases, attribution of credit is also made more complex by the fact that long term developments are difficult to allocate to a specific individual because of the many mutual connections often existing between sometimes fiercely competing, sometimes strongly collaborating groups. Since applications in biology and medicine have been a major driving force in the development of resolution enhancing approaches, we focus on the contribution of enhanced resolution to these fields.

Acoustic Imaging Frequency Dynamics of Ferroelectric Domains by Atomic Force Microscopy

International Nuclear Information System (INIS)

Kun-Yu, Zhao; Hua-Rong, Zeng; Hong-Zhang, Song; Sen-Xing, Hui; Guo-Rong, Li; Qing-Rui, Yin; Shimamura, Kiyoshi; Kannan, Chinna Venkadasamy; Villora, Encarnacion Antonia Garcia; Takekawa, Shunji; Kitamura, Kenji

2008-01-01

We report the acoustic imaging frequency dynamics of ferroelectric domains by low-frequency acoustic probe microscopy based on the commercial atomic force microscopy It is found that ferroelectric domain could be firstly visualized at lower frequency down to 0.5 kHz by AFM-based acoustic microscopy The frequency-dependent acoustic signal revealed a strong acoustic response in the frequency range from 7kHz to 10kHz, and reached maximum at 8.1kHz. The acoustic contrast mechanism can be ascribed to the different elastic response of ferroelectric microstructures to local elastic stress fields, which is induced by the acoustic wave transmitting in the sample when the piezoelectric transducer is vibrating and exciting acoustic wave under ac electric fields due to normal piezoelectric effects. (condensed matter: electronic structure, electrical, magnetic, and optical properties)

Field emission from the surface of highly ordered pyrolytic graphite

Czech Academy of Sciences Publication Activity Database

Knápek, Alexandr; Sobola, D.; Tománek, P.; Pokorná, Zuzana; Urbánek, Michal

2017-01-01

Roč. 395, FEB 15 (2017), s. 157-161 ISSN 0169-4332 R&D Projects: GA TA ČR(CZ) TE01020118 Institutional support: RVO:68081731 Keywords : field emission * HOPG * scanning electron microscopy * scanning near-field optical microscopy Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering OBOR OECD: Nano-processes (applications on nano-scale) Impact factor: 3.387, year: 2016

A novel phase-sensitive scanning near-field optical microscope

International Nuclear Information System (INIS)

Wu Xiao-Yu; Lin Sun; Tan Qiao-Feng; Wang Jia

2015-01-01

Phase is one of the most important parameters of electromagnetic waves. It is the phase distribution that determines the propagation, reflection, refraction, focusing, divergence, and coupling features of light, and further affects the intensity distribution. In recent years, the designs of surface plasmon polariton (SPP) devices have mostly been based on the phase modulation and manipulation. Here we demonstrate a phase sensitive multi-parameter heterodyne scanning near-field optical microscope (SNOM) with an aperture probe in the visible range, with which the near field optical phase and amplitude distributions can be simultaneously obtained. A novel architecture combining a spatial optical path and a fiber optical path is employed for stability and flexibility. Two kinds of typical nano-photonic devices are tested with the system. With the phase-sensitive SNOM, the phase and amplitude distributions of any nano-optical field and localized field generated with any SPP nano-structures and irregular phase modulation surfaces can be investigated. The phase distribution and the interference pattern will help us to gain a better understanding of how light interacts with SPP structures and how SPP waves generate, localize, convert, and propagate on an SPP surface. This will be a significant guidance on SPP nano-structure design and optimization. (paper)

Deep learning enhanced mobile-phone microscopy

KAUST Repository

Rivenson, Yair

2017-12-12

Mobile-phones have facilitated the creation of field-portable, cost-effective imaging and sensing technologies that approach laboratory-grade instrument performance. However, the optical imaging interfaces of mobile-phones are not designed for microscopy and produce spatial and spectral distortions in imaging microscopic specimens. Here, we report on the use of deep learning to correct such distortions introduced by mobile-phone-based microscopes, facilitating the production of high-resolution, denoised and colour-corrected images, matching the performance of benchtop microscopes with high-end objective lenses, also extending their limited depth-of-field. After training a convolutional neural network, we successfully imaged various samples, including blood smears, histopathology tissue sections, and parasites, where the recorded images were highly compressed to ease storage and transmission for telemedicine applications. This method is applicable to other low-cost, aberrated imaging systems, and could offer alternatives for costly and bulky microscopes, while also providing a framework for standardization of optical images for clinical and biomedical applications.

Necrosis and apoptosis pathways of cell death at photodynamic treatment in vitro as revealed by digital holographic microscopy

Science.gov (United States)

Semenova, I. V.; Belashov, A. V.; Belyaeva, T. N.; Kornilova, E. S.; Salova, A. V.; Zhikhoreva, A. A.; Vasyutinskii, O. S.

2018-02-01

Monitoring of variations in morphological characteristics of cultured HeLa cells after photodynamic treatment with Radachlorin photosensitizer is performed by means of digital holographic microscopy. The observed dose-dependent post-treatment variations of phase shift evidence threshold effect of photodynamic treatment and allow for distinguishing between necrotic or apoptotic pathways of cell death. Results obtained by holographic microscopy were confirmed by means of far-field optical microscopy and confocal fluorescence microscopy with commonly used test assays.

Lithographic measurement of EUV flare in the 0.3-NA Micro Exposure Tool optic at the Advanced Light Source

International Nuclear Information System (INIS)

Cain, Jason P.; Naulleau, Patrick; Spanos, Costas J.

2005-01-01

The level of flare present in a 0.3-NA EUV optic (the MET optic) at the Advanced Light Source at Lawrence Berkeley National Laboratory is measured using a lithographic method. Photoresist behavior at high exposure doses makes analysis difficult. Flare measurement analysis under scanning electron microscopy (SEM) and optical microscopy is compared, and optical microscopy is found to be a more reliable technique. In addition, the measured results are compared with predictions based on surface roughness measurement of the MET optical elements. When the fields in the exposure matrix are spaced far enough apart to avoid influence from surrounding fields and the data is corrected for imperfect mask contrast and aerial image proximity effects, the results match predicted values quite well. The amount of flare present in this optic ranges from 4.7% for 2 (micro)m features to 6.8% for 500 nm features

Radon-Wigner transform for optical field analysis

NARCIS (Netherlands)

Alieva, T.; Bastiaans, M.J.; Nijhawan, O.P.; Gupta, A.K.; Musla, A.K.; Singh, Kehar

1998-01-01

The Radon-Wigner transform, associated with the intensity distribution in the fractional Fourier transform system, is used for the analysis of complex structures of coherent as well as partially coherent optical fields. The application of the Radon-Wigner transform to the analysis of fractal fields

Field-Programmable Logic Devices with Optical Input Output

Science.gov (United States)

Szymanski, Ted H.; Saint-Laurent, Martin; Tyan, Victor; Au, Albert; Supmonchai, Boonchuay

2000-02-01

A field-programmable logic device (FPLD) with optical I O is described. FPLD s with optical I O can have their functionality specified in the field by means of downloading a control-bit stream and can be used in a wide range of applications, such as optical signal processing, optical image processing, and optical interconnects. Our device implements six state-of-the-art dynamically programmable logic arrays (PLA s) on a 2 mm 2 mm die. The devices were fabricated through the Lucent Technologies Advanced Research Projects Agency Consortium for Optical and Optoelectronic Technologies in Computing (Lucent ARPA COOP) workshop by use of 0.5- m complementary metal-oxide semiconductor self-electro-optic device technology and were delivered in 1998. All devices are fully functional: The electronic data paths have been verified at 200 MHz, and optical tests are pending. The device has been programmed to implement a two-stage optical switching network with six 4 4 crossbar switches, which can realize more than 190 10 6 unique programmable input output permutations. The same device scaled to a 2 cm 2 cm substrate could support as many as 4000 optical I O and 1 Tbit s of optical I O bandwidth and offer fully programmable digital functionality with approximately 110,000 programmable logic gates. The proposed optoelectronic FPLD is also ideally suited to realizing dense, statically reconfigurable crossbar switches. We describe an attractive application area for such devices: a rearrangeable three-stage optical switch for a wide-area-network backbone, switching 1000 traffic streams at the OC-48 data rate and supporting several terabits of traffic.

Simultaneous topographical, electrical and optical microscopy of optoelectronic devices at the nanoscale

KAUST Repository

Kumar, Naresh

2017-01-12

Novel optoelectronic devices rely on complex nanomaterial systems where the nanoscale morphology and local chemical composition are critical to performance. However, the lack of analytical techniques that can directly probe these structure-property relationships at the nanoscale presents a major obstacle to device development. In this work, we present a novel method for non-destructive, simultaneous mapping of the morphology, chemical composition and photoelectrical properties with <20 nm spatial resolution by combining plasmonic optical signal enhancement with electrical-mode scanning probe microscopy. We demonstrate that this combined approach offers subsurface sensitivity that can be exploited to provide molecular information with a nanoscale resolution in all three spatial dimensions. By applying the technique to an organic solar cell device, we show that the inferred surface and subsurface composition distribution correlates strongly with the local photocurrent generation and explains macroscopic device performance. For instance, the direct measurement of fullerene phase purity can distinguish between high purity aggregates that lead to poor performance and lower purity aggregates (fullerene intercalated with polymer) that result in strong photocurrent generation and collection. We show that the reliable determination of the structure-property relationship at the nanoscale can remove ambiguity from macroscopic device data and support the identification of the best routes for device optimisation. The multi-parameter measurement approach demonstrated herein is expected to play a significant role in guiding the rational design of nanomaterial-based optoelectronic devices, by opening a new realm of possibilities for advanced investigation via the combination of nanoscale optical spectroscopy with a whole range of scanning probe microscopy modes.

Perspective: Differential dynamic microscopy extracts multi-scale activity in complex fluids and biological systems

Science.gov (United States)

Cerbino, Roberto; Cicuta, Pietro

2017-09-01

Differential dynamic microscopy (DDM) is a technique that exploits optical microscopy to obtain local, multi-scale quantitative information about dynamic samples, in most cases without user intervention. It is proving extremely useful in understanding dynamics in liquid suspensions, soft materials, cells, and tissues. In DDM, image sequences are analyzed via a combination of image differences and spatial Fourier transforms to obtain information equivalent to that obtained by means of light scattering techniques. Compared to light scattering, DDM offers obvious advantages, principally (a) simplicity of the setup; (b) possibility of removing static contributions along the optical path; (c) power of simultaneous different microscopy contrast mechanisms; and (d) flexibility of choosing an analysis region, analogous to a scattering volume. For many questions, DDM has also advantages compared to segmentation/tracking approaches and to correlation techniques like particle image velocimetry. The very straightforward DDM approach, originally demonstrated with bright field microscopy of aqueous colloids, has lately been used to probe a variety of other complex fluids and biological systems with many different imaging methods, including dark-field, differential interference contrast, wide-field, light-sheet, and confocal microscopy. The number of adopting groups is rapidly increasing and so are the applications. Here, we briefly recall the working principles of DDM, we highlight its advantages and limitations, we outline recent experimental breakthroughs, and we provide a perspective on future challenges and directions. DDM can become a standard primary tool in every laboratory equipped with a microscope, at the very least as a first bias-free automated evaluation of the dynamics in a system.

Vector electric field measurement via position-modulated Kelvin probe force microscopy

Science.gov (United States)

Dwyer, Ryan P.; Smieska, Louisa M.; Tirmzi, Ali Moeed; Marohn, John A.

2017-10-01

High-quality spatially resolved measurements of electric fields are critical to understanding charge injection, charge transport, and charge trapping in semiconducting materials. Here, we report a variation of frequency-modulated Kelvin probe force microscopy that enables spatially resolved measurements of the electric field. We measure electric field components along multiple directions simultaneously by employing position modulation and lock-in detection in addition to numeric differentiation of the surface potential. We demonstrate the technique by recording linescans of the in-plane electric field vector in the vicinity of a patch of trapped charge in a 2,7-diphenyl[1]benzothieno[3,2-b][1]benzothiophene (DPh-BTBT) organic field-effect transistor. This technique is simple to implement and should be especially useful for studying electric fields in spatially inhomogeneous samples like organic transistors and photovoltaic blends.

Optical Emissions of Sprite Streamers in Weak Electric Fields

Science.gov (United States)

Liu, N.; Pasko, V. P.

2004-12-01

Sprites commonly consist of large numbers of needle-shaped filaments of ionization [e.g., Gerken and Inan, JASTP, 65, 567, 2003] and typically initiate at altitudes 70-75 km in a form of upward and downward propagating streamers [Stanley et al., GRL, 26, 3201, 1999; Stenbaek-Nielsen et al., GRL, 27, 3829, 2000; McHarg et al., JGR, 107, 1364, 2002; Moudry et al., JASTP, 65, 509, 2003]. The strong electric fields E exceeding the conventional breakdown threshold field Ek are needed for initiation of sprite streamers from single electron avalanches and recent modeling studies indicate that streamers propagating in fields E>Ek experience strong acceleration and expansion in good agreement with the above cited observations [Liu and Pasko, JGR, 109, A04301, 2004]. The initiated streamers are capable of propagating in fields substantially lower than Ek [Allen and Ghaffar, J. Phys. D: Appl. Phys., 28, 331, 1995] and it is expected that a significant part of sprite optical output comes from regions with EEk). Additionally, the values of electric fields inside of the streamer channel are always well below Ek and since the excitation coefficients for optical emissions are very sensitive to the driving electric field magnitude most of the optical luminosity of streamers in this case arises from streamer tips, indicating that observed streamer filaments in many cases may be produced by time averaging of optical luminosity coming from localized regions around streamer tips as streamers move through an instrument's field of view. We will discuss pressure dependent differences of optical emissions at different sprite altitudes, and important similarities between observed sprite streamers and recent time resolved (van Veldhuizen et al., IEEE Trans. Plasma Sci., 30, 162, 2002; Yi and Williams, J. Phys. D. Appl. Phys., 35, 205, 2002].

Sub-nanosecond time-resolved near-field scanning magneto-optical microscope.

Science.gov (United States)

Rudge, J; Xu, H; Kolthammer, J; Hong, Y K; Choi, B C

2015-02-01

We report on the development of a new magnetic microscope, time-resolved near-field scanning magneto-optical microscope, which combines a near-field scanning optical microscope and magneto-optical contrast. By taking advantage of the high temporal resolution of time-resolved Kerr microscope and the sub-wavelength spatial resolution of a near-field microscope, we achieved a temporal resolution of ∼50 ps and a spatial resolution of microscope, the magnetic field pulse induced gyrotropic vortex dynamics occurring in 1 μm diameter, 20 nm thick CoFeB circular disks has been investigated. The microscope provides sub-wavelength resolution magnetic images of the gyrotropic motion of the vortex core at a resonance frequency of ∼240 MHz.

Near-field characteristics of highly non-paraxial subwavelength optical fields with hybrid states of polarization

International Nuclear Information System (INIS)

Chen Rui-Pin; Gao Teng-Yue; Chew Khian-Hooi; Dai Chao-Qing; Zhou Guo-Quan; He Sai-Ling

2017-01-01

The vectorial structure of an optical field with hybrid states of polarization (SoP) in the near-field is studied by using the angular spectrum method of an electromagnetic beam. Physical images of the longitudinal components of evanescent waves are illustrated and compared with those of the transverse components from the vectorial structure. Our results indicate that the relative weight integrated over the transverse plane of the evanescent wave depends strongly on the number of the polarization topological charges. The shapes of the intensity profiles of the longitudinal components are different from those of the transverse components, and it can be manipulated by changing the initial SoP of the field cross-section. The longitudinal component of evanescent wave dominates the near-field region. In addition, it also leads to three-dimensional shape variations of the optical field and the optical spin angular momentum flux density distributions. (paper)

Comparative analysis of imaging configurations and objectives for Fourier microscopy.

Science.gov (United States)

Kurvits, Jonathan A; Jiang, Mingming; Zia, Rashid

2015-11-01

Fourier microscopy is becoming an increasingly important tool for the analysis of optical nanostructures and quantum emitters. However, achieving quantitative Fourier space measurements requires a thorough understanding of the impact of aberrations introduced by optical microscopes that have been optimized for conventional real-space imaging. Here we present a detailed framework for analyzing the performance of microscope objectives for several common Fourier imaging configurations. To this end, we model objectives from Nikon, Olympus, and Zeiss using parameters that were inferred from patent literature and confirmed, where possible, by physical disassembly. We then examine the aberrations most relevant to Fourier microscopy, including the alignment tolerances of apodization factors for different objective classes, the effect of magnification on the modulation transfer function, and vignetting-induced reductions of the effective numerical aperture for wide-field measurements. Based on this analysis, we identify an optimal objective class and imaging configuration for Fourier microscopy. In addition, the Zemax files for the objectives and setups used in this analysis have been made publicly available as a resource for future studies.

Electric-field effects in optically generated spin transport

International Nuclear Information System (INIS)

Miah, M. Idrish

2009-01-01

Transport of spin-polarized electrons in semiconductors is studied experimentally. Spins are generated by optical excitation because of the selection rules governing optical transitions from heavy-hole and light-hole states to conduction-band states. Experiments designed for the control of spins in semiconductors investigate the bias-dependent spin transport process and detect the spin-polarized electrons during transport. A strong bias dependence is observed. The electric-field effects on the spin-polarized electron transport are also found to be depended on the excitation photon energy and temperature. Based on a field-dependent spin relaxation mechanism, the electric-field effects in the transport process are discussed.

Electric-field effects in optically generated spin transport

Energy Technology Data Exchange (ETDEWEB)

Miah, M. Idrish [Nanoscale Science and Technology Centre and School of Biomolecular and Physical Sciences, Griffith University, Nathan, Brisbane, QLD 4111 (Australia); Department of Physics, University of Chittagong, Chittagong 4331 (Bangladesh)], E-mail: m.miah@griffith.edu.au

2009-05-25

Transport of spin-polarized electrons in semiconductors is studied experimentally. Spins are generated by optical excitation because of the selection rules governing optical transitions from heavy-hole and light-hole states to conduction-band states. Experiments designed for the control of spins in semiconductors investigate the bias-dependent spin transport process and detect the spin-polarized electrons during transport. A strong bias dependence is observed. The electric-field effects on the spin-polarized electron transport are also found to be depended on the excitation photon energy and temperature. Based on a field-dependent spin relaxation mechanism, the electric-field effects in the transport process are discussed.

French Society of Microscopy, 10. conference; Societe Francaise des Microscopies, 10. colloque

Energy Technology Data Exchange (ETDEWEB)

Thibault-Penisson, J; Cremer, Ch; Susini, J; Kirklanda, A I; Rigneault, H; Renault, O; Bailly, A; Zagonel, L F; Barrett, N; Bogner, A; Gauthier, C; Jouneau, P H; Thollet, G; Fuchs, G; Basset, D; Deconihout, B; Vurpillot, F; Vella, A; Matthieu, G; Cadel, E; Bostel, A; Blavette, D; Baumeister, W; Usson, Y; Zaefferer, St; Laffont, L; Weyland, M; Thomas, J M; Midgley, P; Benlekbir, S; Epicier, Th; Diop, B N; Roux, St; Ou, M; Perriat, P; Bausach, M; Aouine, M; Berhault, G; Idrissi, H; Cottevieille, M; Jonic, S; Larquet, E; Svergun, D; Vannoni, M A; Boisset, N; Ersena, O; Werckmann, J; Ulhaq, C; Hirlimann, Ch; Tihay, F; Cuong, Pham-Huu; Crucifix, C; Schultz, P; Jornsanoha, P; Thollet, G; Masenelli-Varlot, K; Gauthier, C; Ludwig, W; King, A; Johnson, G; Gonzalves-Hoennicke, M; Reischig, P; Messaoudi, C; Ibrahim, R; Marco, S; Klie, R F; Zhao, Y; Yang, G; Zhu, Y; Hue, F; Hytch, M; Hartmann, J M; Bogumilowicz, Y; Claverie, A; Klein, H; Alloyeau, D; Ricolleau, C; Langlois, C; Le Bouar, Y; Loiseau, A; Colliex, C; Stephan, O; Kociak, M; Tence, M; Gloter, A; Imhoff, D; Walls, M; Nelayah, J; March, K; Couillard, M; Ailliot, C; Bertin, F; Cooper, D; Rivallin, P; Dumelie, N; Benhayoune, H; Balossier, G; Cheynet, M; Pokrant, S; Tichelaar, F; Rouviere, J L; Cooper, D; Truche, R; Chabli, A; Debili, M Y; Houdellier, F; Warot-Fonrose, B; Hytch, M J; Snoeck, E; Calmels, L; Serin, V; Schattschneider, P; Jacob, D; Cordier, P

2007-07-01

This document gathers the resumes of some of the presentations made at this conference whose aim was to present the last developments and achievements of the 3 complementary microscopies: optical microscopy, electron microscopy and X-ray microscopy. The contributions have been organized around the following 12 topics: 1) new technical developments, 2) 3-dimensional imaging, 3) quantitative microscopy, 4) technical progress in photon microscopy, 5) synchrotron radiation, 6) measurements of patterns, deformations and strains, 7) materials for energy and transports, 8) nano-structures, 9) virus: structure and infection mechanisms, 10) 3-dimensional imaging for molecules, cells and cellular tissues, 11) nano-particles and colloids, and 12) liquid crystals.

Experimental and theoretical analysis for improved microscope design of optical projection tomographic microscopy.

Science.gov (United States)

Coe, Ryan L; Seibel, Eric J

2013-09-01

We present theoretical and experimental results of axial displacement of objects relative to a fixed condenser focal plane (FP) in optical projection tomographic microscopy (OPTM). OPTM produces three-dimensional, reconstructed images of single cells from two-dimensional projections. The cell rotates in a microcapillary to acquire projections from different perspectives where the objective FP is scanned through the cell while the condenser FP remains fixed at the center of the microcapillary. This work uses a combination of experimental and theoretical methods to improve the OPTM instrument design.

Optical Remote Sensing of Electric Fields Above Thunderstorms

Science.gov (United States)

Burns, B. M.; Carlson, B. E.; Lauben, D.; Cohen, M.; Smith, D.; Inan, U. S.

2010-12-01

Measurement of thunderstorm electric fields typically require balloon-borne measurements in the region of interest. Such measurements are cumbersome and provide limited information at a single point. Remote sensing of electric fields by Kerr-effect induced optical polarization changes of background skylight circumvents many of these difficulties and can in principle provide a high-speed movie of electric field behavior. Above-thundercloud 100 kV/m quasi-static electric fields are predicted to produce polarization changes at above the part in one million level that should be detectable at a ground instrument featuring 1 cm2sr geometric factor and 1 kHz bandwidth (though more sensitivity is nonetheless desired). Currently available optical and electronic components may meet these requirements. We review the principles of this measurement and discuss the current status of a field-ready prototype instrument currently in construction.

An inverse method for determining the interaction force between the probe and sample using scanning near-field optical microscopy

International Nuclear Information System (INIS)

Chang, Win-Jin; Fang, Te-Hua

2006-01-01

This study proposes a means for calculating the interaction force during the scanning process using a scanning near-field optical microscope (SNOM) probe. The determination of the interaction force in the scanning system is regarded as an inverse vibration problem. The conjugate gradient method is applied to treat the inverse problem using available displacement measurements. The results show that the conjugate gradient method is less sensitive to measurement errors and prior information on the functional form of quality was not required. Furthermore, the initial guesses for the interaction force can be arbitrarily chosen for the iteration process

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection

Science.gov (United States)

Mertz, Jerome; Kim, Jinhyun

2010-01-01

It is well known that light-sheet illumination can enable optically sectioned wide-field imaging of macroscopic samples. However, the optical sectioning capacity of a light-sheet macroscope is undermined by sample-induced scattering or aberrations that broaden the thickness of the sheet illumination. We present a technique to enhance the optical sectioning capacity of a scanning light-sheet microscope by out-of-focus background rejection. The technique, called HiLo microscopy, makes use of two images sequentially acquired with uniform and structured sheet illumination. An optically sectioned image is then synthesized by fusing high and low spatial frequency information from both images. The benefits of combining light-sheet macroscopy and HiLo background rejection are demonstrated in optically cleared whole mouse brain samples, using both green fluorescent protein (GFP)-fluorescence and dark-field scattered light contrast.

Optical properties of a multibarrier structure under intense laser fields

Science.gov (United States)

Ospina, D. A.; Akimov, V.; Mora-Ramos, M. E.; Morales, A. L.; Tulupenko, V.; Duque, C. A.

2015-11-01

Using the diagonalization method and within the effective mass and parabolic band approximations, the energy spectrum and the wave functions are investigated in biased multibarrier structure taking into account the effects of nonresonant intense laser fields. We calculated the optical properties from the susceptibility using a nonperturbative formalism recently reported. We study the changes in the intersubband optical absorption coefficients and refraction index for several values of the dressing laser parameter and for some specific values of the electric field applied along the growth direction of the heterostructure. It is concluded from our study that the peaks in the optical absorption spectrum have redshifts or blueshifts as a function of the laser parameter and the electric field. These parameters could be suitable tools for tuning the electronic and optical properties of the multibarrier structure.

3D automatic quantification applied to optically sectioned images to improve microscopy analysis

Directory of Open Access Journals (Sweden)

JE Diaz-Zamboni

2009-08-01

Full Text Available New fluorescence microscopy techniques, such as confocal or digital deconvolution microscopy, allow to easily obtain three-dimensional (3D information from specimens. However, there are few 3D quantification tools that allow extracting information of these volumes. Therefore, the amount of information acquired by these techniques is difficult to manipulate and analyze manually. The present study describes a model-based method, which for the first time shows 3D visualization and quantification of fluorescent apoptotic body signals, from optical serial sections of porcine hepatocyte spheroids correlating them to their morphological structures. The method consists on an algorithm that counts apoptotic bodies in a spheroid structure and extracts information from them, such as their centroids in cartesian and radial coordinates, relative to the spheroid centre, and their integrated intensity. 3D visualization of the extracted information, allowed us to quantify the distribution of apoptotic bodies in three different zones of the spheroid.

Scanning near-field optical microscopy

OpenAIRE

van Hulst, N.F.; ten Wolde, Arthur

1998-01-01

An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today’s science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and temp...

Optical Field-Strength Polarization of Two-Mode Single-Photon States

Science.gov (United States)

Linares, J.; Nistal, M. C.; Barral, D.; Moreno, V.

2010-01-01

We present a quantum analysis of two-mode single-photon states based on the probability distributions of the optical field strength (or position quadrature) in order to describe their quantum polarization characteristics, where polarization is understood as a significative confinement of the optical field-strength values on determined regions of…

Macroscopic self-consistent model for external-reflection near-field microscopy

International Nuclear Information System (INIS)

Berntsen, S.; Bozhevolnaya, E.; Bozhevolnyi, S.

1993-01-01

The self-consistent macroscopic approach based on the Maxwell equations in two-dimensional geometry is developed to describe tip-surface interaction in external-reflection near-field microscopy. The problem is reduced to a single one-dimensional integral equation in terms of the Fourier components of the field at the plane of the sample surface. This equation is extended to take into account a pointlike scatterer placed on the sample surface. The power of light propagating toward the detector as the fiber mode is expressed by using the self-consistent field at the tip surface. Numerical results for trapezium-shaped tips are presented. The authors show that the sharper tip and the more confined fiber mode result in better resolution of the near-field microscope. Moreover, it is found that the tip-surface distance should not be too small so that better resolution is ensured. 14 refs., 10 figs

Integrated Photoacoustic and Fluorescence Confocal Microscopy

OpenAIRE

Wang, Yu; Maslov, Konstantin; Kim, Chulhong; Hu, Song; Wang, Lihong V.

2010-01-01

We have developed a dual-modality imaging system by integrating optical-resolution photoacoustic microscopy and fluorescence confocal microscopy to provide optical absorption and fluorescence contrasts simultaneously. By sharing the same laser source and objective lens, intrinsically registered photoacoustic and fluorescence images are acquired in a single scan. The micrometer resolution allows imaging of both blood and lymphatic vessels down to the capillary level. Simultaneous photoacoustic...

Microscopy and Image Analysis.

Science.gov (United States)

McNamara, George; Difilippantonio, Michael; Ried, Thomas; Bieber, Frederick R

2017-07-11

This unit provides an overview of light microscopy, including objectives, light sources, filters, film, and color photography for fluorescence microscopy and fluorescence in situ hybridization (FISH). We believe there are excellent opportunities for cytogeneticists, pathologists, and other biomedical readers, to take advantage of specimen optical clearing techniques and expansion microscopy-we briefly point to these new opportunities. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Nanoshells for in vivo imaging using two-photon excitation microscopy

Energy Technology Data Exchange (ETDEWEB)

Gao Liang; Nammalvar, Vengadesan [Department of Bioengineering, Rice University, Houston, TX 77005 (United States); Vadakkan, Tegy J, E-mail: lg3@rice.edu, E-mail: venkyn@rice.edu [Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030 (United States)

2011-09-07

Gold nanoshells have been intensively investigated and applied to various biomedical fields because of their flexible optical tunability and biological compatibility. They hold great potential to serve as luminescent contrast agents excitable with near-infrared (NIR) lasers. In this paper, we describe the development of nanoshells with a peak of plasmon resonance at 800 nm and their subsequent use for in vivo blood vessel imaging using two-photon excitation microscopy at an excitation wavelength of 750 nm. We were able to image single nanoshell particles in blood vessels and generate optical contrast for blood vessel structure using luminescent signals. These results confirm the feasibility of engineering nanoshells with controlled optical properties for single-particle-based in vivo imaging.

Field trial of 160 Gb/s all-optical packet switching

NARCIS (Netherlands)

Liu, Y.; Herrera Llorente, J.; Raz, O.; Tangdiongga, E.; Marti, J.; Ramos, F.; Maxwell, G.D.; Poustie, A.; Mulvad, H.C.H.; Hill, M.T.; Waardt, de H.; Khoe, G.D.; Koonen, A.M.J.; Dorren, H.J.S.; Nakano, Y.

2007-01-01

We present the results of a transmission experiment, over 110 km of field installed fiber, for an all-optical 160 Gb/s packet switching system. The system uses in-band optical labels which are processed entirely in the optical domain using a narrow-band all-optical filter. The label decision

Resonant Scanning with Large Field of View Reduces Photobleaching and Enhances Fluorescence Yield in STED Microscopy.

Science.gov (United States)

Wu, Yong; Wu, Xundong; Lu, Rong; Zhang, Jin; Toro, Ligia; Stefani, Enrico

2015-10-01

Photobleaching is a major limitation of superresolution Stimulated Depletion Emission (STED) microscopy. Fast scanning has long been considered an effective means to reduce photobleaching in fluorescence microscopy, but a careful quantitative study of this issue is missing. In this paper, we show that the photobleaching rate in STED microscopy can be slowed down and the fluorescence yield be enhanced by scanning with high speed, enabled by using large field of view in a custom-built resonant-scanning STED microscope. The effect of scanning speed on photobleaching and fluorescence yield is more remarkable at higher levels of depletion laser irradiance, and virtually disappears in conventional confocal microscopy. With ≥6 GW∙cm(-2) depletion irradiance, we were able to extend the fluorophore survival time of Atto 647N and Abberior STAR 635P by ~80% with 8-fold wider field of view. We confirm that STED Photobleaching is primarily caused by the depletion light acting upon the excited fluorophores. Experimental data agree with a theoretical model. Our results encourage further increasing the linear scanning speed for photobleaching reduction in STED microscopy.

In Vivo Confocal Microscopy and Anterior Segment Optic Coherence Tomography Findings in Ocular Ochronosis

Directory of Open Access Journals (Sweden)

Elif Demirkilinc Biler

2015-01-01

Full Text Available Purpose. To report clinical and in vivo confocal microscopy (IVCM findings of two patients with ocular ochronosis secondary due to alkaptonuria. Materials and Methods. Complete ophthalmologic examinations, including IVCM (HRT II/Rostock Cornea Module, Heidelberg, Germany, anterior segment optical coherence tomography (AS-OCT (Topcon 3D spectral-domain OCT 2000, Topcon Medical Systems, Paramus, NJ, USA, corneal topography (Pentacam, OCULUS Optikgeräte GmbH, Wetzlar, Germany, and anterior segment photography, were performed. Results. Biomicroscopic examination showed bilateral darkly pigmented lesions of the nasal and temporal conjunctiva and episclera in both patients. In vivo confocal microscopy of the lesions revealed prominent degenerative changes, including vacuoles and fragmentation of collagen fibers in the affected conjunctival lamina propria and episclera. Hyperreflective pigment granules in different shapes were demonstrated in the substantia propria beneath the basement membrane. AS-OCT of Case 1 demonstrated hyporeflective areas. Fundus examination was within normal limits in both patients, except tilted optic discs with peripapillary atrophy in one of the patients. Corneal topography, thickness, and macular OCT were normal bilaterally in both cases. Conclusion. The degenerative and anatomic changes due to ochronotic pigment deposition in alkaptonuria can be demonstrated in detail with IVCM and AS-OCT. Confocal microscopic analysis in ocular ochronosis may serve as a useful adjunct in diagnosis and monitoring of the disease progression.

X-ray microscopy in Aarhus

International Nuclear Information System (INIS)

Uggerhoej, Erik; Abraham-Peskir, Joanna V.

2000-01-01

The Aarhus imaging soft X-ray microscope is now a busy multi-user facility. The optical set-up will be described and project highlights discussed. a) Metal-induced structural changes in whole cells in solution. The effects of aluminum, copper, nickel and zinc on protozoa investigated by using a combination of light microscopy, confocal scanning laser microscopy and X-ray microscopy. b) Botanical studies by X-ray microscopy used to compliment electron microscopy studies. c) Sludge morphology and iron precipitation in Danish freshwater plants by combining X-ray, scanning electron and transmission electron microscopy

Dynamics of Molecular Gyroscopes Created by Strong Optical Fields

Science.gov (United States)

Mullin, Amy