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.

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.

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.

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.

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.

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.

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.

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...

New scanning technique for the optical vortex microscope.

Science.gov (United States)

Augustyniak, Ireneusz; Popiołek-Masajada, Agnieszka; Masajada, Jan; Drobczyński, Sławomir

2012-04-01

In the optical vortex microscopy the focused Gaussian beam with optical vortex scans a sample. An optical vortex can be introduced into a laser beam with the use of a special optical element--a vortex lens. When moving the vortex lens, the optical vortex changes its position inside the spot formed by a focused laser beam. This effect can be used as a new precise scanning technique. In this paper, we study the optical vortex behavior at the sample plane. We also estimate if the new scanning technique results in observable effects that could be used for a phase object detection.

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)

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.

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.

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.

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.

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...

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

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.

Revealing t-tubules in striated muscle with new optical super-resolution microscopy techniques

Directory of Open Access Journals (Sweden)

Isuru D. Jayasinghe

2014-12-01

Full Text Available The t-tubular system plays a central role in the synchronisation of calcium signalling and excitation-contraction coupling in most striated muscle cells. Light microscopy has been used for imaging t-tubules for well over 100 years and together with electron microscopy (EM, has revealed the three-dimensional complexities of the t-system topology within cardiomyocytes and skeletal muscle fibres from a range of species. The emerging super-resolution single molecule localisation microscopy (SMLM techniques are offering a near 10-fold improvement over the resolution of conventional fluorescence light microscopy methods, with the ability to spectrally resolve nanometre scale distributions of multiple molecular targets. In conjunction with the next generation of electron microscopy, SMLM has allowed the visualisation and quantification of intricate t-tubule morphologies within large areas of muscle cells at an unprecedented level of detail. In this paper, we review recent advancements in the t-tubule structural biology with the utility of various microscopy techniques. We outline the technical considerations in adapting SMLM to study t-tubules and its potential to further our understanding of the molecular processes that underlie the sub-micron scale structural alterations observed in a range of muscle pathologies.

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

Biological applications of near-field scanning optical microscopy

Science.gov (United States)

Moers, Marco H. P.; Ruiter, A. G. T.; Jalocha, Alain; van Hulst, Niko F.; Kalle, W. H. J.; Wiegant, J. C. A. G.; Raap, A. K.

1995-09-01

Near-field Scanning Optical Microscopy (NSOM) is a true optical microscopic technique allowing fluorescence, absorption, reflection and polarization contrast with the additional advantage of nanometer lateral resolution, unlimited by diffraction and operation at ambient conditions. NSOM based on metal coated adiabatically tapered fibers, combined with shear force feedback and operated in illumination mode, has proven to be the most powerful NSOM arrangement, because of its true localization of the optical interaction, its various optical contrast possibilities and its sensitivity down to the single molecular level. In this paper applications of `aperture' NSOM to Fluorescence In Situ Hybridization of human metaphase chromosomes are presented, where the localized fluorescence allows to identify specific DNA sequences. All images are accompanied by the simultaneously acquired force image, enabling direct comparison of the optical contrast with the sample topography on nanometer scale, far beyond the diffraction limit. Thus the unique combination of high resolution, specific optical contrast and ambient operation offers many new direction possibilities in biological studies.

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.

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

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)

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

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.

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.

Wide-field two-dimensional multifocal optical-resolution photoacoustic computed microscopy

Science.gov (United States)

Xia, Jun; Li, Guo; Wang, Lidai; Nasiriavanaki, Mohammadreza; Maslov, Konstantin; Engelbach, John A.; Garbow, Joel R.; Wang, Lihong V.

2014-01-01

Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technique that directly images optical absorption in tissue at high spatial resolution. To date, the majority of OR-PAM systems are based on single focused optical excitation and ultrasonic detection, limiting the wide-field imaging speed. While one-dimensional multifocal OR-PAM (1D-MFOR-PAM) has been developed, the potential of microlens and transducer arrays has not been fully realized. Here, we present the development of two-dimensional multifocal optical-resolution photoacoustic computed microscopy (2D-MFOR-PACM), using a 2D microlens array and a full-ring ultrasonic transducer array. The 10 × 10 mm2 microlens array generates 1800 optical foci within the focal plane of the 512-element transducer array, and raster scanning the microlens array yields optical-resolution photoacoustic images. The system has improved the in-plane resolution of a full-ring transducer array from ≥100 µm to 29 µm and achieved an imaging time of 36 seconds over a 10 × 10 mm2 field of view. In comparison, the 1D-MFOR-PAM would take more than 4 minutes to image over the same field of view. The imaging capability of the system was demonstrated on phantoms and animals both ex vivo and in vivo. PMID:24322226

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.

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

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

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

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.

Optical detection of ultrasound using an apertureless near-field scanning optical microscopy system

Science.gov (United States)

Ahn, Phillip; Zhang, Zhen; Sun, Cheng; Balogun, Oluwaseyi

2013-01-01

Laser ultrasonics techniques are power approaches for non-contact generation and detection of high frequency ultrasound on a local scale. In these techniques, optical diffraction limits the spatial information that can be accessed from a measurement. In order to improve the lateral spatial resolution, we incorporate an apertureless near-field scanning optical microscope (aNSOM) into laser ultrasonics setup for local detection of laser generated ultrasound. The aNSOM technique relies on the measurement of a weak backscattered near-field light intensity resulting from the oblique illumination of a nanoscale probe-tip positioned close to a sample surface. We enhance the optical near-field intensity by coupling light to surface plasmon polaritons (SPPs) on the shaft of an atomic force microscopy (AFM) cantilever. The SPPs propagate down the AFM shaft, localize at the tip apex, and are backscattered to the far-field when the separation distance between the probe tip and the sample surface is comparable to the probe-tip radius. The backscattered near-field intensity is dynamically modulated when an ultrasonic wave arrives at the sample surface leading to a transient change in the tip-sample separation distance. We present experimental results detailing measurement of broadband and narrowband laser generated ultrasound in solids with frequencies reaching up to 180 MHz range.

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.

High-spatial-resolution sub-surface imaging using a laser-based acoustic microscopy technique.

Science.gov (United States)

Balogun, Oluwaseyi; Cole, Garrett D; Huber, Robert; Chinn, Diane; Murray, Todd W; Spicer, James B

2011-01-01

Scanning acoustic microscopy techniques operating at frequencies in the gigahertz range are suitable for the elastic characterization and interior imaging of solid media with micrometer-scale spatial resolution. Acoustic wave propagation at these frequencies is strongly limited by energy losses, particularly from attenuation in the coupling media used to transmit ultrasound to a specimen, leading to a decrease in the depth in a specimen that can be interrogated. In this work, a laser-based acoustic microscopy technique is presented that uses a pulsed laser source for the generation of broadband acoustic waves and an optical interferometer for detection. The use of a 900-ps microchip pulsed laser facilitates the generation of acoustic waves with frequencies extending up to 1 GHz which allows for the resolution of micrometer-scale features in a specimen. Furthermore, the combination of optical generation and detection approaches eliminates the use of an ultrasonic coupling medium, and allows for elastic characterization and interior imaging at penetration depths on the order of several hundred micrometers. Experimental results illustrating the use of the laser-based acoustic microscopy technique for imaging micrometer-scale subsurface geometrical features in a 70-μm-thick single-crystal silicon wafer with a (100) orientation are presented.

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...

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.

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...

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.

A novel self-sensing technique for tapping-mode atomic force microscopy

Energy Technology Data Exchange (ETDEWEB)

Ruppert, Michael G.; Moheimani, S. O. Reza [The University of Newcastle, University Drive, Callaghan NSW 2308 (Australia)

2013-12-15

This work proposes a novel self-sensing tapping-mode atomic force microscopy operation utilizing charge measurement. A microcantilever coated with a single piezoelectric layer is simultaneously used for actuation and deflection sensing. The cantilever can be batch fabricated with existing micro electro mechanical system processes. The setup enables the omission of the optical beam deflection technique which is commonly used to measure the cantilever oscillation amplitude. Due to the high amount of capacitive feedthrough in the measured charge signal, a feedforward control technique is employed to increase the dynamic range from less than 1 dB to approximately 35 dB. Experiments show that the conditioned charge signal achieves excellent signal-to-noise ratio and can therefore be used as a feedback signal for atomic force microscopy imaging.

ADVANCED OPTICAL TECHNIQUES TO EXPLORE BRAIN STRUCTURE AND FUNCTION

OpenAIRE

Silvestri, L.; Mascaro, A. L. Allegra; Lotti, J.; Sacconi, L.; Pavone, F. S.

2013-01-01

Understanding brain structure and function, and the complex relationships between them, is one of the grand challenges of contemporary sciences. Thanks to their flexibility, optical techniques could be the key to explore this complex network. In this manuscript, we briefly review recent advancements in optical methods applied to three main issues: anatomy, plasticity and functionality. We describe novel implementations of light-sheet microscopy to resolve neuronal anatomy in whole fixed brain...

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.

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.

Review of advanced imaging techniques

Directory of Open Access Journals (Sweden)

Yu Chen

2012-01-01

Full Text Available Pathology informatics encompasses digital imaging and related applications. Several specialized microscopy techniques have emerged which permit the acquisition of digital images ("optical biopsies" at high resolution. Coupled with fiber-optic and micro-optic components, some of these imaging techniques (e.g., optical coherence tomography are now integrated with a wide range of imaging devices such as endoscopes, laparoscopes, catheters, and needles that enable imaging inside the body. These advanced imaging modalities have exciting diagnostic potential and introduce new opportunities in pathology. Therefore, it is important that pathology informaticists understand these advanced imaging techniques and the impact they have on pathology. This paper reviews several recently developed microscopic techniques, including diffraction-limited methods (e.g., confocal microscopy, 2-photon microscopy, 4Pi microscopy, and spatially modulated illumination microscopy and subdiffraction techniques (e.g., photoactivated localization microscopy, stochastic optical reconstruction microscopy, and stimulated emission depletion microscopy. This article serves as a primer for pathology informaticists, highlighting the fundamentals and applications of advanced optical imaging techniques.

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.

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.

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.

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)

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

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.

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

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.

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.

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.

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.

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

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.

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

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.

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.

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.

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.

Digital holographic microscopy as a technique to monitor macrophages infected by leishmania

Science.gov (United States)

Mendoza-Rodríguez, E.; Organista-Castelblanco, C.; Camacho, M.; Monroy-Ramírez, F.

2017-06-01

The Digital Holographic Microscopy in Transmission technique (DHM) is considered a useful tool in the noninvasive quantifying of transparent biological objects like living cells. In this work, we propose this technique to study and to monitor control macrophages infected by Leishmania (mouse lineJ774.A1). When the promastigotes enter in contact with healthy macrophages, they got phagocytosed and latterly confined in the formed parasitophorous vacuole. These processes change the morphology and density of the host macrophage. Both parameters can be measured in a label-free analysis of cells with the aid of the DHM technique. Our technique begins with the optical record of the holograms using a modified Mach-Zehnder interferometer and the reconstruction of the complex optical field transmitted by macrophages. In the latter point, we employ the angular spectrum algorithm. With the complex optical field reconstruction, we compute the field amplitude and the phase difference maps, which leads to describe one morphological characterization for the samples. Using phase difference maps is possible to measure internal variations for the integral refractive index, estimating the infection level of macrophages. Through the changes in the integral refractive index, it is also possible to describe and quantify in two different states the evolution of the infection. With these results some parameters of cells have been quantified, making the DHM technique a viable tool for diagnosis of biological samples under the presence of some pathogen.

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

New Technique for Fabrication of Scanning Single-Electron Transistor Microscopy Tips

Science.gov (United States)

Goodwin, Eric; Tessmer, Stuart

Fabrication of glass tips for Scanning Single-Electron Transistor Microscopy (SSETM) can be expensive, time consuming, and inconsistent. Various techniques have been tried, with varying levels of success in regards to cost and reproducibility. The main requirement for SSETM tips is to have a sharp tip ending in a micron-scale flat face to allow for deposition of a quantum dot. Drawing inspiration from methods used to create tips from optical fibers for Near-Field Scanning Optical Microscopes, our group has come up with a quick and cost effective process for creating SSETM tips. By utilizing hydrofluoric acid to etch the tips and oleic acid to guide the etch profile, optical fiber tips with appropriate shaping can be rapidly prepared. Once etched, electric leads are thermally evaporated onto each side of the tip, while an aluminum quantum dot is evaporated onto the face. Preliminary results using various metals, oxide layers, and lead thicknesses have proven promising.

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.

Scanning Near-Field Optical Microscopy

Directory of Open Access Journals (Sweden)

Dušan Vobornik

2008-02-01

Full Text Available 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 temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution.

Scanning near-field optical microscopy.

Science.gov (United States)

Vobornik, Dusan; Vobornik, Slavenka

2008-02-01

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 temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM) is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution.

Transmission-type angle deviation microscopy

International Nuclear Information System (INIS)

Chiu, M.-H.; Lai, C.-W.; Tan, C.-T.; Lai, C.-F.

2008-01-01

We present a new microscopy technique that we call transmission angle deviation microscopy (TADM). It is based on common-path heterodyne interferometry and geometrical optics. An ultrahigh sensitivity surface plasmon resonance (SPR) angular sensor is used to expand dynamic measurement ranges and to improve the axial resolution in three-dimensional optical microscopy. When transmitted light is incident upon a specimen, the beam converges or diverges because of refractive and/or surface height variations. Advantages include high axial resolution (∼32 nm), nondestructive and noncontact measurement, and larger measurement ranges (± 80 μm) for a numerical aperture of 0.21in a transparent measurement medium. The technique can be used without conductivity and pretreatment

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

A SIMULTANEOUS MULTI-PROBE DETECTION LABEL-FREE OPTICAL-RESOLUTION PHOTOACOUSTIC MICROSCOPY TECHNIQUE BASED ON MICROCAVITY TRANSDUCER

Directory of Open Access Journals (Sweden)

YONGBO WU

2013-07-01

Full Text Available We demonstrate the feasibility of simultaneous multi-probe detection for an optical-resolution photoacoustic microscopy (OR-PAM system. OR-PAM has elicited the attention of biomedical imaging researchers because of its optical absorption contrast and high spatial resolution with great imaging depth. OR-PAM allows label-free and noninvasive imaging by maximizing the optical absorption of endogenous biomolecules. However, given the inadequate absorption of some biomolecules, detection sensitivity at the same incident intensity requires improvement. In this study, a modulated continuous wave with power density less than 3 mW/cm2 (1/4 of the ANSI safety limit excited the weak photoacoustic (PA signals of biological cells. A microcavity transducer is developed based on the bulk modulus of gas five orders of magnitude lower than that of solid; air pressure variation is inversely proportional to cavity volume at the same temperature increase. Considering that a PA wave expands in various directions, detecting PA signals from different positions and adding them together can increase detection sensitivity and signal-to-noise ratio. Therefore, we employ four detectors to acquire tiny PA signals simultaneously. Experimental results show that the developed OR-PAM system allows the label-free imaging of cells with weak optical absorption.

Observation of self-assembled fluorescent beads by scanning near-field optical microscopy and atomic force microscopy

International Nuclear Information System (INIS)

Oh, Y.J.; Jo, W.; Kim, Min-Gon; Kyu Park, Hyun; Hyun Chung, Bong

2006-01-01

Optical response and topography of fluorescent latex beads both on flat self-assembled monolayer and on a micron-patterned surface with poly(dimethylsiloxane) are studied. Scanning near-field optical microscopy and atomic force microscopy were utilized together for detecting fluorescence and imaging topography of the patterned latex beads, respectively. As a result, the micro-patterned latex beads where a specific chemical binding occurred show a strong signal, whereas no signals are observed in the case of nonspecific binding. With fluorescein isothiocyanate (FITC), it is convenient to measure fluorescence signal from the patterned beads allowing us to monitor the small balls of fluorescent latex

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.

Developing new optical imaging techniques for single particle and molecule tracking in live cells

Energy Technology Data Exchange (ETDEWEB)

Sun, Wei [Iowa State Univ., Ames, IA (United States)

2010-01-01

Differential interference contrast (DIC) microscopy is a far-field as well as wide-field optical imaging technique. Since it is non-invasive and requires no sample staining, DIC microscopy is suitable for tracking the motion of target molecules in live cells without interfering their functions. In addition, high numerical aperture objectives and condensers can be used in DIC microscopy. The depth of focus of DIC is shallow, which gives DIC much better optical sectioning ability than those of phase contrast and dark field microscopies. In this work, DIC was utilized to study dynamic biological processes including endocytosis and intracellular transport in live cells. The suitability of DIC microscopy for single particle tracking in live cells was first demonstrated by using DIC to monitor the entire endocytosis process of one mesoporous silica nanoparticle (MSN) into a live mammalian cell. By taking advantage of the optical sectioning ability of DIC, we recorded the depth profile of the MSN during the endocytosis process. The shape change around the nanoparticle due to the formation of a vesicle was also captured. DIC microscopy was further modified that the sample can be illuminated and imaged at two wavelengths simultaneously. By using the new technique, noble metal nanoparticles with different shapes and sizes were selectively imaged. Among all the examined metal nanoparticles, gold nanoparticles in rod shapes were found to be especially useful. Due to their anisotropic optical properties, gold nanorods showed as diffraction-limited spots with disproportionate bright and dark parts that are strongly dependent on their orientation in the 3D space. Gold nanorods were developed as orientation nanoprobes and were successfully used to report the self-rotation of gliding microtubules on kinesin coated substrates. Gold nanorods were further used to study the rotational motions of cargoes during the endocytosis and intracellular transport processes in live mammalian

Fabrication and characterization of optical-fiber nanoprobes for scanning near-field optical microscopy.

Science.gov (United States)

Essaidi, N; Chen, Y; Kottler, V; Cambril, E; Mayeux, C; Ronarch, N; Vieu, C

1998-02-01

The current scanning near-field optical microscopy has been developed with optical-fiber probes obtained by use of either laser-heated pulling or chemical etching. For high-resolution near-field imaging, the detected signal is rapidly attenuated as the aperture size of the probe decreases. It is thus important to fabricate probes optimized for both spot size and optical transmission. We present a two-step fabrication that allowed us to achieve an improved performance of the optical-fiber probes. Initially, a CO(2) laser-heated pulling was used to produce a parabolic transitional taper ending with a top thin filament. Then, a rapid chemical etching with 50% buffered hydrofluoric acid was used to remove the thin filament and to result in a final conical tip on the top of the parabolic transitional taper. Systematically, we obtained optical-fiber nanoprobes with the apex size as small as 10 nm and the final cone angle varying from 15 degrees to 80 degrees . It was found that the optical transmission efficiency increases rapidly as the taper angle increases from 15 degrees to 50 degrees , but a further increase in the taper angle gives rise to important broadening of the spot size. Finally, the fabricated nanoprobes were used in photon-scanning tunneling microscopy, which allowed observation of etched double lines and grating structures with periods as small as 200 nm.

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.

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.

Structured illumination microscopy and its new developments

Directory of Open Access Journals (Sweden)

Jianling Chen

2016-05-01

Full Text Available Optical microscopy allows us to observe the biological structures and processes within living cells. However, the spatial resolution of the optical microscopy is limited to about half of the wavelength by the light diffraction. Structured illumination microscopy (SIM, a type of new emerging super-resolution microscopy, doubles the spatial resolution by illuminating the specimen with a patterned light, and the sample and light source requirements of SIM are not as strict as the other super-resolution microscopy. In addition, SIM is easier to combine with the other imaging techniques to improve their imaging resolution, leading to the developments of diverse types of SIM. SIM has great potential to meet the various requirements of living cells imaging. Here, we review the recent developments of SIM and its combination with other imaging techniques.

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.

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.

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.

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.

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.

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

Scanning near-field optical microscopy and near-field optical probes: properties, fabrication, and control of parameters

International Nuclear Information System (INIS)

Dryakhlushin, V F; Veiko, V P; Voznesenskii, N B

2007-01-01

A brief review of modern applications of scanning near-field optical (SNO) devices in microscopy, spectroscopy, and lithography is presented in the introduction. The problem of the development of SNO probes, as the most important elements of SNO devices determining their resolution and efficiency, is discussed. Based on the works of the authors, two different methods for fabricating SNO probes by using the adiabatic tapering of an optical fibre are considered: the laser-heated mechanical drawing and chemical etching. A nondestructive optical method for controlling the nanometre aperture of SNO probes is proposed, substantiated, and tested experimentally. The method is based on the reconstruction of a near-field source with the help of a theoretical algorithm of the inverse problem from the experimental far-filed intensity distribution. Some prospects for a further refinement of the construction and technology of SNO probes are discussed. (optical microscopy)

Occlusal overload investigations by noninvasive technology: fluorescence microscopy and en-face optical coherence tomography

Science.gov (United States)

Marcauteanu, Corina; Negrutiu, Meda; Sinescu, Cosmin; Demjan, Enikö; Hughes, Michael; Bradu, Adrian; Dobre, George; Podoleanu, Adrian G.

2009-07-01

The aim of this study is the early detection and monitoring of occlusal overload in bruxing patients. En-Face Optical coherence tomography (eF-OCT) and fluorescence microscopy (FM) were used for the imaging of several anterior teeth extracted from patients with light active bruxism. We found a characteristic pattern of enamel cracks, that reached the tooth surface. We concluded that the combination of the en-Face OCT and FM is a promising non-invasive alternative technique for reliable monitoring of occlusal overload.

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

Image-based overlay and alignment metrology through optically opaque media with sub-surface probe microscopy

Science.gov (United States)

van Es, Maarten H.; Mohtashami, Abbas; Piras, Daniele; Sadeghian, Hamed

2018-03-01

Nondestructive subsurface nanoimaging through optically opaque media is considered to be extremely challenging and is essential for several semiconductor metrology applications including overlay and alignment and buried void and defect characterization. The current key challenge in overlay and alignment is the measurement of targets that are covered by optically opaque layers. Moreover, with the device dimensions moving to the smaller nodes and the issue of the so-called loading effect causing offsets between between targets and product features, it is increasingly desirable to perform alignment and overlay on product features or so-called on-cell overlay, which requires higher lateral resolution than optical methods can provide. Our recently developed technique known as SubSurface Ultrasonic Resonance Force Microscopy (SSURFM) has shown the capability for high-resolution imaging of structures below a surface based on (visco-)elasticity of the constituent materials and as such is a promising technique to perform overlay and alignment with high resolution in upcoming production nodes. In this paper, we describe the developed SSURFM technique and the experimental results on imaging buried features through various layers and the ability to detect objects with resolution below 10 nm. In summary, the experimental results show that the SSURFM is a potential solution for on-cell overlay and alignment as well as detecting buried defects or voids and generally metrology through optically opaque layers.

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.

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.

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

Interferometric and nonlinear-optical spectral-imaging techniques for outer space and live cells

Science.gov (United States)

Itoh, Kazuyoshi

2015-12-01

Multidimensional signals such as the spectral images allow us to have deeper insights into the natures of objects. In this paper the spectral imaging techniques that are based on optical interferometry and nonlinear optics are presented. The interferometric imaging technique is based on the unified theory of Van Cittert-Zernike and Wiener-Khintchine theorems and allows us to retrieve a spectral image of an object in the far zone from the 3D spatial coherence function. The retrieval principle is explained using a very simple object. The promising applications to space interferometers for astronomy that are currently in progress will also be briefly touched on. An interesting extension of interferometric spectral imaging is a 3D and spectral imaging technique that records 4D information of objects where the 3D and spectral information is retrieved from the cross-spectral density function of optical field. The 3D imaging is realized via the numerical inverse propagation of the cross-spectral density. A few techniques suggested recently are introduced. The nonlinear optical technique that utilizes stimulated Raman scattering (SRS) for spectral imaging of biomedical targets is presented lastly. The strong signals of SRS permit us to get vibrational information of molecules in the live cell or tissue in real time. The vibrational information of unstained or unlabeled molecules is crucial especially for medical applications. The 3D information due to the optical nonlinearity is also the attractive feature of SRS spectral microscopy.

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.

Electrical characterization of grain boundaries of CZTS thin films using conductive atomic force microscopy techniques

Energy Technology Data Exchange (ETDEWEB)

Muhunthan, N.; Singh, Om Pal [Compound Semiconductor Solar Cell, Physics of Energy Harvesting Division, New Delhi 110012 (India); Toutam, Vijaykumar, E-mail: toutamvk@nplindia.org [Quantum Phenomena and Applications Division, CSIR-National Physical Laboratory, Dr. K.S. Krishnan Marg, New Delhi 110012 (India); Singh, V.N., E-mail: singhvn@nplindia.org [Compound Semiconductor Solar Cell, Physics of Energy Harvesting Division, New Delhi 110012 (India)

2015-10-15

Graphical abstract: Experimental setup for conducting AFM (C-AFM). - Highlights: • Cu{sub 2}ZnSnS{sub 4} (CZTS) thin film was grown by reactive co-sputtering. • The electronic properties were probed using conducting atomic force microscope, scanning Kelvin probe microscopy and scanning capacitance microscopy. • C-AFM current flow mainly through grain boundaries rather than grain interiors. • SKPM indicated higher potential along the GBs compared to grain interiors. • The SCM explains that charge separation takes place at the interface of grain and grain boundary. - Abstract: Electrical characterization of grain boundaries (GB) of Cu-deficient CZTS (Copper Zinc Tin Sulfide) thin films was done using atomic force microscopic (AFM) techniques like Conductive atomic force microscopy (CAFM), Kelvin probe force microscopy (KPFM) and scanning capacitance microscopy (SCM). Absorbance spectroscopy was done for optical band gap calculations and Raman, XRD and EDS for structural and compositional characterization. Hall measurements were done for estimation of carrier mobility. CAFM and KPFM measurements showed that the currents flow mainly through grain boundaries (GB) rather than grain interiors. SCM results showed that charge separation mainly occurs at the interface of grain and grain boundaries and not all along the grain boundaries.

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.

Three-ring filters increase the effective NA up to 1.46 in optical sectioning fluorescence microscopy

International Nuclear Information System (INIS)

Martinez-Corral, M; Ibanez-Lopez, C; Caballero, M T; Munoz-Escriva, L; Saavedra, G

2003-01-01

Single-photon fluorescence confocal microscopy techniques can be combined with the use of specific binary filters in order to increase their optical sectioning capability. We present a novel class of axially super-resolving binary pupil filters specially designed to reach this aim. These filters let us to obtain a relevant compression of the z-response together with the reduction of the photo-bleaching effect typically inherent to apodization techniques. The fact of joining both the three-ring filters we propose in the illumination path, and the confocal detection gives rise to an important effective increase of lenses of effective numerical aperture

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.

Near-field scanning optical microscopy using polymethylmethacrylate optical fiber probes

International Nuclear Information System (INIS)

Chibani, H.; Dukenbayev, K.; Mensi, M.; Sekatskii, S.K.; Dietler, G.

2010-01-01

We report the first use of polymethylmethacrylate (PMMA) optical fiber-made probes for scanning near-field optical microscopy (SNOM). The sharp tips were prepared by chemical etching of the fibers in ethyl acetate, and the probes were prepared by proper gluing of sharpened fibers onto the tuning fork in the conditions of the double resonance (working frequency of a tuning fork coincides with the resonance frequency of dithering of the free-standing part of the fiber) reported earlier for the case of glass fibers. Quality factors of the probes in the range 2000-6000 were obtained, which enables the realization of an excellent topographical resolution including state-of-art imaging of single DNA molecules. Near-field optical performance of the microscope is illustrated by the Photon Scanning Tunneling Microscope images of fluorescent beads with a diameter of 100 nm. The preparation of these plastic fiber probes proved to be easy, needs no hazardous material and/or procedures, and typical lifetime of a probe essentially exceeds that characteristic for the glass fiber probe.

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.

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.

Multimodal imaging of vascular network and blood microcirculation by optical diagnostic techniques

International Nuclear Information System (INIS)

Kuznetsov, Yu L; Kalchenko, V V; Meglinski, I V

2011-01-01

We present a multimodal optical diagnostic approach for simultaneous non-invasive in vivo imaging of blood and lymphatic microvessels, utilising a combined use of fluorescence intravital microscopy and a method of dynamic light scattering. This approach makes it possible to renounce the use of fluorescent markers for visualisation of blood vessels and, therefore, significantly (tenfold) reduce the toxicity of the technique and minimise side effects caused by the use of contrast fluorescent markers. We demonstrate that along with the ability to obtain images of lymph and blood microvessels with a high spatial resolution, current multimodal approach allows one to observe in real time permeability of blood vessels. This technique appears to be promising in physiology studies of blood vessels, and especially in the study of peripheral cardiovascular system in vivo. (optical technologies in biophysics and medicine)

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.

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,…

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.

Nonlinear adaptive optics: aberration correction in three photon fluorescence microscopy for mouse brain imaging

Science.gov (United States)

Sinefeld, David; Paudel, Hari P.; Wang, Tianyu; Wang, Mengran; Ouzounov, Dimitre G.; Bifano, Thomas G.; Xu, Chris

2017-02-01

Multiphoton fluorescence microscopy is a well-established technique for deep-tissue imaging with subcellular resolution. Three-photon microscopy (3PM) when combined with long wavelength excitation was shown to allow deeper imaging than two-photon microscopy (2PM) in biological tissues, such as mouse brain, because out-of-focus background light can be further reduced due to the higher order nonlinear excitation. As was demonstrated in 2PM systems, imaging depth and resolution can be improved by aberration correction using adaptive optics (AO) techniques which are based on shaping the scanning beam using a spatial light modulator (SLM). In this way, it is possible to compensate for tissue low order aberration and to some extent, to compensate for tissue scattering. Here, we present a 3PM AO microscopy system for brain imaging. Soliton self-frequency shift is used to create a femtosecond source at 1675 nm and a microelectromechanical (MEMS) SLM serves as the wavefront shaping device. We perturb the 1020 segment SLM using a modified nonlinear version of three-point phase shifting interferometry. The nonlinearity of the fluorescence signal used for feedback ensures that the signal is increasing when the spot size decreases, allowing compensation of phase errors in an iterative optimization process without direct phase measurement. We compare the performance for different orders of nonlinear feedback, showing an exponential growth in signal improvement as the nonlinear order increases. We demonstrate the impact of the method by applying the 3PM AO system for in-vivo mouse brain imaging, showing improvement in signal at 1-mm depth inside the brain.

[Getting an insight into the brain - new optical clearing techniques and imaging using light-sheet microscope].

Science.gov (United States)

Pawłowska, Monika; Legutko, Diana; Stefaniuk, Marzena

2017-01-01

One of the biggest challenges in neuroscience is to understand how brain operates. For this, it would be the best to image the whole brain with at least cellular resolution, preserving the three-dimensional structure in order to capture the connections between different areas. Most currently available high-resolution imaging techniques are based on preparing thin brain sections that are next photographed one by one and subsequently bigger structures are reconstructed. These techniques are laborious and create artifacts. Recent optical clearing methods allow to obtain literally transparent brains that can be imaged using light-sheet microscope. The present review summarizes the most popular optical clearing techniques, describing their different mechanisms and comparing advantages and disadvantages of different approaches, and presents the principle of light-sheet microscopy and its use in imaging. Finally, it gives examples of application of optical tissue clearing and light-sheet imaging in neuroscience and beyond it.

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.

Atomic force and scanning near-field optical microscopy study of carbocyanine dye J-aggregates

Czech Academy of Sciences Publication Activity Database

Prokhorov, V.V.; Petrova, M.G.; Kovaleva, Natalia; Demikhov, E.I.

2014-01-01

Roč. 10, č. 5 (2014), s. 700-704 ISSN 1573-4137 Institutional support: RVO:68378271 Keywords : carbocyanine dye * elementary fibri * high-resolution atomic force microscopy * J-aggregate * probe microscopy * scanning near-field optical microscopy Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.096, year: 2014

Comparative study of quantitative phase imaging techniques for refractometry of optical fibers

Science.gov (United States)

de Dorlodot, Bertrand; Bélanger, Erik; Bérubé, Jean-Philippe; Vallée, Réal; Marquet, Pierre

2018-02-01

The refractive index difference profile of optical fibers is the key design parameter because it determines, among other properties, the insertion losses and propagating modes. Therefore, an accurate refractive index profiling method is of paramount importance to their development and optimization. Quantitative phase imaging (QPI) is one of the available tools to retrieve structural characteristics of optical fibers, including the refractive index difference profile. Having the advantage of being non-destructive, several different QPI methods have been developed over the last decades. Here, we present a comparative study of three different available QPI techniques, namely the transport-of-intensity equation, quadriwave lateral shearing interferometry and digital holographic microscopy. To assess the accuracy and precision of those QPI techniques, quantitative phase images of the core of a well-characterized optical fiber have been retrieved for each of them and a robust image processing procedure has been applied in order to retrieve their refractive index difference profiles. As a result, even if the raw images for all the three QPI methods were suffering from different shortcomings, our robust automated image-processing pipeline successfully corrected these. After this treatment, all three QPI techniques yielded accurate, reliable and mutually consistent refractive index difference profiles in agreement with the accuracy and precision of the refracted near-field benchmark measurement.

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

Superresolution Microscopy of the Nuclear Envelope and Associated Proteins.

Science.gov (United States)

Xie, Wei; Horn, Henning F; Wright, Graham D

2016-01-01

Superresolution microscopy is undoubtedly one of the most exciting technologies since the invention of the optical microscope. Capable of nanometer-scale resolution to surpass the diffraction limit and coupled with the versatile labeling techniques available, it is revolutionizing the study of cell biology. Our understanding of the nucleus, the genetic and architectural center of the cell, has gained great advancements through the application of various superresolution microscopy techniques. This chapter describes detailed procedures of multichannel superresolution imaging of the mammalian nucleus, using structured illumination microscopy and single-molecule localization microscopy.

Optical coherence tomography: Technique and applications

DEFF Research Database (Denmark)

Thomsen, Jakob Borup; Sander, Birgit; Mogensen, Mette

2009-01-01

Optical coherence tomography (OCT) is a noninvasive optical imaging modality providing real-time video rate images in two and three dimensions of biological tissues with micrometer resolution. OCT fills the gap between ultrasound and confocal microscopy, since it has a higher resolution than...... of retinal diseases. The potential of OCT in many other applications is currently being explored, such as in developmental biology, skin cancer diagnostics, vulnerable plaque detection in cardiology, esophageal diagnostics and a number of other applications within oncology....

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.

Extending the methodology of X-ray crystallography to allow X-ray microscopy without X-ray optics

International Nuclear Information System (INIS)

Miao Jianwei; Kirz, Janos; Sayre, David; Charalambous, Pambos

2000-01-01

We demonstrate that the soft X-ray diffraction pattern from a micron-size noncrystalline specimen can be recorded and inverted to form a high-resolution image. The phase problem is overcome by oversampling the diffraction pattern. The image is obtained using an iterative algorithm. The technique provides a method for X-ray microscopy requiring no high-resolution X-ray optical elements or detectors. In the present work, a resolution of approximately 60 nm was obtained, but we believe that considerably higher resolution can be achieved

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.

Gold nanocone probes for near-field scanning optical microscopy

Energy Technology Data Exchange (ETDEWEB)

Zeeb, Bastian; Schaefer, Christian; Nill, Peter; Fleischer, Monika; Kern, Dieter P. [Institute of Applied Physics, University of Tuebingen, Auf der Morgenstelle 10, 72076 Tuebingen (Germany)

2010-07-01

Apertureless near-field scanning optical microscopy (ANSOM) provides the possibility to collect simultaneously high-resolution topographical and sub-diffraction limited optical information from a surface. When optically excited, the scanning probes act as optical antennae with a strong near-field enhancement near the tip apex. Spatial resolution and optical near-field enhancement depend strongly on the properties and geometry of the scanning probe - in particular on very sharp tip radii. Various possibilities for fabricating good antennae have been pursued. Most commonly, scanning probes consist of electrochemically etched gold wires which are sharp but not well-defined in geometry. We present two different approaches for ultra sharp and well-defined antennae based upon fabricating gold nanocones with a tip radius smaller than 10 nm which can be used in ANSOM. A transfer process is presented that can be used to attach single gold nanocones to non-metallic probes such as sharp glass fiber tips. Alternatively, new processes are presented to fabricate cones directly on pillars of different materials such as silicon or bismuth, which can be applied to cantilever tips for ANSOM scanning applications.

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

Application of oblique plane microscopy to high speed live cell imaging

Science.gov (United States)

Kumar, Sunil; Wilding, Dean; Sikkel, Markus B.; Lyon, Alexander R.; MacLeod, Ken T.; Dunsby, Chris

2011-07-01

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. We present high speed 2D and 3D optically sectioned OPM imaging of live cells using a high NA water immersion lens.

Evanescent field characterisation for a d-shaped optical fibre using scanning near-field optical microscopy

International Nuclear Information System (INIS)

Huntington, S.T.; Nugent, K.A.; Roberts, A.; Mulvaney, P.; Lo, K.M.

1997-01-01

Scanning near field optical microscopy is used to measure the evanescent filed and mode profile of a Ge-doped D-shaped optical fibre. The structure of the fibre is determined by differential etching followed by an investigation of the resultant topography with an atomic force microscope. This information is then used to theoretically model the expected behaviour of the fibre and it is shown that the theoretically model the expected behaviour of the fibre and it is shown that the theoretical results are in excellent agreement with the experimentally observed fields

Differential-interference-contrast digital in-line holography microscopy based on a single-optical-element.

Science.gov (United States)

Zhang, Yuchao; Xie, Changqing

2015-11-01

Both digital in-line holography (DIH) and zone plate-based microscopy have received considerable interest as powerful imaging tools. However, the former suffers from a twin-image noise problem. The latter suffers from low efficiency and difficulty in fabrication. Here, we present an effective and efficient phase-contrast imaging approach, named differential-interference-contrast digital in-line holography (DIC-DIH), by using a single optical element to split the incident light into a plane wave and a converging spherical wave and generate a two-dimensional (2D) DIC effect simultaneously. Specifically, to improve image contrast, we present a new single optical element, termed 2D DIC compound photon sieves, by combining two overlaid binary gratings and a compound photon sieve through two logical XOR operations. The proof-of-concept experiments demonstrate that the proposed technique can eliminate the twin-image noise problem and improve image contrast with high efficiency. Additionally, we present an example of the phase-contrast imaging nonuniform thick photoresist development process.

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...

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 techniques for mechanical characterization at nanoscale

International Nuclear Information System (INIS)

Passeri, D.; Anastasiadis, P.; Tamburri, E.; Gugkielmotti, V.; Rossi, M.

2013-01-01

Three atomic force microscopy (AFM)-based techniques are reviewed that allow one to conduct accurate measurements of mechanical properties of either stiff or compliant materials at a nanometer scale. Atomic force acoustic microscopy, AFM-based depth sensing indentation, and torsional harmonic AFM are briefly described. Examples and results of quantitative characterization of stiff (an ultrathin SeSn film), soft polymeric (polyaniline fibers doped with detonation nanodiamond) and biological (collagen fibers) materials are reported.

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

Investigation of ceramic devices by analytical electron microscopy techniques

International Nuclear Information System (INIS)

Shiojiri, M.; Saijo, H.; Isshiki, T.; Kawasaki, M.; Yoshioka, T.; Sato, S.; Nomura, T.

1999-01-01

Ceramics are widely used as capacitors and varistors. Their electrical properties depend on the structure, which is deeply influenced not only by the composition of raw materials and additives but also by heating treatments in the production process. This paper reviews our investigations of SrTiO 3 ceramic devices, which have been performed using various microscopy techniques such as high-resolution transmission electron microscopy (HRTEM), cathodoluminescence scanning electron microscopy (CLSEM), field emission SEM (FE-SEM), energy dispersive X-ray spectroscopy (EDS), electron energy-loss spectroscopy (EELS) and high angle annular dark field (HAADF) imaging method in a FE-(scanning) transmission electron microscope(FE-(S)TEM). (author)

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.

Ex vivo confocal microscopy: a new diagnostic technique for mucormycosis.

Science.gov (United States)

Leclercq, A; Cinotti, E; Labeille, B; Perrot, J L; Cambazard, F

2016-05-01

Skin-dedicated ex vivo confocal microscopy (EVCM) has so far mainly been employed to identify cutaneous tumours on freshly excised samples. We present two cases where EVCM has been used to diagnose cutaneous mucormycosis. The skin biopsies were evaluated by the skin-dedicated ex vivo confocal microscope VivaScope 2500(®) (MAVIG GmbH, Munich Germany) under both reflectance and fluorescence mode. Conventional direct optical examination on skin scraping and histological examination were later performed. Mucormycetes observed by EVCM presented as hyper-reflective elongated 20 μm in diameter structures with perpendicular ramifications. Fungi were found both under reflectance and fluorescence mode and were better visible with acridine orange under fluorescence EVCM. Conventional direct optical examination on skin scraping and histological examination found the same elongated and branching structures confirming the presence of Mucormycetes. Ex vivo confocal microscopy has both the advantages of being fast as the direct optical examination, and to be able to show the localisation of the fungi in the tissue like the histological examination. In our cases, EVCM allowed to rapidly confirm the clinical diagnosis of mucormycosis, which is essential for the treatment of this fungal infection. Further studies are needed to compare the performance of EVCM with the findings of conventional histological and mycological examinations. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Possibilities and Challenges of Scanning Hard X-ray Spectro-microscopy Techniques in Material Sciences

Directory of Open Access Journals (Sweden)

Andrea Somogyi

2015-06-01

Full Text Available Scanning hard X-ray spectro-microscopic imaging opens unprecedented possibilities in the study of inhomogeneous samples at different length-scales. It gives insight into the spatial variation of the major and minor components, impurities and dopants of the sample, and their chemical and electronic states at micro- and nano-meter scales. Measuring, modelling and understanding novel properties of laterally confined structures are now attainable. The large penetration depth of hard X-rays (several keV to several 10 keV beam energy makes the study of layered and buried structures possible also in in situ and in operando conditions. The combination of different X-ray analytical techniques complementary to scanning spectro-microscopy, such as X-ray diffraction, X-ray excited optical luminescence, secondary ion mass spectrometry (SIMS and nano-SIMS, provides access to optical characteristics and strain and stress distributions. Complex sample environments (temperature, pressure, controlled atmosphere/vacuum, chemical environment are also possible and were demonstrated, and allow as well the combination with other analysis techniques (Raman spectroscopy, infrared imaging, mechanical tensile devices, etc. on precisely the very same area of the sample. The use of the coherence properties of X-rays from synchrotron sources is triggering emerging experimental imaging approaches with nanometer lateral resolution. New fast analytical possibilities pave the way towards statistically significant studies at multi- length-scales and three dimensional tomographic investigations. This paper gives an overview of these techniques and their recent achievements in the field of material sciences.

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

Fluorescence confocal polarizing microscopy

Indian Academy of Sciences (India)

Much of the modern understanding of orientational order in liquid crystals (LCs) is based on polarizing microscopy (PM). A PM image bears only two-dimensional (2D) information, integrating the 3D pattern of optical birefringence over the path of light. Recently, we proposed a technique to image 3D director patterns by ...

Cathodoluminescence-activated nanoimaging: noninvasive near-field optical microscopy in an electron microscope.

Science.gov (United States)

Bischak, Connor G; Hetherington, Craig L; Wang, Zhe; Precht, Jake T; Kaz, David M; Schlom, Darrell G; Ginsberg, Naomi S

2015-05-13

We demonstrate a new nanoimaging platform in which optical excitations generated by a low-energy electron beam in an ultrathin scintillator are used as a noninvasive, near-field optical scanning probe of an underlying sample. We obtain optical images of Al nanostructures with 46 nm resolution and validate the noninvasiveness of this approach by imaging a conjugated polymer film otherwise incompatible with electron microscopy due to electron-induced damage. The high resolution, speed, and noninvasiveness of this "cathodoluminescence-activated" platform also show promise for super-resolution bioimaging.

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.

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.

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.

Ultra-precise measurement of optical aberrations for sub-Aangstroem transmission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Barthel, J.

2008-06-15

Quantitative investigations of material structures on an atomic scale by means of highresolution transmission electron microscopy (HRTEM) impose not only extreme demands on the mechanic and electromagnetic stability of the applied instruments but require also their precise electron-optical adjustment. Today a physical resolution well below one Aangstroem can be achieved with commercially available microscopes on a daily basis. However, the achieved resolution can often not be reliably exploited for the interpretation of the resulting microscopical data due to the presence of so-called higher-order lens aberrations. At the starting time of this work, a sufficiently accurate procedure to measure higher-order aberrations was urgently missing. Since aberration measurement is a mandatory prerequisite for any technique of aberration control enabling quantitative high-resolution microscopy, the goal of this work is to develop such a measurement procedure for the Sub-Aangstroem regime. The measurement procedures developed in the course of this work are based on the numerical evaluation of a series of images taken from an amorphous object under electron-beam illumination with varying tilt. New techniques have been developed for the evaluation of single images as well as for the optimised evaluation of the whole series. These procedures allow microscope users to perform quantitative HRTEM even at a resolution of 0.5 Aangstroem. The precision reached with the newly developed measurement procedures is unprecedented and surpasses existing solutions by at least one order of magnitude in any respect. All the concepts and procedures for aberration measurement developed in this work have been implemented in a software package which satisfies professional demands with respect to robustness, precision, speed and user-friendliness. The new automatic aberrationmeasurement procedures are suitable to establish HRTEM as a quantitative technique for material science investigations in the

Ultra-precise measurement of optical aberrations for sub-Aangstroem transmission electron microscopy

International Nuclear Information System (INIS)

Barthel, J.

2008-06-01

Quantitative investigations of material structures on an atomic scale by means of highresolution transmission electron microscopy (HRTEM) impose not only extreme demands on the mechanic and electromagnetic stability of the applied instruments but require also their precise electron-optical adjustment. Today a physical resolution well below one Aangstroem can be achieved with commercially available microscopes on a daily basis. However, the achieved resolution can often not be reliably exploited for the interpretation of the resulting microscopical data due to the presence of so-called higher-order lens aberrations. At the starting time of this work, a sufficiently accurate procedure to measure higher-order aberrations was urgently missing. Since aberration measurement is a mandatory prerequisite for any technique of aberration control enabling quantitative high-resolution microscopy, the goal of this work is to develop such a measurement procedure for the Sub-Aangstroem regime. The measurement procedures developed in the course of this work are based on the numerical evaluation of a series of images taken from an amorphous object under electron-beam illumination with varying tilt. New techniques have been developed for the evaluation of single images as well as for the optimised evaluation of the whole series. These procedures allow microscope users to perform quantitative HRTEM even at a resolution of 0.5 Aangstroem. The precision reached with the newly developed measurement procedures is unprecedented and surpasses existing solutions by at least one order of magnitude in any respect. All the concepts and procedures for aberration measurement developed in this work have been implemented in a software package which satisfies professional demands with respect to robustness, precision, speed and user-friendliness. The new automatic aberrationmeasurement procedures are suitable to establish HRTEM as a quantitative technique for material science investigations in the

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

Cement paste surface roughness analysis using coherence scanning interferometry and confocal microscopy

Energy Technology Data Exchange (ETDEWEB)

Apedo, K.L., E-mail: apedo@unistra.fr [ICube, Université de Strasbourg, CNRS, 2 rue Boussingault, 67000 Strasbourg (France); Munzer, C.; He, H. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France); Montgomery, P. [ICube, Université de Strasbourg, CNRS, 23 rue du Loess, 67037 Strasbourg (France); Serres, N. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France); Fond, C. [ICube, Université de Strasbourg, CNRS, 2 rue Boussingault, 67000 Strasbourg (France); Feugeas, F. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France)

2015-02-15

Scanning electron microscopy and scanning probe microscopy have been used for several decades to better understand the microstructure of cementitious materials. Very limited work has been performed to date to study the roughness of cementitious materials by optical microscopy such as coherence scanning interferometry (CSI) and chromatic confocal sensing (CCS). The objective of this paper is to better understand how CSI can be used as a tool to analyze surface roughness and topography of cement pastes. Observations from a series of images acquired using this technique on both polished and unpolished samples are described. The results from CSI are compared with those from a STIL confocal microscopy technique (SCM). Comparison between both optical techniques demonstrates the ability of CSI to measure both polished and unpolished cement pastes. - Highlights: • Coherence scanning interferometry (CSI) was used to analyze cement paste surfaces. • The results from the CSI were compared with those from a confocal microscopy. • 3D roughness parameters were obtained using the window resizing method. • Polished and unpolished cement pastes were studied.

Cement paste surface roughness analysis using coherence scanning interferometry and confocal microscopy

International Nuclear Information System (INIS)

Apedo, K.L.; Munzer, C.; He, H.; Montgomery, P.; Serres, N.; Fond, C.; Feugeas, F.

2015-01-01

Scanning electron microscopy and scanning probe microscopy have been used for several decades to better understand the microstructure of cementitious materials. Very limited work has been performed to date to study the roughness of cementitious materials by optical microscopy such as coherence scanning interferometry (CSI) and chromatic confocal sensing (CCS). The objective of this paper is to better understand how CSI can be used as a tool to analyze surface roughness and topography of cement pastes. Observations from a series of images acquired using this technique on both polished and unpolished samples are described. The results from CSI are compared with those from a STIL confocal microscopy technique (SCM). Comparison between both optical techniques demonstrates the ability of CSI to measure both polished and unpolished cement pastes. - Highlights: • Coherence scanning interferometry (CSI) was used to analyze cement paste surfaces. • The results from the CSI were compared with those from a confocal microscopy. • 3D roughness parameters were obtained using the window resizing method. • Polished and unpolished cement pastes were studied

Digital processing optical transmission and coherent receiving techniques

CERN Document Server

Binh, Le Nguyen

2013-01-01

With coherent mixing in the optical domain and processing in the digital domain, advanced receiving techniques employing ultra-high speed sampling rates have progressed tremendously over the last few years. These advances have brought coherent reception systems for lightwave-carried information to the next stage, resulting in ultra-high capacity global internetworking. Digital Processing: Optical Transmission and Coherent Receiving Techniques describes modern coherent receiving techniques for optical transmission and aspects of modern digital optical communications in the most basic lines. The

Spatiotemporal closure of fractional laser-ablated channels imaged by optical coherence tomography and reflectance confocal microscopy

DEFF Research Database (Denmark)

Banzhaf, Christina A.; Wind, Bas S.; Mogensen, Mette

2016-01-01

Background and Objective Optical coherence tomography (OCT) and reflectance confocal microscopy (RCM) offer high-resolution optical imaging of the skin, which may provide benefit in the context of laser-assisted drug delivery. We aimed to characterize postoperative healing of ablative fractional...... laser (AFXL)-induced channels and dynamics in their spatiotemporal closure using in vivo OCT and RCM techniques. Study design/Materials and Methods The inner forearm of healthy subjects (n = 6) was exposed to 10,600 nm fractional CO2 laser using 5 and 25% densities, 120 μm beam diameter, 5, 15, and 25 m......J/microbeam. Treatment sites were scanned with OCT to evaluate closure of AFXL-channels and RCM to evaluate subsequent re-epithelialization. Results OCT and RCM identified laser channels in epidermis and upper dermis as black, ablated tissue defects surrounded by characteristic hyper-and hyporeflective zones. OCT imaged...

Group velocity measurement using spectral interference in near-field scanning optical microscopy

International Nuclear Information System (INIS)

Mills, John D.; Chaipiboonwong, Tipsuda; Brocklesby, William S.; Charlton, Martin D. B.; Netti, Caterina; Zoorob, Majd E.; Baumberg, Jeremy J.

2006-01-01

Near-field scanning optical microscopy provides a tool for studying the behavior of optical fields inside waveguides. In this experiment the authors measure directly the variation of group velocity between different modes of a planar slab waveguide as the modes propagate along the guide. The measurement is made using the spectral interference between pulses propagating inside the waveguide with different group velocities, collected using a near-field scanning optical microscope at different points down the guide and spectrally resolved. The results are compared to models of group velocities in simple guides

Machine learning techniques for optical communication system optimization

DEFF Research Database (Denmark)

Zibar, Darko; Wass, Jesper; Thrane, Jakob

In this paper, machine learning techniques relevant to optical communication are presented and discussed. The focus is on applying machine learning tools to optical performance monitoring and performance prediction.......In this paper, machine learning techniques relevant to optical communication are presented and discussed. The focus is on applying machine learning tools to optical performance monitoring and performance prediction....

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.

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.

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.

Optical measurements for scientists and engineers a practical guide

CERN Document Server

McClelland, Arthur

2018-01-01

With this accessible, introductory guide, you will quickly learn how to use and apply optical spectroscopy and optical microscopy techniques. Focusing on day-to-day implementation and offering practical lab tips throughout, it provides step-by-step instructions on how to select the best technique for a particular application, how to set up and customize new optical systems, and how to analyze optical data. You will gain an intuitive understanding of the full range of standard optical techniques, from fluorescence and Raman spectroscopy to super resolution microscopy. Understand how to navigate around an optics lab with clear descriptions of the most common optical components and tools. Including explanations of basic optics and photonics, and easy-to-understand mathematics, this is an invaluable resource for graduate students, instructors, researchers and professionals who use or teach optical measurements in laboratories.

Near-field and far-field modeling of scattered surface waves. Application to the apertureless scanning near-field optical microscopy

International Nuclear Information System (INIS)

Muller, J.; Parent, G.; Fumeron, S.; Jeandel, G.; Lacroix, D.

2011-01-01

The detection of surface waves through scanning near-field optical microscopy (SNOM) is a promising technique for thermal measurements at very small scales. Recent studies have shown that electromagnetic waves, in the vicinity of a scattering structure such as an atomic force microscopy (AFM) tip, can be scattered from near to far-field and thus detected. In the present work, a model based on the finite difference time domain (FDTD) method and the near-field to far-field (NFTFF) transformation for electromagnetic waves propagation is presented. This model has been validated by studying the electromagnetic field of a dipole in vacuum and close to a dielectric substrate. Then simulations for a tetrahedral tip close to an interface are presented and discussed.

Coherent anti-stokes Raman scattering (CARS) microscopy: a novel technique for imaging the retina.

Science.gov (United States)

Masihzadeh, Omid; Ammar, David A; Kahook, Malik Y; Lei, Tim C

2013-05-01

To image the cellular and noncellular structures of the retina in an intact mouse eye without the application of exogenous fluorescent labels using noninvasive, nondestructive techniques. Freshly enucleated mouse eyes were imaged using two nonlinear optical techniques: coherent anti-Stokes Raman scattering (CARS) and two-photon autofluorescence (TPAF). Cross sectional transverse sections and sequential flat (en face) sagittal sections were collected from a region of sclera approximately midway between the limbus and optic nerve. Imaging proceeded from the surface of the sclera to a depth of ∼60 μm. The fluorescent signal from collagen fibers within the sclera was evident in the TPAF channel; the scleral collagen fibers showed no organization and appeared randomly packed. The sclera contained regions lacking TPAF and CARS fluorescence of ∼3 to 15 μm in diameter that could represent small vessels or scleral fibroblasts. Intense punctate CARS signals from the retinal pigment epithelial layer were of a size and shape of retinyl storage esters. Rod outer segments could be identified by the CARS signal from their lipid-rich plasma membranes. CARS microscopy can be used to image the outer regions of the mammalian retina without the use of a fluorescent dye or exogenously expressed recombinant protein. With technical advancements, CARS/TPAF may represent a new avenue for noninvasively imaging the retina and might complement modalities currently used in clinical practice.

Channeling contrast microscopy: a new technique for microanalysis of semiconductors

International Nuclear Information System (INIS)

McCallum, J.C.

1985-01-01

The technique of channeling contrast microscopy has been developed over the past few years for use with the Melbourne microprobe. It has been used for several profitable analyses of small-scale structures in semiconductor materials. This paper outlines the basic features of the technique and examples of its applications are given

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.

An integrated optical coherence microscopy imaging and optical stimulation system for optogenetic pacing in Drosophila melanogaster (Conference Presentation)

Science.gov (United States)

Alex, Aneesh; Li, Airong; Men, Jing; Jerwick, Jason; Tanzi, Rudolph E.; Zhou, Chao

2016-03-01

Electrical stimulation is the clinical standard for cardiac pacing. Although highly effective in controlling cardiac rhythm, the invasive nature, non-specificity to cardiac tissues and possible tissue damage limits its applications. Optogenetic pacing of the heart is a promising alternative, which is non-invasive and more specific, has high spatial and temporal precision, and avoids the shortcomings in electrical stimulation. Drosophila melanogaster, which is a powerful model organism with orthologs of nearly 75% of human disease genes, has not been studied for optogenetic pacing in the heart. Here, we developed a non-invasive integrated optical pacing and optical coherence microscopy (OCM) imaging system to control the heart rhythm of Drosophila at different developmental stages using light. The OCM system is capable of providing high imaging speed (130 frames/s) and ultrahigh imaging resolutions (1.5 μm and 3.9 μm for axial and transverse resolutions, respectively). A light-sensitive pacemaker was developed in Drosophila by specifically expressing the light-gated cation channel, channelrhodopsin-2 (ChR2) in transgenic Drosophila heart. We achieved non-invasive and specific optical control of the Drosophila heart rhythm throughout the fly's life cycle (larva, pupa, and adult) by stimulating the heart with 475 nm pulsed laser light. Heart response to stimulation pulses was monitored non-invasively with OCM. This integrated non-invasive optogenetic control and in vivo imaging technique provides a novel platform for performing research studies in developmental cardiology.

Exploring Neural Cell Dynamics with Digital Holographic Microscopy

KAUST Repository

Marquet, Pierre; Jourdain, Pascal; Boss, Daniel; Depeursinge, Christian D.; Magistretti, Pierre J.

2013-01-01

In this talk, I will present how digital holographic microscopy, as a powerful quantitative phase technique, can non-invasively measure cell dynamics and especially resolve local neuronal network activity through simultaneous multiple site optical recording.

Exploring Neural Cell Dynamics with Digital Holographic Microscopy

KAUST Repository

Marquet, Pierre

2013-04-21

In this talk, I will present how digital holographic microscopy, as a powerful quantitative phase technique, can non-invasively measure cell dynamics and especially resolve local neuronal network activity through simultaneous multiple site optical recording.

Refractometry of melanocyte cell nuclei using optical scatter images recorded by digital Fourier microscopy.

Science.gov (United States)

Seet, Katrina Y T; Nieminen, Timo A; Zvyagin, Andrei V

2009-01-01

The cell nucleus is the dominant optical scatterer in the cell. Neoplastic cells are characterized by cell nucleus polymorphism and polychromism-i.e., the nuclei exhibits an increase in the distribution of both size and refractive index. The relative size parameter, and its distribution, is proportional to the product of the nucleus size and its relative refractive index and is a useful discriminant between normal and abnormal (cancerous) cells. We demonstrate a recently introduced holographic technique, digital Fourier microscopy (DFM), to provide a sensitive measure of this relative size parameter. Fourier holograms were recorded and optical scatter of individual scatterers were extracted and modeled with Mie theory to determine the relative size parameter. The relative size parameter of individual melanocyte cell nuclei were found to be 16.5+/-0.2, which gives a cell nucleus refractive index of 1.38+/-0.01 and is in good agreement with previously reported data. The relative size parameters of individual malignant melanocyte cell nuclei are expected to be greater than 16.5.

Improving the visualization of electron-microscopy data through optical flow interpolation

KAUST Repository

Carata, Lucian

2013-01-01

Technical developments in neurobiology have reached a point where the acquisition of high resolution images representing individual neurons and synapses becomes possible. For this, the brain tissue samples are sliced using a diamond knife and imaged with electron-microscopy (EM). However, the technique achieves a low resolution in the cutting direction, due to limitations of the mechanical process, making a direct visualization of a dataset difficult. We aim to increase the depth resolution of the volume by adding new image slices interpolated from the existing ones, without requiring modifications to the EM image-capturing method. As classical interpolation methods do not provide satisfactory results on this type of data, the current paper proposes a re-framing of the problem in terms of motion volumes, considering the depth axis as a temporal axis. An optical flow method is adapted to estimate the motion vectors of pixels in the EM images, and this information is used to compute and insert multiple new images at certain depths in the volume. We evaluate the visualization results in comparison with interpolation methods currently used on EM data, transforming the highly anisotropic original dataset into a dataset with a larger depth resolution. The interpolation based on optical flow better reveals neurite structures with realistic undistorted shapes, and helps to easier map neuronal connections. © 2011 ACM.

Nonlinear optical techniques for surface studies

International Nuclear Information System (INIS)

Shen, Y.R.

1981-09-01

Recent effort in developing nonlinear optical techniques for surface studies is reviewed. Emphasis is on monolayer detection of adsorbed molecules on surfaces. It is shown that surface coherent antiStokes Raman scattering (CARS) with picosecond pulses has the sensitivity of detecting submonolayer of molecules. On the other hand, second harmonic or sum-frequency generation is also sensitive enough to detect molecular monolayers. Surface-enhanced nonlinear optical effects on some rough metal surfaces have been observed. This facilitates the detection of molecular monolayers on such surfaces, and makes the study of molecular adsorption at a liquid-metal interface feasible. Advantages and disadvantages of the nonlinear optical techniques for surface studies are discussed

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.

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

Nonlinear optical microscopy reveals invading endothelial cells anisotropically alter three-dimensional collagen matrices

International Nuclear Information System (INIS)

Lee, P.-F.; Yeh, Alvin T.; Bayless, Kayla J.

2009-01-01

The interactions between endothelial cells (ECs) and the extracellular matrix (ECM) are fundamental in mediating various steps of angiogenesis, including cell adhesion, migration and sprout formation. Here, we used a noninvasive and non-destructive nonlinear optical microscopy (NLOM) technique to optically image endothelial sprouting morphogenesis in three-dimensional (3D) collagen matrices. We simultaneously captured signals from collagen fibers and endothelial cells using second harmonic generation (SHG) and two-photon excited fluorescence (TPF), respectively. Dynamic 3D imaging revealed EC interactions with collagen fibers along with quantifiable alterations in collagen matrix density elicited by EC movement through and morphogenesis within the matrix. Specifically, we observed increased collagen density in the area between bifurcation points of sprouting structures and anisotropic increases in collagen density around the perimeter of lumenal structures, but not advancing sprout tips. Proteinase inhibition studies revealed membrane-associated matrix metalloproteinase were utilized for sprout advancement and lumen expansion. Rho-associated kinase (p160ROCK) inhibition demonstrated that the generation of cell tension increased collagen matrix alterations. This study followed sprouting ECs within a 3D matrix and revealed that the advancing structures recognize and significantly alter their extracellular environment at the periphery of lumens as they progress

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.

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....

Application of Optical Coherence Tomography Freeze-Drying Microscopy for Designing Lyophilization Process and Its Impact on Process Efficiency and Product Quality.

Science.gov (United States)

Korang-Yeboah, Maxwell; Srinivasan, Charudharshini; Siddiqui, Akhtar; Awotwe-Otoo, David; Cruz, Celia N; Muhammad, Ashraf

2018-01-01

Optical coherence tomography freeze-drying microscopy (OCT-FDM) is a novel technique that allows the three-dimensional imaging of a drug product during the entire lyophilization process. OCT-FDM consists of a single-vial freeze dryer (SVFD) affixed with an optical coherence tomography (OCT) imaging system. Unlike the conventional techniques, such as modulated differential scanning calorimetry (mDSC) and light transmission freeze-drying microscopy, used for predicting the product collapse temperature (Tc), the OCT-FDM approach seeks to mimic the actual product and process conditions during the lyophilization process. However, there is limited understanding on the application of this emerging technique to the design of the lyophilization process. In this study, we investigated the suitability of OCT-FDM technique in designing a lyophilization process. Moreover, we compared the product quality attributes of the resulting lyophilized product manufactured using Tc, a critical process control parameter, as determined by OCT-FDM versus as estimated by mDSC. OCT-FDM analysis revealed the absence of collapse even for the low protein concentration (5 mg/ml) and low solid content formulation (1%w/v) studied. This was confirmed by lab scale lyophilization. In addition, lyophilization cycles designed using Tc values obtained from OCT-FDM were more efficient with higher sublimation rate and mass flux than the conventional cycles, since drying was conducted at higher shelf temperature. Finally, the quality attributes of the products lyophilized using Tc determined by OCT-FDM and mDSC were similar, and product shrinkage and cracks were observed in all the batches of freeze-dried products irrespective of the technique employed in predicting Tc.

Three-dimensional live microscopy beyond the diffraction limit

International Nuclear Information System (INIS)

Fiolka, Reto

2013-01-01

In fluorescence microscopy it has become possible to fundamentally overcome the diffraction limited resolution in all three spatial dimensions. However, to have the most impact in biological sciences, new optical microscopy techniques need to be compatible with live cell imaging: image acquisition has to be fast enough to capture cellular dynamics at the new resolution limit while light exposure needs to be minimized to prevent photo-toxic effects. With increasing spatial resolution, these requirements become more difficult to meet, even more so when volumetric imaging is performed. In this review, techniques that have been successfully applied to three-dimensional, super-resolution live microscopy are presented and their relative strengths and weaknesses are discussed. (special issue article)

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)

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

Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.

Science.gov (United States)

Ma, Qian; Khademhosseinieh, Bahar; Huang, Eric; Qian, Haoliang; Bakowski, Malina A; Troemel, Emily R; Liu, Zhaowei

2016-08-16

The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume.

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.

Dental diagnostics using optical coherence techniques

Energy Technology Data Exchange (ETDEWEB)

Nathel, H. [Lawrence Livermore National Lab., CA (United States); Colston, B. [Univ. of California, San Francisco, CA (United States); Armitage, G. [Univ. of California, Davis, CA (United States)] [and others

1994-11-15

Optical radiation can be used for diagnostic purposes in oral medicine. However, due to the turbid, amorphous, and inhomogeneous nature of dental tissue conventional techniques used to transilluminate materials are not well suited to dental tissues. Optical coherence techniques either in the time- of frequency-domain offer the capabilities of discriminating scattered from unscattered light, thus allowing for imaging through turbid tissue. Currently, using optical time-domain reflectometry we are able to discriminate specular from diffuse reflections occurring at tissue boundaries. We have determined the specular reflectivity of enamel and dentin to be approximately 6.6 x 10{sup -5} and 1.3 x 10{sup -6}, respectively. Implications to periodontal imaging will be discussed.

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

Tablet surface characterisation by various imaging techniques

DEFF Research Database (Denmark)

Seitavuopio, Paulus; Rantanen, Jukka; Yliruusi, Jouko

2003-01-01

The aim of this study was to characterise tablet surfaces using different imaging and roughness analytical techniques including optical microscopy, scanning electron microscopy (SEM), laser profilometry and atomic force microscopy (AFM). The test materials compressed were potassium chloride (KCl......) and sodium chloride (NaCl). It was found that all methods used suggested that the KCl tablets were smoother than the NaCl tablets and higher compression pressure made the tablets smoother. Imaging methods like optical microscopy and SEM can give useful information about the roughness of the sample surface...

Optical rangefinding applications using communications modulation technique

Science.gov (United States)

Caplan, William D.; Morcom, Christopher John

2010-10-01

A novel range detection technique combines optical pulse modulation patterns with signal cross-correlation to produce an accurate range estimate from low power signals. The cross-correlation peak is analyzed by a post-processing algorithm such that the phase delay is proportional to the range to target. This technique produces a stable range estimate from noisy signals. The advantage is higher accuracy obtained with relatively low optical power transmitted. The technique is useful for low cost, low power and low mass sensors suitable for tactical use. The signal coding technique allows applications including IFF and battlefield identification systems.

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.

Four-Dimensional Ultrafast Electron Microscopy: Insights into an Emerging Technique

KAUST Repository

Adhikari, Aniruddha; Eliason, Jeffrey K.; Sun, Jingya; Bose, Riya; Flannigan, David J.; Mohammed, Omar F.

2016-01-01

Four-dimensional ultrafast electron microscopy (4D-UEM) is a novel analytical technique that aims to fulfill the long-held dream of researchers to investigate materials at extremely short spatial and temporal resolutions by integrating the excellent

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.

Fluorescence confocal polarizing microscopy: Three-dimensional ...

Indian Academy of Sciences (India)

Much of the modern understanding of orientational order in liquid crystals (LCs) is based on polarizing microscopy (PM). A PM image bears only two-dimensional (2D) information, integrating the 3D pattern of optical birefringence over the path of light. Recently, we proposed a technique to image 3D director patterns by ...

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.)

A dynamic plasmonic manipulation technique assisted by phase modulation of an incident optical vortex beam

International Nuclear Information System (INIS)

Yuan, G H; Wang, Q; Tan, P S; Lin, J; Yuan, X-C

2012-01-01

A novel phase modulation method for dynamic manipulation of surface plasmon polaritons (SPPs) with a phase engineered optical vortex (OV) beam illuminating on nanoslits is experimentally demonstrated. Because of the unique helical phase carried by an OV beam, dynamic control of SPP multiple focusing and standing wave generation is realized by changing the OV beam’s topological charge constituent with the help of a liquid-crystal spatial light modulator. Measurement of SPP distributions with near-field scanning optical microscopy showed an excellent agreement with numerical predictions. The proposed phase modulation technique for manipulating SPPs features has seemingly dynamic and reconfigurable advantages, with profound potential for development of SPP coupling, routing, multiplexing and high-resolution imaging devices on plasmonic chips. (paper)

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

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.

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)

Multilayer mounting for long-term light sheet microscopy of zebrafish.

Science.gov (United States)

Weber, Michael; Mickoleit, Michaela; Huisken, Jan

2014-02-27

Light sheet microscopy is the ideal imaging technique to study zebrafish embryonic development. Due to minimal photo-toxicity and bleaching, it is particularly suited for long-term time-lapse imaging over many hours up to several days. However, an appropriate sample mounting strategy is needed that offers both confinement and normal development of the sample. Multilayer mounting, a new embedding technique using low-concentration agarose in optically clear tubes, now overcomes this limitation and unleashes the full potential of light sheet microscopy for real-time developmental biology.

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)

The necessity of microscopy to characterize the optical properties of size-selected, nonspherical aerosol particles.

Science.gov (United States)

Veghte, Daniel P; Freedman, Miriam A

2012-11-06

It is currently unknown whether mineral dust causes a net warming or cooling effect on the climate system. This uncertainty stems from the varied and evolving shape and composition of mineral dust, which leads to diverse interactions of dust with solar and terrestrial radiation. To investigate these interactions, we have used a cavity ring-down spectrometer to study the optical properties of size-selected calcium carbonate particles, a reactive component of mineral dust. The size selection of nonspherical particles like mineral dust can differ from spherical particles in the polydispersity of the population selected. To calculate the expected extinction cross sections, we use Mie scattering theory for monodisperse spherical particles and for spherical particles with the polydispersity observed in transmission electron microscopy images. Our results for calcium carbonate are compared to the well-studied system of ammonium sulfate. While ammonium sulfate extinction cross sections agree with Mie scattering theory for monodisperse spherical particles, the results for calcium carbonate deviate at large and small particle sizes. We find good agreement for both systems, however, between the calculations performed using the particle images and the cavity ring-down data, indicating that both ammonium sulfate and calcium carbonate can be treated as polydisperse spherical particles. Our results indicate that having an independent measure of polydispersity is essential for understanding the optical properties of nonspherical particles measured with cavity ring-down spectroscopy. Our combined spectroscopy and microscopy techniques demonstrate a novel method by which cavity ring-down spectroscopy can be extended for the study of more complex aerosol particles.

Performance Monitoring Techniques Supporting Cognitive Optical Networking

DEFF Research Database (Denmark)

Caballero Jambrina, Antonio; Borkowski, Robert; Zibar, Darko

2013-01-01

High degree of heterogeneity of future optical networks, such as services with different quality-of-transmission requirements, modulation formats and switching techniques, will pose a challenge for the control and optimization of different parameters. Incorporation of cognitive techniques can help...... to solve this issue by realizing a network that can observe, act, learn and optimize its performance, taking into account end-to-end goals. In this letter we present the approach of cognition applied to heterogeneous optical networks developed in the framework of the EU project CHRON: Cognitive...... Heterogeneous Reconfigurable Optical Network. We focus on the approaches developed in the project for optical performance monitoring, which enable the feedback from the physical layer to the cognitive decision system by providing accurate description of the performance of the established lightpaths....

In Depth Analyses of LEDs by a Combination of X-ray Computed Tomography (CT) and Light Microscopy (LM) Correlated with Scanning Electron Microscopy (SEM).

Science.gov (United States)

Meyer, Jörg; Thomas, Christian; Tappe, Frank; Ogbazghi, Tekie

2016-06-16

In failure analysis, device characterization and reverse engineering of light emitting diodes (LEDs), and similar electronic components of micro-characterization, plays an important role. Commonly, different techniques like X-ray computed tomography (CT), light microscopy (LM) and scanning electron microscopy (SEM) are used separately. Similarly, the results have to be treated for each technique independently. Here a comprehensive study is shown which demonstrates the potentials leveraged by linking CT, LM and SEM. In depth characterization is performed on a white emitting LED, which can be operated throughout all characterization steps. Major advantages are: planned preparation of defined cross sections, correlation of optical properties to structural and compositional information, as well as reliable identification of different functional regions. This results from the breadth of information available from identical regions of interest (ROIs): polarization contrast, bright and dark-field LM images, as well as optical images of the LED cross section in operation. This is supplemented by SEM imaging techniques and micro-analysis using energy dispersive X-ray spectroscopy.

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.

Fluctuation microscopy: a probe of medium range order

International Nuclear Information System (INIS)

Treacy, M M J; Gibson, J M; Fan, L; Paterson, D J; McNulty, I

2005-01-01

Fluctuation microscopy is a hybrid diffraction-imaging technique that detects medium range order in amorphous materials by examining spatial fluctuations in coherent scattering. These fluctuations appear as speckle in images and diffraction patterns. The volume of material contributing to the speckle is determined by the point-spread function (the resolution) of the imaging optics and the sample thickness. The spatial periodicities being probed are related to the diffraction vector. Statistical analysis of the speckle allows the random and non-random (ordered) contributions to be discriminated. The image resolution that gives the maximum speckle contrast, as determined by the normalized variance of the image intensity, is determined by the characteristic length scale of the ordering. Because medium range ordering length scales can extend out to about the tenth coordination shell, fluctuation microscopy tends to be a low image resolution technique. This review presents the kinematical scattering theory underpinning fluctuation microscopy and a description of fluctuation electron microscopy as it has been employed in the transmission electron microscope for studying amorphous materials. Recent results using soft x-rays for studying nanoscale materials are also presented. We summarize outstanding issues and point to possible future directions for fluctuation microscopy as a technique

Recent achievements in multi-keV x-ray microscopy

International Nuclear Information System (INIS)

Susini, J.; Barrett, R.; Salome, M.; Kaulich, B.

2002-01-01

X-ray microscopy (XRM) techniques are emerging as powerful and complementary tools for sub-micron investigations. Soft XRM traditionally offers the possibility to form direct images of thick hydrated biological material in near-native environments, at a spatial resolution well beyond that achievable with visible light microscopy. Natural contrast is available in the soft X-ray region, in the so-called 'water-window', due to the presence of absorption edges of the major constituents (C,N,O). Recent advances in manufacturing techniques have enlarged the accessible energy range of micro-focusing optics and offer new applications in a broad range of disciplines. XRM in the 1-20 keV energy range is better suited to map trace elements in fluorescence yield, 3-D tomographic imaging and in micro-diffraction. After a brief introduction to the principles and methods, the main attributes of X-ray microscopy will be presented. This presentation will be biased towards sub-micron microscopy developed at the ESRF in the 2-10 keV energy. Strengths and weaknesses of X-ray microscopy and spectro-microscopy techniques will be discussed and illustrated by examples in biology, materials sciences and geology. (authors)

CCD-based thermoreflectance microscopy: principles and applications

International Nuclear Information System (INIS)

Farzaneh, M; Maize, K; Shakouri, A; Lueerssen, D; Summers, J A; Hudgings, Janice A; Mayer, P M; Ram, R J; Raad, P E; Pipe, K P

2009-01-01

CCD-based thermoreflectance microscopy has emerged as a high resolution, non-contact imaging technique for thermal profiling and performance and reliability analysis of numerous electronic and optoelectronic devices at the micro-scale. This thermography technique, which is based on measuring the relative change in reflectivity of the device surface as a function of change in temperature, provides high-resolution thermal images that are useful for hot spot detection and failure analysis, mapping of temperature distribution, measurement of thermal transient, optical characterization of photonic devices and measurement of thermal conductivity in thin films. In this paper we review the basic physical principle behind thermoreflectance as a thermography tool, discuss the experimental setup, resolutions achieved, signal processing procedures and calibration techniques, and review the current applications of CCD-based thermoreflectance microscopy in various devices. (topical review)

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

Towards phonon photonics: scattering-type near-field optical microscopy reveals phonon-enhanced near-field interaction

International Nuclear Information System (INIS)

Hillenbrand, Rainer

2004-01-01

Diffraction limits the spatial resolution in classical microscopy or the dimensions of optical circuits to about half the illumination wavelength. Scanning near-field microscopy can overcome this limitation by exploiting the evanescent near fields existing close to any illuminated object. We use a scattering-type near-field optical microscope (s-SNOM) that uses the illuminated metal tip of an atomic force microscope (AFM) to act as scattering near-field probe. The presented images are direct evidence that the s-SNOM enables optical imaging at a spatial resolution on a 10 nm scale, independent of the wavelength used (λ=633 nm and 10 μm). Operating the microscope at specific mid-infrared frequencies we found a tip-induced phonon-polariton resonance on flat polar crystals such as SiC and Si 3 N 4 . Being a spectral fingerprint of any polar material such phonon-enhanced near-field interaction has enormous applicability in nondestructive, material-specific infrared microscopy at nanoscale resolution. The potential of s-SNOM to study eigenfields of surface polaritons in nanostructures opens the door to the development of phonon photonics--a proposed infrared nanotechnology that uses localized or propagating surface phonon polaritons for probing, manipulating and guiding infrared light in nanoscale devices, analogous to plasmon photonics

Optical frequency upconversion technique for transmission of wireless MIMO-type signals over optical fiber.

Science.gov (United States)

Shaddad, R Q; Mohammad, A B; Al-Gailani, S A; Al-Hetar, A M

2014-01-01

The optical fiber is well adapted to pass multiple wireless signals having different carrier frequencies by using radio-over-fiber (ROF) technique. However, multiple wireless signals which have the same carrier frequency cannot propagate over a single optical fiber, such as wireless multi-input multi-output (MIMO) signals feeding multiple antennas in the fiber wireless (FiWi) system. A novel optical frequency upconversion (OFU) technique is proposed to solve this problem. In this paper, the novel OFU approach is used to transmit three wireless MIMO signals over a 20 km standard single mode fiber (SMF). The OFU technique exploits one optical source to produce multiple wavelengths by delivering it to a LiNbO3 external optical modulator. The wireless MIMO signals are then modulated by LiNbO3 optical intensity modulators separately using the generated optical carriers from the OFU process. These modulators use the optical single-sideband with carrier (OSSB+C) modulation scheme to optimize the system performance against the fiber dispersion effect. Each wireless MIMO signal is with a 2.4 GHz or 5 GHz carrier frequency, 1 Gb/s data rate, and 16-quadrature amplitude modulation (QAM). The crosstalk between the wireless MIMO signals is highly suppressed, since each wireless MIMO signal is carried on a specific optical wavelength.

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.

Cell tracking with gadophrin-2: a bifunctional contrast agent for MR imaging, optical imaging, and fluorescence microscopy

International Nuclear Information System (INIS)

Daldrup-Link, Heike E.; Rudelius, Martina; Piontek, Guido; Schlegel, Juergen; Metz, Stephan; Settles, Marcus; Rummeny, Ernst J.; Pichler, Bernd; Heinzmann, Ulrich; Oostendorp, Robert A.J.

2004-01-01

The purpose of this study was to assess the feasibility of use of gadophrin-2 to trace intravenously injected human hematopoietic cells in athymic mice, employing magnetic resonance (MR) imaging, optical imaging (OI), and fluorescence microscopy. Mononuclear peripheral blood cells from GCSF-primed patients were labeled with gadophrin-2 (Schering AG, Berlin, Germany), a paramagnetic and fluorescent metalloporphyrin, using established transfection techniques with cationic liposomes. The labeled cells were evaluated in vitro with electron microscopy and inductively coupled plasma atomic emission spectrometry. Then, 1 x 10 6 -3 x 10 8 labeled cells were injected into 14 nude Balb/c mice and the in vivo cell distribution was evaluated with MR imaging and OI before and 4, 24, and 48 h after intravenous injection (p.i.). Five additional mice served as controls: three mice were untreated controls and two mice were investigated after injection of unlabeled cells. The contrast agent effect was determined quantitatively for MR imaging by calculating signal-to-noise-ratio (SNR) data. After completion of in vivo imaging studies, fluorescence microscopy of excised organs was performed. Intracellular cytoplasmatic uptake of gadophrin-2 was confirmed by electron microscopy. Spectrometry determined an uptake of 31.56 nmol Gd per 10 6 cells. After intravenous injection, the distribution of gadophrin-2 labeled cells in nude mice could be visualized by MR, OI, and fluorescence microscopy. At 4 h p.i., the transplanted cells mainly distributed to lung, liver, and spleen, and 24 h p.i. they also distributed to the bone marrow. Fluorescence microscopy confirmed the distribution of gadophrin-2 labeled cells to these target organs. Gadophrin-2 is suited as a bifunctional contrast agent for MR imaging, OI, and fluorescence microscopy and may be used to combine the advantages of each individual imaging modality for in vivo tracking of intravenously injected hematopoietic cells. (orig.)

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.

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.

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

Decoupled illumination detection in light sheet microscopy for fast volumetric imaging

OpenAIRE

Olarte, Omar; Andilla, Jordi; Artigas García, David; Loza-Alvarez, Pablo

2015-01-01

Current microscopy demands the visualization of large three-dimensional samples with increased sensitivity, higher resolution, and faster speed. Several imaging techniques based on widefield, point-scanning, and light-sheet strategies have been designed to tackle some of these demands. Although successful, all these require the illuminated volumes to be tightly coupled with the detection optics to accomplish efficient optical sectioning. Here, we break this paradigm and produce optical sectio...

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.

Multiphoton Microscopy for Ophthalmic Imaging

Directory of Open Access Journals (Sweden)

Emily A. Gibson

2011-01-01

Full Text Available We review multiphoton microscopy (MPM including two-photon autofluorescence (2PAF, second harmonic generation (SHG, third harmonic generation (THG, fluorescence lifetime (FLIM, and coherent anti-Stokes Raman Scattering (CARS with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the particular strength that each has for functional tissue imaging. MPM is compared with current clinical ophthalmological imaging techniques such as reflectance confocal microscopy, optical coherence tomography, and fluorescence imaging. In addition, we discuss the future prospects for MPM in disease detection and clinical monitoring of disease progression, understanding fundamental disease mechanisms, and real-time monitoring of drug delivery.

Re-scan confocal microscopy: scanning twice for better resolution.

Science.gov (United States)

De Luca, Giulia M R; Breedijk, Ronald M P; Brandt, Rick A J; Zeelenberg, Christiaan H C; de Jong, Babette E; Timmermans, Wendy; Azar, Leila Nahidi; Hoebe, Ron A; Stallinga, Sjoerd; Manders, Erik M M

2013-01-01

We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required.

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.

Nanohybrids Near-Field Optical Microscopy: From Image Shift to Biosensor Application

Directory of Open Access Journals (Sweden)

Nayla El-Kork

2016-01-01

Full Text Available Near-Field Optical Microscopy is a valuable tool for the optical and topographic study of objects at a nanometric scale. Nanoparticles constitute important candidates for such type of investigations, as they bear an important weight for medical, biomedical, and biosensing applications. One, however, has to be careful as artifacts can be easily reproduced. In this study, we examined hybrid nanoparticles (or nanohybrids in the near-field, while in solution and attached to gold nanoplots. We found out that they can be used for wavelength modulable near-field biosensors within conditions of artifact free imaging. In detail, we refer to the use of topographic/optical image shift and the imaging of Local Surface Plasmon hot spots to validate the genuineness of the obtained images. In summary, this study demonstrates a new way of using simple easily achievable comparative methods to prove the authenticity of near-field images and presents nanohybrid biosensors as an application.

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.

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

Single molecule mapping of the optical field distribution of probes for near-field microscopy

NARCIS (Netherlands)

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

1999-01-01

The most difficult task in near-field scanning optical microscopy (NSOM) is to make a high quality subwavelength aperture probe, Recently we have developed high definition NSOM probes by focused ion beam (FIB) milling. These probes have a higher brightness, better polarization characteristics,

2. Brazilian Congress on Cell Biology and 7. Brazilian Colloquium on Electron Microscopy - Abstracts

International Nuclear Information System (INIS)

1980-01-01

Immunology, virology, bacteriology, genetics and protozoology are some of the subjects treated in the 2. Brazilian Congress on Cell Biology. Studies using radioisotopic techniques and ultrastructural cytological studies are presented. Use of optical - and electron microscopy in some of these studies is discussed. In the 7. Brazilian Colloquium on Electron Microscopy, the application of this technique to materials science is discussed (failure analysis in metallurgy, energy dispersion X-ray analysis, etc). (I.C.R.) [pt

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.

Reciprocity theory of apertureless scanning near-field optical microscopy with point-dipole probes.

Science.gov (United States)

Esslinger, Moritz; Vogelgesang, Ralf

2012-09-25

Near-field microscopy offers the opportunity to reveal optical contrast at deep subwavelength scales. In scanning near-field optical microscopy (SNOM), the diffraction limit is overcome by a nanoscopic probe in close proximity to the sample. The interaction of the probe with the sample fields necessarily perturbs the bare sample response, and a critical issue is the interpretation of recorded signals. For a few specific SNOM configurations, individual descriptions have been modeled, but a general and intuitive framework is still lacking. Here, we give an exact formulation of the measurable signals in SNOM which is easily applicable to experimental configurations. Our results are in close analogy with the description Tersoff and Hamann have derived for the tunneling currents in scanning tunneling microscopy. For point-like scattering probe tips, such as used in apertureless SNOM, the theory simplifies dramatically to a single scalar relation. We find that the measured signal is directly proportional to the field of the coupled tip-sample system at the position of the tip. For weakly interacting probes, the model thus verifies the empirical findings that the recorded signal is proportional to the unperturbed field of the bare sample. In the more general case, it provides guidance to an intuitive and faithful interpretation of recorded images, facilitating the characterization of tip-related distortions and the evaluation of novel SNOM configurations, both for aperture-based and apertureless SNOM.

Optical properties of WO3 thin films using surface plasmon resonance technique

International Nuclear Information System (INIS)

Paliwal, Ayushi; Sharma, Anjali; Gupta, Vinay; Tomar, Monika

2014-01-01

Indigenously assembled surface plasmon resonance (SPR) technique has been exploited to study the thickness dependent dielectric properties of WO 3 thin films. WO 3 thin films (80 nm to 200 nm) have been deposited onto gold (Au) coated glass prism by sputtering technique. The structural, optical properties and surface morphology of the deposited WO 3 thin films were studied using X-ray diffraction, UV-visible spectrophotometer, Raman spectroscopy, and Scanning electron microscopy (SEM). XRD analysis shows that all the deposited WO 3 thin films are exhibiting preferred (020) orientation and Raman data indicates that the films possess single phase monoclinic structure. SEM images reveal the variation in grain size with increase in thickness. The SPR reflectance curves of the WO 3 /Au/prism structure were utilized to estimate the dielectric properties of WO 3 thin films at optical frequency (λ = 633 nm). As the thickness of WO 3 thin film increases from 80 nm to 200 nm, the dielectric constant is seen to be decreasing from 5.76 to 3.42, while the dielectric loss reduces from 0.098 to 0.01. The estimated value of refractive index of WO 3 film is in agreement to that obtained from UV-visible spectroscopy studies. The strong dispersion in refractive index is observed with wavelength of incident laser light

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

Multimodal reconstruction of microvascular-flow distributions using combined two-photon microscopy and Doppler optical coherence tomography.

Science.gov (United States)

Gagnon, Louis; Sakadžić, Sava; Lesage, Fréderic; Mandeville, Emiri T; Fang, Qianqian; Yaseen, Mohammad A; Boas, David A

2015-01-01

Computing microvascular cerebral blood flow ([Formula: see text]) in real cortical angiograms is challenging. Here, we investigated whether the use of Doppler optical coherence tomography (DOCT) flow measurements in individual vessel segments can help in reconstructing [Formula: see text] across the entire vasculature of a truncated cortical angiogram. A [Formula: see text] computational framework integrating DOCT measurements is presented. Simulations performed on a synthetic angiogram showed that the addition of DOCT measurements, especially close to large inflowing or outflowing vessels, reduces the impact of pressure boundary conditions and estimated vessel resistances resulting in a more accurate reconstruction of [Formula: see text]. Our technique was then applied to reconstruct microvascular flow distributions in the mouse cortex down to [Formula: see text] by combining two-photon laser scanning microscopy angiography with DOCT.

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...

Depth-profiling by confocal Raman microscopy (CRM): data correction by numerical techniques.

Science.gov (United States)

Tomba, J Pablo; Eliçabe, Guillermo E; Miguel, María de la Paz; Perez, Claudio J

2011-03-01

The data obtained in confocal Raman microscopy (CRM) depth profiling experiments with dry optics are subjected to significant distortions, including an artificial compression of the depth scale, due to the combined influence of diffraction, refraction, and instrumental effects that operate on the measurement. This work explores the use of (1) regularized deconvolution and (2) the application of simple rescaling of the depth scale as methodologies to obtain an improved, more precise, confocal response. The deconvolution scheme is based on a simple predictive model for depth resolution and the use of regularization techniques to minimize the dramatic oscillations in the recovered response typical of problem inversion. That scheme is first evaluated using computer simulations on situations that reproduce smooth and sharp sample transitions between two materials and finally it is applied to correct genuine experimental data, obtained in this case from a sharp transition (planar interface) between two polymeric materials. It is shown that the methodology recovers very well most of the lost profile features in all the analyzed situations. The use of simple rescaling appears to be only useful for correcting smooth transitions, particularly those extended over distances larger than those spanned by the operative depth resolution, which limits the strategy to the study of profiles near the sample surface. However, through computer simulations, it is shown that the use of water immersion objectives may help to reduce optical distortions and to expand the application window of this simple methodology, which could be useful, for instance, to safely monitor Fickean sorption/desorption of penetrants in polymer films/coatings in a nearly noninvasive way.

Optical Axis Identification Technique for Free Space Optics Transmission

Directory of Open Access Journals (Sweden)

Yuki Tashiro

2017-07-01

Full Text Available This paper describes optical axis adjustment technique for an active free space optics transmission system. This system precisely controls the direction of a collimated thin laser beam using a motor driven laser emitting mechanism and positioning photodiodes. Before beginning laser beam feedback control, it is required to guide the laser beam within the range of the positioning photodiodes for initial laser beam alignment. This paper proposes an arrival position presumption method of laser beam traveling along the long distance from transmitter. A positioning sensor containing several photodiodes measures laser luminescence distribution, and analytically calculates the optical axis of laser beam according to the modified Gaussian beam optics based on four or five distributed local intensity of laser luminescence. Experiments are conducted to evaluate the accuracy of the presumption, and results reveal that the method is effective in leading the laser beam onto a distant receiver.

Numerical tilting compensation in microscopy based on wavefront sensing using transport of intensity equation method

Science.gov (United States)

Hu, Junbao; Meng, Xin; Wei, Qi; Kong, Yan; Jiang, Zhilong; Xue, Liang; Liu, Fei; Liu, Cheng; Wang, Shouyu

2018-03-01

Wide-field microscopy is commonly used for sample observations in biological research and medical diagnosis. However, the tilting error induced by the oblique location of the image recorder or the sample, as well as the inclination of the optical path often deteriorates the imaging quality. In order to eliminate the tilting in microscopy, a numerical tilting compensation technique based on wavefront sensing using transport of intensity equation method is proposed in this paper. Both the provided numerical simulations and practical experiments prove that the proposed technique not only accurately determines the tilting angle with simple setup and procedures, but also compensates the tilting error for imaging quality improvement even in the large tilting cases. Considering its simple systems and operations, as well as image quality improvement capability, it is believed the proposed method can be applied for tilting compensation in the optical microscopy.

Functional Micrococcus lysodeikticus layers deposited by laser technique for the optical sensing of lysozyme.

Science.gov (United States)

Dinca, Valentina; Zaharie-Butucel, Diana; Stanica, Luciana; Brajnicov, Simona; Marascu, Valentina; Bonciu, Anca; Cristocea, Andra; Gaman, Laura; Gheorghiu, Mihaela; Astilean, Simion; Vasilescu, Alina

2018-02-01

Whole cell optical biosensors, made by immobilizing whole algal, bacterial or mammalian cells on various supports have found applications in several fields, from ecology and ecotoxicity testing to biopharmaceutical production or medical diagnostics. We hereby report the deposition of functional bacterial layers of Micrococcus lysodeikticus (ML) via Matrix-Assisted Pulsed Laser Evaporation (MAPLE) on poly(diallyldimethylamonium) (PDDA)-coated-glass slides and their application as an optical biosensor for the detection of lysozyme in serum. Lysozyme is an enzyme upregulated in inflammatory diseases and ML is an enzymatic substrate for this enzyme. The MAPLE-deposited bacterial interfaces were characterised by Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), Fourier-Transformed Infrared Spectroscopy (FTIR), Raman and optical microscopy and were compared with control interfaces deposited via layer-by-layer on the same substrate. After MAPLE deposition and coating with graphene oxide (GO), ML-modified interfaces retained their functionality and sensitivity to lysozyme's lytic action. The optical biosensor detected lysozyme in undiluted serum in the clinically relevant range up to 10μgmL -1 , in a fast and simple manner. Copyright © 2017 Elsevier B.V. All rights reserved.

Measurement of replication structures at the nanometer scale using super-resolution light microscopy.

Science.gov (United States)

Baddeley, D; Chagin, V O; Schermelleh, L; Martin, S; Pombo, A; Carlton, P M; Gahl, A; Domaing, P; Birk, U; Leonhardt, H; Cremer, C; Cardoso, M C

2010-01-01

DNA replication, similar to other cellular processes, occurs within dynamic macromolecular structures. Any comprehensive understanding ultimately requires quantitative data to establish and test models of genome duplication. We used two different super-resolution light microscopy techniques to directly measure and compare the size and numbers of replication foci in mammalian cells. This analysis showed that replication foci vary in size from 210 nm down to 40 nm. Remarkably, spatially modulated illumination (SMI) and 3D-structured illumination microscopy (3D-SIM) both showed an average size of 125 nm that was conserved throughout S-phase and independent of the labeling method, suggesting a basic unit of genome duplication. Interestingly, the improved optical 3D resolution identified 3- to 5-fold more distinct replication foci than previously reported. These results show that optical nanoscopy techniques enable accurate measurements of cellular structures at a level previously achieved only by electron microscopy and highlight the possibility of high-throughput, multispectral 3D analyses.

Near-infrared multispectral photoacoustic microscopy using a graded-index fiber amplifier

Directory of Open Access Journals (Sweden)

Takashi Buma

2016-09-01

Full Text Available We demonstrate optical resolution photoacoustic microscopy (OR-PAM of lipid-rich tissue using a multi-wavelength pulsed laser based on nonlinear fiber optics. 1047 nm laser pulses are converted to 1098, 1153, 1215, and 1270 nm pulses via stimulated Raman scattering in a graded-index multimode fiber. Multispectral PAM of a lipid phantom is demonstrated with our low-cost and simple technique.

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.

Optical metrology for analysis of lobster-eye x-ray optics

International Nuclear Information System (INIS)

Irving, Thomas H.K.; Peele, Andrew G.; Nugent, Keith A.

2003-01-01

A new method that uses optical microscopy to determine the physical structure of lobster-eye x-ray optics is described. This approach offers the ability to predict x-ray performance without having to take an x-ray measurement. An overlapping series of images of the entrance and exit faces of an optic are obtained and examined by purpose-built software. A 24-parameter description of each channel is obtained from which a quantitative analysis of all the major optic defects, except surface roughness, is performed. Results for a planar lobster-eye optic are used to illustrate this technique and discuss its abilities as well as directions for future enhancements

Fundamentals and techniques of nonimaging optics research

Science.gov (United States)

Winston, R.; Ogallagher, J.

1987-07-01

Nonimaging Optics differs from conventional approaches in its relaxation of unnecessary constraints on energy transport imposed by the traditional methods for optimizing image formation and its use of more broadly based analytical techniques such as phase space representations of energy flow, radiative transfer analysis, thermodynamic arguments, etc. Based on these means, techniques for designing optical elements which approach and in some cases attain the maximum concentration permitted by the Second Law of Thermodynamics were developed. The most widely known of these devices are the family of Compound Parabolic Concentrators (CPC's) and their variants and the so called Flow-Line or trumpet concentrator derived from the geometric vector flux formalism developed under this program. Applications of these and other such ideal or near-ideal devices permits increases of typically a factor of four (though in some cases as much as an order of magnitude) in the concentration above that possible with conventional means. Present efforts can be classed into two main areas: (1) classical geometrical nonimaging optics, and (2) logical extensions of nonimaging concepts to the physical optics domain.

Improving the visualization of electron-microscopy data through optical flow interpolation

KAUST Repository

Carata, Lucian; Shao, Dan; Hadwiger, Markus; Grö eller, Eduard

2013-01-01

with electron-microscopy (EM). However, the technique achieves a low resolution in the cutting direction, due to limitations of the mechanical process, making a direct visualization of a dataset difficult. We aim to increase the depth resolution of the volume

The thin layer technique and its application to electron microscopy; La technique des couches minces et son application a la microscopie electronique

Energy Technology Data Exchange (ETDEWEB)

Ranc, G. [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

1957-10-15

This work deals with the technique of thin layers obtained by evaporation under vacuum, in the thickness range extending from a few monoatomic layers to several hundred angstroms. The great theoretical and practical interest of these layers has, it is well known, given rise to many investigations from Faraday onwards. Within the necessarily restricted limits of this study, we shall approach the problem more particularly from the point of view of: - their production; - their use in electron microscopy. A critical appraisal is made, in the light of present-day knowledge, based on our personal experience and on an extensive bibliography which we have collected on the subject. (author) [French] Le present travail concerne la technique des couches minces obtenues par evaporation sous vide, dans le domaine d'epaisseur qui s'etend de quelques couches monoatomiques a plusieurs centaines d'angstroms. L'interet theorique et pratique considerable de ces couches a suscite, comme on sait, de nombreux travaux depuis Faraday. Dans le cadre necessairement restreint de cette these, nous aborderons plus particulierement le point de vue de: - leur obtention; - leur utilisation en microscopie electronique. Il s'agit d'une mise au point critique, a la lumiere des connaissances actuelles, appuyee sur notre experience personnelle et sur une importante bibliographie, qu'il nous a ete donne de reunir a ce sujet. (auteur)

The thin layer technique and its application to electron microscopy; La technique des couches minces et son application a la microscopie electronique

Energy Technology Data Exchange (ETDEWEB)

Ranc, G [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

1957-10-15

This work deals with the technique of thin layers obtained by evaporation under vacuum, in the thickness range extending from a few monoatomic layers to several hundred angstroms. The great theoretical and practical interest of these layers has, it is well known, given rise to many investigations from Faraday onwards. Within the necessarily restricted limits of this study, we shall approach the problem more particularly from the point of view of: - their production; - their use in electron microscopy. A critical appraisal is made, in the light of present-day knowledge, based on our personal experience and on an extensive bibliography which we have collected on the subject. (author) [French] Le present travail concerne la technique des couches minces obtenues par evaporation sous vide, dans le domaine d'epaisseur qui s'etend de quelques couches monoatomiques a plusieurs centaines d'angstroms. L'interet theorique et pratique considerable de ces couches a suscite, comme on sait, de nombreux travaux depuis Faraday. Dans le cadre necessairement restreint de cette these, nous aborderons plus particulierement le point de vue de: - leur obtention; - leur utilisation en microscopie electronique. Il s'agit d'une mise au point critique, a la lumiere des connaissances actuelles, appuyee sur notre experience personnelle et sur une importante bibliographie, qu'il nous a ete donne de reunir a ce sujet. (auteur)

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)

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.

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.

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...

Developing a New Biophysical Tool to Combine Magneto-Optical Tweezers with Super-Resolution Fluorescence Microscopy

Directory of Open Access Journals (Sweden)

Zhaokun Zhou

2015-06-01

Full Text Available We present a novel experimental setup in which magnetic and optical tweezers are combined for torque and force transduction onto single filamentous molecules in a transverse configuration to allow simultaneous mechanical measurement and manipulation. Previously we have developed a super-resolution imaging module which, in conjunction with advanced imaging techniques such as Blinking assisted Localisation Microscopy (BaLM, achieves localisation precision of single fluorescent dye molecules bound to DNA of ~30 nm along the contour of the molecule; our work here describes developments in producing a system which combines tweezing and super-resolution fluorescence imaging. The instrument also features an acousto-optic deflector that temporally divides the laser beam to form multiple traps for high throughput statistics collection. Our motivation for developing the new tool is to enable direct observation of detailed molecular topological transformation and protein binding event localisation in a stretching/twisting mechanical assay that previously could hitherto only be deduced indirectly from the end-to-end length variation of DNA. Our approach is simple and robust enough for reproduction in the lab without the requirement of precise hardware engineering, yet is capable of unveiling the elastic and dynamic properties of filamentous molecules that have been hidden using traditional tools.

Towards automated segmentation of cells and cell nuclei in nonlinear optical microscopy.

Science.gov (United States)

Medyukhina, Anna; Meyer, Tobias; Schmitt, Michael; Romeike, Bernd F M; Dietzek, Benjamin; Popp, Jürgen

2012-11-01

Nonlinear optical (NLO) imaging techniques based e.g. on coherent anti-Stokes Raman scattering (CARS) or two photon excited fluorescence (TPEF) show great potential for biomedical imaging. In order to facilitate the diagnostic process based on NLO imaging, there is need for an automated calculation of quantitative values such as cell density, nucleus-to-cytoplasm ratio, average nuclear size. Extraction of these parameters is helpful for the histological assessment in general and specifically e.g. for the determination of tumor grades. This requires an accurate image segmentation and detection of locations and boundaries of cells and nuclei. Here we present an image processing approach for the detection of nuclei and cells in co-registered TPEF and CARS images. The algorithm developed utilizes the gray-scale information for the detection of the nuclei locations and the gradient information for the delineation of the nuclear and cellular boundaries. The approach reported is capable for an automated segmentation of cells and nuclei in multimodal TPEF-CARS images of human brain tumor samples. The results are important for the development of NLO microscopy into a clinically relevant diagnostic tool. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Digital Fourier microscopy for soft matter dynamics

International Nuclear Information System (INIS)

Giavazzi, Fabio; Cerbino, Roberto

2014-01-01

Soft matter is studied with a large portfolio of methods. Light scattering and video microscopy are the most employed at optical wavelengths. Light scattering provides ensemble-averaged information on soft matter in the reciprocal space. The wave-vectors probed correspond to length scales ranging from a few nanometers to fractions of millimetre. Microscopy probes the sample directly in the real space, by offering a unique access to the local properties. However, optical resolution issues limit the access to length scales smaller than approximately 200 nm. We describe recent work that bridges the gap between scattering and microscopy. Several apparently unrelated techniques are found to share a simple basic idea: the correlation properties of the sample can be characterized in the reciprocal space via spatial Fourier analysis of images collected in the real space. We describe the main features of such digital Fourier microscopy (DFM), by providing examples of several possible experimental implementations of it, some of which not yet realized in practice. We also provide an overview of experimental results obtained with DFM for the study of the dynamics of soft materials. Finally, we outline possible future developments of DFM that would ease its adoption as a standard laboratory method. (topical review)

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.

Optical coherence techniques for plasma doppler spectroscopy

International Nuclear Information System (INIS)

Howard, J.; Michael, C.; Glass, F.; Cheetham, A.D.

2000-01-01

A new electro-optically Modulated Optical Solid-State (MOSS) interferometer has been constructed for measurement of the low order spectral moments of line emission from optically thin radiant media. The instrument, which is based on the principle of the Fourier transform spectrometer, has high etendue and is rugged, compact and inexpensive. By employing electro-optical path-length modulation techniques, the spectral information is transferred to the temporal frequency domain and can be obtained using a single photodetector. Specifically, the zeroth moment (brightness) is given by the average signal level, the first moment (shift) by the modulation phase and the second moment (line width) by the modulation amplitude. (author)

Cognitive Heterogeneous Reconfigurable Optical Networks (CHRON): Enabling Technologies and Techniques

DEFF Research Database (Denmark)

Tafur Monroy, Idelfonso; Zibar, Darko; Guerrero Gonzalez, Neil

2011-01-01

We present the approach of cognition applied to heterogeneous optical networks developed in the framework of the EU project CHRON: Cognitive Heterogeneous Reconfigurable Optical Network. We introduce and discuss in particular the technologies and techniques that will enable a cognitive optical...... network to observe, act, learn and optimizes its performance, taking into account its high degree of heterogeneity with respect to quality of service, transmission and switching techniques....

Optical modulation techniques for analog signal processing and CMOS compatible electro-optic modulation

Science.gov (United States)

Gill, Douglas M.; Rasras, Mahmoud; Tu, Kun-Yii; Chen, Young-Kai; White, Alice E.; Patel, Sanjay S.; Carothers, Daniel; Pomerene, Andrew; Kamocsai, Robert; Beattie, James; Kopa, Anthony; Apsel, Alyssa; Beals, Mark; Mitchel, Jurgen; Liu, Jifeng; Kimerling, Lionel C.

2008-02-01

Integrating electronic and photonic functions onto a single silicon-based chip using techniques compatible with mass-production CMOS electronics will enable new design paradigms for existing system architectures and open new opportunities for electro-optic applications with the potential to dramatically change the management, cost, footprint, weight, and power consumption of today's communication systems. While broadband analog system applications represent a smaller volume market than that for digital data transmission, there are significant deployments of analog electro-optic systems for commercial and military applications. Broadband linear modulation is a critical building block in optical analog signal processing and also could have significant applications in digital communication systems. Recently, broadband electro-optic modulators on a silicon platform have been demonstrated based on the plasma dispersion effect. The use of the plasma dispersion effect within a CMOS compatible waveguide creates new challenges and opportunities for analog signal processing since the index and propagation loss change within the waveguide during modulation. We will review the current status of silicon-based electrooptic modulators and also linearization techniques for optical modulation.

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...

Activated sludge characterization through microscopy: A review on quantitative image analysis and chemometric techniques

Energy Technology Data Exchange (ETDEWEB)

Mesquita, Daniela P. [IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal); Amaral, A. Luís [IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal); Instituto Politécnico de Coimbra, ISEC, DEQB, Rua Pedro Nunes, Quinta da Nora, 3030-199 Coimbra (Portugal); Ferreira, Eugénio C., E-mail: ecferreira@deb.uminho.pt [IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal)

2013-11-13

Graphical abstract: -- Highlights: •Quantitative image analysis shows potential to monitor activated sludge systems. •Staining techniques increase the potential for detection of operational problems. •Chemometrics combined with quantitative image analysis is valuable for process monitoring. -- Abstract: In wastewater treatment processes, and particularly in activated sludge systems, efficiency is quite dependent on the operating conditions, and a number of problems may arise due to sludge structure and proliferation of specific microorganisms. In fact, bacterial communities and protozoa identification by microscopy inspection is already routinely employed in a considerable number of cases. Furthermore, quantitative image analysis techniques have been increasingly used throughout the years for the assessment of aggregates and filamentous bacteria properties. These procedures are able to provide an ever growing amount of data for wastewater treatment processes in which chemometric techniques can be a valuable tool. However, the determination of microbial communities’ properties remains a current challenge in spite of the great diversity of microscopy techniques applied. In this review, activated sludge characterization is discussed highlighting the aggregates structure and filamentous bacteria determination by image analysis on bright-field, phase-contrast, and fluorescence microscopy. An in-depth analysis is performed to summarize the many new findings that have been obtained, and future developments for these biological processes are further discussed.

Optical properties of WO{sub 3} thin films using surface plasmon resonance technique

Energy Technology Data Exchange (ETDEWEB)

Paliwal, Ayushi; Sharma, Anjali; Gupta, Vinay, E-mail: drguptavinay@gmail.com, E-mail: vgupta@physics.du.ac.in [Department of Physics and Astrophysics, University of Delhi, Delhi 110007 (India); Tomar, Monika [Department of Physics, Miranda House, University of Delhi, Delhi 110007 (India)

2014-01-28

Indigenously assembled surface plasmon resonance (SPR) technique has been exploited to study the thickness dependent dielectric properties of WO{sub 3} thin films. WO{sub 3} thin films (80 nm to 200 nm) have been deposited onto gold (Au) coated glass prism by sputtering technique. The structural, optical properties and surface morphology of the deposited WO{sub 3} thin films were studied using X-ray diffraction, UV-visible spectrophotometer, Raman spectroscopy, and Scanning electron microscopy (SEM). XRD analysis shows that all the deposited WO{sub 3} thin films are exhibiting preferred (020) orientation and Raman data indicates that the films possess single phase monoclinic structure. SEM images reveal the variation in grain size with increase in thickness. The SPR reflectance curves of the WO{sub 3}/Au/prism structure were utilized to estimate the dielectric properties of WO{sub 3} thin films at optical frequency (λ = 633 nm). As the thickness of WO{sub 3} thin film increases from 80 nm to 200 nm, the dielectric constant is seen to be decreasing from 5.76 to 3.42, while the dielectric loss reduces from 0.098 to 0.01. The estimated value of refractive index of WO{sub 3} film is in agreement to that obtained from UV-visible spectroscopy studies. The strong dispersion in refractive index is observed with wavelength of incident laser light.

Fundamentals of fluorescence microscopy exploring life with light

CERN Document Server

Mondal, Partha Pratim

2014-01-01

This book starts at an introductory level and leads reader to the most advanced developments in fluorescence imaging and super-resolution techniques that have enabled the emergence of new disciplines such as nanobioimaging, multiphoton microscopy, photodynamic therapy, nanometrology and nanosensors. The interdisciplinary subject of fluorescence microscopy and imaging requires complete knowledge of imaging optics and molecular physics. So, this book approaches the subject by introducing optical imaging concepts before going deep into the advanced imaging systems and their applications. Molecular orbital theory forms the basis for understanding fluorescent molecules and thereby facilitates complete explanation of light-matter interaction at the geometrical focus. The two disciplines have some overlap since light controls the states of molecules and conversely, molecular states control the emitted light. These two mechanisms together determine essential fluorescence  factors and phenomena such as, molecular cro...

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

Frequency-Shifted Interferometry — A Versatile Fiber-Optic Sensing Technique

Directory of Open Access Journals (Sweden)

Fei Ye

2014-06-01

Full Text Available Fiber-optic sensing is a field that is developing at a fast pace. Novel fiber-optic sensor designs and sensing principles constantly open doors for new opportunities. In this paper, we review a fiber-optic sensing technique developed in our research group called frequency-shifted interferometry (FSI. This technique uses a continuous-wave light source, an optical frequency shifter, and a slow detector. We discuss the operation principles of several FSI implementations and show their applications in fiber length and dispersion measurement, locating weak reflections along a fiber link, fiber-optic sensor multiplexing, and high-sensitivity cavity ring-down measurement. Detailed analysis of FSI system parameters is also presented.

New directions in point-contact spectroscopy based on scanning tunneling microscopy techniques (Review Article)

International Nuclear Information System (INIS)

Tartaglini, E.; Verhagen, T.G.A.; Galli, F.; Trouwborst, M.L.; Aarts, J.; Van-Ruitebbeek, J.M.; Muller, R.; Shiota, T.

2013-01-01

Igor Yanson showed 38 years ago for the first time a point-contact measurement where he probed the energy resolved spectroscopy of the electronic scattering inside the metal. Since this first measurement, the pointcontact spectroscopy (PCS) technique improved enormously. The application of the scanning probe microscopy (SPM) techniques in the late 1980s allowed achieving contacts with a diameter of a single atom. With the introduction of the mechanically controlled break junction technique, even spectroscopy on freely suspended chains of atoms could be performed. In this paper, we briefly review the current developments of PCS and show recent experiments in advanced scanning PCS based on SPM techniques. We describe some results obtained with both needle-anvil type of point contacts and scanning tunneling microscopy (STM). We also show our first attempt to lift up with a STM a chain of single gold atoms from a Au(110) surface.

Computational adaptive optics for broadband optical interferometric tomography of biological tissue.

Science.gov (United States)

Adie, Steven G; Graf, Benedikt W; Ahmad, Adeel; Carney, P Scott; Boppart, Stephen A

2012-05-08

Aberrations in optical microscopy reduce image resolution and contrast, and can limit imaging depth when focusing into biological samples. Static correction of aberrations may be achieved through appropriate lens design, but this approach does not offer the flexibility of simultaneously correcting aberrations for all imaging depths, nor the adaptability to correct for sample-specific aberrations for high-quality tomographic optical imaging. Incorporation of adaptive optics (AO) methods have demonstrated considerable improvement in optical image contrast and resolution in noninterferometric microscopy techniques, as well as in optical coherence tomography. Here we present a method to correct aberrations in a tomogram rather than the beam of a broadband optical interferometry system. Based on Fourier optics principles, we correct aberrations of a virtual pupil using Zernike polynomials. When used in conjunction with the computed imaging method interferometric synthetic aperture microscopy, this computational AO enables object reconstruction (within the single scattering limit) with ideal focal-plane resolution at all depths. Tomographic reconstructions of tissue phantoms containing subresolution titanium-dioxide particles and of ex vivo rat lung tissue demonstrate aberration correction in datasets acquired with a highly astigmatic illumination beam. These results also demonstrate that imaging with an aberrated astigmatic beam provides the advantage of a more uniform depth-dependent signal compared to imaging with a standard gaussian beam. With further work, computational AO could enable the replacement of complicated and expensive optical hardware components with algorithms implemented on a standard desktop computer, making high-resolution 3D interferometric tomography accessible to a wider group of users and nonspecialists.

Analysis technique for controlling system wavefront error with active/adaptive optics

Science.gov (United States)

Genberg, Victor L.; Michels, Gregory J.

2017-08-01

The ultimate goal of an active mirror system is to control system level wavefront error (WFE). In the past, the use of this technique was limited by the difficulty of obtaining a linear optics model. In this paper, an automated method for controlling system level WFE using a linear optics model is presented. An error estimate is included in the analysis output for both surface error disturbance fitting and actuator influence function fitting. To control adaptive optics, the technique has been extended to write system WFE in state space matrix form. The technique is demonstrated by example with SigFit, a commercially available tool integrating mechanical analysis with optical analysis.

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.

High-resolution intravital microscopy.

Directory of Open Access Journals (Sweden)

Volker Andresen

Full Text Available Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy--the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and

High-Resolution Intravital Microscopy

Science.gov (United States)

Andresen, Volker; Pollok, Karolin; Rinnenthal, Jan-Leo; Oehme, Laura; Günther, Robert; Spiecker, Heinrich; Radbruch, Helena; Gerhard, Jenny; Sporbert, Anje; Cseresnyes, Zoltan; Hauser, Anja E.; Niesner, Raluca

2012-01-01

Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy - the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning) while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs) of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and developmental biology

3D Image Analysis of Geomaterials using Confocal Microscopy

Science.gov (United States)

Mulukutla, G.; Proussevitch, A.; Sahagian, D.

2009-05-01

Confocal microscopy is one of the most significant advances in optical microscopy of the last century. It is widely used in biological sciences but its application to geomaterials lingers due to a number of technical problems. Potentially the technique can perform non-invasive testing on a laser illuminated sample that fluoresces using a unique optical sectioning capability that rejects out-of-focus light reaching the confocal aperture. Fluorescence in geomaterials is commonly induced using epoxy doped with a fluorochrome that is impregnated into the sample to enable discrimination of various features such as void space or material boundaries. However, for many geomaterials, this method cannot be used because they do not naturally fluoresce and because epoxy cannot be impregnated into inaccessible parts of the sample due to lack of permeability. As a result, the confocal images of most geomaterials that have not been pre-processed with extensive sample preparation techniques are of poor quality and lack the necessary image and edge contrast necessary to apply any commonly used segmentation techniques to conduct any quantitative study of its features such as vesicularity, internal structure, etc. In our present work, we are developing a methodology to conduct a quantitative 3D analysis of images of geomaterials collected using a confocal microscope with minimal amount of prior sample preparation and no addition of fluorescence. Two sample geomaterials, a volcanic melt sample and a crystal chip containing fluid inclusions are used to assess the feasibility of the method. A step-by-step process of image analysis includes application of image filtration to enhance the edges or material interfaces and is based on two segmentation techniques: geodesic active contours and region competition. Both techniques have been applied extensively to the analysis of medical MRI images to segment anatomical structures. Preliminary analysis suggests that there is distortion in the

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.

Review of quantitative phase-digital holographic microscopy: promising novel imaging technique to resolve neuronal network activity and identify cellular biomarkers of psychiatric disorders

KAUST Repository

Marquet, Pierre

2014-09-22

Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed.

Investigating the mesostructure of ordered porous silica nanocomposites by transmission electron microscopy techniques

Energy Technology Data Exchange (ETDEWEB)

Bullita, S.; Casula, M. F., E-mail: casulaf@unica.it [INSTM and Department of Chemical and Geological Science, University of Cagliari, Monserrato (Canada) (Italy); Piludu, M. [Department of Biomedical Sciences, University of Cagliari, Monserrato (Canada) (Italy); Falqui, A. [INSTM and Department of Chemical and Geological Science, University of Cagliari, Monserrato (Canada) Italy and KAUST-King Abdullah University of Science and Technology, Jeddah (Saudi Arabia); Carta, D. [INSTM and Department of Chemical and Geological Science, University of Cagliari, Monserrato (Canada), Italy and Faculty of Physical Sciences and Engineering, University of Southampton, Southampton (United Kingdom); Corrias, A. [INSTM and Department of Chemical and Geological Science, University of Cagliari, Monserrato (Canada) Italy and School of Physical Sciences, Ingram Building, University of Kent, Canterbury (United Kingdom)

2014-10-21

Nanocomposites made out of FeCo alloy nanocrystals supported onto pre-formed mesoporous ordered silica which features a cubic arrangement of pores (SBA-16) were investigated. Information on the effect of the nanocrystals on the mesostructure (i.e. pore arrangement symmetry, pore size, and shape) were deduced by a multitechnique approach including N2 physisorption, low angle X-ray diffraction, and Transmission electron microscopy. It is shown that advanced transmission electron microscopy techniques are required, however, to gain direct evidence on key compositional and textural features of the nanocomposites. In particular, electron tomography and microtomy techniques make clear that the FeCo nanocrystals are located within the pores of the SBA-16 silica, and that the ordered mesostructure of the nanocomposite is retained throughout the observed specimen.

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.

Physical, optical and electrical properties of copper selenide (CuSe) thin films deposited by solution growth technique at room temperature

International Nuclear Information System (INIS)

Gosavi, S.R.; Deshpande, N.G.; Gudage, Y.G.; Sharma, Ramphal

2008-01-01

Copper selenide (CuSe) thin films are grown onto amorphous glass substrate from an aqueous alkaline medium using solution growth technique (SGT) at room temperature. The preparative parameters were optimized to obtain good quality of thin films. The as-deposited films were characterized for physical, optical and electrical properties. X-ray diffraction (XRD) pattern reveals that the films are polycrystalline in nature. Energy dispersive analysis by X-ray (EDAX) shows formation of stoichiometric CuSe compound. Uniform deposition of CuSe thin films on glass substrate was observed from scanning electron microscopy (SEM) and atomic force microscopy (AFM) micrographs. Average grain size was determined to 144.53 ± 10 nm using atomic force microscopy. The band gap was found to be 2.03 eV with direct band-to-band transition. Semi-conducting behaviour was observed from resistivity measurements. Ohmic behaviour was seen from I-V curve with good electrical conductivity

Invited Review Article: Pump-probe microscopy

Science.gov (United States)

Wilson, Jesse W.; Robles, Francisco E.; Warren, Warren S.

2016-01-01

Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications. PMID:27036751

Invited Review Article: Pump-probe microscopy

Energy Technology Data Exchange (ETDEWEB)

Fischer, Martin C., E-mail: Martin.Fischer@duke.edu; Wilson, Jesse W.; Robles, Francisco E. [Department of Chemistry, Duke University, Durham, North Carolina 27708 (United States); Warren, Warren S. [Departments of Chemistry, Biomedical Engineering, Physics, and Radiology, Duke University, Durham, North Carolina 27708 (United States)

2016-03-15

Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications.

Compact Optical Technique for Streak Camera Calibration

International Nuclear Information System (INIS)

Curt Allen; Terence Davies; Frans Janson; Ronald Justin; Bruce Marshall; Oliver Sweningsen; Perry Bell; Roger Griffith; Karla Hagans; Richard Lerche

2004-01-01

The National Ignition Facility is under construction at the Lawrence Livermore National Laboratory for the U.S. Department of Energy Stockpile Stewardship Program. Optical streak cameras are an integral part of the experimental diagnostics instrumentation. To accurately reduce data from the streak cameras a temporal calibration is required. This article describes a technique for generating trains of precisely timed short-duration optical pulses that are suitable for temporal calibrations

Noninvasive determination of optical lever sensitivity in atomic force microscopy

International Nuclear Information System (INIS)

Higgins, M.J.; Proksch, R.; Sader, J.E.; Polcik, M.; Mc Endoo, S.; Cleveland, J.P.; Jarvis, S.P.

2006-01-01

Atomic force microscopes typically require knowledge of the cantilever spring constant and optical lever sensitivity in order to accurately determine the force from the cantilever deflection. In this study, we investigate a technique to calibrate the optical lever sensitivity of rectangular cantilevers that does not require contact to be made with a surface. This noncontact approach utilizes the method of Sader et al. [Rev. Sci. Instrum. 70, 3967 (1999)] to calibrate the spring constant of the cantilever in combination with the equipartition theorem [J. L. Hutter and J. Bechhoefer, Rev. Sci. Instrum. 64, 1868 (1993)] to determine the optical lever sensitivity. A comparison is presented between sensitivity values obtained from conventional static mode force curves and those derived using this noncontact approach for a range of different cantilevers in air and liquid. These measurements indicate that the method offers a quick, alternative approach for the calibration of the optical lever sensitivity

Noninvasive determination of optical lever sensitivity in atomic force microscopy

Science.gov (United States)

Higgins, M. J.; Proksch, R.; Sader, J. E.; Polcik, M.; Mc Endoo, S.; Cleveland, J. P.; Jarvis, S. P.

2006-01-01

Atomic force microscopes typically require knowledge of the cantilever spring constant and optical lever sensitivity in order to accurately determine the force from the cantilever deflection. In this study, we investigate a technique to calibrate the optical lever sensitivity of rectangular cantilevers that does not require contact to be made with a surface. This noncontact approach utilizes the method of Sader et al. [Rev. Sci. Instrum. 70, 3967 (1999)] to calibrate the spring constant of the cantilever in combination with the equipartition theorem [J. L. Hutter and J. Bechhoefer, Rev. Sci. Instrum. 64, 1868 (1993)] to determine the optical lever sensitivity. A comparison is presented between sensitivity values obtained from conventional static mode force curves and those derived using this noncontact approach for a range of different cantilevers in air and liquid. These measurements indicate that the method offers a quick, alternative approach for the calibration of the optical lever sensitivity.

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.

Spectroscopic ellipsometric modeling of a Bi–Te–Se write layer of an optical data storage device as guided by atomic force microscopy, scanning electron microscopy, and X-ray diffraction

Energy Technology Data Exchange (ETDEWEB)

Wang, Hao; Madaan, Nitesh; Bagley, Jacob; Diwan, Anubhav [Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602 (United States); Liu, Yiqun [Department of Chemistry, Lehigh University, Bethlehem, PA 18015 (United States); Davis, Robert C. [Department of Physics and Astronomy, Brigham Young University, Provo, UT 84602 (United States); Lunt, Barry M. [Department of Information Technology, Brigham Young University, Provo, UT 84602 (United States); Smith, Stacey J., E-mail: ssmith@chem.byu.edu [Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602 (United States); Linford, Matthew R., E-mail: mrlinford@chem.byu.edu [Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602 (United States)

2014-10-31

Conventional magnetic tape is the most widely used medium for archival data storage. However, data stored on it need to be migrated every ca. 5 years. Recently, optical discs that store information for hundreds, or even more than 1000 years, have been introduced to the market. We recently proposed that technology in these optical discs be used to make an optical tape that would show greater permanence than its magnetic counterpart. Here we provide a detailed optical characterization of a sputtered thin film of bismuth, tellurium, and selenium (BTS) that is a proposed data storage layer for these devices. The methodology described herein should be useful in the future development of related materials. Spectroscopic ellipsometry (SE) data are obtained using interference enhancement, and the modeling of this data is guided by results from atomic force microscopy (AFM), scanning electron microscopy (SEM), X-ray diffraction (XRD), and X-ray reflectivity (XRR). By AFM, ca. 40 nm BTS films show ca. 10 nm roughness. SEM images also suggest considerable roughness in the films and indicate that they are composed of 13.1 ± 5.9 nm grains. XRD confirms that the films are crystalline and predicts a grain size of 17 ± 2 nm. XRD results are consistent with the composition of the films — a mildly oxidized BTS material. Three models of increasing complexity are investigated to explain the SE data. The first model consists of a smooth, homogeneous BTS film. The second model adds a roughness layer to the previous model. The third model also has two layers. The bottom layer is modeled as a mixture of BTS and void using a Bruggeman effective medium approximation. The upper layer is similarly modeled, but with a gradient. The first model was unable to adequately model the SE data. The second model was an improvement — lower MSE (4.4) and good agreement with step height measurements. The third model was even better — very low MSE (2.6) and good agreement with AFM results. The

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.

Ex vivo confocal microscopy: an emerging technique in dermatology

Science.gov (United States)

Perrot, Jean Luc; Labeille, Bruno; Cambazard, Frédéric; Rubegni, Pietro

2018-01-01

This review aims to give an overview of the current available applications of ex vivo confocal microscopy (EVCM) in dermatology. EVCM is a relatively new imaging technique that allows microscopic examination of freshly excised unfixed tissue. It enables a rapid examination of the skin sample directly in the surgery room and thus represents an alternative to the intraoperative micrographic control of the surgical margins of cutaneous tumors by standard microscopic examination on cryopreserved sections during Mohs surgery. Although this technique has mainly been developed for the margin’s control of basal cell carcinoma, many other skin tumors have been studied, including melanoma. Use of EVCM is continuing to evolve, and many possible applications are under investigation, such as the study of nails and hair diseases and the diagnosis of skin infections. PMID:29785327

Optical Properties of Rotationally Twinned Nanowire Superlattices

DEFF Research Database (Denmark)

Bao, Jiming; Bell, David C.; Capasso, Federico

2008-01-01

We have developed a technique so that both transmission electron microscopy and microphotoluminescence can be performed on the same semiconductor nanowire over a large range of optical power, thus allowing us to directly correlate structural and optical properties of rotationally twinned zinc...... a heterostructure in a chemically homogeneous nanowire material and alter in a major way its optical properties opens new possibilities for band-structure engineering....

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.

Optical techniques for in-core measurements

International Nuclear Information System (INIS)

Brichard, B.

2007-01-01

The in-situ measurement of dimensional changes is a key issue for advanced irradiation programs in Material Test Reactors. It is for example crucial to monitor the changes of the dimensions of nuclear fuel assemblies as well as those of mechanically stressed structural material samples during in-pile irradiations. Different techniques already exist to carry out such measurements but they all come with a number of drawbacks. SCK-CEN and CEA have therefore decided to share the development of a measurement system that was never applied before in the core of a nuclear reactor. It relies on optical dimensional measurements and brings along unprecedented non-intrusiveness combined with high resolution. A clear advantage in using compact optical sensors results in a more efficient occupation of the irradiation volume available for target testings as well as a significant reduction of the gamma-heating associated with the in-pile instrumentation. The objectives of these shared studies are to design, develop, test and qualify an in-pile dimensional measurement system based on optical techniques, with the goal to implement this system in future MTR irradiation experiments. In 2006, we focussed our activities on sensor analysis, selection of the sensor prototypes, procurement and first irradiation experiment

Optical metrology techniques for dimensional stability measurements

NARCIS (Netherlands)

Ellis, Jonathan David

2010-01-01

This thesis work is optical metrology techniques to determine material stability. In addition to displacement interferometry, topics such as periodic nonlinearity, Fabry-Perot interferometry, refractometry, and laser stabilization are covered.

Comparison of mouse mammary gland imaging techniques and applications: Reflectance confocal microscopy, GFP Imaging, and ultrasound

International Nuclear Information System (INIS)

Tilli, Maddalena T; Parrish, Angela R; Cotarla, Ion; Jones, Laundette P; Johnson, Michael D; Furth, Priscilla A

2008-01-01

Genetically engineered mouse models of mammary gland cancer enable the in vivo study of molecular mechanisms and signaling during development and cancer pathophysiology. However, traditional whole mount and histological imaging modalities are only applicable to non-viable tissue. We evaluated three techniques that can be quickly applied to living tissue for imaging normal and cancerous mammary gland: reflectance confocal microscopy, green fluorescent protein imaging, and ultrasound imaging. In the current study, reflectance confocal imaging offered the highest resolution and was used to optically section mammary ductal structures in the whole mammary gland. Glands remained viable in mammary gland whole organ culture when 1% acetic acid was used as a contrast agent. Our application of using green fluorescent protein expressing transgenic mice in our study allowed for whole mammary gland ductal structures imaging and enabled straightforward serial imaging of mammary gland ducts in whole organ culture to visualize the growth and differentiation process. Ultrasound imaging showed the lowest resolution. However, ultrasound was able to detect mammary preneoplastic lesions 0.2 mm in size and was used to follow cancer growth with serial imaging in living mice. In conclusion, each technique enabled serial imaging of living mammary tissue and visualization of growth and development, quickly and with minimal tissue preparation. The use of the higher resolution reflectance confocal and green fluorescent protein imaging techniques and lower resolution ultrasound were complementary

Scanning second-harmonic optical microscopy of self-assembled InAlGaAs quantum dots

DEFF Research Database (Denmark)

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

2001-01-01

Microscopy provides a suitable technique for local probing of small ensembles of (or even individual) QD's, and when combined with the detection of second-harmonic (SH) generation the technique becomes suitable to reveal tiny changes of symmetry originating either in the material structures or in...

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...

Electrochemical Design of Optical Nanoantennas

Directory of Open Access Journals (Sweden)

Vasilchenko V.E.

2015-01-01

Full Text Available Electrochemical techniques for fabricating tapered gold nanoantennas (tips are discussed. In the paper, the tunable design of nanoantennas is demonstrated. Tip parameters such as a tip apex curvature, mesoscopic morphology, aspect ratio and enhancement factor can be varied with etching electrolyte and applied voltage. The low-cost method makes tipehnahced optical spectroscopy and microscopy feasible for routine optical measurements beyond the diffraction limit.

Magnetic force microscopy: advanced technique for the observation of magnetic domains

International Nuclear Information System (INIS)

Asenjo, A.; Garcia, J. M.; Vazquez, M.

2001-01-01

An overview on the Magnetic Force Microscopy, MFM, as an advanced technique to observe magnetic domains and walls is displayed. Basic concepts are first introduced on the domain structure formation as well as on other techniques to observe magnetic domains. Afterwards, the MFM instrumentation is described making also an emphasis in micro magnetic consideration to interpret the images. Finally, a set of selected advanced magnetic materials with different domain structures is chosen to show the wide possibilities of this techniques to characterise the surface magnetic behaviour. The domain structure of materials as commercial magnetic recording media, thin films and multilayers, amorphous micro tubes, nanocrystalline ribbons, perovskites or magnetic nano wires is shown. (Author) 16 refs

Coherent anti-Stokes Raman scattering microscopy (CARS): Instrumentation and applications

International Nuclear Information System (INIS)

Djaker, Nadia; Lenne, Pierre-Francois; Marguet, Didier; Colonna, Anne; Hadjur, Christophe; Rigneault, Herve

2007-01-01

Recent advances in laser physics have permitted the development of a new kind of microscopy based on stimulated Raman scattering. This new technique known as Coherent anti-Stokes Raman scattering (CARS) microscopy allows vibrational imaging with high sensitivity, high spectral resolution and three-dimensional sectioning capabilities. We review recent advances in CARS microscopy, with applications to chemical and biological systems. We also present an application of CARS microscopy with high optical resolution and spectral selectivity, in resolving structures in surface ex vivo stratum corneum by looking at the CH 2 stretching vibrational band. A strong CARS signal is backscattered from an intense forward generated CARS signal in thick samples. This makes noninvasive imaging of deep structures possible, without labeling or chemical treatments

Laboratory soft x-ray microscopy and tomography

International Nuclear Information System (INIS)

Bertilson, Michael

2011-01-01

Soft x-ray microscopy in the water-window (λ = 2.28 nm - 4.36 nm) is based on zone-plate optics and allows high-resolution imaging of, e.g., cells and soils in their natural or near-natural environment. Three-dimensional imaging is provided via tomographic techniques, soft x-ray cryo tomography. However, soft x-ray microscopes with such capabilities have been based on large-scale synchrotron x-ray facilities, thereby limiting their accessibility for a wider scientific community. This Thesis describes the development of the Stockholm laboratory soft x-ray microscope to three-dimensional cryo tomography and to new optics-based contrast mechanisms. The microscope relies on a methanol or nitrogen liquid-jet laser-plasma source, normal-incidence multilayer or zone-plate condenser optics, in-house fabricated zone-plate objectives, and allows operation at two wavelengths in the water-window, λ = 2.48 nm and λ = 2.48 nm. With the implementation of a new state-of-the-art normal-incidence multilayer condenser for operation at λ = 2.48 nm and a tiltable cryogenic sample stage the microscope now allows imaging of dry, wet or cryo-fixed samples. This arrangement was used for the first demonstration of laboratory soft x-ray cryo microscopy and tomography. The performance of the microscope has been demonstrated in a number of experiments described in this Thesis, including, tomographic imaging with a resolution of 140 nm, cryo microscopy and tomography of various cells and parasites, and for studies of aqueous soils and clays. The Thesis also describes the development and implementation of single-element differential-interference and Zernike phase-contrast zone-plate objectives. The enhanced contrast provided by these optics reduce exposure times or lowers the dose in samples and are of major importance for harder x-ray microscopy. The implementation of a high-resolution 50 nm compound zone-plate objective for sub-25-nm resolution imaging is also described. All experiments

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

A STUDY ON OPTICAL CHARACTER RECOGNITION TECHNIQUES

OpenAIRE

Narendra Sahu; Manoj Sonkusare

2018-01-01

Optical Character Recognition (OCR) is the process which enables a system to without human intervention identifies the scripts or alphabets written into the users’ verbal communication. Optical Character identification has grown to be individual of the mainly flourishing applications of knowledge in the field of pattern detection and artificial intelligence. In our survey we study on the various OCR techniques. In this paper we resolve and examine the hypothetical and numerical models of Opti...

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.

Multiple speckle illumination for optical-resolution photoacoustic imaging

Science.gov (United States)

Poisson, Florian; Stasio, Nicolino; Moser, Christophe; Psaltis, Demetri; Bossy, Emmanuel

2017-03-01

Optical-resolution photoacoustic microscopy offers exquisite and specific contrast to optical absorption. Conventional approaches generally involves raster scanning a focused spot over the sample. Here, we demonstrate that a full-field illumination approach with multiple speckle illumination can also provide diffraction-limited optical-resolution photoacoustic images. Two different proof-of-concepts are demonstrated with micro-structured test samples. The first approach follows the principle of correlation/ghost imaging,1, 2 and is based on cross-correlating photoacoustic signals under multiple speckle illumination with known speckle patterns measured during a calibration step. The second approach is a speckle scanning microscopy technique, which adapts the technique proposed in fluorescence microscopy by Bertolotti and al.:3 in our work, spatially unresolved photoacoustic measurements are performed for various translations of unknown speckle patterns. A phase-retrieval algorithm is used to reconstruct the object from the knowledge of the modulus of its Fourier Transform yielded by the measurements. Because speckle patterns naturally appear in many various situations, including propagation through biological tissue or multi-mode fibers (for which focusing light is either very demanding if not impossible), speckle-illumination-based photoacoustic microscopy provides a powerful framework for the development of novel reconstruction approaches, well-suited to compressed sensing approaches.2

Monitoring techniques for the manufacture of tapered optical fibers.

Science.gov (United States)

Mullaney, Kevin; Correia, Ricardo; Staines, Stephen E; James, Stephen W; Tatam, Ralph P

2015-10-01

The use of a range of optical techniques to monitor the process of fabricating optical fiber tapers is investigated. Thermal imaging was used to optimize the alignment of the optical system; the transmission spectrum of the fiber was monitored to confirm that the tapers had the required optical properties and the strain induced in the fiber during tapering was monitored using in-line optical fiber Bragg gratings. Tapers were fabricated with diameters down to 5 μm and with waist lengths of 20 mm using single-mode SMF-28 fiber.

Possibilities for Advanced Encoding Techniques at Signal Transmission in the Optical Transmission Medium

Directory of Open Access Journals (Sweden)

Filip Čertík

2016-01-01

Full Text Available This paper presents a possible simulation of negative effects in the optical transmission medium and an analysis for the utilization of different signal processing techniques at the optical signal transmission. An attention is focused on the high data rate signal transmission in the optical fiber influenced by linear and nonlinear environmental effects presented by the prepared simulation model. The analysis includes possible utilization of OOK, BPSK, DBPSK, BFSK, QPSK, DQPSK, 8PSK, and 16QAM modulation techniques together with RS, BCH, and LDPC encoding techniques for the signal transmission in the optical fiber. Moreover, the prepared simulation model is compared with real optical transmission systems. In the final part, a comparison of the selected modulation techniques with different encoding techniques and their implementation in real transmission systems is shown.

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

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.

Second harmonic generation microscopy

DEFF Research Database (Denmark)

Brüggemann, Dagmar Adeline; Brewer, Jonathan R.; Risbo, Jens

2010-01-01

Myofibers and collagen show non-linear optical properties enabling imaging using second harmonic generation (SHG) microscopy. The technique is evaluated for use as a tool for real-time studies of thermally induced changes in thin samples of unfixed and unstained pork. The forward and the backward...... scattered SHG light reveal complementary features of the structures of myofibers and collagen fibers. Upon heating the myofibers show no structural changes before reaching a temperature of 53 °C. At this temperature the SHG signal becomes extinct. The extinction of the SHG at 53 °C coincides with a low......-temperature endotherm peak observable in the differential scanning calorimetry (DSC) thermograms. DSC analysis of epimysium, the connective tissue layer that enfold skeletal muscles, produces one large endotherm starting at 57 °C and peaking at 59.5 °C. SHG microscopy of collagen fibers reveals a variability of thermal...

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.

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

Nonlinear Polarimetric Microscopy for Biomedical Imaging

Science.gov (United States)

Samim, Masood

susceptibilities. The developed nonlinear optical polarimetric microscopy is applicable to a wide variety of structural studies on ordered materials, and provides a non-invasive possibility to study the structural organization and dynamics within biological samples. For example, the technique is well suited for studies of a muscle contraction, histopathology of collagen structure for cancer tissue diagnostics, investigations of the polysacharide structural organization within a starch granule of a plant, or developmental study of the retina in an eye, among other applications.

Handheld Fluorescence Microscopy based Flow Analyzer.

Science.gov (United States)

Saxena, Manish; Jayakumar, Nitin; Gorthi, Sai Siva

2016-03-01

Fluorescence microscopy has the intrinsic advantages of favourable contrast characteristics and high degree of specificity. Consequently, it has been a mainstay in modern biological inquiry and clinical diagnostics. Despite its reliable nature, fluorescence based clinical microscopy and diagnostics is a manual, labour intensive and time consuming procedure. The article outlines a cost-effective, high throughput alternative to conventional fluorescence imaging techniques. With system level integration of custom-designed microfluidics and optics, we demonstrate fluorescence microscopy based imaging flow analyzer. Using this system we have imaged more than 2900 FITC labeled fluorescent beads per minute. This demonstrates high-throughput characteristics of our flow analyzer in comparison to conventional fluorescence microscopy. The issue of motion blur at high flow rates limits the achievable throughput in image based flow analyzers. Here we address the issue by computationally deblurring the images and show that this restores the morphological features otherwise affected by motion blur. By further optimizing concentration of the sample solution and flow speeds, along with imaging multiple channels simultaneously, the system is capable of providing throughput of about 480 beads per second.

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.

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.

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...

Optical supervised filtering technique based on Hopfield neural network

Science.gov (United States)

Bal, Abdullah

2004-11-01

Hopfield neural network is commonly preferred for optimization problems. In image segmentation, conventional Hopfield neural networks (HNN) are formulated as a cost-function-minimization problem to perform gray level thresholding on the image histogram or the pixels' gray levels arranged in a one-dimensional array [R. Sammouda, N. Niki, H. Nishitani, Pattern Rec. 30 (1997) 921-927; K.S. Cheng, J.S. Lin, C.W. Mao, IEEE Trans. Med. Imag. 15 (1996) 560-567; C. Chang, P. Chung, Image and Vision comp. 19 (2001) 669-678]. In this paper, a new high speed supervised filtering technique is proposed for image feature extraction and enhancement problems by modifying the conventional HNN. The essential improvement in this technique is to use 2D convolution operation instead of weight-matrix multiplication. Thereby, neural network based a new filtering technique has been obtained that is required just 3 × 3 sized filter mask matrix instead of large size weight coefficient matrix. Optical implementation of the proposed filtering technique is executed easily using the joint transform correlator. The requirement of non-negative data for optical implementation is provided by bias technique to convert the bipolar data to non-negative data. Simulation results of the proposed optical supervised filtering technique are reported for various feature extraction problems such as edge detection, corner detection, horizontal and vertical line extraction, and fingerprint enhancement.

Turbo Equalization Techniques Toward Robust PDM 16-QAM Optical Fiber Transmission

DEFF Research Database (Denmark)

Arlunno, Valeria; Caballero Jambrina, Antonio; Borkowski, Robert

2014-01-01

In this paper, we show numerically and experimentally that turbo equalization (TE) is an efficient technique to mitigate performance degradations stemming from optical fiber propagation effects in both optical fiber dispersion managed and unmanaged coherent detection links. The effectiveness....... As TE can be included in the current coherent detection transceiver technologies and complement other equalization techniques, it has prospects for application in next-generation high-capacity and long-reach optical transmission links....

Validation and qualification of surface-applied fibre optic strain sensors using application-independent optical techniques

International Nuclear Information System (INIS)

Schukar, Vivien G; Kadoke, Daniel; Kusche, Nadine; Münzenberger, Sven; Gründer, Klaus-Peter; Habel, Wolfgang R

2012-01-01

Surface-applied fibre optic strain sensors were investigated using a unique validation facility equipped with application-independent optical reference systems. First, different adhesives for the sensor's application were analysed regarding their material properties. Measurements resulting from conventional measurement techniques, such as thermo-mechanical analysis and dynamic mechanical analysis, were compared with measurements resulting from digital image correlation, which has the advantage of being a non-contact technique. Second, fibre optic strain sensors were applied to test specimens with the selected adhesives. Their strain-transfer mechanism was analysed in comparison with conventional strain gauges. Relative movements between the applied sensor and the test specimen were visualized easily using optical reference methods, digital image correlation and electronic speckle pattern interferometry. Conventional strain gauges showed limited opportunities for an objective strain-transfer analysis because they are also affected by application conditions. (paper)

Real-time high dynamic range laser scanning microscopy

Science.gov (United States)

Vinegoni, C.; Leon Swisher, C.; Fumene Feruglio, P.; Giedt, R. J.; Rousso, D. L.; Stapleton, S.; Weissleder, R.

2016-04-01

In conventional confocal/multiphoton fluorescence microscopy, images are typically acquired under ideal settings and after extensive optimization of parameters for a given structure or feature, often resulting in information loss from other image attributes. To overcome the problem of selective data display, we developed a new method that extends the imaging dynamic range in optical microscopy and improves the signal-to-noise ratio. Here we demonstrate how real-time and sequential high dynamic range microscopy facilitates automated three-dimensional neural segmentation. We address reconstruction and segmentation performance on samples with different size, anatomy and complexity. Finally, in vivo real-time high dynamic range imaging is also demonstrated, making the technique particularly relevant for longitudinal imaging in the presence of physiological motion and/or for quantification of in vivo fast tracer kinetics during functional imaging.

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

Characterization of Byzantine pottery from Oltina (Constanţa County), Romania, using PIXE and Optical Microscopy

Science.gov (United States)

Bugoi, Roxana; Talmaţchi, Cristina; Haitǎ, Constantin; Ceccato, Daniele

2018-02-01

An assemblage of 58 ceramic shards discovered in archaeological excavations at Oltina, Romania, dated to the 10th-11th century CE, was subjected to archaeometric investigations in order to reveal the raw materials and manufacturing techniques employed by the potters from the Lower Danube zone during the Byzantine ruling. The initial grouping of the shards according to stylistic criteria was refined by the subsequent petrographic study. Optical Microscopy (OM) detailed the general mineralogy and the pottery fabric, i.e. the textural characteristics, porosity and microstructure, surface treatments and firing. The PIXE analyses of potteries performed at AN2000 accelerator of LNL, INFN, Italy led to the identification of the chemical composition of the ceramic shards. The Hierarchical Cluster Analysis of the PIXE data evidenced several categories of shards with distinct compositional signatures, the main division being the one separating the ceramic fragments made of kaolinitic clays from the rest of the Oltina potteries.

Use of Kelvin probe force microscopy for identification of CVD grown graphene flakes on copper foil

Science.gov (United States)

Kumar, Rakesh; Mehta, B. R.; Kanjilal, D.

2017-05-01

Graphene flakes have been grown by chemical vapour deposition (CVD) method on Cu foils. The obtained graphene flakes have been characterized by optical microscopy, field emission scanning electron microscopy, Kelvin probe force microscopy (KPFM) and Raman spectroscopy. The graphene flakes grown on Cu foil comprise mainly single layer graphene and confirm that the nucleation for graphene growth starts very quickly. Moreover, KPFM has been found to be a valuable technique to differentiate between covered and uncovered portion of Cu foil by graphene flakes deposited for shorter duration. The results show that KPFM can be a very useful technique in understanding the mechanism of graphene growth.

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.

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.

Spin-filter scanning tunneling microscopy : a novel technique for the analysis of spin polarization on magnetic surfaces and spintronic devices

NARCIS (Netherlands)

Vera Marun, I.J.

2010-01-01

This thesis deals with the development of a versatile technique to measure spin polarization with atomic resolution. A microscopy technique that can measure electronic spin polarization is relevant for characterization of magnetic nanostructures and spintronic devices. Scanning tunneling microscopy

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.

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.

All-optical delay technique for supporting multiple antennas in a hybrid optical - wireless transmission system

DEFF Research Database (Denmark)

Prince, Kamau; Chiuchiarelli, A; Presi, M

2008-01-01

We introduce a novel continuously-variable optical delay technique to support beam-forming wireless communications systems using antenna arrays. We demonstrate delay with 64-QAM modulated signals at a rate of 15 Msymbol/sec with 2.5 GHz carrier frequency.......We introduce a novel continuously-variable optical delay technique to support beam-forming wireless communications systems using antenna arrays. We demonstrate delay with 64-QAM modulated signals at a rate of 15 Msymbol/sec with 2.5 GHz carrier frequency....

Technique to measure contact angle of micro/nanodroplets using atomic force microscopy

International Nuclear Information System (INIS)

Jung, Yong Chae; Bhushan, Bharat

2008-01-01

Contact angle is the primary parameter that characterizes wetting; however, the measurement techniques have been limited to droplets with a diameter as low as about 50 μm. The authors developed an atomic force microscopy-based technique to measure the contact angle of micro- and nanodroplets deposited using a modified nanoscale dispensing tip. The obtained contact angle results were compared with those of a macrodroplet (2.1 mm diameter). It was found that the contact angle on various surfaces decreases with decreasing the droplet size

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

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.

Enhancement of Lithium Niobate nanophotonic structures via spin-coating technique for optical waveguides application

Directory of Open Access Journals (Sweden)

Fakhri Makram A.

2017-01-01

Full Text Available This work is dedicated to investigation of temperature effects in Lithium Niobate (LiNbO3 nanostructures. The LiNbO3 nanostructures were deposited on glass substrate by spin-coating technique. LiNbO3 was set down at 3000 rpm for 30 sec and annealed from 100 to 600 °C. The structures were characterized and analyzed by scanning electron microscopy (SEM and ultra-violet visible (UV-vis spectrophotometer. The measured results have showed that by increasing annealing temperatures, the structures start to be more crystallized and be more homogenized until the optimum arrangement was achieved. Once this was accomplished, it's applicable for optical waveguides development. Eventually, it starts to be less crystallization and non-homogeneous. Energy gap was recorded to be at average value of 3.9 eV.

X-ray optics and X-ray microscopes: new challenges

International Nuclear Information System (INIS)

Susini, J.

2004-01-01

Soon after the discovery of X-rays in 1895 by W. Roentgen, it became rapidly clear that the methods traditionally used in the visible light regime, namely refraction, diffraction and reflection were difficult to apply for X-ray optics. The physical origins of these difficulties are closely linked to the very nature of interaction of X-rays with matter. The small deviation δ of the refractive index of condensed matter from unity makes it difficult to extend refraction-based optics from the optical spectral region to the X-ray region because the refraction angle is proportional to δ. Similarly it is very challenging to extend diffraction-based focusing techniques to X-rays because the diffraction angle scales inversely with wavelength. Finally, the use of reflection-based optics is also limited by the very small critical angle for total reflection. All those fundamental limitations prevented for almost one century, the development of X-ray microscopy whereas electron microscopy became a standard tool. In the past twenty years, interests for X-ray microscopy revived, mainly because of several major advances in X-ray sources and X-ray optics. X-ray microscopy techniques are now emerging as powerful and complementary tools for submicron investigations. Soft X-ray microscopes offer traditionally the possibility to form direct images of thick hydrated biological material in near-native environment, at a spatial resolution well beyond that achievable with visible light microscopy. Natural contrast is available in the soft X-ray region, in the so-called ''water-window'', due to the presence of absorption edges of the major constituents (C,N,O). Recent advances in manufacturing techniques have enlarged the accessible energy range of micro-focussing optics and offer new applications in a broad range of disciplines. X-ray microscopy in the 1 - 30 keV energy range is better suited for fluorescence to map trace elements, tomography for 3D imaging and micro-diffraction. The

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.

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.

LIBS, Raman spectroscopy, and optical microscopy analyses of superficial encrustations on ancient tesserae in Lebanon

Science.gov (United States)

Tomkowska, Anna; Chmielewski, Krzysztof; Skrzyczanowski, Wojciech; Mularczyk-Oliwa, Monika; Ostrowski, Roman; Strzelec, Marek

2017-07-01

The aim of research was determination of composition and nature of superficial deposits, cumulated at the selected mosaic's tesserae from Lebanon. Selected were three series of objects from different locations, namely from the seaside and mountain archaeological sites as well as from the mosaics exposed in the city center. Analyzed were stone and ceramic tesserae. The selection of objects was dictated by wide diversification of factors influencing the state of preservation and composition of deposits in given location. Investigations were performed including LIBS, FT-IR, Raman spectroscopy and optical 3D microscopy. The experimental results included composition and kind of deposit at the tesserae surfaces, and composition of tesserae itself. Compounds in the superficial deposits were identified. Confirmed was occurrence of different encrustations in dependence on geographic localization of a given sample. The interpretation of results was supported by multivariate statistical techniques, especially by the factor analysis. Performed analyses constitute the pioneer realization in terms of determination of deposits composition at the surface of mosaics from the Lebanon territory.

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.

Analyzing speckle contrast for HiLo microscopy optimization

Science.gov (United States)

Mazzaferri, J.; Kunik, D.; Belisle, J. M.; Singh, K.; Lefrançois, S.; Costantino, S.

2011-07-01

HiLo microscopy is a recently developed technique that provides both optical sectioning and fast imaging with a simple implementation and at a very low cost. The methodology combines widefield and speckled illumination images to obtain one optically sectioned image. Hence, the characteristics of such speckle illumination ultimately determine the quality of HiLo images and the overall performance of the method. In this work, we study how speckle contrast influence local variations of fluorescence intensity and brightness profiles of thick samples. We present this article as a guide to adjust the parameters of the system for optimizing the capabilities of this novel technology.

Electro-optic techniques for VLSI interconnect

Science.gov (United States)

Neff, J. A.

1985-03-01

A major limitation to achieving significant speed increases in very large scale integration (VLSI) lies in the metallic interconnects. They are costly not only from the charge transport standpoint but also from capacitive loading effects. The Defense Advanced Research Projects Agency, in pursuit of the fifth generation supercomputer, is investigating alternatives to the VLSI metallic interconnects, especially the use of optical techniques to transport the information either inter or intrachip. As the on chip performance of VLSI continues to improve via the scale down of the logic elements, the problems associated with transferring data off and onto the chip become more severe. The use of optical carriers to transfer the information within the computer is very appealing from several viewpoints. Besides the potential for gigabit propagation rates, the conversion from electronics to optics conveniently provides a decoupling of the various circuits from one another. Significant gains will also be realized in reducing cross talk between the metallic routings, and the interconnects need no longer be constrained to the plane of a thin film on the VLSI chip. In addition, optics can offer an increased programming flexibility for restructuring the interconnect network.

Field results of antifouling techniques for optical instruments

Science.gov (United States)

Strahle, W.J.; Hotchkiss, F.S.; Martini, Marinna A.

1998-01-01

An anti-fouling technique is developed for the protection of optical instruments from biofouling which leaches a bromide compound into a sample chamber and pumps new water into the chamber prior to measurement. The primary advantage of using bromide is that it is less toxic than the metal-based antifoulants. The drawback of the bromide technique is also discussed.

SNOM and AFM microscopy techniques to study the effect of non-ionizing radiation on the morphological and biochemical properties of human keratinocytes cell line (HaCaT).

Science.gov (United States)

Rieti, S; Manni, V; Lisi, A; Giuliani, L; Sacco, D; D'Emilia, E; Cricenti, A; Generosi, R; Luce, M; Grimaldi, S

2004-01-01

In this study we have employed atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM) techniques to study the effect of the interaction between human keratinocytes (HaCaT) and electromagnetic fields at low frequency. HaCaT cells were exposed to a sinusoidal magnetic field at a density of 50 Hz, 1 mT. AFM analysis revealed modification in shape and morphology in exposed cells with an increase in the areas of adhesion between cells. This latter finding was confirmed by SNOM indirect immunofluorescence analysis performed with a fluorescent antibody against the adhesion marker beta4 integrin, which revealed an increase of beta4 integrin segregation in the cell membrane of 50-Hz exposed cells, suggesting that a higher percentage of these cells shows a modified pattern of this adhesion marker.

Study of Pictorial technique in a colonial wall painting from El Padre coffee plantation using XRF and complementary technique

International Nuclear Information System (INIS)

Rodriguez, G.; Mendoza, A.

2001-01-01

This investigation is based on the study of the pictorial technique employed in a wall painting from El Padre coffee plantation situation in Madruga, Havana, and built at beginning of XIX century. The knowledge about pictorical techniques that was used in a painting is a typical research in Archaeometry, because it is so important the art historical information as scientific results to conclude about the real materials and procedures that were used in the past by the artists . The prediction of a wall painting pictorical technique is carrying out trough the knowledge of pigments, mortars layers and binding substance composition. The non destructive inorganic microanalysis of mortars layers and pigments was determined by a portable XRF system, which is very useful because of the most of pigments are of inorganic nature. A pinpoint samples microanalysis of each color from the painting were done by complementary and traditional technique in stratigraphic studies such as: optical microscopy, micro- chemical drop test, Scanning Electron Microscopy with Energy Dispersive X-Ray attachment (SEM-EDX), gas chromatography and Fourier Transformed Infra Red Spectroscopy (FTIR). The overlapping of layers was obtained by optical microscopy, The mortars composition was investigated by micro-chemical drop test and SEM/EDX and further confirmed by FTIR. The presence of binding substances in colors layers was identified by specific staining test and Gas Chromatography

High-Throughput Light Sheet Microscopy for the Automated Live Imaging of Larval Zebrafish

Science.gov (United States)

Baker, Ryan; Logan, Savannah; Dudley, Christopher; Parthasarathy, Raghuveer

The zebrafish is a model organism with a variety of useful properties; it is small and optically transparent, it reproduces quickly, it is a vertebrate, and there are a large variety of transgenic animals available. Because of these properties, the zebrafish is well suited to study using a variety of optical technologies including light sheet fluorescence microscopy (LSFM), which provides high-resolution three-dimensional imaging over large fields of view. Research progress, however, is often not limited by optical techniques but instead by the number of samples one can examine over the course of an experiment, which in the case of light sheet imaging has so far been severely limited. Here we present an integrated fluidic circuit and microscope which provides rapid, automated imaging of zebrafish using several imaging modes, including LSFM, Hyperspectral Imaging, and Differential Interference Contrast Microscopy. Using this system, we show that we can increase our imaging throughput by a factor of 10 compared to previous techniques. We also show preliminary results visualizing zebrafish immune response, which is sensitive to gut microbiota composition, and which shows a strong variability between individuals that highlights the utility of high throughput imaging. National Science Foundation, Award No. DBI-1427957.

Characterization of nanocrystalline zirconia powders by electron optical techniques

International Nuclear Information System (INIS)

Bursill, L.A.

1989-01-01

Electron optical techniques are described for the characterization of the size distribution of agglomerates, aggregates and primary micro- and nanocrystallites of as-processed zirconia powders. These techniques allow for direct identification of individual crystallites as tetragonal or monoclinic, by optical transform of high-resolution electron micrographs. The latter also permit surface morphology to be examined with atomic resolution. Applications to a range of pure and doped zirconia powders, of recent commercial interest, are presented, which enable the results of concurrent studies by sedimentation, surface specific area measurements, porosity and sinterability to be correctly interpreted. 18 figs

Comparative study of nano copper aluminate spinel prepared by sol–gel and modified sol–gel techniques: Structural, electrical, optical and catalytic studies

International Nuclear Information System (INIS)

Kumar, R. Thinesh; Suresh, P.; Selvam, N. Clament Sagaya; Kennedy, L. John; Vijaya, J. Judith

2012-01-01

Highlights: ► A modified sol–gel method for the preparation of nano CuAl 2 O 4 spinel using ethylenediamine was studied. ► Role of ethylenediamine in enhancing the structural, electrical, optical and catalytic properties of copper aluminate is highlighted. ► Effect of preparation method on the activity and selectivity of the samples on the oxidation of benzyl alcohol by CuAl 2 O 4 is studied. - Abstract: The effect of ethylenediamine addition in the sol–gel method for the preparation of nano CuAl 2 O 4 spinel for the enhancement in their structural, electrical, optical and catalytic properties was investigated. The samples were prepared by two different methods: sol–gel and modified sol–gel technique using ethylenediamine. X-ray diffraction (XRD), Fourier transform infrared spectra (FT-IR), scanning electron microscopy (SEM), high resolution-transmission electron microscopy (HR-TEM), energy dispersive X-ray analysis (EDX), nitrogen adsorption/desorption isotherms, temperature dependent conductance measurements, thermoelectric power (TEP) measurements and UV–visible diffuse reflectance (UV–vis-DRS) spectra were used to characterize the samples prepared. CuAl 2 O 4 prepared by modified sol–gel technique was found to possess a higher surface area, lower crystallite size, lower activation energy and high porosity than the one prepared by sol–gel method which in turn lead to the improved performance of it towards the selective oxidation of benzyl alcohol to benzaldehyde. Effect of solvent on the catalytic oxidation of benzyl alcohol by the nano CuAl 2 O 4 prepared by modified sol–gel technique was also investigated.

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)

Study on the crystallization of the metal glass with the ferromagnetic resonance and transmission electron microscopy techniques

International Nuclear Information System (INIS)

Biasi, R.S. de; Rodrigues, R.W.D.; Pascual, R.; Pessoa, C.S.

1983-01-01

The crystallization of the metal glass METGLAS 2826A has been studied with the ferromagnetic resonance and electron transmission microscopy techniques. The first-derivative linewidth of the absorption curve was measured for several times of isothermal treatments at 375 0 C. After an initial decrease, attributed to stress relaxation, the linewidth increases linearly with the transformed fraction of the first crystallization phase. Comparison with the electron microscopy results shows that the ferromagnetic resonance technique is particularly useful for short and medium aging times. (Author) [pt

Characterization of duplex stainless steels by TEM [transmission electron microscopy], SANS [small-angle neutron scattering], and APFIM [atom-probe field ion microscopy] techniques

International Nuclear Information System (INIS)

Chung, H.M.; Chopra, O.K.

1987-06-01

Results are presented of complementary characterization of aged duplex stainless steels by advanced metallographic techniques, including transmission and high-voltage electron microscopies; small-angle neutron scattering; and atom-probe field ion microscopy. On the basis of the characterization, the mechanisms of aging embrittlement have been shown to be associated with the precipitation of Ni- and Si-rich G phase and Cr-rich α' in the ferrite, and M 23 C 6 carbides on the austenite-ferrite phase boundaries. 19 refs., 19 figs., 1 tab

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

Optical and digital techniques for information security

CERN Document Server

2005-01-01

Optical and Digital Techniques for Information Security is the first book in a series focusing on Advanced Sciences and Technologies for Security Applications. This book encompases the results of research investigation and technologies used to secure, verify, recognize, track, and authenticate objects and information from theft, counterfeiting, and manipulation by unauthorized persons and agencies. This Information Security book will draw on the diverse expertise in optical sciences and engineering, digital image processing, imaging systems, information processing, computer based information systems, sensors, detectors, and biometrics to report innovative technologies that can be applied to information security issues. The Advanced Sciences and Technologies for Security Applications series focuses on research monographs in the areas of: -Recognition and identification (including optical imaging, biometrics, authentication, verification, and smart surveillance systems) -Biological and chemical threat detection...

Handbook of coherent domain optical methods biomedical diagnostics, environmental and material science

CERN Document Server

2004-01-01

For the first time in one set of books, coherent-domain optical methods are discussed in the framework of various applications, which are characterized by a strong light scattering. A few chapters describe basic research containing the updated results on coherent and polarized light non-destructive interactions with a scattering medium, in particular, diffraction, interference, and speckle formation at multiple scattering. These chapters allow for understanding coherent-domain diagnostic techniques presented in later chapters. A large portion of Volume I is dedicated to analysis of various aspects of optical coherence tomography (OCT) - a very new and growing field of coherent optics. Two chapters on laser scanning confocal microscopy give insight to recent extraordinary results on in vivo imaging and compare the possibilities and achievements of confocol, excitation multiphoton, and OCT microscopy. This two volume reference contains descriptions of holography, interferometry and optical heterodyning techniqu...

Application condition of optical communication technique in the nuclear power plants

International Nuclear Information System (INIS)

Sakurai, Jun

1999-01-01

As the optical communication technique can process rapidly a lot of information and exclude perfectly error action due to noise, it is adopted gradually to commercial and company communications (containing operational managements in large scale facilities) in worldwide scale in stead of conventional communication technique (containing operational controls and measurements). In application to the nuclear power plants, as forming not only change in properties but also deterioration due to radiation damage in many cases of exposure to various types of radiations such as neutron, gamma-ray, and so forth in difference with conventional using environment, its using range is limited at present. In future, development of optical fibers or elements with excellent high temperature and radiation resistances usable stably at reactor core for a long time is essential. The regular application of the optical communication technique at the nuclear power plants begins just now, which is an expected field for future large development. And, for the old nuclear power plant in present operation, substitution to the optical communication technique in accompany with replace of appliances at periodical inspections will also be conducted. Its response is already required rapidly in the Tokyo Electric Power Co., Ltd.. (G.K.)

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.

Super-resolution and super-localization microscopy: A novel tool for imaging chemical and biological processes

Energy Technology Data Exchange (ETDEWEB)

Dong, Bin [Iowa State Univ., Ames, IA (United States)

2015-01-01

Optical microscopy imaging of single molecules and single particles is an essential method for studying fundamental biological and chemical processes at the molecular and nanometer scale. The best spatial resolution (~ λ/2) achievable in traditional optical microscopy is governed by the diffraction of light. However, single molecule-based super-localization and super-resolution microscopy imaging techniques have emerged in the past decade. Individual molecules can be localized with nanometer scale accuracy and precision for studying of biological and chemical processes.This work uncovered the heterogeneous properties of the pore structures. In this dissertation, the coupling of molecular transport and catalytic reaction at the single molecule and single particle level in multilayer mesoporous nanocatalysts was elucidated. Most previous studies dealt with these two important phenomena separately. A fluorogenic oxidation reaction of non-fluorescent amplex red to highly fluorescent resorufin was tested. The diffusion behavior of single resorufin molecules in aligned nanopores was studied using total internal reflection fluorescence microscopy (TIRFM).

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.

High speed low power optical detection of sub-wavelength scatterer

NARCIS (Netherlands)

Roy, S.; Bouwens, M.A.J.; Wei, L.; Pereira, S.F.; Urbach, H.P.; Walle, P. van der

2015-01-01

Optical detection of scatterers on a flat substrate, generally done using dark field microscopy technique, is challenging since it requires high power illumination to obtain sufficient SNR (Signal to Noise Ratio) to be able to detect sub-wavelength particles. We developed a bright field technique,

Detection and Diagnosis of Oral Neoplasia with an Optical Coherence Microscope

National Research Council Canada - National Science Library

Clark, Anne

2003-01-01

.... The technique combines the sub-cellular resolution of high numerical aperture (NA) confocal microscopy with the increased sensitivity and penetration depth of optical coherence tomography (OCT...

Optical Measurement Techniques Innovations for Industry and the Life Sciences

CERN Document Server

Peiponen, Kai-Erik; Priezzhev, Alexander V

2009-01-01

Devoted to novel optical measurement techniques that are applied both in industry and life sciences, this book contributes a fresh perspective on the development of modern optical sensors. These sensors are often essential in detecting and controlling parameters that are important for both industrial and biomedical applications. The book provides easy access for beginners wishing to gain familiarity with the innovations of modern optics.

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane.

Science.gov (United States)

Stone, Matthew B; Shelby, Sarah A; Veatch, Sarah L

2017-06-14

Lipids and the membranes they form are fundamental building blocks of cellular life, and their geometry and chemical properties distinguish membranes from other cellular environments. Collective processes occurring within membranes strongly impact cellular behavior and biochemistry, and understanding these processes presents unique challenges due to the often complex and myriad interactions between membrane components. Super-resolution microscopy offers a significant gain in resolution over traditional optical microscopy, enabling the localization of individual molecules even in densely labeled samples and in cellular and tissue environments. These microscopy techniques have been used to examine the organization and dynamics of plasma membrane components, providing insight into the fundamental interactions that determine membrane functions. Here, we broadly introduce the structure and organization of the mammalian plasma membrane and review recent applications of super-resolution microscopy to the study of membranes. We then highlight some inherent challenges faced when using super-resolution microscopy to study membranes, and we discuss recent technical advancements that promise further improvements to super-resolution microscopy and its application to the plasma membrane.

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...

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

Advanced Microscopy of Microbial Cells

DEFF Research Database (Denmark)

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

2011-01-01

Growing awareness of heterogeneity in cells of microbial populations has emphasized the importance of advanced microscopy for visualization and understanding of the molecular mechanisms underlying cell-to-cell variation. In this review, we highlight some of the recent advances in confocal...... 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...

Description of a neural sheath tumor of the trigeminal nerve: immunohistochemical and electron microscopy study

OpenAIRE

Khademi, Bijan; Owji, Seied Mohammad; Khosh, Khadije Jamshidi; Mohammadianpanah, Mohammad; Gandomi, Behrooz

2006-01-01

CONTEXT: Malignant neural sheath tumors of the trigeminal nerve affecting the nasal cavity and the paranasal sinuses are extremely rare. With conventional optical microscopy, their identification is difficult, and it is necessary to confirm them by means of electron microscopy and immunohistochemical techniques. CASE REPORT: The patient was a 41-year-old woman with a ten-month progressive history of pain followed by painful edema in the left facial region, and with symptoms of bleeding, secre...

Electron microscopy of surfaces

International Nuclear Information System (INIS)

Venables, J.A.

1981-01-01

Electron beam techniques used to study clean surfaces and surface processes on a microscopic scale are reviewed. Recent experimental examples and possible future developments are discussed. Special emphasis is given to (i) transmission diffraction and microscopy techniques, including atomic imaging; (ii) Auger microscopy on bulk and thin film samples; (iii) secondary electron microscopy, especially low energy secondaries for work-function imaging and photoelectron imaging; and (iv) reflection electron microscopy and diffraction. (orig.)

Z-scan: A simple technique for determination of third-order optical nonlinearity

Energy Technology Data Exchange (ETDEWEB)

Singh, Vijender, E-mail: chahal-gju@rediffmail.com [Department of Applied Science, N.C. College of Engineering, Israna, Panipat-132107, Haryana (India); Aghamkar, Praveen, E-mail: p-aghamkar@yahoo.co.in [Department of Physics, Chaudhary Devi Lal University, Sirsa-125055, Haryana (India)

2015-08-28

Z-scan is a simple experimental technique to measure intensity dependent nonlinear susceptibilities of third-order nonlinear optical materials. This technique is used to measure the sign and magnitude of both real and imaginary part of the third order nonlinear susceptibility (χ{sup (3)}) of nonlinear optical materials. In this paper, we investigate third-order nonlinear optical properties of Ag-polymer composite film by using single beam z-scan technique with Q-switched, frequency doubled Nd: YAG laser (λ=532 nm) at 5 ns pulse. The values of nonlinear absorption coefficient (β), nonlinear refractive index (n{sub 2}) and third-order nonlinear optical susceptibility (χ{sup (3)}) of permethylazine were found to be 9.64 × 10{sup −7} cm/W, 8.55 × 10{sup −12} cm{sup 2}/W and 5.48 × 10{sup −10} esu, respectively.

A Review of Domain Modelling and Domain Imaging Techniques in Ferroelectric Crystals

Directory of Open Access Journals (Sweden)

John E. Huber

2011-02-01

Full Text Available The present paper reviews models of domain structure in ferroelectric crystals, thin films and bulk materials. Common crystal structures in ferroelectric materials are described and the theory of compatible domain patterns is introduced. Applications to multi-rank laminates are presented. Alternative models employing phase-field and related techniques are reviewed. The paper then presents methods of observing ferroelectric domain structure, including optical, polarized light, scanning electron microscopy, X-ray and neutron diffraction, atomic force microscopy and piezo-force microscopy. Use of more than one technique for unambiguous identification of the domain structure is also described.

Confocal filtering in cathodoluminescence microscopy of nanostructures

Science.gov (United States)

Narváez, Angela C.; Weppelman, I. Gerward C.; Moerland, Robert J.; Hoogenboom, Jacob P.; Kruit, Pieter

2014-06-01

Cathodoluminescence (CL) microscopy allows optical characterization of nanostructures at high spatial resolution. At the nanoscale, a main challenge of the technique is related to the background CL generated within the sample substrate. Here, we implement confocal detection of the CL signal to minimize the background contribution to the measurement. Nano-phosphors were used as point sources to evaluate the filtering capabilities of our confocal CL system, obtaining an axial intensity profile with 2.7 μm full width at half maximum for the central peak, in good correspondence with theoretical expectations. Considering the electron interaction volume, we found that the confocal filter becomes effective for electron energies above 20 keV, when using a 25 μm pinhole (0.86 Airy units). To illustrate our approach, we present confocal CL imaging of gold nanowires and triangular shaped plates deposited on an indium-tin oxide covered glass substrate, comparing the images with those obtained in standard unfiltered CL detection. The results show that confocal CL microscopy is a valuable tool for the investigation of nanostructures on highly cathodoluminescent substrates, widely used in biological and optical applications.

Confocal filtering in cathodoluminescence microscopy of nanostructures

Energy Technology Data Exchange (ETDEWEB)

Narváez, Angela C., E-mail: a.c.narvaez@tudelft.nl, E-mail: j.p.hoogenboom@tudelft.nl; Weppelman, I. Gerward C.; Moerland, Robert J.; Hoogenboom, Jacob P., E-mail: a.c.narvaez@tudelft.nl, E-mail: j.p.hoogenboom@tudelft.nl; Kruit, Pieter [Imaging Physics, Faculty of Applied Sciences, Delft University of Technology, Lorentzweg 1, 2628CJ Delft (Netherlands)

2014-06-23

Cathodoluminescence (CL) microscopy allows optical characterization of nanostructures at high spatial resolution. At the nanoscale, a main challenge of the technique is related to the background CL generated within the sample substrate. Here, we implement confocal detection of the CL signal to minimize the background contribution to the measurement. Nano-phosphors were used as point sources to evaluate the filtering capabilities of our confocal CL system, obtaining an axial intensity profile with 2.7 μm full width at half maximum for the central peak, in good correspondence with theoretical expectations. Considering the electron interaction volume, we found that the confocal filter becomes effective for electron energies above 20 keV, when using a 25 μm pinhole (0.86 Airy units). To illustrate our approach, we present confocal CL imaging of gold nanowires and triangular shaped plates deposited on an indium-tin oxide covered glass substrate, comparing the images with those obtained in standard unfiltered CL detection. The results show that confocal CL microscopy is a valuable tool for the investigation of nanostructures on highly cathodoluminescent substrates, widely used in biological and optical applications.

Fiber break location technique utilizing stimulated Brillouin scattering effects in optical fiber

International Nuclear Information System (INIS)

Bakar, A A A; Al-Mansoori, M H; Mahdi, M A; Mohd Azau, M A; Zainal Abidin, M S

2009-01-01

A new technique of fiber break detection system in optical communication networks is proposed and experimentally demonstrated in this paper. This technique is based-on continuous wave light source rather than pulsed source that is commonly deployed in existing techniques. The nonlinear effect of stimulated Brillouin scattering is manipulated to locate the fiber-break position in optical communication networks. This technique enables the utilization of a less-sensitive photodetector to detect the Brillouin Stokes line since its intensity increases with the fiber length in the detectable region. The fiber break location can be determined with accuracy of more than 98% for fiber length less than 50 km using this technique

Effect of complexing agent TEA: The structural, morphological, topographical and optical properties of FexSx nano thin films deposited by SILAR technique

International Nuclear Information System (INIS)

Manikandan, K.; Mani, P.; Surendra Dilip, C.; Valli, S.; Fermi Hilbert Inbaraj, P.; Joseph Prince, J.

2014-01-01

Iron sulfide thin films (Fe x S x ) (x = 0.05 M, 0.10 M, 0.15 M, 0.20 M and 0.25 M) were deposited by SILAR method from equimolar and equivolume aqueous solutions of ferrous nitrate and sodium sulfide with the addition of complexing agent TEA. The structural, morphological and optical characteristics of the films were derived from X-ray diffraction (XRD), scanning electron microscopy (SEM), atomic force microscopy (AFM) and UV–vis spectral techniques. The mixed characteristics (crystalline and amorphous) of the deposited films and the increasing crystalline qualities with the concentrations were understood from the XRD analysis. The grain sizes and roughness of the films were decreases with the increasing concentration and also at the higher concentration films are shown by the same images presence of hexagonal like crystallite structure. The influence of complexing agent TEA on the surface roughness and morphological properties are confirmed by the atomic force microscope (AFM) results. The effect of increasing substrate concentration on the absorption and transmission measurements and its impact on the optical band-gap energy were enumerated from the UV–vis analysis.

Printed organic smart devices characterized by nonlinear optical

DEFF Research Database (Denmark)

Pastorelli, Francesco; Accanto, Nicolo; Jørgensen, Mikkel

2017-01-01

In this study, we demonstrate that nonlinear optical microscopy is a promising technique to characterize organic printed electronics. Using ultrashort laser pulses we stimulate two-photon absorption in a roll coated polymer semiconductor and map the resulting two-photon induced photoluminescence...

High-definition optical coherence tomography

DEFF Research Database (Denmark)

Boone, Marc; Norrenberg, Sarah; Jemec, Gregor

2013-01-01

to those described for reflectance confocal microscopy but with the advantages not only to visualize individual cells up to a depth of 570 μm but also in both slice and en face mode. An adapted algorithmic method for pattern analysis of common inflammatory skin diseases could be proposed. This new......High-definition optical coherence tomography (HD-OCT) is a non-invasive technique for morphological investigation of tissue with cellular resolution filling the imaging gap between reflectance confocal microscopy and conventional optical coherence tomography. The aim of this study is first...... dermatitis. Additional studies to test the sensitivity and specificity of the proposed algorithm for pattern analysis are essential. The other categories of Ackerman's pattern recognition need to be evaluated. This study provides a set of morphological features generated by HD-OCT imaging very similar...

Shedding new light on lipid functions with CARS and SRS microscopy

Science.gov (United States)

Yu, Yong; Ramachandran, Prasanna V.; Wang, Meng C.

2014-01-01

Modern optical microscopy has granted biomedical scientists unprecedented access to the inner workings of a cell, and revolutionized our understanding of the molecular mechanisms underlying physiological and disease states. In spite of these advances, however, visualization of certain classes of molecules (e.g. lipids) at the sub-cellular level has remained elusive. Recently developed chemical imaging modalities – Coherent Anti-Stokes Raman Scattering (CARS) microscopy and Stimulated Raman Scattering (SRS) microscopy – have helped bridge this gap. By selectively imaging the vibration of a specific chemical group, these non-invasive techniques allow high-resolution imaging of individual molecules in vivo, and circumvent the need for potentially perturbative extrinsic labels. These tools have already been applied to the study of fat metabolism, helping uncover novel regulators of lipid storage. Here we review the underlying principle of CARS and SRS microscopy, and discuss the advantages and caveats of each technique. We also review recent applications of these tools in the study of lipids as well as other biomolecules, and conclude with a brief guide for interested researchers to build and use CARS/SRS systems for their own research. PMID:24576891

Investigation into local cell mechanics by atomic force microscopy mapping and optical tweezer vertical indentation.

Science.gov (United States)

Coceano, G; Yousafzai, M S; Ma, W; Ndoye, F; Venturelli, L; Hussain, I; Bonin, S; Niemela, J; Scoles, G; Cojoc, D; Ferrari, E

2016-02-12

Investigating the mechanical properties of cells could reveal a potential source of label-free markers of cancer progression, based on measurable viscoelastic parameters. The Young's modulus has proved to be the most thoroughly studied so far, however, even for the same cell type, the elastic modulus reported in different studies spans a wide range of values, mainly due to the application of different experimental conditions. This complicates the reliable use of elasticity for the mechanical phenotyping of cells. Here we combine two complementary techniques, atomic force microscopy (AFM) and optical tweezer microscopy (OTM), providing a comprehensive mechanical comparison of three human breast cell lines: normal myoepithelial (HBL-100), luminal breast cancer (MCF-7) and basal breast cancer (MDA-MB-231) cells. The elastic modulus was measured locally by AFM and OTM on single cells, using similar indentation approaches but different measurement parameters. Peak force tapping AFM was employed at nanonewton forces and high loading rates to draw a viscoelastic map of each cell and the results indicated that the region on top of the nucleus provided the most meaningful results. OTM was employed at those locations at piconewton forces and low loading rates, to measure the elastic modulus in a real elastic regime and rule out the contribution of viscous forces typical of AFM. When measured by either AFM or OTM, the cell lines' elasticity trend was similar for the aggressive MDA-MB-231 cells, which were found to be significantly softer than the other two cell types in both measurements. However, when comparing HBL-100 and MCF-7 cells, we found significant differences only when using OTM.

Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells

Science.gov (United States)

Hofemeier, Arne D.; Hachmeister, Henning; Pilger, Christian; Schürmann, Matthias; Greiner, Johannes F. W.; Nolte, Lena; Sudhoff, Holger; Kaltschmidt, Christian; Huser, Thomas; Kaltschmidt, Barbara

2016-05-01

Tissue engineering by stem cell differentiation is a novel treatment option for bone regeneration. Most approaches for the detection of osteogenic differentiation are invasive or destructive and not compatible with live cell analysis. Here, non-destructive and label-free approaches of Raman spectroscopy, coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) microscopy were used to detect and image osteogenic differentiation of human neural crest-derived inferior turbinate stem cells (ITSCs). Combined CARS and SHG microscopy was able to detect markers of osteogenesis within 14 days after osteogenic induction. This process increased during continued differentiation. Furthermore, Raman spectroscopy showed significant increases of the PO43- symmetric stretch vibrations at 959 cm-1 assigned to calcium hydroxyapatite between days 14 and 21. Additionally, CARS microscopy was able to image calcium hydroxyapatite deposits within 14 days following osteogenic induction, which was confirmed by Alizarin Red-Staining and RT- PCR. Taken together, the multimodal label-free analysis methods Raman spectroscopy, CARS and SHG microscopy can monitor osteogenic differentiation of adult human stem cells into osteoblasts with high sensitivity and spatial resolution in three dimensions. Our findings suggest a great potential of these optical detection methods for clinical applications including in vivo observation of bone tissue-implant-interfaces or disease diagnosis.

X-ray Tomographic Microscopy at TOMCAT

Energy Technology Data Exchange (ETDEWEB)

Marone, F; Hintermueller, C; McDonald, S; Abela, R; Mikuljan, G; Isenegger, A; Stampanoni, M, E-mail: federica.marone@psi.c [Swiss Light Source, Paul Scherrer Institut, 5232 Villigen (Switzerland)

2009-09-01

Synchrotron-based X-ray Tomographic Microscopy is a powerful technique for fast non-destructive, high resolution quantitative volumetric investigations on diverse samples. At the TOMCAT (TOmographic Microscopy and Coherent rAdiology experimenTs) beamline at the Swiss Light Source, synchrotron light is delivered by a 2.9 T superbend. The main optical component, a Double Crystal Multilayer Monochromator, covers an energy range between 8 and 45 keV. The standard TOMCAT detector offers field of views ranging from 0.75x0.75 mm{sup 2} up to 12.1x12.1 mm{sup 2} with a pixel size of 0.37 {mu}m and 5.92 {mu}m, respectively. In addition to routine measurements, which exploit the absorption contrast, the high coherence of the source also enables phase contrast tomography, implemented with two complementary techniques (Modified Transport of Intensity approach and Grating Interferometry). Typical acquisition times for a tomogram are in the order of few minutes, ensuring high throughput and allowing for semi-dynamical investigations. Raw data are automatically post-processed online and full reconstructed volumes are available shortly after a scan with minimal user intervention.

Imaging of Au nanoparticles deeply buried in polymer matrix by various atomic force microscopy techniques

International Nuclear Information System (INIS)

Kimura, Kuniko; Kobayashi, Kei; Matsushige, Kazumi; Yamada, Hirofumi

2013-01-01

Recently, some papers reported successful imaging of subsurface features using atomic force microscopy (AFM). Some theoretical studies have also been presented, however the imaging mechanisms are not fully understood yet. In the preceeding papers, imaging of deeply buried nanometer-scale features has been successful only if they were buried in a soft matrix. In this paper, subsurface features (Au nanoparticles) buried in a soft polymer matrix were visualized. To elucidate the imaging mechanisms, various AFM techniques; heterodyne force microscopy, ultrasonic atomic force microscopy (UAFM), 2nd-harmonic UAFM and force modulation microscopy (FMM) were employed. The particles buried under 960 nm from the surface were successfully visualized which has never been achieved. The results elucidated that it is important for subsurface imaging to choose a cantilever with a suitable stiffness range for a matrix. In case of using the most suitable cantilever, the nanoparticles were visualized using every technique shown above except for FMM. The experimental results suggest that the subsurface features buried in a soft matrix with a depth of at least 1 µm can affect the local viscoelasticity (mainly viscosity) detected as the variation of the amplitude and phase of the tip oscillation on the surface. This phenomenon presumably makes it possible to visualize such deeply buried nanometer-scale features in a soft matrix. - Highlights: • We visualized subsurface features buried in soft matrix, and investigated its imaging mechanism. • AFM techniques; UAFM, FMM, HFM and 2nd-harmonic UAFM were applied to elucidate the mechanism. • Au nanoparticles buried under 960 nm from surface were visualized, which has never been achieved. • Imaging at contact resonance using a cantilever of suitable stiffness is important. • Subsurface features in a soft matrix affect surface viscoelasticity, which are detected by AFM

An integrated electron and optical metallographic procedure for the identification of precipitate phases in type 316 stainless steel

International Nuclear Information System (INIS)

Slattery, G.F.; O'Riordan, P.; Lambert, M.E.; Green, S.M.

1981-01-01

A sequential and integrated metallographic procedure has been developed and successfully employed to differentiate between carbide, sigma, chi, Laves and ferrite phases which are commonly encountered in type 316 austenitic steel. The experimental techniques of optical and electron microscopy to identify these phases have been outlined and provide a rapid and convenient method of characterizing the microstructure of the steel. The techniques sequence involves selective metallographic etching, Nomarski interference microscopy, scanning electron microscopy, energy dispersive microanalysis, transmission electron microscopy and electron diffraction. (author)

Infrared spectroscopy of molecular submonolayers on surfaces by infrared scanning tunneling microscopy: tetramantane on Au111.

Science.gov (United States)

Pechenezhskiy, Ivan V; Hong, Xiaoping; Nguyen, Giang D; Dahl, Jeremy E P; Carlson, Robert M K; Wang, Feng; Crommie, Michael F

2013-09-20

We have developed a new scanning-tunneling-microscopy-based spectroscopy technique to characterize infrared (IR) absorption of submonolayers of molecules on conducting crystals. The technique employs a scanning tunneling microscope as a precise detector to measure the expansion of a molecule-decorated crystal that is irradiated by IR light from a tunable laser source. Using this technique, we obtain the IR absorption spectra of [121]tetramantane and [123]tetramantane on Au(111). Significant differences between the IR spectra for these two isomers show the power of this new technique to differentiate chemical structures even when single-molecule-resolved scanning tunneling microscopy (STM) images look quite similar. Furthermore, the new technique was found to yield significantly better spectral resolution than STM-based inelastic electron tunneling spectroscopy, and to allow determination of optical absorption cross sections. Compared to IR spectroscopy of bulk tetramantane powders, infrared scanning tunneling microscopy (IRSTM) spectra reveal narrower and blueshifted vibrational peaks for an ordered tetramantane adlayer. Differences between bulk and surface tetramantane vibrational spectra are explained via molecule-molecule interactions.

Optical Techniques for Millimeter-Wave Phased Array Communications Antennas

National Research Council Canada - National Science Library

Edge, Colin

1998-01-01

The scope of this program was to study the application of optical techniques to signal distribution and beamforming networks in phased array antennas for Army mobile tactical communications systems...

In vivo oral imaging with integrated portable photoacoustic microscopy and optical coherence tomography

Science.gov (United States)

Qin, Wei; Qi, Weizhi; Jin, Tian; Guo, Heng; Xi, Lei

2017-12-01

Oral diseases, especially oral cancers, are becoming serious health problems in humans. To image vasculatures and structures simultaneously in the human oral cavity which are tightly associated with various oral diseases, we develop a dual-modality portable optical resolution photoacoustic microscopy (ORPAM) and optical coherence tomography (OCT) system. This system utilizes a new rotary scanning mechanism and a compact design of the imaging head, making it portable and free of translation of the imaging interface or samples. Through the phantom experiments, both modalities yield high lateral resolutions of 8.1 μm (ORPAM) and 8.56 μm (OCT), respectively. The axial resolutions are measured to be 116.5 μm for ORPAM and 6.1 μm for OCT. In vivo imaging of a mouse ear was carried out to evaluate the performance of the system in biological tissues. In addition, in vivo oral imaging of a healthy human lip and monitoring recovery progress of a lip ulcer demonstrate the clinical potential of this system.

A digital transducer and digital microphone using an optical technique

Science.gov (United States)

Ghelmansarai, F. A.

1996-09-01

A transducer is devised to measure pressure, displacements or angles by optical means. This transducer delivers a digital output without relying on interferometry techniques or analogue-to-digital converters. This device is based on an optical scanner and an optical detector. An inter-digital photoconductive detector (IDPC) is employed that delivers a series of pulses, whose number depends on the scan length. A pre-objective scanning configuration is used that allows for the possibility of a flat image plane. The optical scanner provides scanning of IDPC and the generated scan length is proportional to the measurand.

Four-Dimensional Ultrafast Electron Microscopy: Insights into an Emerging Technique

KAUST Repository

Adhikari, Aniruddha

2016-12-15

Four-dimensional ultrafast electron microscopy (4D-UEM) is a novel analytical technique that aims to fulfill the long-held dream of researchers to investigate materials at extremely short spatial and temporal resolutions by integrating the excellent spatial resolution of electron microscopes with the temporal resolution of ultrafast femtosecond laser-based spectroscopy. The ingenious use of pulsed photoelectrons to probe surfaces and volumes of materials enables time-resolved snapshots of the dynamics to be captured in a way hitherto impossible by other conventional techniques. The flexibility of 4D-UEM lies in the fact that it can be used in both the scanning (S-UEM) and transmission (UEM) modes depending upon the type of electron microscope involved. While UEM can be employed to monitor elementary structural changes and phase transitions in samples using real-space mapping, diffraction, electron energy-loss spectroscopy, and tomography, S-UEM is well suited to map ultrafast dynamical events on materials surfaces in space and time. This review provides an overview of the unique features that distinguish these techniques and also illustrates the applications of both S-UEM and UEM to a multitude of problems relevant to materials science and chemistry.

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.

Transfer doping of single isolated nanodiamonds, studied by scanning probe microscopy techniques.

Science.gov (United States)

Bolker, Asaf; Saguy, Cecile; Kalish, Rafi

2014-09-26

The transfer doping of diamond surfaces has been applied in various novel two-dimensional electronic devices. Its extension to nanodiamonds (ND) is essential for ND-based applications in many fields. In particular, understanding the influence of the crystallite size on transfer doping is desirable. Here, we report the results of a detailed study of the electronic energetic band structure of single, isolated transfer-doped nanodiamonds with nanometric resolution using a combination of scanning tunneling spectroscopy and Kelvin force microscopy measurements. The results show how the band gap, the valence band maximum, the electron affinity and the work function all depend on the ND's size and nanoparticle surface properties. The present analysis, which combines information from both scanning tunneling spectroscopy and Kelvin force microscopy, should be applicable to any nanoparticle or surface that can be measured with scanning probe techniques.

Transfer doping of single isolated nanodiamonds, studied by scanning probe microscopy techniques

Science.gov (United States)

Bolker, Asaf; Saguy, Cecile; Kalish, Rafi

2014-09-01

The transfer doping of diamond surfaces has been applied in various novel two-dimensional electronic devices. Its extension to nanodiamonds (ND) is essential for ND-based applications in many fields. In particular, understanding the influence of the crystallite size on transfer doping is desirable. Here, we report the results of a detailed study of the electronic energetic band structure of single, isolated transfer-doped nanodiamonds with nanometric resolution using a combination of scanning tunneling spectroscopy and Kelvin force microscopy measurements. The results show how the band gap, the valence band maximum, the electron affinity and the work function all depend on the ND’s size and nanoparticle surface properties. The present analysis, which combines information from both scanning tunneling spectroscopy and Kelvin force microscopy, should be applicable to any nanoparticle or surface that can be measured with scanning probe techniques.

Transfer doping of single isolated nanodiamonds, studied by scanning probe microscopy techniques

International Nuclear Information System (INIS)

Bolker, Asaf; Kalish, Rafi; Saguy, Cecile

2014-01-01

The transfer doping of diamond surfaces has been applied in various novel two-dimensional electronic devices. Its extension to nanodiamonds (ND) is essential for ND-based applications in many fields. In particular, understanding the influence of the crystallite size on transfer doping is desirable. Here, we report the results of a detailed study of the electronic energetic band structure of single, isolated transfer-doped nanodiamonds with nanometric resolution using a combination of scanning tunneling spectroscopy and Kelvin force microscopy measurements. The results show how the band gap, the valence band maximum, the electron affinity and the work function all depend on the ND’s size and nanoparticle surface properties. The present analysis, which combines information from both scanning tunneling spectroscopy and Kelvin force microscopy, should be applicable to any nanoparticle or surface that can be measured with scanning probe techniques. (paper)

Multimodal microscopy of immune cells and melanoma for longitudinal studies

Science.gov (United States)

Entenberg, David; Aranda, Iana; Li, Yongbiao; Toledo-Crow, Ricardo; Schaer, David; Li, Yanyun

2006-02-01

Intravital microscopy of cancer is a well established tool that provides direct visualization of the tumor cycle. It traditionally involves one of several strategies: invasive subcutaneous (SC) implantation of tumors followed by surgical opening of skin flaps for imaging, techniques utilizing skin fold chambers and implanted optical windows or intradermal injections under 200μm from the skin surface. All of these techniques allow the use of fluorescent proteins as markers for biologically significant constituents. However, observation methods utilizing skin-flaps, skin-fold chambers and optical windows are invasive and tend to alter the immune environment of the tissue and/or limit the duration of studies that can be performed. If implanted correctly, intradermally injected tumors can be minimally invasive, will not require biopsies or surgical intervention to observe and are accessible for direct transdermal imaging with a number of in vivo modalities. We present our work in the development of a small animal intravital microscopy workstation that allows the acquisition of different contrast imaging modalities: reflectance confocal, wide field epifluorescence, multiphoton and second harmonic generation (SHG). The images are acquired pair-wise simultaneously and sequentially in time. The aim of our instrumentation is to gather all information generated by the single probing beam via the reflected or back-scattered signal, SHG signal and various fluorescence signals. Additionally, we also present our development of a microscopic tissue navigation technique to mark, label and track sites of interest. This technique enables us to revisit sites periodically and record, with different imaging contrasts, their biological changes over time.

Actively stabilized optical fiber interferometry technique for online/in-process surface measurement

International Nuclear Information System (INIS)

Wang Kaiwei; Martin, Haydn; Jiang Xiangqian

2008-01-01

In this paper, we report the recent progress in optical-beam scanning fiber interferometry for potential online nanoscale surface measurement based on the previous research. It attempts to generate a robust and miniature measurement device for future development into a multiprobe array measurement system. In this research, both fiber-optic-interferometry and the wavelength-division-multiplexing techniques have been used, so that the optical probe and the optical interferometer are well spaced and fast surface scanning can be carried out, allowing flexibility for online measurement. In addition, this system provides a self-reference signal to stabilize the optical detection with high common-mode noise suppression by adopting an active phase tracking and stabilization technique. Low-frequency noise was significantly reduced compared with unstabilized result. The measurement of a sample surface shows an attained repeatability of 3.3 nm

The thin layer technique and its application to electron microscopy

International Nuclear Information System (INIS)

Ranc, G.

1957-10-01

This work deals with the technique of thin layers obtained by evaporation under vacuum, in the thickness range extending from a few monoatomic layers to several hundred angstroms. The great theoretical and practical interest of these layers has, it is well known, given rise to many investigations from Faraday onwards. Within the necessarily restricted limits of this study, we shall approach the problem more particularly from the point of view of: - their production; - their use in electron microscopy. A critical appraisal is made, in the light of present-day knowledge, based on our personal experience and on an extensive bibliography which we have collected on the subject. (author) [fr

Effect of iron doping on structural and optical properties of TiO2 thin film by sol–gel routed spin coating technique

Directory of Open Access Journals (Sweden)

Stephen Lourduraj

2017-08-01

Full Text Available Thin films of iron (Fe-doped titanium dioxide (Fe:TiO2 were prepared by sol–gel spin coating technique and further calcined at 450∘C. The structural and optical properties of Fe-doped TiO2 thin films were investigated by X-ray diffraction (XRD, scanning electron microscopy (SEM, ultraviolet–visible spectroscopy (UV–vis and atomic force microscopic (AFM techniques. The XRD results confirm the nanostructured TiO2 thin films having crystalline nature with anatase phase. The characterization results show that the calcined thin films having high crystallinity and the effect of iron substitution lead to decreased crystallinity. The SEM investigations of Fe-doped TiO2 films also gave evidence that the films were continuous spherical shaped particles with a nanometric range of grain size and film was porous in nature. AFM analysis establishes that the uniformity of the TiO2 thin film with average roughness values. The optical measurements show that the films having high transparency in the visible region and the optical band gap energy of Fe-doped TiO2 film with iron (Fe decrease with increase in iron content. These important requirements for the Fe:TiO2 films are to be used as window layers in solar cells.

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)

All-optical photoacoustic microscopy using a MEMS scanning mirror

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Chen, Sung-Liang; Xie, Zhixing; Ling, Tao; Wei, Xunbin; Guo, L. Jay; Wang, Xueding

2013-03-01

It has been studied that a potential marker to obtain prognostic information about bladder cancer is tumor neoangiogenesis, which can be quantified by morphometric characteristics such as microvascular density. Photoacoustic microscopy (PAM) can render sensitive three-dimensional (3D) mapping of microvasculature, providing promise to evaluate the neoangiogenesis that is closely related to the diagnosis of bladder cancer. To ensure good image quality, it is desired to acquire bladder PAM images from its inside via the urethra, like conventional cystoscope. Previously, we demonstrated all-optical PAM systems using polymer microring resonators to detect photoacoustic signals and galvanometer mirrors for laser scanning. In this work, we build a miniature PAM system using a microelectromechanical systems (MEMS) scanning mirror, demonstrating a prototype of an endoscopic PAM head capable of high imaging quality of the bladder. The system has high resolutions of 17.5 μm in lateral direction and 19 μm in the axial direction at a distance of 5.4 mm. Images of printed grids and the 3D structure of microvasculature in animal bladders ex vivo by the system are demonstrated.

Hierarchical super-structure identified by polarized light microscopy, electron microscopy and nanoindentation: Implications for the limits of biological control over the growth mode of abalone sea shells

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Schneider Andreas S

2012-09-01

Full Text Available Abstract Background Mollusc shells are commonly investigated using high-resolution imaging techniques based on cryo-fixation. Less detailed information is available regarding the light-optical properties. Sea shells of Haliotis pulcherina were embedded for polishing in defined orientations in order to investigate the interface between prismatic calcite and nacreous aragonite by standard materialographic methods. A polished thin section of the interface was prepared with a defined thickness of 60 μm for quantitative birefringence analysis using polarized light and LC-PolScope microscopy. Scanning electron microscopy images were obtained for comparison. In order to study structural-mechanical relationships, nanoindentation experiments were performed. Results Incident light microscopy revealed a super-structure in semi-transparent regions of the polished cross-section under a defined angle. This super-structure is not visible in transmitted birefringence analysis due to the blurred polarization of small nacre platelets and numerous organic interfaces. The relative orientation and homogeneity of calcite prisms was directly identified, some of them with their optical axes exactly normal to the imaging plane. Co-oriented "prism colonies" were identified by polarized light analyses. The nacreous super-structure was also visualized by secondary electron imaging under defined angles. The domains of the super-structure were interpreted to consist of crystallographically aligned platelet stacks. Nanoindentation experiments showed that mechanical properties changed with the same periodicity as the domain size. Conclusions In this study, we have demonstrated that insights into the growth mechanisms of nacre can be obtained by conventional light-optical methods. For example, we observed super-structures formed by co-oriented nacre platelets as previously identified using X-ray Photo-electron Emission Microscopy (X-PEEM [Gilbert et al., Journal of the

Applications Of Measurement Techniques To Develop Small-Diameter, Undersea Fiber Optic Cables

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Kamikawa, Neil T.; Nakagawa, Arthur T.

1984-12-01

Attenuation, strain, and optical time domain reflectometer (OTDR) measurement techniques were applied successfully in the development of a minimum-diameter, electro-optic sea floor cable. Temperature and pressure models for excess attenuation in polymer coated, graded-index fibers were investigated analytically and experimentally using these techniques in the laboratory. The results were used to select a suitable fiber for the cable. Measurements also were performed on these cables during predeployment and sea-trial testing to verify laboratory results. Application of the measurement techniques and results are summarized in this paper.

Optimizing Nanoscale Quantitative Optical Imaging of Subfield Scattering Targets

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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

Optical Polarizationin Biomedical Applications

CERN Document Server

Tuchin, Valery V; Zimnyakov, Dmitry A

2006-01-01

Optical Polarization in Biomedical Applications introduces key developments in optical polarization methods for quantitative studies of tissues, while presenting the theory of polarization transfer in a random medium as a basis for the quantitative description of polarized light interaction with tissues. This theory uses the modified transfer equation for Stokes parameters and predicts the polarization structure of multiple scattered optical fields. The backscattering polarization matrices (Jones matrix and Mueller matrix) important for noninvasive medical diagnostic are introduced. The text also describes a number of diagnostic techniques such as CW polarization imaging and spectroscopy, polarization microscopy and cytometry. As a new tool for medical diagnosis, optical coherent polarization tomography is analyzed. The monograph also covers a range of biomedical applications, among them cataract and glaucoma diagnostics, glucose sensing, and the detection of bacteria.

Enhancement of fluorescence confocal scanning microscopy lateral resolution by use of structured illumination

International Nuclear Information System (INIS)

Kim, Taejoong; Gweon, DaeGab; Lee, Jun-Hee

2009-01-01

Confocal microscopy is an optical imaging technique used to reconstruct three-dimensional images without physical sectioning. As with other optical microscopes, the lateral resolution of the confocal microscope cannot surpass the diffraction limit. This paper presents a novel imaging system, structured illumination confocal scanning microscopy (SICSM), that uses structured illumination to improve the lateral resolution of the confocal microscope. The SICSM can easily be implemented by introducing a structured illumination generating optics to conventional line-scanning fluorescence confocal microscopy. In this paper, we report our analysis of the lateral and axial resolutions of the SICSM by use of mathematical imaging theory. Numerical simulation results show that the lateral resolution of the SICSM is 1.43-fold better than that of the confocal microscope. In the axial direction, however, the resolution of the SICSM is ∼15% poorer than that of the confocal microscope. This deterioration arises because of a decrease in the axial cut-off frequency caused by the process of generating structured illumination. We propose the use of imaging conditions under which a compromise between the axial and lateral resolutions is chosen. Finally, we show simulated images of diversely shaped test objects to demonstrate the lateral and axial resolution performance of the SICSM

Electronically tunable femtosecond all-fiber optical parametric oscillator for multi-photon microscopy

Science.gov (United States)

Hellwig, Tim; Brinkmann, Maximilian; Fallnich, Carsten

2018-02-01

We present a femtosecond fiber-based optical parametric oscillator (FOPO) for multiphoton microscopy with wavelength tuning by electronic repetition rate tuning in combination with a dispersive filter in the FOPO cavity. The all-spliced, all-fiber FOPO cavity is based on polarization-maintaining fibers and a broadband output coupler, allowing to get access to the resonant signal pulses as well as the idler pulses simultaneously. The system was pumped by a gain-switched fiber-coupled laser diode emitting pulses at a central wavelength of 1030 nm and an electronically tunable repetition frequency of about 2 MHz. The pump pulses were amplified in an Ytterbium fiber amplifier system with a pulse duration after amplification of 13 ps. Tuning of the idler (1140 nm - 1300 nm) and signal wavelengths (850 nm - 940 nm) was achieved by changing the repetition frequency of the pump laser by about 4 kHz. The generated signal pulses reached a pulse energy of up to 9.2 nJ at 920 nm and were spectrally broadened to about 6 nm in the FOPO by a combination of self-phase and cross-phase modulation. We showed external compression of the idler pulses at 920 nm to about 430 fs and appleid them to two-photon excitation microscopy with green fluorescent dyes. The presented system constitutes an important step towards a fully fiber-integrated all-electronically tunable and, thereby, programmable light source and already embodies a versatile and flexible light source for applications, e.g., for smart microscopy.

Microscopy of thin polymer blend films of polystyrene and poly-n-butyl-methacrylate

International Nuclear Information System (INIS)

Schmitt, T.; Guttmann, P.; Schmahl, G.; Schmidt, O.; Schoenhense, G.; Mueller-Buschbaum, P.; Stamm, M.

2000-01-01

The structure of thin polymer blend films of polystyrene (PS) and poly-n-butyl-methacrylate (PnBMA) was examined with Transmission X-ray Microscopy (TXM), Scanning Force Microscopy (SFM), X-Ray Photoemission Electron Microscopy (X-PEEM) and Optical Microscopy (OM). Thin films were prepared by spin casting of a toluene solution of the polymer mixture onto silicon wafers retaining the native oxide. Depending on blend composition and annealing conditions smooth films with and without holes or films with well pronounced surface features (ribbons or islands) were produced. By TXM measurements a high lateral resolution study of the as cast and the annealed polymer blend samples was performed. The contrast in TXM is due to different absorption of x-radiation of the used polymers and due to variation in thickness. With X-PEEM the lateral distribution of the two polymers near the surface was mapped by employing the characteristic Near Edge X-ray Absorption Fine Structure (NEXAFS) spectra of the polymers. The TXM technique is a microscopic method integrating over the total film thickness, whereas the X-PEEM technique is a highly surface sensitive method. TXM and X-PEEM are therefore complementary methods which provide important information on the structure of thin polymer blend films additional to the standard techniques SFM and OM

Liquidus temperature and optical properties measurement by containerless techniques

Science.gov (United States)

Anderson, Collin D.

1993-01-01

Reactive alloy liquidus temperatures measured by conventional, contained techniques are often in error due to reactions with containers and gaseous impurities. This paper describes a new liquidus temperature measurement technique that avoids these problems by employing containerless processing. This technique relies on precise and accurate noncontact temperature measurements (NCTM), which are made possible by spectral emissivity values. The spectral emissivities, epsilon(sub lambda), are measured along with the optical properties (real, n, and imaginary, k, components of the index of refraction) using polarimetric techniques on electromagnetically levitated specimens. Results from work done at Vanderbilt University and Intersonics on the Ti-Al system are presented to demonstrate the above techniques.

Quantitative high dynamic range beam profiling for fluorescence microscopy

International Nuclear Information System (INIS)

Mitchell, T. J.; Saunter, C. D.; O’Nions, W.; Girkin, J. M.; Love, G. D.

2014-01-01

Modern developmental biology relies on optically sectioning fluorescence microscope techniques to produce non-destructive in vivo images of developing specimens at high resolution in three dimensions. As optimal performance of these techniques is reliant on the three-dimensional (3D) intensity profile of the illumination employed, the ability to directly record and analyze these profiles is of great use to the fluorescence microscopist or instrument builder. Though excitation beam profiles can be measured indirectly using a sample of fluorescent beads and recording the emission along the microscope detection path, we demonstrate an alternative approach where a miniature camera sensor is used directly within the illumination beam. Measurements taken using our approach are solely concerned with the illumination optics as the detection optics are not involved. We present a miniature beam profiling device and high dynamic range flux reconstruction algorithm that together are capable of accurately reproducing quantitative 3D flux maps over a large focal volume. Performance of this beam profiling system is verified within an optical test bench and demonstrated for fluorescence microscopy by profiling the low NA illumination beam of a single plane illumination microscope. The generality and success of this approach showcases a widely flexible beam amplitude diagnostic tool for use within the life sciences

DotLens smartphone microscopy for biological and biomedical applications (Conference Presentation)

Science.gov (United States)

Sung, Yu-Lung; Zhao, Fusheng; Shih, Wei-Chuan

2017-02-01

Recent advances in inkjet-printed optics have created a new class of lens fabrication technique. Lenses with a tunable geometry, magnification, and focal length can be fabricated by dispensing controlled amounts of liquid polymer onto a heated surface. This fabrication technique is highly cost-effective, and can achieve optically smooth surface finish. Dubbed DotLens, a single of which weighs less than 50 mg and occupies a volume less than 50 μL. DotLens can be attached onto any smartphone camera akin to a contact lens, and enable smartphones to obtain image resolution as fine as 1 µm. The surface curvature modifies the optical path of light to the image sensor, and enables the camera to focus as close as 2 mm. This enables microscopic imaging on a smartphone without any additional attachments, and has shown great potential in mobile point-of-care diagnostic systems, particularly for histology of tissue sections and cytology of blood cells. DotLens Smartphone Microscopy represents an innovative approach fundamentally different from other smartphone microscopes. In this paper, we describe the application and performance of DotLens smartphone microscopy in biological and biomedical research. In particular, we show recent results from images collected from pathology tissue slides with cancer features. In addition, we show performance in cytological analysis of blood smear. This tool has empowered Citizen Science investigators to collect microscopic images from various interesting objects.

Advances in Retinal Optical Imaging

Directory of Open Access Journals (Sweden)

Yanxiu Li

2018-04-01

Full Text Available Retinal imaging has undergone a revolution in the past 50 years to allow for better understanding of the eye in health and disease. Significant improvements have occurred both in hardware such as lasers and optics in addition to software image analysis. Optical imaging modalities include optical coherence tomography (OCT, OCT angiography (OCTA, photoacoustic microscopy (PAM, scanning laser ophthalmoscopy (SLO, adaptive optics (AO, fundus autofluorescence (FAF, and molecular imaging (MI. These imaging modalities have enabled improved visualization of retinal pathophysiology and have had a substantial impact on basic and translational medical research. These improvements in technology have translated into early disease detection, more accurate diagnosis, and improved management of numerous chorioretinal diseases. This article summarizes recent advances and applications of retinal optical imaging techniques, discusses current clinical challenges, and predicts future directions in retinal optical imaging.

Atomic Force Microscopy Techniques for Nanomechanical Characterization: A Polymeric Case Study

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Reggente, Melania; Rossi, Marco; Angeloni, Livia; Tamburri, Emanuela; Lucci, Massimiliano; Davoli, Ivan; Terranova, Maria Letizia; Passeri, Daniele

2015-04-01

Atomic force microscopy (AFM) is a versatile tool to perform mechanical characterization of surface samples at the nanoscale. In this work, we review two of such methods, namely contact resonance AFM (CR-AFM) and torsional harmonics AFM (TH-AFM). First, such techniques are illustrated and their applicability on materials with elastic moduli in different ranges are discussed, together with their main advantages and limitations. Then, a case study is presented in which we report the mechanical characterization using both CR-AFM and TH-AFM of polyaniline and polyaniniline doped with nanodiamond particles tablets prepared by a pressing process. We determined the indentation modulus values of their surfaces, which were found in fairly good agreement, thus demonstrating the accuracy of the techniques. Finally, the determined surface elastic moduli have been compared with the bulk ones measured through standard indentation testing.

Fundamentals and techniques of nonimaging optics, volume 2

Science.gov (United States)

Winston, R.; Ogallagher, J. J.

1983-06-01

The formalism describing the operation is being defined, nonimaging concentrators and are being designed, and experiments and analytical studies are being conducted to evaluate the performance of new designs. In addition, components and materials particularly suited for use with these techniques were studied, and applications with an emphasis on solar energy concentration were explored. The motivation for the basic principles of nonimaging optics as they developed before 1978 are surveyed. The present status of the subdiscipline of nonimaging optics is summarized and an overview of the potential for future developments which is just beginning to emerge is presented.

Characterization of patinas by means of microscopic techniques

International Nuclear Information System (INIS)

Vazquez-Calvo, C.; Alvarez de Buergo, M.; Fort, R.; Varas, M.J.

2007-01-01

Many stone-made historic buildings have a yellowish layer called 'patina' on their external surface. In some cases, it is due to the natural ageing of the stone caused by chemical-physical reactions between the surface of the stone and the environment, and in other cases it is the result of biological activity. The origin of these patinas can be also be due to ancient protective treatments. The use of organic additives, such as protein-based compounds, in lime or gypsum-based patinas is a traditional technique, which has been used in past centuries for the conservation and protection of stone materials. The thinness of the patinas ensures that microscopic techniques are irreplaceable for their analysis. Optical Microscopy, Fluorescence Microscopy, Scanning Electron Microscopy together with an Energy Dispersive X-ray Spectrometer, and Electron Microprobe are the microscopic techniques used for the characterization of these coverings, providing very useful information on their composition, texture and structure