Optical fabrication and testing; Proceedings of the Meeting, Singapore, Oct. 22-27, 1990

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lorenzen, M.; Campbell, D.R.; Johnson, C.W.

1991-01-01

Various papers on optical fabrication and testing are presented. Individual topics addressed include: interferometry with laser diodes, new methods for economic production of prisms and lenses, interferometer accuracy and precision, optical testing with wavelength scanning interferometer, digital Talbot interferometer, high-sensitivity interferometric technique for strain measurements, absolute interferometric testing of spherical surfaces, contouring using gratings created on an LCD panel, three-dimensional inspection using laser-based dynamic fringe projection, noncontact optical microtopography, laser scan microscope and infrared laser scan microscope, photon scanning tunneling microscopy. Also discussed are: combination-matching problems in the layout design of minilaser rangefinder, design and testing of a cube-corner arraymore » for laser ranging, mode and far-field pattern of diode laser-phased arrays, new glasses for optics and optoelectronics, optical properties of Li-doped ZnO films, application and machining of Zerodur for optical purposes, finish machining of optical components in mass production.« less

Solid state optical microscope

DOEpatents

Young, I.T.

1983-08-09

A solid state optical microscope wherein wide-field and high-resolution images of an object are produced at a rapid rate by utilizing conventional optics with a charge-coupled photodiode array. A galvanometer scanning mirror, for scanning in one of two orthogonal directions is provided, while the charge-coupled photodiode array scans in the other orthogonal direction. Illumination light from the object is incident upon the photodiodes, creating packets of electrons (signals) which are representative of the illuminated object. The signals are then processed, stored in a memory, and finally displayed as a video signal. 2 figs.

Solid state optical microscope

DOEpatents

Young, Ian T.

1983-01-01

A solid state optical microscope wherein wide-field and high-resolution images of an object are produced at a rapid rate by utilizing conventional optics with a charge-coupled photodiode array. A galvanometer scanning mirror, for scanning in one of two orthogonal directions is provided, while the charge-coupled photodiode array scans in the other orthogonal direction. Illumination light from the object is incident upon the photodiodes, creating packets of electrons (signals) which are representative of the illuminated object. The signals are then processed, stored in a memory, and finally displayed as a video signal.

Atmospheric scanning electron microscope for correlative microscopy.

PubMed

Morrison, Ian E G; Dennison, Clare L; Nishiyama, Hidetoshi; Suga, Mitsuo; Sato, Chikara; Yarwood, Andrew; O'Toole, Peter J

2012-01-01

The JEOL ClairScope is the first truly correlative scanning electron and optical microscope. An inverted scanning electron microscope (SEM) column allows electron images of wet samples to be obtained in ambient conditions in a biological culture dish, via a silicon nitride film window in the base. A standard inverted optical microscope positioned above the dish holder can be used to take reflected light and epifluorescence images of the same sample, under atmospheric conditions that permit biochemical modifications. For SEM, the open dish allows successive staining operations to be performed without moving the holder. The standard optical color camera used for fluorescence imaging can be exchanged for a high-sensitivity monochrome camera to detect low-intensity fluorescence signals, and also cathodoluminescence emission from nanophosphor particles. If these particles are applied to the sample at a suitable density, they can greatly assist the task of perfecting the correlation between the optical and electron images. Copyright © 2012 Elsevier Inc. All rights reserved.

Evaluation of a completely robotized neurosurgical operating microscope.

PubMed

Kantelhardt, Sven R; Finke, Markus; Schweikard, Achim; Giese, Alf

2013-01-01

Operating microscopes are essential for most neurosurgical procedures. Modern robot-assisted controls offer new possibilities, combining the advantages of conventional and automated systems. We evaluated the prototype of a completely robotized operating microscope with an integrated optical coherence tomography module. A standard operating microscope was fitted with motors and control instruments, with the manual control mode and balance preserved. In the robot mode, the microscope was steered by a remote control that could be fixed to a surgical instrument. External encoders and accelerometers tracked microscope movements. The microscope was additionally fitted with an optical coherence tomography-scanning module. The robotized microscope was tested on model systems. It could be freely positioned, without forcing the surgeon to take the hands from the instruments or avert the eyes from the oculars. Positioning error was about 1 mm, and vibration faded in 1 second. Tracking of microscope movements, combined with an autofocus function, allowed determination of the focus position within the 3-dimensional space. This constituted a second loop of navigation independent from conventional infrared reflector-based techniques. In the robot mode, automated optical coherence tomography scanning of large surface areas was feasible. The prototype of a robotized optical coherence tomography-integrated operating microscope combines the advantages of a conventional manually controlled operating microscope with a remote-controlled positioning aid and a self-navigating microscope system that performs automated positioning tasks such as surface scans. This demonstrates that, in the future, operating microscopes may be used to acquire intraoperative spatial data, volume changes, and structural data of brain or brain tumor tissue.

Analytical model of the optical vortex microscope.

PubMed

Płocinniczak, Łukasz; Popiołek-Masajada, Agnieszka; Masajada, Jan; Szatkowski, Mateusz

2016-04-20

This paper presents an analytical model of the optical vortex scanning microscope. In this microscope the Gaussian beam with an embedded optical vortex is focused into the sample plane. Additionally, the optical vortex can be moved inside the beam, which allows fine scanning of the sample. We provide an analytical solution of the whole path of the beam in the system (within paraxial approximation)-from the vortex lens to the observation plane situated on the CCD camera. The calculations are performed step by step from one optical element to the next. We show that at each step, the expression for light complex amplitude has the same form with only four coefficients modified. We also derive a simple expression for the vortex trajectory of small vortex displacements.

Structured illumination 3D microscopy using adaptive lenses and multimode fibers

NASA Astrophysics Data System (ADS)

Czarske, Jürgen; Philipp, Katrin; Koukourakis, Nektarios

2017-06-01

Microscopic techniques with high spatial and temporal resolution are required for in vivo studying biological cells and tissues. Adaptive lenses exhibit strong potential for fast motion-free axial scanning. However, they also lead to a degradation of the achievable resolution because of aberrations. This hurdle can be overcome by digital optical technologies. We present a novel High-and-Low-frequency (HiLo) 3D-microscope using structured illumination and an adaptive lens. Uniform illumination is used to obtain optical sectioning for the high-frequency (Hi) components of the image, and nonuniform illumination is needed to obtain optical sectioning for the low-frequency (Lo) components of the image. Nonuniform illumination is provided by a multimode fiber. It ensures robustness against optical aberrations of the adaptive lens. The depth-of-field of our microscope can be adjusted a-posteriori by computational optics. It enables to create flexible scans, which compensate for irregular axial measurement positions. The adaptive HiLo 3D-microscope provides an axial scanning range of 1 mm with an axial resolution of about 4 microns and sub-micron lateral resolution over the full scanning range. In result, volumetric measurements with high temporal and spatial resolution are provided. Demonstration measurements of zebrafish embryos with reporter gene-driven fluorescence in the thyroid gland are presented.

Atmospheric scanning electron microscope observes cells and tissues in open medium through silicon nitride film.

PubMed

Nishiyama, Hidetoshi; Suga, Mitsuo; Ogura, Toshihiko; Maruyama, Yuusuke; Koizumi, Mitsuru; Mio, Kazuhiro; Kitamura, Shinichi; Sato, Chikara

2010-03-01

Direct observation of subcellular structures and their characterization is essential for understanding their physiological functions. To observe them in open environment, we have developed an inverted scanning electron microscope with a detachable, open-culture dish, capable of 8 nm resolution, and combined with a fluorescence microscope quasi-simultaneously observing the same area from the top. For scanning electron microscopy from the bottom, a silicon nitride film window in the base of the dish maintains a vacuum between electron gun and open sample dish while allowing electrons to pass through. Electrons are backscattered from the sample and captured by a detector under the dish. Cells cultured on the open dish can be externally manipulated under optical microscopy, fixed, and observed using scanning electron microscopy. Once fine structures have been revealed by scanning electron microscopy, their component proteins may be identified by comparison with separately prepared fluorescence-labeled optical microscopic images of the candidate proteins, with their heavy-metal-labeled or stained ASEM images. Furthermore, cell nuclei in a tissue block stained with platinum-blue were successfully observed without thin-sectioning, which suggests the applicability of this inverted scanning electron microscope to cancer diagnosis. This microscope visualizes mesoscopic-scale structures, and is also applicable to non-bioscience fields including polymer chemistry. (c) 2010 Elsevier Inc. All rights reserved.

Hard X-Ray Scanning Microscope with Multilayer Laue Lens Nanofocusing Optics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nazaretski, Evgeny

Evgeny Nazaretski, a physicist at Brookhaven Lab’s National Synchrotron Light Source II, spearheaded the development of a one-of-a-kind x-ray microscope with novel nanofocusing optics called multilayer Laue lenses.

A compact CCD-monitored atomic force microscope with optical vision and improved performances.

PubMed

Mingyue, Liu; Haijun, Zhang; Dongxian, Zhang

2013-09-01

A novel CCD-monitored atomic force microscope (AFM) with optical vision and improved performances has been developed. Compact optical paths are specifically devised for both tip-sample microscopic monitoring and cantilever's deflection detecting with minimized volume and optimal light-amplifying ratio. The ingeniously designed AFM probe with such optical paths enables quick and safe tip-sample approaching, convenient and effective tip-sample positioning, and high quality image scanning. An image stitching method is also developed to build a wider-range AFM image under monitoring. Experiments show that this AFM system can offer real-time optical vision for tip-sample monitoring with wide visual field and/or high lateral optical resolution by simply switching the objective; meanwhile, it has the elegant performances of nanometer resolution, high stability, and high scan speed. Furthermore, it is capable of conducting wider-range image measurement while keeping nanometer resolution. Copyright © 2013 Wiley Periodicals, Inc.

Acoustical nanometre-scale vibrations of live cells detected by a near-field optical setup

NASA Astrophysics Data System (ADS)

Piga, Rosaria; Micheletto, Ruggero; Kawakami, Yoichi

2007-04-01

The Scanning Near-field Optical Microscope (SNOM) is able to detect tiny vertical movement on the cell membrane in the range of only 1 nanometer or less, about 3 orders of magnitude better than conventional optical microscopes. Here we show intriguing data of cell membrane nanometer-scale dynamics associated to different phenomena of the cell’s The Scanning Near-field Optical Microscope (SNOM) is able to detect tiny vertical movement on the cell membrane in the range of only 1 nanometer or less, about 3 orders of magnitude better than conventional optical microscopes. Here we show intriguing data of cell membrane nanometer-scale dynamics associated to different phenomena of the cell’s life, such as cell cycle and cell death, on rat pheochromocytoma line PC12. Working in culture medium with alive and unperturbed samples, we could detect nanometer-sized movements; Fourier components revealed a clear distinct behavior associated to regulation of neurite outgrowth and changes on morphology after necrotic stimulus.

Optical method for distance and displacement measurements of the probe-sample separation in a scanning near-field optical microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Santamaria, L.; Siller, H. R.; Garcia-Ortiz, C. E., E-mail: cegarcia@cicese.mx

In this work, we present an alternative optical method to determine the probe-sample separation distance in a scanning near-field optical microscope. The experimental method is based in a Lloyd’s mirror interferometer and offers a measurement precision deviation of ∼100 nm using digital image processing and numerical analysis. The technique can also be strategically combined with the characterization of piezoelectric actuators and stability evaluation of the optical system. It also opens the possibility for the development of an automatic approximation control system valid for probe-sample distances from 5 to 500 μm.

Hard X-Ray Scanning Microscope with Multilayer Laue Lens Nanofocusing Optics

ScienceCinema

Nazaretski, Evgeny

2018-06-13

Evgeny Nazaretski, a physicist at Brookhaven Lab’s National Synchrotron Light Source II, spearheaded the development of a one-of-a-kind x-ray microscope with novel nanofocusing optics called multilayer Laue lenses.

Method for nanoscale spatial registration of scanning probes with substrates and surfaces

NASA Technical Reports Server (NTRS)

Wade, Lawrence A. (Inventor)

2010-01-01

Embodiments in accordance with the present invention relate to methods and apparatuses for aligning a scanning probe used to pattern a substrate, by comparing the position of the probe to a reference location or spot on the substrate. A first light beam is focused on a surface of the substrate as a spatial reference point. A second light beam then illuminates the scanning probe being used for patterning. An optical microscope images both the focused light beam, and a diffraction pattern, shadow, or light backscattered by the illuminated scanning probe tip of a scanning probe microscope (SPM), which is typically the tip of the scanning probe on an atomic force microscope (AFM). Alignment of the scanning probe tip relative to the mark is then determined by visual observation of the microscope image. This alignment process may be repeated to allow for modification or changing of the scanning probe microscope tip.

A hybrid scanning force and light microscope for surface imaging and three-dimensional optical sectioning in differential interference contrast.

PubMed

Stemmer, A

1995-04-01

The design of a scanned-cantilever-type force microscope is presented which is fully integrated into an inverted high-resolution video-enhanced light microscope. This set-up allows us to acquire thin optical sections in differential interference contrast (DIC) or polarization while the force microscope is in place. Such a hybrid microscope provides a unique platform to study how cell surface properties determine, or are affected by, the three-dimensional dynamic organization inside the living cell. The hybrid microscope presented in this paper has proven reliable and versatile for biological applications. It is the only instrument that can image a specimen by force microscopy and high-power DIC without having either to translate the specimen or to remove the force microscope. Adaptation of the design features could greatly enhance the suitability of other force microscopes for biological work.

Thermal radiation scanning tunnelling microscopy

NASA Astrophysics Data System (ADS)

de Wilde, Yannick; Formanek, Florian; Carminati, Rémi; Gralak, Boris; Lemoine, Paul-Arthur; Joulain, Karl; Mulet, Jean-Philippe; Chen, Yong; Greffet, Jean-Jacques

2006-12-01

In standard near-field scanning optical microscopy (NSOM), a subwavelength probe acts as an optical `stethoscope' to map the near field produced at the sample surface by external illumination. This technique has been applied using visible, infrared, terahertz and gigahertz radiation to illuminate the sample, providing a resolution well beyond the diffraction limit. NSOM is well suited to study surface waves such as surface plasmons or surface-phonon polaritons. Using an aperture NSOM with visible laser illumination, a near-field interference pattern around a corral structure has been observed, whose features were similar to the scanning tunnelling microscope image of the electronic waves in a quantum corral. Here we describe an infrared NSOM that operates without any external illumination: it is a near-field analogue of a night-vision camera, making use of the thermal infrared evanescent fields emitted by the surface, and behaves as an optical scanning tunnelling microscope. We therefore term this instrument a `thermal radiation scanning tunnelling microscope' (TRSTM). We show the first TRSTM images of thermally excited surface plasmons, and demonstrate spatial coherence effects in near-field thermal emission.

Operation of a wet near-field scanning optical microscope in stable zones by minimizing the resonance change of tuning forks.

PubMed

Park, Kyoung-Duck; Park, Doo Jae; Lee, Seung Gol; Choi, Geunchang; Kim, Dai-Sik; Byeon, Clare Chisu; Choi, Soo Bong; Jeong, Mun Seok

2014-02-21

A resonant shift and a decrease of resonance quality of a tuning fork attached to a conventional fiber optic probe in the vicinity of liquid is monitored systematically while varying the protrusion length and immersion depth of the probe. Stable zones where the resonance modification as a function of immersion depth is minimized are observed. A wet near-field scanning optical microscope (wet-NSOM) is operated for a sample within water by using such a stable zone.

Solid-state optical microscope

DOEpatents

Young, I.T.

1981-01-07

A solid state optical microscope is described wherein wide-field and high-resolution images of an object are produced at a rapid rate by utilizing conventional optics with a charge-coupled photodiode array. Means for scanning in one of two orthogonal directions are provided, while the charge-coupled photodiode array scans in the other orthogonal direction. Illumination light from the object is incident upon the photodiodes, creating packets of electrons (signals) which are representative of the illuminated object. The signals are then processed, stored in a memory, and finally displayed as a video signal.

Development and applications of optical interferometric micrometrology in the Angstrom and subangstrom range

NASA Technical Reports Server (NTRS)

Lauer, James L.; Abel, Phillip B.

1988-01-01

The characteristics of the scanning tunneling microscope and atomic force microscope (AFM) are briefly reviewed, and optical methods, mainly interferometry, of sufficient resolution to measure AFM deflections are discussed. The methods include optical resonators, laser interferometry, multiple-beam interferometry, and evanescent wave detection. Experimental results using AFM are reviewed.

The relationship between morphological changes of lens epithelial cells and intraocular lens optic material.

PubMed

Majima, K

1998-01-01

To examine the morphological changes of lens epithelial cells (LECs) occurring directly beneath and at regions contacting various intraocular lens (IOL) optic materials, human LECs were cultured on human anterior lens capsules and were further incubated upon placing above the cells lens optics made of polymethylmethacrylate, silicone, and soft acrylic material. Observations as to the morphological changes of LECs under phase-contrast microscope and scanning electron microscope were performed on the 14th day of incubation. Gatherings of LECs were observed at regions contacting the soft acrylic material under phase-contrast microscope, and gatherings of LECs were observed accurately at the same regions mentioned above under scanning electron microscope. On the other hand, LECs in contact with two other optic materials did not show morphological changes. The results suggest that LECs attached to and proliferated on not only the anterior lens capsules but also the soft acrylic IOL optics. The model used in this study may be useful in studying the relationship between cellular movement of LECs and IOL optic material.

Reprint of: Atmospheric scanning electron microscope observes cells and tissues in open medium through silicon nitride film.

PubMed

Nishiyama, Hidetoshi; Suga, Mitsuo; Ogura, Toshihiko; Maruyama, Yuusuke; Koizumi, Mitsuru; Mio, Kazuhiro; Kitamura, Shinichi; Sato, Chikara

2010-11-01

Direct observation of subcellular structures and their characterization is essential for understanding their physiological functions. To observe them in open environment, we have developed an inverted scanning electron microscope with a detachable, open-culture dish, capable of 8 nm resolution, and combined with a fluorescence microscope quasi-simultaneously observing the same area from the top. For scanning electron microscopy from the bottom, a silicon nitride film window in the base of the dish maintains a vacuum between electron gun and open sample dish while allowing electrons to pass through. Electrons are backscattered from the sample and captured by a detector under the dish. Cells cultured on the open dish can be externally manipulated under optical microscopy, fixed, and observed using scanning electron microscopy. Once fine structures have been revealed by scanning electron microscopy, their component proteins may be identified by comparison with separately prepared fluorescence-labeled optical microscopic images of the candidate proteins, with their heavy-metal-labeled or stained ASEM images. Furthermore, cell nuclei in a tissue block stained with platinum-blue were successfully observed without thin-sectioning, which suggests the applicability of this inverted scanning electron microscope to cancer diagnosis. This microscope visualizes mesoscopic-scale structures, and is also applicable to non-bioscience fields including polymer chemistry. Copyright © 2010 Elsevier Inc. All rights reserved.

The design and construction of a cost-efficient confocal laser scanning microscope

NASA Astrophysics Data System (ADS)

Xi, Peng; Rajwa, Bartlomiej; Jones, James T.; Robinson, J. Paul

2007-03-01

The optical dissection ability of confocal microscopy makes it a powerful tool for biological materials. However, the cost and complexity of confocal scanning laser microscopy hinders its wide application in education. We describe the construction of a simplified confocal scanning laser microscope and demonstrate three-dimensional projection based on cost-efficient commercial hardware, together with available open source software.

Scanning ion-conductance and atomic force microscope with specialized sphere-shaped nanopippettes

NASA Astrophysics Data System (ADS)

Zhukov, M. V.; Sapozhnikov, I. D.; Golubok, A. O.; Chubinskiy-Nadezhdin, V. I.; Komissarenko, F. E.; Lukashenko, S. Y.

2017-11-01

A scanning ion-conductance microscope was designed on the basis of scanning probe microscope NanoTutor. The optimal parameters of nanopipettes fabrication were found according to scanning electron microscopy diagnostics, current-distance I (Z) and current-voltage characteristics. A comparison of images of test objects, including biological samples, was carried out in the modes of optical microscopy, atomic force microscopy and scanning ion-conductance microscopy. Sphere-shaped nanopippettes probes were developed and tested to increase the stability of pipettes, reduce invasiveness and improve image quality of atomic force microscopy in tapping mode. The efficiency of sphere-shaped nanopippettes is shown.

Two-Photon Imaging with Diffractive Optical Elements

PubMed Central

Watson, Brendon O.; Nikolenko, Volodymyr; Yuste, Rafael

2009-01-01

Two-photon imaging has become a useful tool for optical monitoring of neural circuits, but it requires high laser power and serial scanning of each pixel in a sample. This results in slow imaging rates, limiting the measurements of fast signals such as neuronal activity. To improve the speed and signal-to-noise ratio of two-photon imaging, we introduce a simple modification of a two-photon microscope, using a diffractive optical element (DOE) which splits the laser beam into several beamlets that can simultaneously scan the sample. We demonstrate the advantages of DOE scanning by enhancing the speed and sensitivity of two-photon calcium imaging of action potentials in neurons from neocortical brain slices. DOE scanning can easily improve the detection of time-varying signals in two-photon and other non-linear microscopic techniques. PMID:19636390

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

PubMed

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.

Compact variable-temperature scanning force microscope.

PubMed

Chuang, Tien-Ming; de Lozanne, Alex

2007-05-01

A compact design for a cryogenic variable-temperature scanning force microscope using a fiber-optic interferometer to measure cantilever deflection is presented. The tip-sample coarse approach and the lateral tip positioning are performed by piezoelectric positioners in situ. The microscope has been operated at temperatures between 6 and 300 K. It is designed to fit into an 8 T superconducting magnet with the field applied in the out-of-plane direction. The results of scanning in various modes are demonstrated, showing contrast based on magnetic field gradients or surface potentials.

A combined scanning tunnelling microscope and x-ray interferometer

NASA Astrophysics Data System (ADS)

Yacoot, Andrew; Kuetgens, Ulrich; Koenders, Ludger; Weimann, Thomas

2001-10-01

A monolithic x-ray interferometer made from silicon and a scanning tunnelling microscope have been combined and used to calibrate grating structures with periodicities of 100 nm or less. The x-ray interferometer is used as a translation stage which moves in discrete steps of 0.192 nm, the lattice spacing of the silicon (220) planes. Hence, movements are traceable to the definition of the metre and the nonlinearity associated with the optical interferometers used to measure displacement in more conventional metrological scanning probe microscopes (MSPMs) removed.

A high-resolution, confocal laser-scanning microscope and flash photolysis system for physiological studies.

PubMed

Parker, I; Callamaras, N; Wier, W G

1997-06-01

We describe the construction of a high-resolution confocal laser-scanning microscope, and illustrate its use for studying elementary Ca2+ signalling events in cells. An avalanche photodiode module and simple optical path provide a high efficiency system for detection of fluorescence signals, allowing use of a small confocal aperture giving near diffraction-limited spatial resolution (< 300 nm lateral and < 400 nm axial). When operated in line-scan mode, the maximum temporal resolution is 1 ms, and the associated computer software allows complete flexibility to record line-scans continuously for long (minutes) periods or to obtain any desired pixel resolution in x-y scans. An independent UV irradiation system permits simultaneous photolysis of caged compounds over either a uniform, wide field (arc lamp source) or at a tightly focussed spot (frequency-tripled Nd:YAG laser). The microscope thus provides a versatile tool for optical studies of dynamic cellular processes, as well as excellent resolution for morphological studies. The confocal scanner can be added to virtually any inverted microscope for a component cost that is only a small fraction of that of comparable commercial instruments, yet offers better performance and greater versatility.

Re-scan confocal microscopy: scanning twice for better resolution

PubMed Central

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. PMID:24298422

Spectral ophthalmoscopy based on supercontinuum

NASA Astrophysics Data System (ADS)

Cheng, Yueh-Hung; Yu, Jiun-Yann; Wu, Han-Hsuan; Huang, Bo-Jyun; Chu, Shi-Wei

2010-02-01

Confocal scanning laser ophthalmoscope (CSLO) has been established to be an important diagnostic tool for retinopathies like age-related macular degeneration, glaucoma and diabetes. Compared to a confocal laser scanning microscope, CSLO is also capable of providing optical sectioning on retina with the aid of a pinhole, but the microscope objective is replaced by the optics of eye. Since optical spectrum is the fingerprint of local chemical composition, it is attractive to incorporate spectral acquisition into CSLO. However, due to the limitation of laser bandwidth and chromatic/geometric aberration, the scanning systems in current CSLO are not compatible with spectral imaging. Here we demonstrate a spectral CSLO by combining a diffraction-limited broadband scanning system and a supercontinuum laser source. Both optical sectioning capability and sub-cellular resolution are demonstrated on zebrafish's retina. To our knowledge, it is also the first time that CSLO is applied onto the study of fish vision. The versatile spectral CSLO system will be useful to retinopathy diagnosis and neuroscience research.

Nonlinear optical microscopy for immunoimaging: a custom optimized system of high-speed, large-area, multicolor imaging

PubMed Central

Li, Hui; Cui, Quan; Zhang, Zhihong; Luo, Qingming

2015-01-01

Background The nonlinear optical microscopy has become the current state-of-the-art for intravital imaging. Due to its advantages of high resolution, superior tissue penetration, lower photodamage and photobleaching, as well as intrinsic z-sectioning ability, this technology has been widely applied in immunoimaging for a decade. However, in terms of monitoring immune events in native physiological environment, the conventional nonlinear optical microscope system has to be optimized for live animal imaging. Generally speaking, three crucial capabilities are desired, including high-speed, large-area and multicolor imaging. Among numerous high-speed scanning mechanisms used in nonlinear optical imaging, polygon scanning is not only linearly but also dispersion-freely with high stability and tunable rotation speed, which can overcome disadvantages of multifocal scanning, resonant scanner and acousto-optical deflector (AOD). However, low frame rate, lacking large-area or multicolor imaging ability make current polygonbased nonlinear optical microscopes unable to meet the requirements of immune event monitoring. Methods We built up a polygon-based nonlinear optical microscope system which was custom optimized for immunoimaging with high-speed, large-are and multicolor imaging abilities. Results Firstly, we validated the imaging performance of the system by standard methods. Then, to demonstrate the ability to monitor immune events, migration of immunocytes observed by the system based on typical immunological models such as lymph node, footpad and dorsal skinfold chamber are shown. Finally, we take an outlook for the possible advance of related technologies such as sample stabilization and optical clearing for more stable and deeper intravital immunoimaging. Conclusions This study will be helpful for optimizing nonlinear optical microscope to obtain more comprehensive and accurate information of immune events. PMID:25694951

High-resolution microscope for tip-enhanced optical processes in ultrahigh vacuum

NASA Astrophysics Data System (ADS)

Steidtner, Jens; Pettinger, Bruno

2007-10-01

An optical microscope based on tip-enhanced optical processes that can be used for studies on adsorbates as well as thin layers and nanostructures is presented. The microscope provides chemical and topographic informations with a resolution of a few nanometers and can be employed in ultrahigh vacuum as well as gas phase. The construction involves a number of improvements compared to conventional instruments. The central idea is to mount, within an UHV system, an optical platform with all necessary optical elements to a rigid frame that also carries the scanning tunneling microscope unit and to integrate a high numerical aperture parabolic mirror between the scanning probe microscope head and the sample. The parabolic mirror serves to focus the incident light and to collect a large fraction of the scattered light. The first experimental results of Raman measurements on silicon samples as well as brilliant cresyl blue layers on single crystalline gold and platinum surfaces in ultrahigh vacuum are presented. For dye adsorbates a Raman enhancement of ˜106 and a net signal gain of up to 4000 was observed. The focus diameter (˜λ/2) was measured by Raman imaging the focal region on a Si surface. The requirements of the parabolic mirror in terms of alignment accuracy were experimentally determined as well.

Micropaleontological studies of lunar and terrestrial precambrian materials

NASA Technical Reports Server (NTRS)

Schope, J. W.

1974-01-01

Optical microscopic and scanning electron microscopic studies of rock chips and dust returned by Apollo 14, 15, 16, and 17 are analyzed along with optical microscopic studies of petrographic thin sections of breccias and basalts returned by Apollo 14, 15, and 16. Results show no evidence of modern or fossil lunar organisms. The lunar surface is now, and apparently has been throughout the geologic past, inimical to known biologic systems.

Rapid fabrication of micro-nanometric tapered fiber lens and characterization by a novel scanning optical microscope with submicron resolution.

PubMed

Zheng, Shouguo; Zeng, Xinhua; Luo, Wei; Jradi, Safi; Plain, Jérôme; Li, Miao; Renaud-Goud, Philippe; Deturche, Régis; Wang, Zengfu; Kou, Jieting; Bachelot, Renaud; Royer, Pascal

2013-01-14

In numerous applications of optical scanning microscopy, a reference tapered fiber lens with high symmetry at sub-wavelength scale remains a challenge. Here, we demonstrate the ability to manufacture it with a wide range of geometry control, either for the length from several hundred nanometers to several hundred microns, or for the curvature radius from several tens of nanometers to several microns on the endface of a single mode fiber. On this basis, a scanning optical microscope has been developed, which allows for fast characterization of various sub-wavelength tapered fiber lenses. Focal position and depth of microlenses with different geometries have been determined to be ranged from several hundreds of nanometers to several microns. FDTD calculations are consistent with experimental results.

Experimental stress–strain analysis of tapered silica optical fibers with nanofiber waist

DOE Office of Scientific and Technical Information (OSTI.GOV)

Holleis, S.; Hoinkes, T.; Wuttke, C.

2014-04-21

We experimentally determine tensile force–elongation diagrams of tapered optical fibers with a nanofiber waist. The tapered optical fibers are produced from standard silica optical fibers using a heat and pull process. Both, the force–elongation data and scanning electron microscope images of the rupture points indicate a brittle material. Despite the small waist radii of only a few hundred nanometers, our experimental data can be fully explained by a nonlinear stress–strain model that relies on material properties of macroscopic silica optical fibers. This is an important asset when it comes to designing miniaturized optical elements as one can rely on themore » well-founded material characteristics of standard optical fibers. Based on this understanding, we demonstrate a simple and non-destructive technique that allows us to determine the waist radius of the tapered optical fiber. We find excellent agreement with independent scanning electron microscope measurements of the waist radius.« less

Development of an ultrasound microscope combined with optical microscope for multiparametric characterization of a single cell.

PubMed

Arakawa, Mototaka; Shikama, Joe; Yoshida, Koki; Nagaoka, Ryo; Kobayashi, Kazuto; Saijo, Yoshifumi

2015-09-01

Biomechanics of the cell has been gathering much attention because it affects the pathological status in atherosclerosis and cancer. In the present study, an ultrasound microscope system combined with optical microscope for characterization of a single cell with multiple ultrasound parameters was developed. The central frequency of the transducer was 375 MHz and the scan area was 80 × 80 μm with up to 200 × 200 sampling points. An inverted optical microscope was incorporated in the design of the system, allowing for simultaneous optical observations of cultured cells. Two-dimensional mapping of multiple ultrasound parameters, such as sound speed, attenuation, and acoustic impedance, as well as the thickness, density, and bulk modulus of specimen/cell under investigation, etc., was realized by the system. Sound speed and thickness of a 3T3-L1 fibroblast cell were successfully obtained by the system. The ultrasound microscope system combined with optical microscope further enhances our understanding of cellular biomechanics.

Re-evaluation of differential phase contrast (DPC) in a scanning laser microscope using a split detector as an alternative to differential interference contrast (DIC) optics.

PubMed

Amos, W B; Reichelt, S; Cattermole, D M; Laufer, J

2003-05-01

In this paper, differential phase imaging (DPC) with transmitted light is implemented by adding a suitable detection system to a standard commercially available scanning confocal microscope. DPC, a long-established method in scanning optical microscopy, depends on detecting the intensity difference between opposite halves or quadrants of a split photodiode detector placed in an aperture plane. Here, DPC is compared with scanned differential interference contrast (DIC) using a variety of biological specimens and objective lenses of high numerical aperture. While DPC and DIC images are generally similar, DPC seems to have a greater depth of field. DPC has several advantages over DIC. These include low cost (no polarizing or strain-free optics are required), absence of a double scanning spot, electronically variable direction of shading and the ability to image specimens in plastic dishes where birefringence prevents the use of DIC. DPC is also here found to need 20 times less laser power at the specimen than DIC.

Non linear optical investigations of silver nanoparticles synthesised by curcumin reduction

NASA Astrophysics Data System (ADS)

Dhanya, N. P.

2017-11-01

Metal nanoparticles have considerable applications in assorted fields like medicine, biology, photonics, metallurgy etc. Optical applications of Silver nanoparticles are of significant interest among researchers nowadays. In this paper, we report a single step chemical reduction of silver nanoparticles with Curcumin both as a reducing and stabilising agent at room temperature. Structural, plasmonic and non linear optical properties of the prepared nanoparticles are explored using Scanning Electron Microscope, Transmission Electron Microscope, UV absorption spectrometry, Spectroflurometry and Z scan. UV-Vis absorption studies affirm the Surface Plasmon Resonance (SPR) absorption and spectroflurometric studies announce the emission spectrum of the prepared silvernanoparticles at 520 nm. SEM and TEM images uphold the existence of uniform sized, spherical silvernanoparticles. Nonlinear optical studies are accomplished with the open aperture z scan technique in the nanosecond regime. The nonlinearity is in virtue of saturable absorption, two-photon absorption and excited state absorption. The marked nonlinearity and optical limiting of the Curcumin reduced silvernanoparticles enhances its photonic applications.

To boldly glow ... applications of laser scanning confocal microscopy in developmental biology.

PubMed

Paddock, S W

1994-05-01

The laser scanning confocal microscope (LSCM) is now established as an invaluable tool in developmental biology for improved light microscope imaging of fluorescently labelled eggs, embryos and developing tissues. The universal application of the LSCM in biomedical research has stimulated improvements to the microscopes themselves and the synthesis of novel probes for imaging biological structures and physiological processes. Moreover the ability of the LSCM to produce an optical series in perfect register has made computer 3-D reconstruction and analysis of light microscope images a practical option.

Laser scanning confocal microscope with programmable amplitude, phase, and polarization of the illumination beam.

PubMed

Boruah, B R; Neil, M A A

2009-01-01

We describe the design and construction of a laser scanning confocal microscope with programmable beam forming optics. The amplitude, phase, and polarization of the laser beam used in the microscope can be controlled in real time with the help of a liquid crystal spatial light modulator, acting as a computer generated hologram, in conjunction with a polarizing beam splitter and two right angled prisms assembly. Two scan mirrors, comprising an on-axis fast moving scan mirror for line scanning and an off-axis slow moving scan mirror for frame scanning, configured in a way to minimize the movement of the scanned beam over the pupil plane of the microscope objective, form the XY scan unit. The confocal system, that incorporates the programmable beam forming unit and the scan unit, has been implemented to image in both reflected and fluorescence light from the specimen. Efficiency of the system to programmably generate custom defined vector beams has been demonstrated by generating a bottle structured focal volume, which in fact is the overlap of two cross polarized beams, that can simultaneously improve both the lateral and axial resolutions if used as the de-excitation beam in a stimulated emission depletion confocal microscope.

Apertureless near-field scanning optical microscope working with or without laser source.

PubMed

Formanek, F; De Wilde, Y; Aigouy, L; Chen, Y

2004-01-01

An apertureless near-field scanning optical microscope (ANSOM), used indifferent configurations, is presented. Our versatile home-made setup, based on a sharp tungsten tip glued onto a quartz tuning fork and working in tapping mode, allows to perform imaging over a broad spectral range. We have recorded optical images in the visible (wavelength, lambda = 655 nm) and in the infrared (lambda = 10.6 microm), proving that the setup routinely achieves an optical resolution of <50 nm regardless of the illumination wavelength. We have also shown optical images recorded in the visible (lambda = 655 nm) in an inverted configuration where the tip does not perturb the focused spot of the illumination laser. Approach curves as well as image profiles have revealed that on demodulating the optical signal at higher harmonics, we can obtain an effective probe sharpening which results in an improvement of the resolution. Finally, we have presented optical images recorded in the infrared without any illumination, that is, the usual laser source is replaced by a simple heating of the sample. This has shown that the ANSOM can be used as a near-field thermal optical microscope (NTOM) to probe the near field generated by the thermal emission of the sample.

Miniaturized fiber-coupled confocal fluorescence microscope with an electrowetting variable focus lens using no moving parts

PubMed Central

Ozbay, Baris N.; Losacco, Justin T.; Cormack, Robert; Weir, Richard; Bright, Victor M.; Gopinath, Juliet T.; Restrepo, Diego; Gibson, Emily A.

2015-01-01

We report a miniature, lightweight fiber-coupled confocal fluorescence microscope that incorporates an electrowetting variable focus lens to provide axial scanning for full three-dimensional (3D) imaging. Lateral scanning is accomplished by coupling our device to a laser-scanning confocal microscope through a coherent imaging fiber-bundle. The optical components of the device are combined in a custom 3D-printed adapter with an assembled weight of <2 g that can be mounted onto the head of a mouse. Confocal sectioning provides an axial resolution of ~12 µm and an axial scan range of ~80 µm. The lateral field-of-view is 300 µm, and the lateral resolution is 1.8 µm. We determined these parameters by imaging fixed sections of mouse neuronal tissue labeled with green fluorescent protein (GFP) and fluorescent bead samples in agarose gel. To demonstrate viability for imaging intact tissue, we resolved multiple optical sections of ex vivo mouse olfactory nerve fibers expressing yellow fluorescent protein (YFP). PMID:26030555

Scanning laser microscope for imaging nanostructured superconductors

NASA Astrophysics Data System (ADS)

Ishida, Takekazu; Arai, Kohei; Akita, Yukio; Miyanari, Mitsunori; Minami, Yusuke; Yotsuya, Tsutomu; Kato, Masaru; Satoh, Kazuo; Uno, Mayumi; Shimakage, Hisashi; Miki, Shigehito; Wang, Zhen

2010-10-01

The nanofabrication of superconductors yields various interesting features in superconducting properties. A variety of different imaging techniques have been developed for probing the local superconducting profiles. A scanning pulsed laser microscope has been developed by the combination of the XYZ piezo-driven stages and an optical fiber with an aspheric focusing lens. The scanning laser microscope is used to understand the position-dependent properties of a superconducting MgB 2 stripline of length 100 μm and width of 3 μm under constant bias current. Our results show that the superconducting stripline can clearly be seen in the contour image of the scanning laser microscope on the signal voltage. It is suggested from the observed image that the inhomogeneity is relevant in specifying the operating conditions such as detection efficiency of the sensor.

Micro-optical design of a three-dimensional microlens scanner for vertically integrated micro-opto-electro-mechanical systems.

PubMed

Baranski, Maciej; Bargiel, Sylwester; Passilly, Nicolas; Gorecki, Christophe; Jia, Chenping; Frömel, Jörg; Wiemer, Maik

2015-08-01

This paper presents the optical design of a miniature 3D scanning system, which is fully compatible with the vertical integration technology of micro-opto-electro-mechanical systems (MOEMS). The constraints related to this integration strategy are considered, resulting in a simple three-element micro-optical setup based on an afocal scanning microlens doublet and a focusing microlens, which is tolerant to axial position inaccuracy. The 3D scanning is achieved by axial and lateral displacement of microlenses of the scanning doublet, realized by micro-electro-mechanical systems microactuators (the transmission scanning approach). Optical scanning performance of the system is determined analytically by use of the extended ray transfer matrix method, leading to two different optical configurations, relying either on a ball lens or plano-convex microlenses. The presented system is aimed to be a core component of miniature MOEMS-based optical devices, which require a 3D optical scanning function, e.g., miniature imaging systems (confocal or optical coherence microscopes) or optical tweezers.

Failure Analysis of Heavy-Ion-Irradiated Schottky Diodes

NASA Technical Reports Server (NTRS)

Casey, Megan C.; Lauenstein, Jean-Marie; Wilcox, Edward P.; Topper, Alyson D.; Campola, Michael J.; Label, Kenneth A.

2017-01-01

In this work, we use high- and low-magnitude optical microscope images, infrared camera images, and scanning electron microscope images to identify and describe the failure locations in heavy-ion-irradiated Schottky diodes.

A wide field-of-view microscope based on holographic focus grid

NASA Astrophysics Data System (ADS)

Wu, Jigang; Cui, Xiquan; Zheng, Guoan; Lee, Lap Man; Yang, Changhuei

2010-02-01

We have developed a novel microscope technique that can achieve wide field-of-view (FOV) imaging and yet possess resolution that is comparable to conventional microscope. The principle of wide FOV microscope system breaks the link between resolution and FOV magnitude of traditional microscopes. Furthermore, by eliminating bulky optical elements from its design and utilizing holographic optical elements, the wide FOV microscope system is more cost-effective. In our system, a hologram was made to focus incoming collimated beam into a focus grid. The sample is put in the focal plane and the transmissions of the focuses are detected by an imaging sensor. By scanning the incident angle of the incoming beam, the focus grid will scan across the sample and the time-varying transmission can be detected. We can then reconstruct the transmission image of the sample. The resolution of microscopic image is limited by the size of the focus formed by the hologram. The scanning area of each focus spot is determined by the separation of the focus spots and can be made small for fast imaging speed. We have fabricated a prototype system with a 2.4-mm FOV and 1-μm resolution. The prototype system was used to image onion skin cells for a demonstration. The preliminary experiments prove the feasibility of the wide FOV microscope technique, and the possibility of a wider FOV system with better resolution.

Soft x-ray spectromicroscopy using compact scanning transmission x-ray microscope at the photon factory

DOE Office of Scientific and Technical Information (OSTI.GOV)

Takeichi, Yasuo, E-mail: yasuo.takeichi@kek.jp; Inami, Nobuhito; Ono, Kanta

We report the stability and recent performances of a new type of scanning transmission X-ray microscopy. The optics and compact design of the microscope realized mobility and robust performance. Detailed consideration to the vibration control will be described. The insertion device upgraded to elliptical polarization undulator enabled linear dichroism and circular dichroism experiments.

Fractal evaluation of drug amorphicity from optical and scanning electron microscope images

NASA Astrophysics Data System (ADS)

Gavriloaia, Bogdan-Mihai G.; Vizireanu, Radu C.; Neamtu, Catalin I.; Gavriloaia, Gheorghe V.

2013-09-01

Amorphous materials are metastable, more reactive than the crystalline ones, and have to be evaluated before pharmaceutical compound formulation. Amorphicity is interpreted as a spatial chaos, and patterns of molecular aggregates of dexamethasone, D, were investigated in this paper by using fractal dimension, FD. Images having three magnifications of D were taken from an optical microscope, OM, and with eight magnifications, from a scanning electron microscope, SEM, were analyzed. The average FD for pattern irregularities of OM images was 1.538, and about 1.692 for SEM images. The FDs of the two kinds of images are less sensitive of threshold level. 3D images were shown to illustrate dependence of FD of threshold and magnification level. As a result, optical image of single scale is enough to characterize the drug amorphicity. As a result, the OM image at a single scale is enough to characterize the amorphicity of D.

The temperature-dependency of the optical band gap of ZnO measured by electron energy-loss spectroscopy in a scanning transmission electron microscope

NASA Astrophysics Data System (ADS)

Granerød, Cecilie S.; Galeckas, Augustinas; Johansen, Klaus Magnus; Vines, Lasse; Prytz, Øystein

2018-04-01

The optical band gap of ZnO has been measured as a function of temperature using Electron Energy-Loss Spectroscopy (EELS) in a (Scanning) Transmission Electron Microscope ((S)TEM) from approximately 100 K up towards 1000 K. The band gap narrowing shows a close to linear dependency for temperatures above 250 K and is accurately described by Varshni, Bose-Einstein, Pässler and Manoogian-Woolley models. Additionally, the measured band gap is compared with both optical absorption measurements and photoluminescence data. STEM-EELS is here shown to be a viable technique to measure optical band gaps at elevated temperatures, with an available temperature range up to 1500 K and the benefit of superior spatial resolution.

Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons.

PubMed Central

Fan, G Y; Fujisaki, H; Miyawaki, A; Tsay, R K; Tsien, R Y; Ellisman, M H

1999-01-01

A video-rate (30 frames/s) scanning two-photon excitation microscope has been successfully tested. The microscope, based on a Nikon RCM 8000, incorporates a femtosecond pulsed laser with wavelength tunable from 690 to 1050 nm, prechirper optics for laser pulse-width compression, resonant galvanometer for video-rate point scanning, and a pair of nonconfocal detectors for fast emission ratioing. An increase in fluorescent emission of 1.75-fold is consistently obtained with the use of the prechirper optics. The nonconfocal detectors provide another 2.25-fold increase in detection efficiency. Ratio imaging and optical sectioning can therefore be performed more efficiently without confocal optics. Faster frame rates, at 60, 120, and 240 frames/s, can be achieved with proportionally reduced scan lines per frame. Useful two-photon images can be acquired at video rate with a laser power as low as 2.7 mW at specimen with the genetically modified green fluorescent proteins. Preliminary results obtained using this system confirm that the yellow "cameleons" exhibit similar optical properties as under one-photon excitation conditions. Dynamic two-photon images of cardiac myocytes and ratio images of yellow cameleon-2.1, -3.1, and -3.1nu are also presented. PMID:10233058

Nm-scale spatial resolution x-ray imaging with MLL nanofocusing optics: instrumentational requirements and challenges

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nazaretski, E.; Yan, H.; Lauer, K.

2016-08-30

The Hard X-ray Nanoprobe (HXN) beamline at NSLS-II has been designed and constructed to enable imaging experiments with unprecedented spatial resolution and detection sensitivity. The HXN X-ray Microscope is a key instrument for the beamline, providing a suite of experimental capabilities which includes scanning fluorescence, diffraction, differential phase contrast and ptychography utilizing Multilayer Laue Lenses (MLL) and zoneplate (ZP) as nanofocusing optics. In this paper, we present technical requirements for the MLL-based scanning microscope, outline the development concept and present first ~15 x 15 nm 2 spatial resolution x-ray fluorescence images.

Use of scanning near-field optical microscope with an aperture probe for detection of luminescent nanodiamonds

NASA Astrophysics Data System (ADS)

Shershulin, V. A.; Samoylenko, S. R.; Shenderova, O. A.; Konov, V. I.; Vlasov, I. I.

2017-02-01

The suitability of scanning near-field optical microscopy (SNOM) to image photoluminescent diamond nanoparticles with nanoscale resolution is demonstrated. Isolated diamond nanocrystals with an average size of 100 nm, containing negatively charged nitrogen-vacancy (NV-) centers, were chosen as tested material. The NV- luminescence was stimulated by continuous 532 nm laser light. Sizes of analyzed crystallites were monitored by an atomic force microscope. The lateral resolution of the order of 100 nm was reached in SNOM imaging of diamond nanoparticles using 150 nm square aperture of the probe.

Improved Scanners for Microscopic Hyperspectral Imaging

NASA Technical Reports Server (NTRS)

Mao, Chengye

2009-01-01

Improved scanners to be incorporated into hyperspectral microscope-based imaging systems have been invented. Heretofore, in microscopic imaging, including spectral imaging, it has been customary to either move the specimen relative to the optical assembly that includes the microscope or else move the entire assembly relative to the specimen. It becomes extremely difficult to control such scanning when submicron translation increments are required, because the high magnification of the microscope enlarges all movements in the specimen image on the focal plane. To overcome this difficulty, in a system based on this invention, no attempt would be made to move either the specimen or the optical assembly. Instead, an objective lens would be moved within the assembly so as to cause translation of the image at the focal plane: the effect would be equivalent to scanning in the focal plane. The upper part of the figure depicts a generic proposed microscope-based hyperspectral imaging system incorporating the invention. The optical assembly of this system would include an objective lens (normally, a microscope objective lens) and a charge-coupled-device (CCD) camera. The objective lens would be mounted on a servomotor-driven translation stage, which would be capable of moving the lens in precisely controlled increments, relative to the camera, parallel to the focal-plane scan axis. The output of the CCD camera would be digitized and fed to a frame grabber in a computer. The computer would store the frame-grabber output for subsequent viewing and/or processing of images. The computer would contain a position-control interface board, through which it would control the servomotor. There are several versions of the invention. An essential feature common to all versions is that the stationary optical subassembly containing the camera would also contain a spatial window, at the focal plane of the objective lens, that would pass only a selected portion of the image. In one version, the window would be a slit, the CCD would contain a one-dimensional array of pixels, and the objective lens would be moved along an axis perpendicular to the slit to spatially scan the image of the specimen in pushbroom fashion. The image built up by scanning in this case would be an ordinary (non-spectral) image. In another version, the optics of which are depicted in the lower part of the figure, the spatial window would be a slit, the CCD would contain a two-dimensional array of pixels, the slit image would be refocused onto the CCD by a relay-lens pair consisting of a collimating and a focusing lens, and a prism-gratingprism optical spectrometer would be placed between the collimating and focusing lenses. Consequently, the image on the CCD would be spatially resolved along the slit axis and spectrally resolved along the axis perpendicular to the slit. As in the first-mentioned version, the objective lens would be moved along an axis perpendicular to the slit to spatially scan the image of the specimen in pushbroom fashion.

Resolution enhancement in a double-helix phase engineered scanning microscope (RESCH microscope) (Presentation Recording)

NASA Astrophysics Data System (ADS)

Jesacher, Alexander; Ritsch-Marte, Monika; Piestun, Rafael

2015-08-01

Recently we introduced RESCH microscopy [1] - a scanning microscope that allows slightly refocusing the sample after the acquisition has been performed, solely by performing appropriate data post-processing. The microscope features a double-helix phase-engineered emission point spread function in combination with camera-based detection. Based on the principle of transverse resolution enhancement in Image Scanning Microscopy [2,3], we demonstrate similar resolution improvement in RESCH. Furthermore, we outline a pathway for how the collected 3D sample information can be used to construct sharper optical sections. [1] A. Jesacher, M. Ritsch-Marte and R. Piestun, accepted for Optica. [2] C.J.R. Sheppard, "Super-resolution in Confocal imaging," Optik, 80, 53-54 (1988). [3] C.B. Müller and J. Enderlein "Image Scanning Microscopy," Phys. Rev. Lett. 104, 198101 (2010).

Heterodyne Interferometry with a Scanning Optical Microscope.

NASA Astrophysics Data System (ADS)

Hobbs, Philip Charles Danby

The design and implementation of a confocal optical microscope which functions as an electronically scanned heterodyne interferometer are described. Theoretical models based on Fourier optics for general samples and on exact series solution of the scalar Helmholtz equation for a class of trench structures are developed and compared with experimental data. Good agreement is obtained. The associated data acquisition system, also described, enables the system to measure both the amplitude (to 12 bits) and the phase (to 0.1^circ) of a returned optical beam, at a continuous rate of 30,000 points per second. The microscope system uses a wide-band tellurium dioxide acousto-optic cell for electronic scanning, frequency shifting, and beam splitting/combining. It uses a stationary reference beam on the sample for vibration cancellation, which results in a system of great vibration immunity. It can measure relief ranging from a few tenths of a micron down to a few Angstroms, and line widths down to well below 0.4 micron, using light of 0.5 micron wavelength. Angstrom resolution can be achieved in a single full-speed scan, without special vibration isolation equipment, providing that folding mirrors are avoided. A signal processing algorithm based on Fourier deconvolution is presented; it takes advantage of the extra bandwidth of a confocal system and the availability of both amplitude and phase, to improve the lateral resolution by approximately a factor of two. Experimental results are shown, which demonstrate phase edge resolution (10%-90%) of 0.45 lambda (raw data), and 0.18 lambda (after filtering), in excellent agreement with the Fourier optics prediction. The exact scalar theory calculates the response of the microscope as it scans over an infinitely long rectangular trench in a plane boundary on which Dirichlet boundary conditions apply. An expansion in cavity modes inside the trench is used to match the field and its derivatives across the mouth of the trench to get the self-consistent solution. A listing is appended of a program for an HP personal computer which performs the simulation in 1 to 5 minutes' running time for most cases. The trench theory is compared with the Fourier theory and with experimental results for actual metal trenches, with good results.

Multiple beam interference confocal microscopy: a tool for morphological investigation of living cells and tissues

NASA Astrophysics Data System (ADS)

Joshi, Narahari V.; Medina, Honorio

2000-05-01

Multiple beam interference system is used in conjunction with a conventional scanning confocal microscope to examine the morphology and construction of 3D images of Histolytic Ameba and parasite Candida Albicans. The present combination permits to adjoin advantages of both systems, namely the vertical high contrast and optical sectioning. The interference pattern obtained from a multiple internal reflection of a simple, sandwiched between the glass plate and the cover plate, was focussed on an objective of a scanning confocal microscope. According to optical path differences, morphological details were revealed. The combined features, namely improved resolution in z axis, originated from the interference pattern and the optical sectioning of the confocal scanning system, enhance the resolution and contrast dramatically. These features permitted to obtain unprecedented images of Histolytic Ameba and parasite Candida Albicans. Because of the improved contrast, several details like double wall structure of candida, internal structure of ameba are clearly visible.

Fine Metal Mask 3-Dimensional Measurement by using Scanning Digital Holographic Microscope

NASA Astrophysics Data System (ADS)

Shin, Sanghoon; Yu, Younghun

2018-04-01

For three-dimensional microscopy, fast and high axial resolution are very important. Extending the depth of field for digital holographic is necessary for three-dimensional measurements of thick samples. We propose an optical sectioning method for optical scanning digital holography that is performed in the frequency domain by spatial filtering of a reconstructed amplitude image. We established a scanning dual-wavelength off-axis digital holographic microscope to measure samples that exhibit a large amount of coherent noise and a thickness larger than the depth of focus of the objective lens. As a demonstration, we performed a three-dimensional measurement of a fine metal mask with a reconstructed sectional phase image and filtering with a reconstructed amplitude image.

Near-field microscopy and fluorescence spectroscopy: application to chromosomes labelled with different fluorophores.

PubMed

Mahieu-Williame, L; Falgayrettes, P; Nativel, L; Gall-Borrut, P; Costa, L; Salehzada, T; Bisbal, C

2010-04-01

We have coupled a spectrophotometer with a scanning near-field optical microscope to obtain, with a single scan, simultaneously scanning near-field optical microscope fluorescence images at different wavelengths as well as topography and transmission images. Extraction of the fluorescence spectra enabled us to decompose the different wavelengths of the fluorescence signals which normally overlap. We thus obtained images of the different fluorescence emissions of acridine orange bound to single or double stranded nucleic acids in human metaphase chromosomes before and after DNAse I or RNAse A treatment. The analysis of these images allowed us to visualize some specific chromatin areas where RNA is associated with DNA showing that such a technique could be used to identify multiple components within a cell.

Intensity calibration of a laser scanning confocal microscope based on concentrated dyes.

PubMed

Model, Michael A; Blank, James L

2006-10-01

To find water-soluble fluorescent dyes with absorption in various regions of the spectrum and investigate their utility as standards for laser scanning confocal microscopy. Several dyes were found to have characteristics required for fluorescence microscopy standards. The intensity of biological fluorescent specimens was measured against the emission of concentrated dyes. Results using different optics and different microscopes were compared. Slides based on concentrated dyes can be prepared in a highly reproducible manner and are stable under laser scanning. Normalized fluorescence of biological specimens remains consistent with different objective lenses and is tolerant to some mismatch in optical filters or imperfect pinhole alignment. Careful choice of scanning parameters is necessary to ensure linearity of intensity measurements. Concentrated dyes provide a robust and inexpensive intensity standard that can be used in basic research or clinical studies.

A high-resolution combined scanning laser and widefield polarizing microscope for imaging at temperatures from 4 K to 300 K.

PubMed

Lange, M; Guénon, S; Lever, F; Kleiner, R; Koelle, D

2017-12-01

Polarized light microscopy, as a contrast-enhancing technique for optically anisotropic materials, is a method well suited for the investigation of a wide variety of effects in solid-state physics, as, for example, birefringence in crystals or the magneto-optical Kerr effect (MOKE). We present a microscopy setup that combines a widefield microscope and a confocal scanning laser microscope with polarization-sensitive detectors. By using a high numerical aperture objective, a spatial resolution of about 240 nm at a wavelength of 405 nm is achieved. The sample is mounted on a 4 He continuous flow cryostat providing a temperature range between 4 K and 300 K, and electromagnets are used to apply magnetic fields of up to 800 mT with variable in-plane orientation and 20 mT with out-of-plane orientation. Typical applications of the polarizing microscope are the imaging of the in-plane and out-of-plane magnetization via the longitudinal and polar MOKE, imaging of magnetic flux structures in superconductors covered with a magneto-optical indicator film via the Faraday effect, or imaging of structural features, such as twin-walls in tetragonal SrTiO 3 . The scanning laser microscope furthermore offers the possibility to gain local information on electric transport properties of a sample by detecting the beam-induced voltage change across a current-biased sample. This combination of magnetic, structural, and electric imaging capabilities makes the microscope a viable tool for research in the fields of oxide electronics, spintronics, magnetism, and superconductivity.

A high-resolution combined scanning laser and widefield polarizing microscope for imaging at temperatures from 4 K to 300 K

NASA Astrophysics Data System (ADS)

Lange, M.; Guénon, S.; Lever, F.; Kleiner, R.; Koelle, D.

2017-12-01

Polarized light microscopy, as a contrast-enhancing technique for optically anisotropic materials, is a method well suited for the investigation of a wide variety of effects in solid-state physics, as, for example, birefringence in crystals or the magneto-optical Kerr effect (MOKE). We present a microscopy setup that combines a widefield microscope and a confocal scanning laser microscope with polarization-sensitive detectors. By using a high numerical aperture objective, a spatial resolution of about 240 nm at a wavelength of 405 nm is achieved. The sample is mounted on a 4He continuous flow cryostat providing a temperature range between 4 K and 300 K, and electromagnets are used to apply magnetic fields of up to 800 mT with variable in-plane orientation and 20 mT with out-of-plane orientation. Typical applications of the polarizing microscope are the imaging of the in-plane and out-of-plane magnetization via the longitudinal and polar MOKE, imaging of magnetic flux structures in superconductors covered with a magneto-optical indicator film via the Faraday effect, or imaging of structural features, such as twin-walls in tetragonal SrTiO3. The scanning laser microscope furthermore offers the possibility to gain local information on electric transport properties of a sample by detecting the beam-induced voltage change across a current-biased sample. This combination of magnetic, structural, and electric imaging capabilities makes the microscope a viable tool for research in the fields of oxide electronics, spintronics, magnetism, and superconductivity.

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

PubMed Central

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. PMID:23823461

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

PubMed

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.

Versatile optical system for static and dynamic thermomagnetic recording using a scanning laser microscope

NASA Astrophysics Data System (ADS)

Clegg, Warwick W.; Jenkins, David F. L.; Helian, Na; Windmill, James; Windmill, Robert

2001-12-01

Scanning Laser Microscopes (SLM) have been used to characterise the magnetic domain properties of various magnetic and magneto-optical materials. The SLM in our laboratory has been designed to enable both static and dynamic read-write operations to be performed on stationary media. In a conventional (static) SLM, data bits are recorded thermo-magnetically by focusing a pulse of laser light onto the sample surface. If the laser beam has a Gaussian intensity distribution (TEM00) then so will the focused laser spot. The resultant temperature profile will largely mirror the intensity distribution of the focused spot, and in the region where the temperature is sufficiently high for switching to occur, in the presence of bias field, a circular data bit will be recorded. However, in a real magneto-optical drive the bits are written onto non-stationary media, and the resultant bit will be non-circular. A versatile optical system has been developed to facilitate both recording and imaging of data bits. To simulate the action of a Magneto-Optical drive, the laser is pulsed via an Acousto-Optic Modulator, whilst being scanned across the sample using a galvanometer mounted mirror, thus imitating a storage medium rotating above a MO head with high relative velocity between the beam and medium. Static recording is simply achieved by disabling the galvanometer scan mirror. Polar magneto-optic Kerr effect images are acquired using multiple-segment photo-detectors for diffraction-limited scanned spot detection, with either specimen scanning for highest resolution or beam scanning for near real-time image acquisition. Results will be presented to illustrate the systems capabilities.

Two-photon laser scanning microscopy with electrowetting-based prism scanning

PubMed Central

Supekar, Omkar D.; Ozbay, Baris N.; Zohrabi, Mo; Nystrom, Philip D.; Futia, Gregory L.; Restrepo, Diego; Gibson, Emily A.; Gopinath, Juliet T.; Bright, Victor M.

2017-01-01

Laser scanners are an integral part of high resolution biomedical imaging systems such as confocal or 2-photon excitation (2PE) microscopes. In this work, we demonstrate the utility of electrowetting on dielectric (EWOD) prisms as a lateral laser-scanning element integrated in a conventional 2PE microscope. To the best of our knowledge, this is the first such demonstration for EWOD prisms. EWOD devices provide a transmissive, low power consuming, and compact alternative to conventional adaptive optics, and hence this technology has tremendous potential. We demonstrate 2PE microscope imaging of cultured mouse hippocampal neurons with a FOV of 130 × 130 μm2 using EWOD prism scanning. In addition, we show simulations of the optical system with the EWOD prism, to evaluate the effect of propagating a Gaussian beam through the EWOD prism on the imaging quality. Based on the simulation results a beam size of 0.91 mm full width half max was chosen to conduct the imaging experiments, resulting in a numerical aperture of 0.17 of the imaging system. PMID:29296477

Joint Services Electronics Program Annual Progress Report.

DTIC Science & Technology

1987-10-15

polarizability of free carriers in the semiconductor perturb the index of refraction which can be detected in a Nomarski -type optical interferometer. For...interferomters. However, the charge probe relies on a different physical effect and operates by interferometrically detecting the phase change induced in an... Nomarski microscope systems. These techniques will be applied, eventually, in our real-time V.. scanning optical microscope described below. Recently

Scanning electron microscope/energy dispersive x ray analysis of impact residues in LDEF tray clamps

NASA Technical Reports Server (NTRS)

Bernhard, Ronald P.; Durin, Christian; Zolensky, Michael E.

1993-01-01

Detailed optical scanning of tray clamps is being conducted in the Facility for the Optical Inspection of Large Surfaces at JSC to locate and document impacts as small as 40 microns in diameter. Residues from selected impacts are then being characterized by Scanning Electron Microscopy/Energy Dispersive X-ray Analysis at CNES. Results from this analysis will be the initial step to classifying projectile residues into specific sources.

In vivo imaging of the Drosophila Melanogaster heart using a novel optical coherence tomography microscope

NASA Astrophysics Data System (ADS)

Izatt, Susan D.; Choma, Michael A.; Israel, Steven; Wessells, Robert J.; Bodmer, Rolf; Izatt, Joseph A.

2005-03-01

Real time in vivo optical coherence tomography (OCT) imaging of the adult fruit fly Drosophila melanogaster heart using a newly designed OCT microscope allows accurate assessment of cardiac anatomy and function. D. melanogaster has been used extensively in genetic research for over a century, but in vivo evaluation of the heart has been limited by available imaging technology. The ability to assess phenotypic changes with micrometer-scale resolution noninvasively in genetic models such as D. melanogaster is needed in the advancing fields of developmental biology and genetics. We have developed a dedicated small animal OCT imaging system incorporating a state-of-the-art, real time OCT scanner integrated into a standard stereo zoom microscope which allows for simultaneous OCT and video imaging. System capabilities include A-scan, B-scan, and M-scan imaging as well as automated 3D volumetric acquisition and visualization. Transverse and sagittal B-mode scans of the four chambered D. melanogaster heart have been obtained with the OCT microscope and are consistent with detailed anatomical studies from the literature. Further analysis by M-mode scanning is currently under way to assess cardiac function as a function of age and sex by determination of shortening fraction and ejection fraction. These studies create control cardiac data on the wild type D. melanogaster, allowing subsequent evaluation of phenotypic cardiac changes in this model after regulated genetic mutation.

Miniature in vivo MEMS-based line-scanned dual-axis confocal microscope for point-of-care pathology

PubMed Central

Yin, C.; Glaser, A.K.; Leigh, S. Y.; Chen, Y.; Wei, L.; Pillai, P. C. S.; Rosenberg, M. C.; Abeytunge, S.; Peterson, G.; Glazowski, C.; Sanai, N.; Mandella, M. J.; Rajadhyaksha, M.; Liu, J. T. C.

2016-01-01

There is a need for miniature optical-sectioning microscopes to enable in vivo interrogation of tissues as a real-time and noninvasive alternative to gold-standard histopathology. Such devices could have a transformative impact for the early detection of cancer as well as for guiding tumor-resection procedures. Miniature confocal microscopes have been developed by various researchers and corporations to enable optical sectioning of highly scattering tissues, all of which have necessitated various trade-offs in size, speed, depth selectivity, field of view, resolution, image contrast, and sensitivity. In this study, a miniature line-scanned (LS) dual-axis confocal (DAC) microscope, with a 12-mm diameter distal tip, has been developed for clinical point-of-care pathology. The dual-axis architecture has demonstrated an advantage over the conventional single-axis confocal configuration for reducing background noise from out-of-focus and multiply scattered light. The use of line scanning enables fast frame rates (16 frames/sec is demonstrated here, but faster rates are possible), which mitigates motion artifacts of a hand-held device during clinical use. We have developed a method to actively align the illumination and collection beams in a DAC microscope through the use of a pair of rotatable alignment mirrors. Incorporation of a custom objective lens, with a small form factor for in vivo clinical use, enables our device to achieve an optical-sectioning thickness and lateral resolution of 2.0 and 1.1 microns respectively. Validation measurements with reflective targets, as well as in vivo and ex vivo images of tissues, demonstrate the clinical potential of this high-speed optical-sectioning microscopy device. PMID:26977337

Development of a Hybrid Atomic Force Microscopic Measurement System Combined with White Light Scanning Interferometry

PubMed Central

Guo, Tong; Wang, Siming; Dorantes-Gonzalez, Dante J.; Chen, Jinping; Fu, Xing; Hu, Xiaotang

2012-01-01

A hybrid atomic force microscopic (AFM) measurement system combined with white light scanning interferometry for micro/nanometer dimensional measurement is developed. The system is based on a high precision large-range positioning platform with nanometer accuracy on which a white light scanning interferometric module and an AFM head are built. A compact AFM head is developed using a self-sensing tuning fork probe. The head need no external optical sensors to detect the deflection of the cantilever, which saves room on the head, and it can be directly fixed under an optical microscopic interferometric system. To enhance the system’s dynamic response, the frequency modulation (FM) mode is adopted for the AFM head. The measuring data can be traceable through three laser interferometers in the system. The lateral scanning range can reach 25 mm × 25 mm by using a large-range positioning platform. A hybrid method combining AFM and white light scanning interferometry is proposed to improve the AFM measurement efficiency. In this method, the sample is measured firstly by white light scanning interferometry to get an overall coarse morphology, and then, further measured with higher resolution by AFM. Several measuring experiments on standard samples demonstrate the system’s good measurement performance and feasibility of the hybrid measurement method. PMID:22368463

Development of a hybrid atomic force microscopic measurement system combined with white light scanning interferometry.

PubMed

Guo, Tong; Wang, Siming; Dorantes-Gonzalez, Dante J; Chen, Jinping; Fu, Xing; Hu, Xiaotang

2012-01-01

A hybrid atomic force microscopic (AFM) measurement system combined with white light scanning interferometry for micro/nanometer dimensional measurement is developed. The system is based on a high precision large-range positioning platform with nanometer accuracy on which a white light scanning interferometric module and an AFM head are built. A compact AFM head is developed using a self-sensing tuning fork probe. The head need no external optical sensors to detect the deflection of the cantilever, which saves room on the head, and it can be directly fixed under an optical microscopic interferometric system. To enhance the system's dynamic response, the frequency modulation (FM) mode is adopted for the AFM head. The measuring data can be traceable through three laser interferometers in the system. The lateral scanning range can reach 25 mm × 25 mm by using a large-range positioning platform. A hybrid method combining AFM and white light scanning interferometry is proposed to improve the AFM measurement efficiency. In this method, the sample is measured firstly by white light scanning interferometry to get an overall coarse morphology, and then, further measured with higher resolution by AFM. Several measuring experiments on standard samples demonstrate the system's good measurement performance and feasibility of the hybrid measurement method.

A simple but precise method for quantitative measurement of the quality of the laser focus in a scanning optical microscope

PubMed Central

MACRAE, K.; TRAVIS, C.; AMOR, R.; NORRIS, G.; WILSON, S.H.; OPPO, G.‐L.; MCCONNELL, G.

2015-01-01

Summary We report a method for characterizing the focussing laser beam exiting the objective in a laser scanning microscope. This method provides the size of the optical focus, the divergence of the beam, the ellipticity and the astigmatism. We use a microscopic‐scale knife edge in the form of a simple transmission electron microscopy grid attached to a glass microscope slide, and a light‐collecting optical fibre and photodiode underneath the specimen. By scanning the laser spot from a reflective to a transmitting part of the grid, a beam profile in the form of an error function can be obtained and by repeating this with the knife edge at different axial positions relative to the beam waist, the divergence and astigmatism of the postobjective laser beam can be obtained. The measured divergence can be used to quantify how much of the full numerical aperture of the lens is used in practice. We present data of the beam radius, beam divergence, ellipticity and astigmatism obtained with low (0.15, 0.7) and high (1.3) numerical aperture lenses and lasers commonly used in confocal and multiphoton laser scanning microscopy. Our knife‐edge method has several advantages over alternative knife‐edge methods used in microscopy including that the knife edge is easy to prepare, that the beam can be characterized also directly under a cover slip, as necessary to reduce spherical aberrations for objectives designed to be used with a cover slip, and it is suitable for use with commercial laser scanning microscopes where access to the laser beam can be limited. PMID:25864964

Screening small-molecule compound microarrays for protein ligands without fluorescence labeling with a high-throughput scanning microscope.

PubMed

Fei, Yiyan; Landry, James P; Sun, Yungshin; Zhu, Xiangdong; Wang, Xiaobing; Luo, Juntao; Wu, Chun-Yi; Lam, Kit S

2010-01-01

We describe a high-throughput scanning optical microscope for detecting small-molecule compound microarrays on functionalized glass slides. It is based on measurements of oblique-incidence reflectivity difference and employs a combination of a y-scan galvometer mirror and an x-scan translation stage with an effective field of view of 2 cm x 4 cm. Such a field of view can accommodate a printed small-molecule compound microarray with as many as 10,000 to 20,000 targets. The scanning microscope is capable of measuring kinetics as well as endpoints of protein-ligand reactions simultaneously. We present the experimental results on solution-phase protein reactions with small-molecule compound microarrays synthesized from one-bead, one-compound combinatorial chemistry and immobilized on a streptavidin-functionalized glass slide.

Screening small-molecule compound microarrays for protein ligands without fluorescence labeling with a high-throughput scanning microscope

PubMed Central

Fei, Yiyan; Landry, James P.; Sun, Yungshin; Zhu, Xiangdong; Wang, Xiaobing; Luo, Juntao; Wu, Chun-Yi; Lam, Kit S.

2010-01-01

We describe a high-throughput scanning optical microscope for detecting small-molecule compound microarrays on functionalized glass slides. It is based on measurements of oblique-incidence reflectivity difference and employs a combination of a y-scan galvometer mirror and an x-scan translation stage with an effective field of view of 2 cm×4 cm. Such a field of view can accommodate a printed small-molecule compound microarray with as many as 10,000 to 20,000 targets. The scanning microscope is capable of measuring kinetics as well as endpoints of protein-ligand reactions simultaneously. We present the experimental results on solution-phase protein reactions with small-molecule compound microarrays synthesized from one-bead, one-compound combinatorial chemistry and immobilized on a streptavidin-functionalized glass slide. PMID:20210464

Fiber optic light collection system for scanning-tunneling-microscope-induced light emission.

PubMed

Watkins, Neil J; Long, James P; Kafafi, Zakya H; Mäkinen, Antti J

2007-05-01

We report a compact light collection scheme suitable for retrofitting a scanning tunneling microscope (STM) for STM-induced light emission experiments. The approach uses a pair of optical fibers with large core diameters and high numerical apertures to maximize light collection efficiency and to moderate the mechanical precision required for alignment. Bench tests indicate that efficiency reduction is almost entirely due to reflective losses at the fiber ends, while losses due to fiber misalignment have virtually been eliminated. Photon-map imaging with nanometer features is demonstrated on a stepped Au(111) surface with signal rates exceeding 10(4) counts/s.

A novel design of a scanning probe microscope integrated with an ultramicrotome for serial block-face nanotomography

NASA Astrophysics Data System (ADS)

Efimov, Anton E.; Agapov, Igor I.; Agapova, Olga I.; Oleinikov, Vladimir A.; Mezin, Alexey V.; Molinari, Michael; Nabiev, Igor; Mochalov, Konstantin E.

2017-02-01

We present a new concept of a combined scanning probe microscope (SPM)/ultramicrotome apparatus. It enables "slice-and-view" scanning probe nanotomography measurements and 3D reconstruction of the bulk sample nanostructure from series of SPM images after consecutive ultrathin sections. The sample is fixed on a flat XYZ scanning piezostage mounted on the ultramicrotome arm. The SPM measuring head with a cantilever tip and a laser-photodiode tip detection system approaches the sample for SPM measurements of the block-face surface immediately after the ultramicrotome sectioning is performed. The SPM head is moved along guides that are also fixed on the ultramicrotome arm. Thereby, relative dysfunctional displacements of the tip, the sample, and the ultramicrotome knife are minimized. The design of the SPM head enables open frontal optical access to the sample block-face adapted for high-resolution optical lenses for correlative SPM/optical microscopy applications. The new system can be used in a wide range of applications for the study of 3D nanostructures of biological objects, biomaterials, polymer nanocomposites, and nanohybrid materials in various SPM and optical microscopy measuring modes.

Wide-band acousto-optic deflectors for large field of view two-photon microscope.

PubMed

Jiang, Runhua; Zhou, Zhenqiao; Lv, Xiaohua; Zeng, Shaoqun

2012-04-01

Acousto-optic deflector (AOD) is an attractive scanner for two-photon microscopy because it can provide fast and versatile laser scanning and does not involve any mechanical movements. However, due to the small scan range of available AOD, the field of view (FOV) of the AOD-based microscope is typically smaller than that of the conventional galvanometer-based microscope. Here, we developed a novel wide-band AOD to enlarge the scan angle. Considering the maximum acceptable acoustic attenuation in the acousto-optic crystal, relatively lower operating frequencies and moderate aperture were adopted. The custom AOD was able to provide 60 MHz 3-dB bandwidth and 80% peak diffraction efficiency at 840 nm wavelength. Based on a pair of such AOD, a large FOV two-photon microscope was built with a FOV up to 418.5 μm (40× objective). The spatiotemporal dispersion was compensated simultaneously with a single custom-made prism. By means of dynamic power modulation, the variation of laser intensity within the FOV was reduced below 5%. The lateral and axial resolution of the system were 0.58-2.12 μm and 2.17-3.07 μm, respectively. Pollen grain images acquired by this system were presented to demonstrate the imaging capability at different positions across the entire FOV. © 2012 American Institute of Physics

Optical Interferometric Micrometrology

NASA Technical Reports Server (NTRS)

Abel, Phillip B.; Lauer, James R.

1989-01-01

Resolutions in angstrom and subangstrom range sought for atomic-scale surface probes. Experimental optical micrometrological system built to demonstrate calibration of piezoelectric transducer to displacement sensitivity of few angstroms. Objective to develop relatively simple system producing and measuring translation, across surface of specimen, of stylus in atomic-force or scanning tunneling microscope. Laser interferometer used to calibrate piezoelectric transducer used in atomic-force microscope. Electronic portion of calibration system made of commercially available components.

Interferometric scanning optical microscope for surface characterization.

PubMed

Offside, M J; Somekh, M G

1992-11-01

A phase-sensitive scanning optical microscope is described that can measure surface height changes down to 0.1 nm. This is achieved by using two heterodyne Michelson interferometers in parallel. One interferometer probes the sample with a tightly focused beam, and the second has a collimated beam that illuminates a large area of the surface, providing a large area on sample reference. This is facilitated by using a specially constructed objective lens that permits the relative areas illuminated by the two probe beams to be varied both arbitrarily and independently, thus ensuring an accurate absolute phase measurement. We subtracted the phase outputs from each interferometer to provide the sample phase information, canceling the phase noise resulting from microphonics in the process. Results from a prototype version of the microscope are presented that demonstrate the advantages of the system over existing techniques.

Correction of image drift and distortion in a scanning electron microscopy.

PubMed

Jin, P; Li, X

2015-12-01

Continuous research on small-scale mechanical structures and systems has attracted strong demand for ultrafine deformation and strain measurements. Conventional optical microscope cannot meet such requirements owing to its lower spatial resolution. Therefore, high-resolution scanning electron microscope has become the preferred system for high spatial resolution imaging and measurements. However, scanning electron microscope usually is contaminated by distortion and drift aberrations which cause serious errors to precise imaging and measurements of tiny structures. This paper develops a new method to correct drift and distortion aberrations of scanning electron microscope images, and evaluates the effect of correction by comparing corrected images with scanning electron microscope image of a standard sample. The drift correction is based on the interpolation scheme, where a series of images are captured at one location of the sample and perform image correlation between the first image and the consequent images to interpolate the drift-time relationship of scanning electron microscope images. The distortion correction employs the axial symmetry model of charged particle imaging theory to two images sharing with the same location of one object under different imaging fields of view. The difference apart from rigid displacement between the mentioned two images will give distortion parameters. Three-order precision is considered in the model and experiment shows that one pixel maximum correction is obtained for the employed high-resolution electron microscopic system. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Two-photon microscopy and spectroscopy based on a compact confocal scanning head

NASA Astrophysics Data System (ADS)

Diaspro, Alberto; Chirico, Giberto; Federici, Federico; Cannone, Fabio; Beretta, Sabrina; Robello, Mauro; Olivini, Francesca; Ramoino, Paola

2001-07-01

We have combined a confocal laser scanning head modified for TPE (two-photon excitation) microscopy with some spectroscopic modules to study single molecules and molecular aggregates. The behavior of the TPE microscope unit has been characterized by means of point spread function measurements and of the demonstration of its micropatterning abilities. One-photon and two-photon mode can be simply accomplished by switching from a mono-mode optical fiber (one-photon) coupled to conventional laser sources to an optical module that allows IR laser beam (two- photon/TPE) delivery to the confocal laser scanning head. We have then described the characterization of the two-photon microscope for spectroscopic applications: fluorescence correlation, lifetime and fluorescence polarization anisotropy measurements. We describe the measurement of the response of the two-photon microscope to the light polarization and discuss fluorescence polarization anisotropy measurements on Rhodamine 6G as a function of the viscosity and on a globular protein, the Beta-lactoglobulin B labeled with Alexa 532 at very high dilutions. The average rotational and translational diffusion coefficients measured with fluorescence polarization anisotropy and fluorescence correlation methods are in good agreement with the protein size, therefore validating the use of the microscope for two-photon spectroscopy on biomolecules.

Three-dimensional reconstruction of highly complex microscopic samples using scanning electron microscopy and optical flow estimation.

PubMed

Baghaie, Ahmadreza; Pahlavan Tafti, Ahmad; Owen, Heather A; D'Souza, Roshan M; Yu, Zeyun

2017-01-01

Scanning Electron Microscope (SEM) as one of the major research and industrial equipment for imaging of micro-scale samples and surfaces has gained extensive attention from its emerge. However, the acquired micrographs still remain two-dimensional (2D). In the current work a novel and highly accurate approach is proposed to recover the hidden third-dimension by use of multi-view image acquisition of the microscopic samples combined with pre/post-processing steps including sparse feature-based stereo rectification, nonlocal-based optical flow estimation for dense matching and finally depth estimation. Employing the proposed approach, three-dimensional (3D) reconstructions of highly complex microscopic samples were achieved to facilitate the interpretation of topology and geometry of surface/shape attributes of the samples. As a byproduct of the proposed approach, high-definition 3D printed models of the samples can be generated as a tangible means of physical understanding. Extensive comparisons with the state-of-the-art reveal the strength and superiority of the proposed method in uncovering the details of the highly complex microscopic samples.

A novel optical rotary encoder with eccentricity self-detection ability.

PubMed

Li, Xuan; Ye, Guoyong; Liu, Hongzhong; Ban, Yaowen; Shi, Yongsheng; Yin, Lei; Lu, Bingheng

2017-11-01

Eccentricity error is the main error source of optical rotary encoders. Real-time detection and compensation of the eccentricity error is an effective way of improving the accuracy of rotary optical encoders. In this paper, a novel rotary optical encoder is presented to realize eccentricity self-detection. The proposed encoder adopts a spider-web-patterned scale grating as a measuring standard which is scanned by a dual-head scanning unit. Two scanning heads of the dual-head scanning unit, which are arranged orthogonally, have the function of scanning the periodic pattern of the scale grating along the angular and radial directions, respectively. By this means, synchronous measurement of angular and radial displacements of the scale grating is realized. This paper gives the details of the operating principle of the rotary optical encoder, developing and testing work of a prototype. The eccentricity self-detection result agrees well with the result measured by an optical microscope. The experimental result preliminarily proves the feasibility and effectiveness of the proposed optical encoder.

A novel optical rotary encoder with eccentricity self-detection ability

NASA Astrophysics Data System (ADS)

Li, Xuan; Ye, Guoyong; Liu, Hongzhong; Ban, Yaowen; Shi, Yongsheng; Yin, Lei; Lu, Bingheng

2017-11-01

Eccentricity error is the main error source of optical rotary encoders. Real-time detection and compensation of the eccentricity error is an effective way of improving the accuracy of rotary optical encoders. In this paper, a novel rotary optical encoder is presented to realize eccentricity self-detection. The proposed encoder adopts a spider-web-patterned scale grating as a measuring standard which is scanned by a dual-head scanning unit. Two scanning heads of the dual-head scanning unit, which are arranged orthogonally, have the function of scanning the periodic pattern of the scale grating along the angular and radial directions, respectively. By this means, synchronous measurement of angular and radial displacements of the scale grating is realized. This paper gives the details of the operating principle of the rotary optical encoder, developing and testing work of a prototype. The eccentricity self-detection result agrees well with the result measured by an optical microscope. The experimental result preliminarily proves the feasibility and effectiveness of the proposed optical encoder.

High contrast imaging and flexible photomanipulation for quantitative in vivo multiphoton imaging with polygon scanning microscope.

PubMed

Li, Yongxiao; Montague, Samantha J; Brüstle, Anne; He, Xuefei; Gillespie, Cathy; Gaus, Katharina; Gardiner, Elizabeth E; Lee, Woei Ming

2018-02-28

In this study, we introduce two key improvements that overcome limitations of existing polygon scanning microscopes while maintaining high spatial and temporal imaging resolution over large field of view (FOV). First, we proposed a simple and straightforward means to control the scanning angle of the polygon mirror to carry out photomanipulation without resorting to high speed optical modulators. Second, we devised a flexible data sampling method directly leading to higher image contrast by over 2-fold and digital images with 100 megapixels (10 240 × 10 240) per frame at 0.25 Hz. This generates sub-diffraction limited pixels (60 nm per pixels over the FOV of 512 μm) which increases the degrees of freedom to extract signals computationally. The unique combined optical and digital control recorded fine fluorescence recovery after localized photobleaching (r ~10 μm) within fluorescent giant unilamellar vesicles and micro-vascular dynamics after laser-induced injury during thrombus formation in vivo. These new improvements expand the quantitative biological-imaging capacity of any polygon scanning microscope system. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Optical sectioning microscopes with no moving parts using a micro-stripe array light emitting diode.

PubMed

Poher, V; Zhang, H X; Kennedy, G T; Griffin, C; Oddos, S; Gu, E; Elson, D S; Girkin, M; French, P M W; Dawson, M D; Neil, M A

2007-09-03

We describe an optical sectioning microscopy system with no moving parts based on a micro-structured stripe-array light emitting diode (LED). By projecting arbitrary line or grid patterns onto the object, we are able to implement a variety of optical sectioning microscopy techniques such as grid-projection structured illumination and line scanning confocal microscopy, switching from one imaging technique to another without modifying the microscope setup. The micro-structured LED and driver are detailed and depth discrimination capabilities are measured and calculated.

A pragmatic guide to multiphoton microscope design

PubMed Central

Young, Michael D.; Field, Jeffrey J.; Sheetz, Kraig E.; Bartels, Randy A.; Squier, Jeff

2016-01-01

Multiphoton microscopy has emerged as a ubiquitous tool for studying microscopic structure and function across a broad range of disciplines. As such, the intent of this paper is to present a comprehensive resource for the construction and performance evaluation of a multiphoton microscope that will be understandable to the broad range of scientific fields that presently exploit, or wish to begin exploiting, this powerful technology. With this in mind, we have developed a guide to aid in the design of a multiphoton microscope. We discuss source selection, optical management of dispersion, image-relay systems with scan optics, objective-lens selection, single-element light-collection theory, photon-counting detection, image rendering, and finally, an illustrated guide for building an example microscope. PMID:27182429

Note: optical optimization for ultrasensitive photon mapping with submolecular resolution by scanning tunneling microscope induced luminescence.

PubMed

Chen, L G; Zhang, C; Zhang, R; Zhang, X L; Dong, Z C

2013-06-01

We report the development of a custom scanning tunneling microscope equipped with photon collection and detection systems. The optical optimization includes the comprehensive design of aspherical lens for light collimation and condensing, the sophisticated piezo stages for in situ lens adjustment inside ultrahigh vacuum, and the fiber-free coupling of collected photons directly onto the ultrasensitive single-photon detectors. We also demonstrate submolecular photon mapping for the molecular islands of porphyrin on Ag(111) under small tunneling currents down to 10 pA and short exposure time down to 1.2 ms/pixel. A high quantum efficiency up to 10(-2) was also observed.

Photothermal camera port accessory for microscopic thermal diffusivity imaging

NASA Astrophysics Data System (ADS)

Escola, Facundo Zaldívar; Kunik, Darío; Mingolo, Nelly; Martínez, Oscar Eduardo

2016-06-01

The design of a scanning photothermal accessory is presented, which can be attached to the camera port of commercial microscopes to measure thermal diffusivity maps with micrometer resolution. The device is based on the thermal expansion recovery technique, which measures the defocusing of a probe beam due to the curvature induced by the local heat delivered by a focused pump beam. The beam delivery and collecting optics are built using optical fiber technology, resulting in a robust optical system that provides collinear pump and probe beams without any alignment adjustment necessary. The quasiconfocal configuration for the signal collection using the same optical fiber sets very restrictive conditions on the positioning and alignment of the optical components of the scanning unit, and a detailed discussion of the design equations is presented. The alignment procedure is carefully described, resulting in a system so robust and stable that no further alignment is necessary for the day-to-day use, becoming a tool that can be used for routine quality control, operated by a trained technician.

The research progress of metrological 248nm deep ultraviolent microscope inspection device

NASA Astrophysics Data System (ADS)

Wang, Zhi-xin; Li, Qi; Gao, Si-tian; Shi, Yu-shu; Li, Wei; Li, Shi

2016-01-01

In lithography process, the precision of wafer pattern to a large extent depends on the geometric dimensioning and tolerance of photomasks when accuracy of lithography aligner is certain. Since the minimum linewidth (Critical Dimension) of the aligner exposing shrinks to a few tens of nanometers in size, one-tenth of tolerance errors in fabrication may lead to microchip function failure, so it is very important to calibrate these errors of photomasks. Among different error measurement instruments, deep ultraviolent (DUV) microscope because of its high resolution, as well as its advantages compared to scanning probe microscope restrained by measuring range and scanning electron microscope restrained by vacuum environment, makes itself the most suitable apparatus. But currently there is very few DUV microscope adopting 248nm optical system, means it can attain 80nm resolution; furthermore, there is almost no DUV microscope possessing traceable calibration capability. For these reason, the National Institute of Metrology, China is developing a metrological 248nm DUV microscope mainly consists of DUV microscopic components, PZT and air supporting stages as well as interferometer calibration framework. In DUV microscopic component, the Köhler high aperture transmit condenser, DUV splitting optical elements and PMT pinhole scanning elements are built. In PZT and air supporting stages, a novel PZT actuating flexural hinge stage nested separate X, Y direction kinematics and a friction wheel driving long range air supporting stage are researched. In interferometer framework, a heterodyne multi-pass interferometer measures XY axis translation and Z axis rotation through Zerodur mirror mounted on stage. It is expected the apparatus has the capability to calibrate one dimensional linewidths and two dimensional pitches ranging from 200nm to 50μm with expanded uncertainty below 20nm.

Rotary-scanning optical resolution photoacoustic microscopy

NASA Astrophysics Data System (ADS)

Qi, Weizhi; Xi, Lei

2016-10-01

Optical resolution photoacoustic microscopy (ORPAM) is currently one of the fastest evolving photoacoustic imaging modalities. It has a comparable spatial resolution to pure optical microscopic techniques such as epifluorescence microscopy, confocal microscopy, and two-photon microscopy, but also owns a deeper penetration depth. In this paper, we report a rotary-scanning (RS)-ORPAM that utilizes a galvanometer scanner integrated with objective to achieve rotary laser scanning. A 15 MHz cylindrically focused ultrasonic transducer is mounted onto a motorized rotation stage to follow optical scanning traces synchronously. To minimize the loss of signal to noise ratio, the acoustic focus is precisely adjusted to reach confocal with optical focus. Black tapes and carbon fibers are firstly imaged to evaluate the performance of the system, and then in vivo imaging of vasculature networks inside the ears and brains of mice is demonstrated using this system.

The Development of a Scanning Soft X-Ray Microscope.

NASA Astrophysics Data System (ADS)

Rarback, Harvey Miles

We have developed a scanning soft X-ray microscope, which can be used to image natural biological specimens at high resolution and with less damage than electron microscopy. The microscope focuses a monochromatic beam of synchrotron radiation to a nearly diffraction limited spot with the aid of a high resolution Fresnel zone plate, specially fabricated for us at the IBM Watson Research Center. The specimen at one atmosphere is mechanically scanned through the spot and the transmitted radiation is efficiently detected with a flow proportional counter. A computer forms a realtime transmission image of the specimen which is displayed on a color monitor. Our first generation optics have produced images of natural wet specimens at a resolution of 300 nm.

Attainment of 40.5 pm spatial resolution using 300 kV scanning transmission electron microscope equipped with fifth-order aberration corrector.

PubMed

Morishita, Shigeyuki; Ishikawa, Ryo; Kohno, Yuji; Sawada, Hidetaka; Shibata, Naoya; Ikuhara, Yuichi

2018-02-01

The achievement of a fine electron probe for high-resolution imaging in scanning transmission electron microscopy requires technological developments, especially in electron optics. For this purpose, we developed a microscope with a fifth-order aberration corrector that operates at 300 kV. The contrast flat region in an experimental Ronchigram, which indicates the aberration-free angle, was expanded to 70 mrad. By using a probe with convergence angle of 40 mrad in the scanning transmission electron microscope at 300 kV, we attained the spatial resolution of 40.5 pm, which is the projected interatomic distance between Ga-Ga atomic columns of GaN observed along [212] direction.

Imaging properties and its improvements of scanning/imaging x-ray microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Takeuchi, Akihisa, E-mail: take@spring8.or.jp; Uesugi, Kentaro; Suzuki, Yoshio

A scanning / imaging X-ray microscope (SIXM) system has been developed at SPring-8. The SIXM consists of a scanning X-ray microscope with a one-dimensional (1D) X-ray focusing device and an imaging (full-field) X-ray microscope with a 1D X-ray objective. The motivation of the SIXM system is to realize a quantitative and highly-sensitive multimodal 3D X-ray tomography by taking advantages of both the scanning X-ray microscope using multi-pixel detector and the imaging X-ray microscope. Data acquisition process of a 2D image is completely different between in the horizontal direction and in the vertical direction; a 1D signal is obtained with themore » linear-scanning while the other dimensional signal is obtained with the imaging optics. Such condition have caused a serious problem on the imaging properties that the imaging quality in the vertical direction has been much worse than that in the horizontal direction. In this paper, two approaches to solve this problem will be presented. One is introducing a Fourier transform method for phase retrieval from one phase derivative image, and the other to develop and employ a 1D diffuser to produce an asymmetrical coherent illumination.« less

Compensation of temporal and spatial dispersion for multiphoton acousto-optic laser-scanning microscopy

NASA Astrophysics Data System (ADS)

Iyer, Vijay; Saggau, Peter

2003-10-01

In laser-scanning microscopy, acousto-optic (AO) deflection provides a means to quickly position a laser beam to random locations throughout the field-of-view. Compared to conventional laser-scanning using galvanometer-driven mirrors, this approach increases the frame rate and signal-to-noise ratio, and reduces time spent illuminating sites of no interest. However, random-access AO scanning has not yet been combined with multi-photon microscopy, primarily because the femtosecond laser pulses employed are subject to significant amounts of both spatial and temporal dispersion upon propagation through common AO materials. Left uncompensated, spatial dispersion reduces the microscope"s spatial resolution while temporal dispersion reduces the multi-photon excitation efficacy. In previous work, we have demonstrated, 1) the efficacy of a single diffraction grating scheme which reduces the spatial dispersion at least 3-fold throughout the field-of-view, and 2) the use of a novel stacked-prism pre-chirper for compensating the temporal dispersion of a pair of AODs using a shorter mechanical path length (2-4X) than standard prism-pair arrangements. In this work, we demonstrate for the first time the use of these compensation approaches with a custom-made large-area slow-shear TeO2 AOD specifically suited for the development of a high-resolution 2-D random-access AO scanning multi-photon laser-scanning microscope (AO-MPLSM).

Development of an add-on kit for scanning confocal microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Guo, Kaikai; Zheng, Guoan

2017-03-01

Scanning confocal microscopy is a standard choice for many fluorescence imaging applications in basic biomedical research. It is able to produce optically sectioned images and provide acquisition versatility to address many samples and application demands. However, scanning a focused point across the specimen limits the speed of image acquisition. As a result, scanning confocal microscope only works well with stationary samples. Researchers have performed parallel confocal scanning using digital-micromirror-device (DMD), which was used to project a scanning multi-point pattern across the sample. The DMD based parallel confocal systems increase the imaging speed while maintaining the optical sectioning ability. In this paper, we report the development of an add-on kit for high-speed and low-cost confocal microscopy. By adapting this add-on kit to an existing regular microscope, one can convert it into a confocal microscope without significant hardware modifications. Compared with current DMD-based implementations, the reported approach is able to recover multiple layers along the z axis simultaneously. It may find applications in wafer inspection and 3D metrology of semiconductor circuit. The dissemination of the proposed add-on kit under $1000 budget could also lead to new types of experimental designs for biological research labs, e.g., cytology analysis in cell culture experiments, genetic studies on multicellular organisms, pharmaceutical drug profiling, RNA interference studies, investigation of microbial communities in environmental systems, and etc.

Multiplane optical microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Tongcang; Ota, Sadao; Kim, Jeongmin

This disclosure provides systems, methods, and apparatus related to optical microscopy. In one aspect, an apparatus includes a sample holder, a first objective lens, a plurality of optical components, a second objective lens, and a mirror. The apparatus may directly image a cross-section of a sample oblique to or parallel to the optical axis of the first objective lens, without scanning.

Scanning Miniature Microscopes without Lenses

NASA Technical Reports Server (NTRS)

Wang, Yu

2009-01-01

The figure schematically depicts some alternative designs of proposed compact, lightweight optoelectronic microscopes that would contain no lenses and would generate magnified video images of specimens. Microscopes of this type were described previously in Miniature Microscope Without Lenses (NPO - 20218), NASA Tech Briefs, Vol. 22, No. 8 (August 1998), page 43 and Reflective Variants of Miniature Microscope Without Lenses (NPO 20610), NASA Tech Briefs, Vol. 26, No. 9 (September 1999), page 6a. To recapitulate: In the design and construction of a microscope of this type, the focusing optics of a conventional microscope are replaced by a combination of a microchannel filter and a charge-coupled-device (CCD) image detector. Elimination of focusing optics reduces the size and weight of the instrument and eliminates the need for the time-consuming focusing operation. The microscopes described in the cited prior articles contained two-dimensional CCDs registered with two-dimensional arrays of microchannels and, as such, were designed to produce full two-dimensional images, without need for scanning. The microscopes of the present proposal would contain one-dimensional (line image) CCDs registered with linear arrays of microchannels. In the operation of such a microscope, one would scan a specimen along a line perpendicular to the array axis (in other words, one would scan in pushbroom fashion). One could then synthesize a full two-dimensional image of the specimen from the line-image data acquired at one-pixel increments of position along the scan. In one of the proposed microscopes, a beam of unpolarized light for illuminating the specimen would enter from the side. This light would be reflected down onto the specimen by a nonpolarizing beam splitter attached to the microchannels at their lower ends. A portion of the light incident on the specimen would be reflected upward, through the beam splitter and along the microchannels, to form an image on the CCD. If the nonpolarizing beam splitter were replaced by a polarizing one, then the specimen would be illuminated by s-polarized light. Upon reflection from the specimen, some of the s-polarized light would become p-polarized. Only the p-polarized light would contribute to the image on the CCD; in other words, the image would contain information on the polarization rotating characteristic of the specimen.

Mirror-based broadband scanner with minimized aberration

NASA Astrophysics Data System (ADS)

Yu, Jiun-Yann; Tzeng, Yu-Yi; Huang, Chen-Han; Chui, Hsiang-Chen; Chu, Shi-Wei

2009-02-01

To obtain specific biochemical information in optical scanning microscopy, labeling technique is routinely required. Instead of the complex and invasive sample preparation procedures, incorporating spectral acquisition, which commonly requires a broadband light source, provides another mechanism to enhance molecular contrast. But most current optical scanning system is lens-based and thus the spectral bandwidth is limited to several hundred nanometers due to anti-reflection coating and chromatic aberration. The spectral range of interest in biological research covers ultraviolet to infrared. For example, the absorption peak of water falls around 3 μm, while most proteins exhibit absorption in the UV-visible regime. For imaging purpose, the transmission window of skin and cerebral tissues fall around 1300 and 1800 nm, respectively. Therefore, to extend the spectral bandwidth of an optical scanning system from visible to mid-infrared, we propose a system composed of metallic coated mirrors. A common issue in such a mirror-based system is aberrations induced by oblique incidence. We propose to compensate astigmatism by exchanging the sagittal and tangential planes of the converging spherical mirrors in the scanning system. With the aid of an optical design software, we build a diffraction-limited broadband scanning system with wavefront flatness better than λ/4 at focal plane. Combined with a mirror-based objective this microscopic system will exhibit full spectral capability and will be useful in microscopic imaging and therapeutic applications.

Seamless stitching of tile scan microscope images.

PubMed

Legesse, F B; Chernavskaia, O; Heuke, S; Bocklitz, T; Meyer, T; Popp, J; Heintzmann, R

2015-06-01

For diagnostic purposes, optical imaging techniques need to obtain high-resolution images of extended biological specimens in reasonable time. The field of view of an objective lens, however, is often smaller than the sample size. To image the whole sample, laser scanning microscopes acquire tile scans that are stitched into larger mosaics. The appearance of such image mosaics is affected by visible edge artefacts that arise from various optical aberrations which manifest in grey level jumps across tile boundaries. In this contribution, a technique for stitching tiles into a seamless mosaic is presented. The stitching algorithm operates by equilibrating neighbouring edges and forcing the brightness at corners to a common value. The corrected image mosaics appear to be free from stitching artefacts and are, therefore, suited for further image analysis procedures. The contribution presents a novel method to seamlessly stitch tiles captured by a laser scanning microscope into a large mosaic. The motivation for the work is the failure of currently existing methods for stitching nonlinear, multimodal images captured by our microscopic setups. Our method eliminates the visible edge artefacts that appear between neighbouring tiles by taking into account the overall illumination differences among tiles in such mosaics. The algorithm first corrects the nonuniform brightness that exists within each of the tiles. It then compensates for grey level differences across tile boundaries by equilibrating neighbouring edges and forcing the brightness at the corners to a common value. After these artefacts have been removed further image analysis procedures can be applied on the microscopic images. Even though the solution presented here is tailored for the aforementioned specific case, it could be easily adapted to other contexts where image tiles are assembled into mosaics such as in astronomical or satellite photos. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Image Analysis, Microscopic, and Spectrochemical Study of the PVC Dry Blending Process,

DTIC Science & Technology

The dry blending process used in the production of electrical grade pvc formulations has been studies using a combination of image analysis , microscopic...by image analysis techniques. Optical and scanning electron microscopy were used to assess morphological differences. Spectrochemical techniques were used to indicate chemical changes.

Demonstration of a plenoptic microscope based on laser optical feedback imaging.

PubMed

Glastre, Wilfried; Hugon, Olivier; Jacquin, Olivier; Guillet de Chatellus, Hugues; Lacot, Eric

2013-03-25

A new kind of plenoptic imaging system based on Laser Optical Feedback Imaging (LOFI) is presented and is compared to another previously existing device based on microlens array. Improved photometric performances, resolution and depth of field are obtained at the price of a slow point by point scanning. Main properties of plenoptic microscopes such as numerical refocusing on any curved surface or aberrations compensation are both theoretically and experimentally demonstrated with a LOFI-based device.

Three-dimensional imaging of micro-specimen by optical scanning holography

NASA Astrophysics Data System (ADS)

Liu, Jung-Ping; Tsou, Cheng-Hao

2017-04-01

Optical scanning holography (OSH) is a scanning-type digital holographic technique. In OSH, a heterodyne interference pattern is generated to raster scan the object. OSH can be operated in the incoherent mode and thus is able to record a fluorescence hologram. In addition, resolution of the OSH is proportional to the density of the interference pattern. Here we use a high-NA microscope objective to generate a dynamic Fresnel zone plate to record a hologram of micro-specimen. The achieved transverse resolution and longitudinal resolution are 0.78μm and 3.1μm, respectively.

Multiparallel Three-Dimensional Optical Microscopy

NASA Technical Reports Server (NTRS)

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.

Diffracting aperture based differential phase contrast for scanning X-ray microscopy.

PubMed

Kaulich, Burkhard; Polack, Francois; Neuhaeusler, Ulrich; Susini, Jean; di Fabrizio, Enzo; Wilhein, Thomas

2002-10-07

It is demonstrated that in a zone plate based scanning X-ray microscope, used to image low absorbing, heterogeneous matter at a mesoscopic scale, differential phase contrast (DPC) can be implemented without adding any additional optical component to the normal scheme of the microscope. The DPC mode is simply generated by an appropriate positioning and alignment of microscope apertures. Diffraction from the apertures produces a wave front with a non-uniform intensity. The signal recorded by a pinhole photo diode located in the intensity gradient is highly sensitive to phase changes introduced by the specimen to be recorded. The feasibility of this novel DPC technique was proven with the scanning X-ray microscope at the ID21 beamline of the European Synchrotron Radiation facility (ESRF) operated at 6 keV photon energy. We observe a differential phase contrast, similar to Nomarski's differential interference contrast for the light microscope, which results in a tremendous increase in image contrast of up to 20 % when imaging low absorbing specimen.

Virtual k -Space Modulation Optical Microscopy

NASA Astrophysics Data System (ADS)

Kuang, Cuifang; Ma, Ye; Zhou, Renjie; Zheng, Guoan; Fang, Yue; Xu, Yingke; Liu, Xu; So, Peter T. C.

2016-07-01

We report a novel superresolution microscopy approach for imaging fluorescence samples. The reported approach, termed virtual k -space modulation optical microscopy (VIKMOM), is able to improve the lateral resolution by a factor of 2, reduce the background level, improve the optical sectioning effect and correct for unknown optical aberrations. In the acquisition process of VIKMOM, we used a scanning confocal microscope setup with a 2D detector array to capture sample information at each scanned x -y position. In the recovery process of VIKMOM, we first modulated the captured data by virtual k -space coding and then employed a ptychography-inspired procedure to recover the sample information and correct for unknown optical aberrations. We demonstrated the performance of the reported approach by imaging fluorescent beads, fixed bovine pulmonary artery endothelial (BPAE) cells, and living human astrocytes (HA). As the VIKMOM approach is fully compatible with conventional confocal microscope setups, it may provide a turn-key solution for imaging biological samples with ˜100 nm lateral resolution, in two or three dimensions, with improved optical sectioning capabilities and aberration correcting.

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy

PubMed Central

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. PMID:21280920

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy.

PubMed

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.

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy

NASA Astrophysics Data System (ADS)

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.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Attota, Ravikiran, E-mail: Ravikiran.attota@nist.gov; Dixson, Ronald G.

We experimentally demonstrate that the three-dimensional (3-D) shape variations of nanometer-scale objects can be resolved and measured with sub-nanometer scale sensitivity using conventional optical microscopes by analyzing 4-D optical data using the through-focus scanning optical microscopy (TSOM) method. These initial results show that TSOM-determined cross-sectional (3-D) shape differences of 30 nm–40 nm wide lines agree well with critical-dimension atomic force microscope measurements. The TSOM method showed a linewidth uncertainty of 1.22 nm (k = 2). Complex optical simulations are not needed for analysis using the TSOM method, making the process simple, economical, fast, and ideally suited for high volume nanomanufacturing process monitoring.

Review on Microstructure Analysis of Metals and Alloys Using Image Analysis Techniques

NASA Astrophysics Data System (ADS)

Rekha, Suganthini; Bupesh Raja, V. K.

2017-05-01

The metals and alloys find vast application in engineering and domestic sectors. The mechanical properties of the metals and alloys are influenced by their microstructure. Hence the microstructural investigation is very critical. Traditionally the microstructure is studied using optical microscope with suitable metallurgical preparation. The past few decades the computers are applied in the capture and analysis of the optical micrographs. The advent of computer softwares like digital image processing and computer vision technologies are a boon to the analysis of the microstructure. In this paper the literature study of the various developments in the microstructural analysis, is done. The conventional optical microscope is complemented by the use of Scanning Electron Microscope (SEM) and other high end equipments.

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

PubMed

Bullen, A; Patel, S S; Saggau, P

1997-07-01

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging.

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

PubMed Central

Bullen, A; Patel, S S; Saggau, P

1997-01-01

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging. Images FIGURE 6 PMID:9199810

A combined optical, SEM and STM study of growth spirals on the polytypic cadmium iodide crystals

NASA Astrophysics Data System (ADS)

Singh, Rajendra; Samanta, S. B.; Narlikar, A. V.; Trigunayat, G. C.

2000-05-01

Some novel results of a combined sequential study of growth spirals on the basal surface of the richly polytypic CdI 2 crystals by optical microscopy, scanning electron microscopy (SEM) and scanning tunneling microscopy (STM) are presented and discussed. Under the high resolution and magnification achieved in the scanning electron microscope, the growth steps of large heights seen in the optical micrographs are found to have a large number of additional steps of smaller heights existing between any two adjacent large height growth steps. When further seen by a scanning tunneling microscope, which provides still higher resolution, sequences of unit substeps, each of height equal to the unit cell height of the underlying polytype, are revealed to exist on the surface. Several large steps also lie between the unit steps, with heights equal to an integral multiple of either the unit cell height of the underlying polytype or the thickness of a molecular sheet I-Cd-I. It is suggested that initially a giant screw dislocation may form by brittle fracture of the crystal platelet, which may gradually decompose into numerous unit dislocations during subsequent crystal growth.

Confocal fluorescence microscope with dual-axis architecture and biaxial postobjective scanning

PubMed Central

Wang, Thomas D.; Contag, Christopher H.; Mandella, Michael J.; Chan, Ning Y.; Kino, Gordon S.

2007-01-01

We present a novel confocal microscope that has dual-axis architecture and biaxial postobjective scanning for the collection of fluorescence images from biological specimens. This design uses two low-numerical-aperture lenses to achieve high axial resolution and long working distance, and the scanning mirror located distal to the lenses rotates along the orthogonal axes to produce arc-surface images over a large field of view (FOV). With fiber optic coupling, this microscope can potentially be scaled down to millimeter dimensions via microelectromechanical systems (MEMS) technology. We demonstrate a benchtop prototype with a spatial resolution ≤4.4 μm that collects fluorescence images with a high SNR and a good contrast ratio from specimens expressing GFP. Furthermore, the scanning mechanism produces only small differences in aberrations over the image FOV. These results demonstrate proof of concept of the dual-axis confocal architecture for in vivo molecular and cellular imaging. PMID:15250760

Ultra compact multitip scanning tunneling microscope with a diameter of 50 mm.

PubMed

Cherepanov, Vasily; Zubkov, Evgeny; Junker, Hubertus; Korte, Stefan; Blab, Marcus; Coenen, Peter; Voigtländer, Bert

2012-03-01

We present a multitip scanning tunneling microscope (STM) where four independent STM units are integrated on a diameter of 50 mm. The coarse positioning of the tips is done under the control of an optical microscope or scanning electron microscopy in vacuum. The heart of this STM is a new type of piezoelectric coarse approach called KoalaDrive. The compactness of the KoalaDrive allows building a four-tip STM as small as a single-tip STM with a drift of less than 0.2 nm/min at room temperature and lowest resonance frequencies of 2.5 kHz (xy) and 5.5 kHz (z). We present as examples of the performance of the multitip STM four point measurements of silicide nanowires and graphene.

PtyNAMi: ptychographic nano-analytical microscope at PETRA III: interferometrically tracking positions for 3D x-ray scanning microscopy using a ball-lens retroreflector

NASA Astrophysics Data System (ADS)

Schroer, Christian G.; Seyrich, Martin; Kahnt, Maik; Botta, Stephan; Döhrmann, Ralph; Falkenberg, Gerald; Garrevoet, Jan; Lyubomirskiy, Mikhail; Scholz, Maria; Schropp, Andreas; Wittwer, Felix

2017-09-01

In recent years, ptychography has revolutionized x-ray microscopy in that it is able to overcome the diffraction limit of x-ray optics, pushing the spatial resolution limit down to a few nanometers. However, due to the weak interaction of x rays with matter, the detection of small features inside a sample requires a high coherent fluence on the sample, a high degree of mechanical stability, and a low background signal from the x-ray microscope. The x-ray scanning microscope PtyNAMi at PETRA III is designed for high-spatial-resolution 3D imaging with high sensitivity. The design concept is presented with a special focus on real-time metrology of the sample position during tomographic scanning microscopy.

Pupil engineering for a confocal reflectance line-scanning microscope

NASA Astrophysics Data System (ADS)

Patel, Yogesh G.; Rajadhyaksha, Milind; DiMarzio, Charles A.

2011-03-01

Confocal reflectance microscopy may enable screening and diagnosis of skin cancers noninvasively and in real-time, as an adjunct to biopsy and pathology. Current confocal point-scanning systems are large, complex, and expensive. A confocal line-scanning microscope, utilizing a of linear array detector can be simpler, smaller, less expensive, and may accelerate the translation of confocal microscopy in clinical and surgical dermatology. A line scanner may be implemented with a divided-pupil, half used for transmission and half for detection, or with a full-pupil using a beamsplitter. The premise is that a confocal line-scanner with either a divided-pupil or a full-pupil will provide high resolution and optical sectioning that would be competitive to that of the standard confocal point-scanner. We have developed a confocal line-scanner that combines both divided-pupil and full-pupil configurations. This combined-pupil prototype is being evaluated to determine the advantages and limitations of each configuration for imaging skin, and comparison of performance to that of commercially available standard confocal point-scanning microscopes. With the combined configuration, experimental evaluation of line spread functions (LSFs), contrast, signal-to-noise ratio, and imaging performance is in progress under identical optical and skin conditions. Experimental comparisons between divided-pupil and full-pupil LSFs will be used to determine imaging performance. Both results will be compared to theoretical calculations using our previously reported Fourier analysis model and to the confocal point spread function (PSF). These results may lead to a simpler class of confocal reflectance scanning microscopes for clinical and surgical dermatology.

Design and demonstration of multimodal optical scanning microscopy for confocal and two-photon imaging

NASA Astrophysics Data System (ADS)

Chun, Wanhee; Do, Dukho; Gweon, Dae-Gab

2013-01-01

We developed a multimodal microscopy based on an optical scanning system in order to obtain diverse optical information of the same area of a sample. Multimodal imaging researches have mostly depended on a commercial microscope platform, easy to use but restrictive to extend imaging modalities. In this work, the beam scanning optics, especially including a relay lens, was customized to transfer broadband (400-1000 nm) lights to a sample without any optical error or loss. The customized scanning optics guarantees the best performances of imaging techniques utilizing the lights within the design wavelength. Confocal reflection, confocal fluorescence, and two-photon excitation fluorescence images were obtained, through respective implemented imaging channels, to demonstrate imaging feasibility for near-UV, visible, near-IR continuous light, and pulsed light in the scanning optics. The imaging performances for spatial resolution and image contrast were verified experimentally; the results were satisfactory in comparison with theoretical results. The advantages of customization, containing low cost, outstanding combining ability and diverse applications, will contribute to vitalize multimodal imaging researches.

Scanning near-field optical microscopy.

PubMed

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.

Line-scanning, stage scanning confocal microscope

NASA Astrophysics Data System (ADS)

Carucci, John A.; Stevenson, Mary; Gareau, Daniel

2016-03-01

We created a line-scanning, stage scanning confocal microscope as part of a new procedure: video assisted micrographic surgery (VAMS). The need for rapid pathological assessment of the tissue on the surface of skin excisions very large since there are 3.5 million new skin cancers diagnosed annually in the United States. The new design presented here is a confocal microscope without any scanning optics. Instead, a line is focused in space and the sample, which is flattened, is physically translated such that the line scans across its face in a direction perpendicular to the line its self. The line is 6mm long and the stage is capable of scanning 50 mm, hence the field of view is quite large. The theoretical diffraction-limited resolution is 0.7um lateral and 3.7um axial. However, in this preliminary report, we present initial results that are a factor of 5-7 poorer in resolution. The results are encouraging because they demonstrate that the linear array detector measures sufficient signal from fluorescently labeled tissue and also demonstrate the large field of view achievable with VAMS.

Design and evaluation of an intraocular B-scan OCT-guided 36-gauge needle

NASA Astrophysics Data System (ADS)

Shen, Jin H.; Joos, Karen M.

2015-03-01

Optical coherence tomography imaging is widely used in ophthalmology and optometry clinics for diagnosing retinal disorders. External microscope-mounted OCT operating room systems have imaged retinal changes immediately following surgical manipulations. However, the goal is to image critical surgical maneuvers in real time. External microscope-mounted OCT systems have some limitations with problems tracking constantly moving intraocular surgical instruments, and formation of absolute shadows by the metallic surgical instruments upon the underlying tissues of interest. An intraocular OCT-imaging probe was developed to resolve these problems. A disposable 25-gauge probe tip extended beyond the handpiece, with a 36-gauge needle welded to a disposable tip with its end extending an additional 3.5 mm. A sealed 0.35 mm diameter GRIN lens protected the fiber scanner and focused the scanning beam at a 3 to 4 mm distance. The OCT engine was a very high-resolution spectral-domain optical coherence tomography (SDOCT) system (870 nm, Bioptigen, Inc. Durham, NC) which produced 2000 A-scan lines per B-scan image at a frequency of 5 Hz with the fiber optic oscillations matched to this frequency. Real-time imaging of the needle tip as it touched infrared paper was performed. The B-scan OCT-needle was capable of real-time performance and imaging of the phantom material. In the future, the B-scan OCT-guided needle will be used to perform sub-retinal injections.

L-Phenylalanine functionalized silver nanoparticles: Photocatalytic and nonlinear optical applications

NASA Astrophysics Data System (ADS)

Nidya, M.; Umadevi, M.; Sankar, Pranitha; Philip, Reji; Rajkumar, Beulah J. M.

2015-04-01

An extensive study on the behavior of L-Phenylalanine capped silver nanoparticles (Phe-Ag NPs) in the aqueous phase and in a sol-gel thin film showed different UV/Vis, Transmission Electron Microscope (TEM), Dynamic Light Scattering and Zeta potential profiles. Scanning Electron Microscope (SEM) images of the samples in the sol gel film showed Ag embedded in the SiO2 matrix. Surface Enhanced Raman Spectra (SERS) confirmed that both in the aqueous media and in the sol gel film, the attachment of Phe to the Ag NP surface was through the benzene ring, with the sol-gel film showing a better enhancement. Photocatalytic degradation of crystal violet was measured spectrophotometrically using Phe-Ag NPs as a nanocatalyst under visible light illumination. Intensity-dependent nonlinear optical absorption of Phe-Ag measured using the open aperture Z-scan technique revealed that the material is an efficient optical limiter with potential applications.

Distortion Correction for a Brewster Angle Microscope Using an Optical Grating.

PubMed

Sun, Zhe; Zheng, Desheng; Baldelli, Steven

2017-02-21

A distortion-corrected Brewster angle microscope (DC-BAM) is designed, constructed, and tested based on the combination of an optical grating and a relay lens. Avoiding the drawbacks of most conventional BAM instruments, this configuration corrects the image propagation direction and consequently provides an image in focus over the entire field of view without any beam scanning or imaging reconstruction. This new BAM can be applied to both liquid and solid subphases with good spatial resolution in static and dynamic studies.

Destructive Single-Event Effects in Diodes

NASA Technical Reports Server (NTRS)

Casey, Megan C.; Lauenstein, Jean-Marie; Campola, Michael J.; Wilcox, Edward P.; Phan, Anthony M.; Label, Kenneth A.

2017-01-01

In this work, we discuss the observed single-event effects in a variety of types of diodes. In addition, we conduct failure analysis on several Schottky diodes that were heavy-ion irradiated. High- and low-magnitude optical microscope images, infrared camera images, and scanning electron microscope images are used to identify and describe the failure locations.

Smart align -- A new tool for robust non-rigid registration of scanning microscope data

DOE PAGES

Jones, Lewys; Yang, Hao; Pennycook, Timothy J.; ...

2015-07-10

Many microscopic investigations of materials may benefit from the recording of multiple successive images. This can include techniques common to several types of microscopy such as frame averaging to improve signal-to-noise ratios (SNR) or time series to study dynamic processes or more specific applications. In the scanning transmission electron microscope, this might include focal series for optical sectioning or aberration measurement, beam damage studies or camera-length series to study the effects of strain; whilst in the scanning tunnelling microscope, this might include bias voltage series to probe local electronic structure. Whatever the application, such investigations must begin with the carefulmore » alignment of these data stacks, an operation that is not always trivial. In addition, the presence of low-frequency scanning distortions can introduce intra-image shifts to the data. Here, we describe an improved automated method of performing non-rigid registration customised for the challenges unique to scanned microscope data specifically addressing the issues of low-SNR data, images containing a large proportion of crystalline material and/or local features of interest such as dislocations or edges. Careful attention has been paid to artefact testing of the non-rigid registration method used, and the importance of this registration for the quantitative interpretation of feature intensities and positions is evaluated.« less

Smart align -- A new tool for robust non-rigid registration of scanning microscope data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jones, Lewys; Yang, Hao; Pennycook, Timothy J.

Many microscopic investigations of materials may benefit from the recording of multiple successive images. This can include techniques common to several types of microscopy such as frame averaging to improve signal-to-noise ratios (SNR) or time series to study dynamic processes or more specific applications. In the scanning transmission electron microscope, this might include focal series for optical sectioning or aberration measurement, beam damage studies or camera-length series to study the effects of strain; whilst in the scanning tunnelling microscope, this might include bias voltage series to probe local electronic structure. Whatever the application, such investigations must begin with the carefulmore » alignment of these data stacks, an operation that is not always trivial. In addition, the presence of low-frequency scanning distortions can introduce intra-image shifts to the data. Here, we describe an improved automated method of performing non-rigid registration customised for the challenges unique to scanned microscope data specifically addressing the issues of low-SNR data, images containing a large proportion of crystalline material and/or local features of interest such as dislocations or edges. Careful attention has been paid to artefact testing of the non-rigid registration method used, and the importance of this registration for the quantitative interpretation of feature intensities and positions is evaluated.« less

Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues.

PubMed

Sung, Kung-Bin; Liang, Chen; Descour, Michael; Collier, Tom; Follen, Michele; Richards-Kortum, Rebecca

2002-10-01

We have built a fiber-optic confocal reflectance microscope capable of imaging human tissues in near real time. Miniaturization of the objective lens and the mechanical components for positioning and axially scanning the objective enables the device to be used in inner organs of the human body. The lateral resolution is 2 micrometers and axial resolution is 10 micrometers. Confocal images of fixed tissue biopsies and the human lip in vivo have been obtained at 15 frames/s without any fluorescent stains. Both cell morphology and tissue architecture can be appreciated from images obtained with this microscope.

Integration of a high-NA light microscope in a scanning electron microscope.

PubMed

Zonnevylle, A C; Van Tol, R F C; Liv, N; Narvaez, A C; Effting, A P J; Kruit, P; Hoogenboom, J P

2013-10-01

We present an integrated light-electron microscope in which an inverted high-NA objective lens is positioned inside a scanning electron microscope (SEM). The SEM objective lens and the light objective lens have a common axis and focal plane, allowing high-resolution optical microscopy and scanning electron microscopy on the same area of a sample simultaneously. Components for light illumination and detection can be mounted outside the vacuum, enabling flexibility in the construction of the light microscope. The light objective lens can be positioned underneath the SEM objective lens during operation for sub-10 μm alignment of the fields of view of the light and electron microscopes. We demonstrate in situ epifluorescence microscopy in the SEM with a numerical aperture of 1.4 using vacuum-compatible immersion oil. For a 40-nm-diameter fluorescent polymer nanoparticle, an intensity profile with a FWHM of 380 nm is measured whereas the SEM performance is uncompromised. The integrated instrument may offer new possibilities for correlative light and electron microscopy in the life sciences as well as in physics and chemistry. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

Use of digital micromirror devices as dynamic pinhole arrays for adaptive confocal fluorescence microscopy

NASA Astrophysics Data System (ADS)

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

2018-02-01

In this work, we present a new confocal laser scanning microscope capable to perform sensorless wavefront optimization in real time. The device is a parallelized laser scanning microscope in which the excitation light is structured in a lattice of spots by a spatial light modulator, while a deformable mirror provides aberration correction and scanning. A binary DMD is positioned in an image plane of the detection optical path, acting as a dynamic array of reflective confocal pinholes, images by a high performance cmos camera. A second camera detects images of the light rejected by the pinholes for sensorless aberration correction.

Tip-enhanced near-field Raman spectroscopy with a scanning tunneling microscope and side-illumination optics.

PubMed

Yi, K J; He, X N; Zhou, Y S; Xiong, W; Lu, Y F

2008-07-01

Conventional Raman spectroscopy (RS) suffers from low spatial resolution and low detection sensitivity due to the optical diffraction limit and small interaction cross sections. It has been reported that a highly localized and significantly enhanced electromagnetic field could be generated in the proximity of a metallic tip illuminated by a laser beam. In this study, a tip-enhanced RS system was developed to both improve the resolution and enhance the detection sensitivity using the tip-enhanced near-field effects. This instrument, by combining RS with a scanning tunneling microscope and side-illumination optics, demonstrated significant enhancement on both optical sensitivity and spatial resolution using either silver (Ag)-coated tungsten (W) tips or gold (Au) tips. The sensitivity improvement was verified by observing the enhancement effects on silicon (Si) substrates. Lateral resolution was verified to be below 100 nm by mapping Ag nanostructures. By deploying the depolarization technique, an apparent enhancement of 175% on Si substrates was achieved. Furthermore, the developed instrument features fast and reliable optical alignment, versatile sample adaptability, and effective suppression of far-field signals.

Vibration sensitivity of the scanning near-field optical microscope with a tapered optical fiber probe.

PubMed

Chang, Win-Jin; Fang, Te-Hua; Lee, Haw-Long; Yang, Yu-Ching

2005-01-01

In this paper the Rayleigh-Ritz method was used to study the scanning near-field optical microscope (SNOM) with a tapered optical fiber probe's flexural and axial sensitivity to vibration. Not only the contact stiffness but also the geometric parameters of the probe can influence the flexural and axial sensitivity to vibration. According to the analysis, the lateral and axial contact stiffness had a significant effect on the sensitivity of vibration of the SNOM's probe, each mode had a different level of sensitivity and in the first mode the tapered optical fiber probe was the most acceptive to higher levels of flexural and axial vibration. Generally, when the contact stiffness was lower, the tapered probe was more sensitive to higher levels of both axial and flexural vibration than the uniform probe. However, the situation was reversed when the contact stiffness was larger. Furthermore, the effect that the probe's length and its tapered angle had on the SNOM's probe axial and flexural vibration were significant and these two conditions should be incorporated into the design of new SNOM probes.

Volumetric HiLo microscopy employing an electrically tunable lens.

PubMed

Philipp, Katrin; Smolarski, André; Koukourakis, Nektarios; Fischer, Andreas; Stürmer, Moritz; Wallrabe, Ulrike; Czarske, Jürgen W

2016-06-27

Electrically tunable lenses exhibit strong potential for fast motion-free axial scanning in a variety of microscopes. However, they also lead to a degradation of the achievable resolution because of aberrations and misalignment between illumination and detection optics that are induced by the scan itself. Additionally, the typically nonlinear relation between actuation voltage and axial displacement leads to over- or under-sampled frame acquisition in most microscopic techniques because of their static depth-of-field. To overcome these limitations, we present an Adaptive-Lens-High-and-Low-frequency (AL-HiLo) microscope that enables volumetric measurements employing an electrically tunable lens. By using speckle-patterned illumination, we ensure stability against aberrations of the electrically tunable lens. Its depth-of-field can be adjusted a-posteriori and hence enables to create flexible scans, which compensates for irregular axial measurement positions. The adaptive HiLo microscope provides an axial scanning range of 1 mm with an axial resolution of about 4 μm and sub-micron lateral resolution over the full scanning range. Proof of concept measurements at home-built specimens as well as zebrafish embryos with reporter gene-driven fluorescence in the thyroid gland are shown.

Surface characterization and testing II; Proceedings of the Meeting, San Diego, CA, Aug. 10, 11, 1989

NASA Technical Reports Server (NTRS)

Greivenkamp, John E. (Editor); Young, Matt (Editor)

1989-01-01

Various papers on surface characterization and testing are presented. Individual topics addressed include: simple Hartmann test data interpretation, optimum configuration of the Offner null corrector, system for phase-shifting interferometry in the presence of vibration, fringe variation and visibility in speckle-shearing interferometry, functional integral representation of rough surfaces, calibration of surface heights in an interferometric optical profiler, image formation in common path differential profilometers, SEM of optical surfaces, measuring surface profiles with scanning tunneling microscopes, surface profile measurements of curved parts, high-resolution optical profiler, scanning heterodyne interferometer with immunity from microphonics, real-time crystal axis measurements of semiconductor materials, radial metrology with a panoramic annular lens, surface analysis for the characterization of defects in thin-film processes, Spacelab Optical Viewport glass assembly optical test program for the Starlab mission, scanning differential intensity and phase system for optical metrology.

Fluorescence microscopy.

PubMed

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.

Fluorescence Microscopy

PubMed Central

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

2016-01-01

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. PMID:25275114

Excitation of propagating surface plasmons with a scanning tunnelling microscope.

PubMed

Wang, T; Boer-Duchemin, E; Zhang, Y; Comtet, G; Dujardin, G

2011-04-29

Inelastic electron tunnelling excitation of propagating surface plasmon polaritons (SPPs) on a thin gold film is demonstrated. This is done by combining a scanning tunnelling microscope (STM) with an inverted optical microscope. Analysis of the leakage radiation in both the image and Fourier planes unambiguously shows that the majority (up to 99.5%) of the detected photons originate from propagating SPPs with propagation lengths of the order of 10  µm. The remaining photon emission is localized under the STM tip and is attributed to a tip-gold film coupled plasmon resonance as evidenced by the bimodal spectral distribution and enhanced emission intensity observed using a silver STM tip for excitation.

Quantitative and structural analyses of the in vitro and ex vivo biofilm-forming ability of dermatophytes.

PubMed

Brilhante, Raimunda Sâmia Nogueira; Correia, Edmilson Emanuel Monteiro; Guedes, Glaucia Morgana de Melo; Pereira, Vandbergue Santos; Oliveira, Jonathas Sales de; Bandeira, Silviane Praciano; Alencar, Lucas Pereira de; Andrade, Ana Raquel Colares de; Castelo-Branco, Débora de Souza Collares Maia; Cordeiro, Rossana de Aguiar; Pinheiro, Adriana de Queiroz; Chaves, Lúcio Jackson Queiroz; Pereira Neto, Waldemiro de Aquino; Sidrim, José Júlio Costa; Rocha, Marcos Fábio Gadelha

2017-07-01

The aim of this study was to evaluate the in vitro and ex vivo biofilm-forming ability of dermatophytes on a nail fragment. Initially, four isolates of Trichophyton rubrum, six of Trichophyton tonsurans, three of Trichophyton mentagrophytes, ten of Microsporum canis and three of Microsporum gypseum were tested for production biomass by crystal violet assay. Then, one strain per species presenting the best biofilm production was chosen for further studies by optical microscopy (Congo red staining), confocal laser scanning (LIVE/DEAD staining) and scanning electron (secondary electron) microscopy. Biomass quantification by crystal violet assay, optical microscope images of Congo red staining, confocal microscope and scanning electron microscope images revealed that all species studied are able to form biofilms both in vitro and ex vivo, with variable density and architecture. M. gypseum, T. rubrum and T. tonsurans produced robust biofilms, with abundant matrix and biomass, while M. canis produced the weakest biofilms compared to other species. This study sheds light on biofilms of different dermatophyte species, which will contribute to a better understanding of the pathophysiology of dermatophytosis. Further studies of this type are necessary to investigate the processes involved in the formation and composition of dermatophyte biofilms.

75 FR 13486 - Application(s) for Duty-Free Entry of Scientific Instruments

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-22

... nanostructures. This instrument combines an optical microscope with a scanning probe imaging system. Specifically... soft materials than other instruments, as it detects the probe coming close to the sample surface by... conventional AFM type silicon cantilevers as well as cantilevered optical fiber probes with exposed probe...

SCANNING NEAR-FIELD OPTICAL MICROSCOPY

PubMed Central

Vobornik, Dušan; Vobornik, Slavenka

2008-01-01

An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today’s science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and 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. PMID:18318675

Results of examination of the returned Surveyor 3 samples for particulate impacts

NASA Technical Reports Server (NTRS)

Cour-Palais, B. G.; Flaherty, R. E.; High, R. W.; Kessler, D. J.; Mckay, D. S.; Zook, H. A.

1972-01-01

The television housing and a section of the strut of the radar altimeter and Doppler velocity sensor were examined optically and with a scanning electron microscope for particulate impacts. The white surface of the camera was discolored during the months the Surveyor 3 was on the moon; however, most of the craters must have occurred as a result of lunar dust sandblasted by the LM exhaust. The polished section of the strut exhibits contamination which appears brown and seems to be partially composed of crystals. Electron microscopic analysis of the strut section indicated no craters of hypervelocity impact origin, confirmed pitting density results of the optical scans, and indicated that material in the craters is of lunar origin. No meteorite impacts larger than 25 microns were detected on the tubing section.

Highly sensitive mode mapping of whispering-gallery modes by scanning thermocouple-probe microscopy.

PubMed

Klein, Angela E; Schmidt, Carsten; Liebsch, Mattes; Janunts, Norik; Dobynde, Mikhail; Tünnermann, Andreas; Pertsch, Thomas

2014-03-01

We propose a method for mapping optical near-fields with the help of a thermocouple scanning-probe microscope tip. As the tip scans the sample surface, its apex is heated by light absorption, generating a thermovoltage. The thermovoltage map represents the intensity distribution of light at the sample surface. The measurement technique has been employed to map optical whispering-gallery modes in fused silica microdisk resonators operating at near-infrared wavelengths. The method could potentially be employed for near-field imaging of a variety of systems in the near-infrared and visible spectral range.

Analysis of improvement in performance and design parameters for enhancing resolution in an atmospheric scanning electron microscope.

PubMed

Yoon, Yeo Hun; Kim, Seung Jae; Kim, Dong Hwan

2015-12-01

The scanning electron microscope is used in various fields to go beyond diffraction limits of the optical microscope. However, the electron pathway should be conducted in a vacuum so as not to scatter electrons. The pretreatment of the sample is needed for use in the vacuum. To directly observe large and fully hydrophilic samples without pretreatment, the atmospheric scanning electron microscope (ASEM) is needed. We developed an electron filter unit and an electron detector unit for implementation of the ASEM. The key of the electron filter unit is that electrons are transmitted while air molecules remain untransmitted through the unit. The electron detector unit collected the backscattered electrons. We conducted experiments using the selected materials with Havar foil, carbon film and SiN film. © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Optical monitoring of neuronal activity at high frame rate with a digital random-access multiphoton (RAMP) microscope.

PubMed

Otsu, Yo; Bormuth, Volker; Wong, Jerome; Mathieu, Benjamin; Dugué, Guillaume P; Feltz, Anne; Dieudonné, Stéphane

2008-08-30

Two-photon microscopy offers the promise of monitoring brain activity at multiple locations within intact tissue. However, serial sampling of voxels has been difficult to reconcile with millisecond timescales characteristic of neuronal activity. This is due to the conflicting constraints of scanning speed and signal amplitude. The recent use of acousto-optic deflector scanning to implement random-access multiphoton microscopy (RAMP) potentially allows to preserve long illumination dwell times while sampling multiple points-of-interest at high rates. However, the real-life abilities of RAMP microscopy regarding sensitivity and phototoxicity issues, which have so far impeded prolonged optical recordings at high frame rates, have not been assessed. Here, we describe the design, implementation and characterisation of an optimised RAMP microscope. We demonstrate the application of the microscope by monitoring calcium transients in Purkinje cells and cortical pyramidal cell dendrites and spines. We quantify the illumination constraints imposed by phototoxicity and show that stable continuous high-rate recordings can be obtained. During these recordings the fluorescence signal is large enough to detect spikes with a temporal resolution limited only by the calcium dye dynamics, improving upon previous techniques by at least an order of magnitude.

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

PubMed

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

2006-09-01

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

DESIGN NOTE: From nanometre to millimetre: a feasibility study of the combination of scanning probe microscopy and combined optical and x-ray interferometry

NASA Astrophysics Data System (ADS)

Yacoot, Andrew; Koenders, Ludger

2003-09-01

This feasibility study investigates the potential combination of an x-ray interferometer and optical interferometer as a one-dimensional long range high resolution scanning stage for an atomic force microscope (AFM) in order to overcome the problems of non-linearity associated with conventional AFMs and interferometers. Preliminary results of measurements of the uniformity of the period of a grating used as a transfer standards show variations in period at the nanometre level.

Surface profile measurement by using the integrated Linnik WLSI and confocal microscope system

NASA Astrophysics Data System (ADS)

Wang, Wei-Chung; Shen, Ming-Hsing; Hwang, Chi-Hung; Yu, Yun-Ting; Wang, Tzu-Fong

2017-06-01

The white-light scanning interferometer (WLSI) and confocal microscope (CM) are the two major optical inspection systems for measuring three-dimensional (3D) surface profile (SP) of micro specimens. Nevertheless, in practical applications, WLSI is more suitable for measuring smooth and low-slope surfaces. On the other hand, CM is more suitable for measuring uneven-reflective and low-reflective surfaces. As for aspect of surface profiles to be measured, the characteristics of WLSI and CM are also different. WLSI is generally used in semiconductor industry while CM is more popular in printed circuit board industry. In this paper, a self-assembled multi-function optical system was integrated to perform Linnik white-light scanning interferometer (Linnik WLSI) and CM. A connecting part composed of tubes, lenses and interferometer was used to conjunct finite and infinite optical systems for Linnik WLSI and CM in the self-assembled optical system. By adopting the flexibility of tubes and lenses, switching to perform two different optical measurements can be easily achieved. Furthermore, based on the shape from focus method with energy of Laplacian filter, the CM was developed to enhance the on focal information of each pixel so that the CM can provide all-in-focus image for performing the 3D SP measurement and analysis simultaneously. As for Linnik WLSI, eleven-step phase shifting algorithm was used to analyze vertical scanning signals and determine the 3D SP.

MEMS-tunable dielectric metasurface lens.

PubMed

Arbabi, Ehsan; Arbabi, Amir; Kamali, Seyedeh Mahsa; Horie, Yu; Faraji-Dana, MohammadSadegh; Faraon, Andrei

2018-02-23

Varifocal lenses, conventionally implemented by changing the axial distance between multiple optical elements, have a wide range of applications in imaging and optical beam scanning. The use of conventional bulky refractive elements makes these varifocal lenses large, slow, and limits their tunability. Metasurfaces, a new category of lithographically defined diffractive devices, enable thin and lightweight optical elements with precisely engineered phase profiles. Here we demonstrate tunable metasurface doublets, based on microelectromechanical systems (MEMS), with more than 60 diopters (about 4%) change in the optical power upon a 1-μm movement of one metasurface, and a scanning frequency that can potentially reach a few kHz. They can also be integrated with a third metasurface to make compact microscopes (~1 mm thick) with a large corrected field of view (~500 μm or 40 degrees) and fast axial scanning for 3D imaging. This paves the way towards MEMS-integrated metasurfaces as a platform for tunable and reconfigurable optics.

Remote focusing in confocal microscopy by means of a modified Alvarez lens.

PubMed

Bawart, M; Jesacher, A; Bernet, S; Ritsch-Marte, M

2018-06-22

Alvarez lenses are actuated lens-pairs which allow one to tune the optical power by mechanical displacement of subelements. Here, we show that a recently realized modified Alvarez lens design which does not require mechanical actuation can be integrated into a confocal microscope. Instead of mechanically moving them, the sublenses are imaged onto each other in a 4f-configuration, where the lateral image shift leading to a change in optical power is created by a galvo-mirror. The avoidance of mechanical lens shifts leads to a large speed gain for axial (and hence also 3D) image scans compared to classical Alvarez lenses. We demonstrate that the suggested operation principle is compatible with confocal microscopy. In order to optimize the system, we have drawn advantage of the flexibility a liquid-crystal spatial light modulator offers for the implementation. For given specifications, dedicated diffractive optical elements or freeform elements can be used in combination with resonant galvo-scanners or acousto-optic beam deflectors, to achieve even faster z-scans than reported here, reaching video rate. © 2018 The Authors Journal of Microscopy © 2018 Royal Microscopical Society.

Neuroanatomy from Mesoscopic to Nanoscopic Scales: An Improved Method for the Observation of Semithin Sections by High-Resolution Scanning Electron Microscopy

PubMed Central

Rodríguez, José-Rodrigo; Turégano-López, Marta; DeFelipe, Javier; Merchán-Pérez, Angel

2018-01-01

Semithin sections are commonly used to examine large areas of tissue with an optical microscope, in order to locate and trim the regions that will later be studied with the electron microscope. Ideally, the observation of semithin sections would be from mesoscopic to nanoscopic scales directly, instead of using light microscopy and then electron microscopy (EM). Here we propose a method that makes it possible to obtain high-resolution scanning EM images of large areas of the brain in the millimeter to nanometer range. Since our method is compatible with light microscopy, it is also feasible to generate hybrid light and electron microscopic maps. Additionally, the same tissue blocks that have been used to obtain semithin sections can later be used, if necessary, for transmission EM, or for focused ion beam milling and scanning electron microscopy (FIB-SEM). PMID:29568263

Neuroanatomy from Mesoscopic to Nanoscopic Scales: An Improved Method for the Observation of Semithin Sections by High-Resolution Scanning Electron Microscopy.

PubMed

Rodríguez, José-Rodrigo; Turégano-López, Marta; DeFelipe, Javier; Merchán-Pérez, Angel

2018-01-01

Semithin sections are commonly used to examine large areas of tissue with an optical microscope, in order to locate and trim the regions that will later be studied with the electron microscope. Ideally, the observation of semithin sections would be from mesoscopic to nanoscopic scales directly, instead of using light microscopy and then electron microscopy (EM). Here we propose a method that makes it possible to obtain high-resolution scanning EM images of large areas of the brain in the millimeter to nanometer range. Since our method is compatible with light microscopy, it is also feasible to generate hybrid light and electron microscopic maps. Additionally, the same tissue blocks that have been used to obtain semithin sections can later be used, if necessary, for transmission EM, or for focused ion beam milling and scanning electron microscopy (FIB-SEM).

Characterization of a subwavelength-scale 3D void structure using the FDTD-based confocal laser scanning microscopic image mapping technique.

PubMed

Choi, Kyongsik; Chon, James W; Gu, Min; Lee, Byoungho

2007-08-20

In this paper, a simple confocal laser scanning microscopic (CLSM) image mapping technique based on the finite-difference time domain (FDTD) calculation has been proposed and evaluated for characterization of a subwavelength-scale three-dimensional (3D) void structure fabricated inside polymer matrix. The FDTD simulation method adopts a focused Gaussian beam incident wave, Berenger's perfectly matched layer absorbing boundary condition, and the angular spectrum analysis method. Through the well matched simulation and experimental results of the xz-scanned 3D void structure, we first characterize the exact position and the topological shape factor of the subwavelength-scale void structure, which was fabricated by a tightly focused ultrashort pulse laser. The proposed CLSM image mapping technique based on the FDTD can be widely applied from the 3D near-field microscopic imaging, optical trapping, and evanescent wave phenomenon to the state-of-the-art bio- and nanophotonics.

A portable confocal hyperspectral microscope without any scan or tube lens and its application in fluorescence and Raman spectral imaging

NASA Astrophysics Data System (ADS)

Li, Jingwei; Cai, Fuhong; Dong, Yongjiang; Zhu, Zhenfeng; Sun, Xianhe; Zhang, Hequn; He, Sailing

2017-06-01

In this study, a portable confocal hyperspectral microscope is developed. In traditional confocal laser scanning microscopes, scan lens and tube lens are utilized to achieve a conjugate relationship between the galvanometer and the back focal plane of the objective, in order to achieve a better resolution. However, these lenses make it difficult to scale down the volume of the system. In our portable confocal hyperspectral microscope (PCHM), the objective is placed directly next to the galvomirror. Thus, scan lens and tube lens are not included in our system and the size of this system is greatly reduced. Furthermore, the resolution is also acceptable in many biomedical and food-safety applications. Through reducing the optical length of the system, the signal detection efficiency is enhanced. This is conducive to realizing both the fluorescence and Raman hyperspectral imaging. With a multimode fiber as a pinhole, an improved image contrast is also achieved. Fluorescent spectral images for HeLa cells/fingers and Raman spectral images of kumquat pericarp are present. The spectral resolution and spatial resolutions are about 0.4 nm and 2.19 μm, respectively. These results demonstrate that this portable hyperspectral microscope can be used in in-vivo fluorescence imaging and in situ Raman spectral imaging.

Ultrafast scanning probe microscopy

DOEpatents

Weiss, S.; Chemla, D.S.; Ogletree, D.F.; Botkin, D.

1995-05-16

An ultrafast scanning probe microscopy method is described for achieving subpicosecond-temporal resolution and submicron-spatial resolution of an observation sample. In one embodiment of the present claimed invention, a single short optical pulse is generated and is split into first and second pulses. One of the pulses is delayed using variable time delay means. The first pulse is then directed at an observation sample located proximate to the probe of a scanning probe microscope. The scanning probe microscope produces probe-sample signals indicative of the response of the probe to characteristics of the sample. The second pulse is used to modulate the probe of the scanning probe microscope. The time delay between the first and second pulses is then varied. The probe-sample response signal is recorded at each of the various time delays created between the first and second pulses. The probe-sample response signal is then plotted as a function of time delay to produce a cross-correlation of the probe sample response. In so doing, the present invention provides simultaneous subpicosecond-temporal resolution and submicron-spatial resolution of the sample. 6 Figs.

Ultrafast scanning probe microscopy

DOEpatents

Weiss, Shimon; Chemla, Daniel S.; Ogletree, D. Frank; Botkin, David

1995-01-01

An ultrafast scanning probe microscopy method for achieving subpicosecond-temporal resolution and submicron-spatial resolution of an observation sample. In one embodiment of the present claimed invention, a single short optical pulse is generated and is split into first and second pulses. One of the pulses is delayed using variable time delay means. The first pulse is then directed at an observation sample located proximate to the probe of a scanning probe microscope. The scanning probe microscope produces probe-sample signals indicative of the response of the probe to characteristics of the sample. The second pulse is used to modulate the probe of the scanning probe microscope. The time delay between the first and second pulses is then varied. The probe-sample response signal is recorded at each of the various time delays created between the first and second pulses. The probe-sample response signal is then plotted as a function of time delay to produce a cross-correlation of the probe sample response. In so doing, the present invention provides simultaneous subpicosecond-temporal resolution and submicron-spatial resolution of the sample.

Simple fiber-optic confocal microscopy with nanoscale depth resolution beyond the diffraction barrier.

PubMed

Ilev, Ilko; Waynant, Ronald; Gannot, Israel; Gandjbakhche, Amir

2007-09-01

A novel fiber-optic confocal approach for ultrahigh depth-resolution (

Analysis of artificial opals by scanning near field optical microscopy

NASA Astrophysics Data System (ADS)

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

2011-04-01

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

Optical track width measurements below 100 nm using artificial neural networks

NASA Astrophysics Data System (ADS)

Smith, R. J.; See, C. W.; Somekh, M. G.; Yacoot, A.; Choi, E.

2005-12-01

This paper discusses the feasibility of using artificial neural networks (ANNs), together with a high precision scanning optical profiler, to measure very fine track widths that are considerably below the conventional diffraction limit of a conventional optical microscope. The ANN is trained using optical profiles obtained from tracks of known widths, the network is then assessed by applying it to test profiles. The optical profiler is an ultra-stable common path scanning interferometer, which provides extremely precise surface measurements. Preliminary results, obtained with a 0.3 NA objective lens and a laser wavelength of 633 nm, show that the system is capable of measuring a 50 nm track width, with a standard deviation less than 4 nm.

Morphology-controllable of Sn doped ZnO nanorods prepared by spray pyrolysis for transparent electrode application

NASA Astrophysics Data System (ADS)

Hameed, M. Shahul; Princice, J. Joseph; Babu, N. Ramesh; Zahirullah, S. Syed; Deshmukh, Sampat G.; Arunachalam, A.

2018-05-01

Transparent conductive Sn doped ZnO nanorods have been deposited at various doping level by spray pyrolysis technique on glass substrate. The structural, surface morphological and optical properties of these films have been investigated with the help of X-ray diffraction (XRD), scanning electron microscope (SEM), atomic force microscope (AFM) and UV-Vis spectrophotometer respectively. XRD patterns revealed a successful high quality growth of single crystal ZnO nanorods with hexagonal wurtzite structure having (002) preferred orientation. The scanning electron microscope (SEM) image of the prepared films exposed the uniform distribution of Sn doped ZnO nanorod shaped grains. All these films were highly transparent in the visible region with average transmittance of 90%.

Hyper-spectral imaging in scanning-confocal-fluorescence microscopy using a novel broadband diffractive optic

NASA Astrophysics Data System (ADS)

Wang, Peng; Ebeling, Carl G.; Gerton, Jordan; Menon, Rajesh

In this paper, we demonstrate hyper-spectral imaging of fluorescent microspheres in a scanning-confocal-fluorescence microscope by spatially dispersing the spectra using a novel broadband diffractive optic, and applying a nonlinear optimization technique to extract the full-incident spectra. This broadband diffractive optic has a designed optical efficiency of over 90% across the entire visible spectrum. We used this technique to create two-color images of two fluorophores and also extracted their emission spectra with good fidelity. This method can be extended to image both spatially and spectrally overlapping fluorescent samples. Full control in the number of emission spectra and the feasibility of enhanced imaging speed are demonstrated as well.

Ultrahigh resolution multicolor colocalization of single fluorescent probes

DOEpatents

Weiss, Shimon; Michalet, Xavier; Lacoste, Thilo D.

2005-01-18

A novel optical ruler based on ultrahigh-resolution colocalization of single fluorescent probes is described. Two unique families of fluorophores are used, namely energy-transfer fluorescent beads and semiconductor nanocrystal (NC) quantum dots, that can be excited by a single laser wavelength but emit at different wavelengths. A novel multicolor sample-scanning confocal microscope was constructed which allows one to image each fluorescent light emitter, free of chromatic aberrations, by scanning the sample with nanometer scale steps using a piezo-scanner. The resulting spots are accurately localized by fitting them to the known shape of the excitation point-spread-function of the microscope.

[Evaluation of the surface of the new intraocular lenses in the scanning electron microscope].

PubMed

Kałuzny, B J; Szatkowski, J; Kałuzny, J J

2001-01-01

To evaluate the surface of the new PC IOLs commercially available in Poland in 2000. Representative samples of new posterior chamber IOLs produced by 6 different companies (Alcon, Lensita, Medicontur, Opsia, Rayner, Storz), 5 of each, underwent surface examination with Novoscan 30 scanning electron microscope. Although, in general, smooth surface of optic and haptic parts were observed, three samples with irregularities were found. Comparing to previous evaluation performed in 1994, significant improvement in quality of IOLs surface was noted. No considerable differences in this field between above mentioned producers were observed.

Note: Automated electrochemical etching and polishing of silver scanning tunneling microscope tips.

PubMed

Sasaki, Stephen S; Perdue, Shawn M; Rodriguez Perez, Alejandro; Tallarida, Nicholas; Majors, Julia H; Apkarian, V Ara; Lee, Joonhee

2013-09-01

Fabrication of sharp and smooth Ag tips is crucial in optical scanning probe microscope experiments. To ensure reproducible tip profiles, the polishing process is fully automated using a closed-loop laminar flow system to deliver the electrolytic solution to moving electrodes mounted on a motorized translational stage. The repetitive translational motion is controlled precisely on the μm scale with a stepper motor and screw-thread mechanism. The automated setup allows reproducible control over the tip profile and improves smoothness and sharpness of tips (radius 27 ± 18 nm), as measured by ultrafast field emission.

Compact divided-pupil line-scanning confocal microscope for investigation of human tissues

NASA Astrophysics Data System (ADS)

Glazowski, Christopher; Peterson, Gary; Rajadhyaksha, Milind

2013-03-01

Divided-pupil line-scanning confocal microscopy (DPLSCM) can provide a simple and low-cost approach for imaging of human tissues with pathology-like nuclear and cellular detail. Using results from a multidimensional numerical model of DPLSCM, we found optimal pupil configurations for improved axial sectioning, as well as control of speckle noise in the case of reflectance imaging. The modeling results guided the design and construction of a simple (10 component) microscope, packaged within the footprint of an iPhone, and capable of cellular resolution. We present the optical design with experimental video-images of in-vivo human tissues.

Optical computing.

NASA Technical Reports Server (NTRS)

Stroke, G. W.

1972-01-01

Applications of the optical computer include an approach for increasing the sharpness of images obtained from the most powerful electron microscopes and fingerprint/credit card identification. The information-handling capability of the various optical computing processes is very great. Modern synthetic-aperture radars scan upward of 100,000 resolvable elements per second. Fields which have assumed major importance on the basis of optical computing principles are optical image deblurring, coherent side-looking synthetic-aperture radar, and correlative pattern recognition. Some examples of the most dramatic image deblurring results are shown.

Traceable X,Y self-calibration at single nm level of an optical microscope used for coherence scanning interferometry

NASA Astrophysics Data System (ADS)

Ekberg, Peter; Mattsson, Lars

2018-03-01

Coherence scanning interferometry used in optical profilers are typically good for Z-calibration at nm-levels, but the X,Y accuracy is often left without further notice than typical resolution limits of the optics, i.e. of the order of ~1 µm. For the calibration of metrology tools we rely on traceable artefacts, e.g. gauge blocks for traditional coordinate measurement machines, and lithographically mask made artefacts for microscope calibrations. In situations where the repeatability and accuracy of the measurement tool is much better than the uncertainty of the traceable artefact, we are bound to specify the uncertainty based on the calibration artefact rather than on the measurement tool. This is a big drawback as the specified uncertainty of a calibrated measurement may shrink the available manufacturing tolerance. To improve the uncertainty in X,Y we can use self-calibration. Then, we do not need to know anything more than that the artefact contains a pattern with some nominal grid. This also gives the opportunity to manufacture the artefact in-house, rather than buying a calibrated and expensive artefact. The self-calibration approach we present here is based on an iteration algorithm, rather than the traditional mathematical inversion, and it leads to much more relaxed constrains on the input measurements. In this paper we show how the X,Y errors, primarily optical distortions, within the field of view (FOV) of an optical coherence scanning interferometry microscope, can be reduced with a large factor. By self-calibration we achieve an X,Y consistency in the 175  ×  175 µm2 FOV of ~2.3 nm (1σ) using the 50×  objective. Besides the calibrated coordinate X,Y system of the microscope we also receive, as a bonus, the absolute positions of the pattern in the artefact with a combined uncertainty of 6 nm (1σ) by relying on a traceable 1D linear measurement of a twin artefact at NIST.

Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices

PubMed Central

Losavio, Bradley E.; Iyer, Vijay; Saggau, Peter

2009-01-01

We developed a two-photon microscope optimized for physiologically manipulating single neurons through their postsynaptic receptors. The optical layout fulfills the stringent design criteria required for high-speed, high-resolution imaging in scattering brain tissue with minimal photodamage. We detail the practical compensation of spectral and temporal dispersion inherent in fast laser beam scanning with acousto-optic deflectors, as well as a set of biological protocols for visualizing nearly diffraction-limited structures and delivering physiological synaptic stimuli. The microscope clearly resolves dendritic spines and evokes electrophysiological transients in single neurons that are similar to endogenous responses. This system enables the study of multisynaptic integration and will assist our understanding of single neuron function and dendritic computation. PMID:20059271

Multimodal nonlinear microscope based on a compact fiber-format laser source

NASA Astrophysics Data System (ADS)

Crisafi, Francesco; Kumar, Vikas; Perri, Antonio; Marangoni, Marco; Cerullo, Giulio; Polli, Dario

2018-01-01

We present a multimodal non-linear optical (NLO) laser-scanning microscope, based on a compact fiber-format excitation laser and integrating coherent anti-Stokes Raman scattering (CARS), stimulated Raman scattering (SRS) and two-photon-excitation fluorescence (TPEF) on a single platform. We demonstrate its capabilities in simultaneously acquiring CARS and SRS images of a blend of 6-μm poly(methyl methacrylate) beads and 3-μm polystyrene beads. We then apply it to visualize cell walls and chloroplast of an unprocessed fresh leaf of Elodea aquatic plant via SRS and TPEF modalities, respectively. The presented NLO microscope, developed in house using off-the-shelf components, offers full accessibility to the optical path and ensures its easy re-configurability and flexibility.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Henn, T.; Kiessling, T., E-mail: tobias.kiessling@physik.uni-wuerzburg.de; Ossau, W.

We describe a two-color pump-probe scanning magneto-optical Kerr effect microscope which we have developed to investigate electron spin phenomena in semiconductors at cryogenic temperatures with picosecond time and micrometer spatial resolution. The key innovation of our microscope is the usage of an ultrafast “white light” supercontinuum fiber-laser source which provides access to the whole visible and near-infrared spectral range. Our Kerr microscope allows for the independent selection of the excitation and detection energy while avoiding the necessity to synchronize the pulse trains of two separate picosecond laser systems. The ability to independently tune the pump and probe wavelength enables themore » investigation of the influence of excitation energy on the optically induced electron spin dynamics in semiconductors. We demonstrate picosecond real-space imaging of the diffusive expansion of optically excited electron spin packets in a (110) GaAs quantum well sample to illustrate the capabilities of the instrument.« less

Automated Image Analysis Corrosion Working Group Update: February 1, 2018

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wendelberger, James G.

These are slides for the automated image analysis corrosion working group update. The overall goals were: automate the detection and quantification of features in images (faster, more accurate), how to do this (obtain data, analyze data), focus on Laser Scanning Confocal Microscope (LCM) data (laser intensity, laser height/depth, optical RGB, optical plus laser RGB).

SEM, optical, and Moessbauer studies of submicrometer chromite in Allende

NASA Technical Reports Server (NTRS)

Housley, R. M.

1982-01-01

New scanning electron and optical microscope results are presented showing that sub-micrometer chromite is abundant along healed cracks and grain boundaries in Allende chondrule olivine. Some wider healed cracks also contain pentlandite and euhedral Ni3Fe grains. Also reported are Moessbauer measurements on Allende HF-HCl residues confirming a high Fe(+++)/Fe(++) ratio.

Design and performance of an X-ray scanning microscope at the Hard X-ray Nanoprobe beamline of NSLS-II

DOE PAGES

Nazaretski, E.; Yan, H.; Lauer, K.; ...

2017-10-05

A hard X-ray scanning microscope installed at the Hard X-ray Nanoprobe beamline of the National Synchrotron Light Source II has been designed, constructed and commissioned. The microscope relies on a compact, high stiffness, low heat dissipation approach and utilizes two types of nanofocusing optics. It is capable of imaging with ~15 nm × 15 nm spatial resolution using multilayer Laue lenses and 25 nm × 26 nm resolution using zone plates. Fluorescence, diffraction, absorption, differential phase contrast, ptychography and tomography are available as experimental techniques. The microscope is also equipped with a temperature regulation system which allows the temperature ofmore » a sample to be varied in the range between 90 K and 1000 K. The constructed instrument is open for general users and offers its capabilities to the material science, battery research and bioscience communities.« less

Design and performance of an X-ray scanning microscope at the Hard X-ray Nanoprobe beamline of NSLS-II

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nazaretski, E.; Yan, H.; Lauer, K.

A hard X-ray scanning microscope installed at the Hard X-ray Nanoprobe beamline of the National Synchrotron Light Source II has been designed, constructed and commissioned. The microscope relies on a compact, high stiffness, low heat dissipation approach and utilizes two types of nanofocusing optics. It is capable of imaging with ~15 nm × 15 nm spatial resolution using multilayer Laue lenses and 25 nm × 26 nm resolution using zone plates. Fluorescence, diffraction, absorption, differential phase contrast, ptychography and tomography are available as experimental techniques. The microscope is also equipped with a temperature regulation system which allows the temperature ofmore » a sample to be varied in the range between 90 K and 1000 K. The constructed instrument is open for general users and offers its capabilities to the material science, battery research and bioscience communities.« less

An investigation of nitride precipitates in archaeological iron artefacts from Poland.

PubMed

Kedzierski, Z; Stepiński, J; Zielińska-Lipiec, A

2010-03-01

The paper describes the investigations of nitride precipitates in a spearhead and a sword found in the territory of Poland, in cremation graveyards of the Przeworsk Culture, dated to the Roman Period. Three different techniques of the examination of nitride precipitates were employed: optical microscope, scanning electron microscope (scanning electron microscope with energy dispersive X-ray spectrometer) and transmission electron microscope. Two types of precipitates have been observed, and their plate-like shape was demonstrated. The large precipitate has been confirmed to be gamma'-Fe(4)N, whereas the small one has been identified as alpha''-Fe(16)N(2). The origin of nitride precipitates in archaeological iron artefacts from Poland is probably a result of the manufacturing process or cremation as part of burial rites. An examination of available iron artefacts indicates that nitride precipitates (have only limited effect on mechanical properties) influence the hardness of metal only to a very limited degree.

Modern technologies for retinal scanning and imaging: an introduction for the biomedical engineer

PubMed Central

2014-01-01

This review article is meant to help biomedical engineers and nonphysical scientists better understand the principles of, and the main trends in modern scanning and imaging modalities used in ophthalmology. It is intended to ease the communication between physicists, medical doctors and engineers, and hopefully encourage “classical” biomedical engineers to generate new ideas and to initiate projects in an area which has traditionally been dominated by optical physics. Most of the methods involved are applicable to other areas of biomedical optics and optoelectronics, such as microscopic imaging, spectroscopy, spectral imaging, opto-acoustic tomography, fluorescence imaging etc., all of which are with potential biomedical application. Although all described methods are novel and important, the emphasis of this review has been placed on three technologies introduced in the 1990’s and still undergoing vigorous development: Confocal Scanning Laser Ophthalmoscopy, Optical Coherence Tomography, and polarization-sensitive retinal scanning. PMID:24779618

Provenance study through analysis of microstructural characteristics using an optical microscope and scanning electron microscopy for Goryeo celadon excavated from the seabed.

PubMed

Min-su, Han

2013-08-01

This paper aims at identifying the provenance of Goryeo celadons by understanding its microstructural characteristics, such as particles, blisters, forms and amount of pores, and the presence of crystal formation, bodies, and glazes and its boundary, using an optical microscope and scanning electron microscopy (SEM). The analysis of the reproduced samples shows that the glazed layer of the sherd fired at higher temperatures has lower viscosity and therefore it encourages the blisters to be combined together and the layer to become more transparent. In addition, the result showed that the vitrification and melting process of clay minerals such as feldspars and quartzs on the bodies was accelerated for those samples. To factor such characteristics of the microstructure and apply it to the sherds, the samples could be divided into six categories based on status, such as small particles with many small pores or mainly large and small circular pores in the bodies, only a limited number of varied sized blisters in the glazes, and a few blisters and needle-shaped crystals on the boundary surface. In conclusion, the analysis of the microstructural characteristics using an optical microscope and SEM have proven to be useful as a categorizing reference factor in a provenance study on Goryeo celadons.

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

NASA Astrophysics Data System (ADS)

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.

Two-Photon Fluorescence Microscope for Microgravity Research

NASA Technical Reports Server (NTRS)

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

2005-01-01

A two-photon fluorescence microscope has been developed for the study of biophysical phenomena. Two-photon microscopy is a novel form of laser-based scanning microscopy that enables three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon optical microscopy, two-photon microscopy utilizes the simultaneous nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption, so an ultra-fast pulsed laser source is typically employed. On the other hand, the critical energy threshold for two-photon absorption leads to fluorophore excitation that is intrinsically localized to the focal volume. Consequently, two-photon microscopy enables optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction (relative to one-photon optical microscopy) in photon-induced damage because of the longer excitation wavelength. This reduction is especially advantageous for in vivo studies. Relative to confocal microscopy, there is also a reduction in background fluorescence, and, because of a reduction in Rayleigh scattering, there is a 4 increase of penetration depth. The prohibitive cost of a commercial two-photon fluorescence-microscope system, as well as a need for modularity, has led to the construction of a custom-built system (see Figure 1). This system includes a coherent mode-locked titanium: sapphire laser emitting 120-fs-duration pulses at a repetition rate of 80 MHz. The pulsed laser has an average output power of 800 mW and a wavelength tuning range of 700 to 980 nm, enabling the excitation of a variety of targeted fluorophores. The output from the laser is attenuated, spatially filtered, and then directed into a confocal scanning head that has been modified to provide for side entry of the laser beam. The laser output coupler has been replaced with a dichroic filter that reflects the longer-wavelength excitation light and passes the shorter-wavelength fluorescence light. Also, the confocal pinhole has been removed to increase the signal strength. The laser beam is scanned by a twoperpendicular- axis pair of galvanometer mirrors through a pupil transfer lens into the side port of an inverted microscope. Finally, the beam is focused by a 63-magnification, 1.3-numerical- aperture oil-immersion objective lens onto a specimen. The pupil transfer lens serves to match the intermediate image planes of the scanning head and the microscope, and its location is critical. In order to maximize the quality of the image, (that is, the point spread function of the objective lens for all scan positions), the entire system was modeled in optical-design software, and the various free design parameters (the parameters of the spatial-filter components as well as the separations of all of the system components) were determined through an iterative optimization process. A modular design was chosen to facilitate access to the optical train for future fluorescence correlation spectroscopy and fluorescence-lifetime experiments.

Scanning Microscopes Using X Rays and Microchannels

NASA Technical Reports Server (NTRS)

Wang, Yu

2003-01-01

Scanning microscopes that would be based on microchannel filters and advanced electronic image sensors and that utilize x-ray illumination have been proposed. Because the finest resolution attainable in a microscope is determined by the wavelength of the illumination, the xray illumination in the proposed microscopes would make it possible, in principle, to achieve resolutions of the order of nanometers about a thousand times as fine as the resolution of a visible-light microscope. Heretofore, it has been necessary to use scanning electron microscopes to obtain such fine resolution. In comparison with scanning electron microscopes, the proposed microscopes would likely be smaller, less massive, and less expensive. Moreover, unlike in scanning electron microscopes, it would not be necessary to place specimens under vacuum. The proposed microscopes are closely related to the ones described in several prior NASA Tech Briefs articles; namely, Miniature Microscope Without Lenses (NPO-20218), NASA Tech Briefs, Vol. 22, No. 8 (August 1998), page 43; and Reflective Variants of Miniature Microscope Without Lenses (NPO-20610), NASA Tech Briefs, Vol. 26, No. 9 (September 2002) page 6a. In all of these microscopes, the basic principle of design and operation is the same: The focusing optics of a conventional visible-light microscope are replaced by a combination of a microchannel filter and a charge-coupled-device (CCD) image detector. A microchannel plate containing parallel, microscopic-cross-section holes much longer than they are wide is placed between a specimen and an image sensor, which is typically the CCD. The microchannel plate must be made of a material that absorbs the illuminating radiation reflected or scattered from the specimen. The microchannels must be positioned and dimensioned so that each one is registered with a pixel on the image sensor. Because most of the radiation incident on the microchannel walls becomes absorbed, the radiation that reaches the image sensor consists predominantly of radiation that was launched along the longitudinal direction of the microchannels. Therefore, most of the radiation arriving at each pixel on the sensor must have traveled along a straight line from a corresponding location on the specimen. Thus, there is a one-to-one mapping from a point on a specimen to a pixel in the image sensor, so that the output of the image sensor contains image information equivalent to that from a microscope.

Simultaneous photoacoustic and optically mediated ultrasound microscopy: an in vivo study

PubMed Central

Orlova, Anna; Shirmanova, Marina; Postnikova, Anna; Turchin, Ilya

2015-01-01

We propose the use of thermoelastic (TE) excitation of an ultrasonic (US) detector by backscattered laser radiation as a means of upgrading a single-modality photoacoustic (PA) microscope to dual-modality PA/US imaging at minimal cost. The upgraded scanning head of our dual-modality microscope consists of a fiber bundle with 14 output arms and a 32MHz polyvinylidene difluoride (PVDF) detector with a 34 MHz bandwidth (−6 dB level), 12.7 mm focal length, and a 0.25 numerical aperture. A single optical pulse delivered through the fiber bundle to the biotissue being investigated, in combination with a metalized surface on the PVDF detector allows us to obtain both PA and US A-scans. To demonstrate the in vivo capabilities of the proposed method we present the results of bimodal imaging of the brain of a newborn rat, a mouse tail and a mouse tumor. PMID:25780752

Design of a cathodoluminescence image generator using a Raspberry Pi coupled to a scanning electron microscope.

PubMed

Benítez, Alfredo; Santiago, Ulises; Sanchez, John E; Ponce, Arturo

2018-01-01

In this work, an innovative cathodoluminescence (CL) system is coupled to a scanning electron microscope and synchronized with a Raspberry Pi computer integrated with an innovative processing signal. The post-processing signal is based on a Python algorithm that correlates the CL and secondary electron (SE) images with a precise dwell time correction. For CL imaging, the emission signal is collected through an optical fiber and transduced to an electrical signal via a photomultiplier tube (PMT). CL Images are registered in a panchromatic mode and can be filtered using a monochromator connected between the optical fiber and the PMT to produce monochromatic CL images. The designed system has been employed to study ZnO samples prepared by electrical arc discharge and microwave methods. CL images are compared with SE images and chemical elemental mapping images to correlate the emission regions of the sample.

Design of a cathodoluminescence image generator using a Raspberry Pi coupled to a scanning electron microscope

NASA Astrophysics Data System (ADS)

Benítez, Alfredo; Santiago, Ulises; Sanchez, John E.; Ponce, Arturo

2018-01-01

In this work, an innovative cathodoluminescence (CL) system is coupled to a scanning electron microscope and synchronized with a Raspberry Pi computer integrated with an innovative processing signal. The post-processing signal is based on a Python algorithm that correlates the CL and secondary electron (SE) images with a precise dwell time correction. For CL imaging, the emission signal is collected through an optical fiber and transduced to an electrical signal via a photomultiplier tube (PMT). CL Images are registered in a panchromatic mode and can be filtered using a monochromator connected between the optical fiber and the PMT to produce monochromatic CL images. The designed system has been employed to study ZnO samples prepared by electrical arc discharge and microwave methods. CL images are compared with SE images and chemical elemental mapping images to correlate the emission regions of the sample.

An ultrahigh-vacuum cryostat for simultaneous scanning tunneling microscopy and magneto-transport measurements down to 400 mK

NASA Astrophysics Data System (ADS)

Liebmann, Marcus; Bindel, Jan Raphael; Pezzotta, Mike; Becker, Stefan; Muckel, Florian; Johnsen, Tjorven; Saunus, Christian; Ast, Christian R.; Morgenstern, Markus

2017-12-01

We present the design and calibration measurements of a scanning tunneling microscope setup in a 3He ultrahigh-vacuum cryostat operating at 400 mK with a hold time of 10 days. With 2.70 m in height and 4.70 m free space needed for assembly, the cryostat fits in a one-story lab building. The microscope features optical access, an xy table, in situ tip and sample exchange, and enough contacts to facilitate atomic force microscopy in tuning fork operation and simultaneous magneto-transport measurements on the sample. Hence, it enables scanning tunneling spectroscopy on microstructured samples which are tuned into preselected transport regimes. A superconducting magnet provides a perpendicular field of up to 14 T. The vertical noise of the scanning tunneling microscope amounts to 1 pmrms within a 700 Hz bandwidth. Tunneling spectroscopy using one superconducting electrode revealed an energy resolution of 120 μeV. Data on tip-sample Josephson contacts yield an even smaller feature size of 60 μeV, implying that the system operates close to the physical noise limit.

An ultrahigh-vacuum cryostat for simultaneous scanning tunneling microscopy and magneto-transport measurements down to 400 mK.

PubMed

Liebmann, Marcus; Bindel, Jan Raphael; Pezzotta, Mike; Becker, Stefan; Muckel, Florian; Johnsen, Tjorven; Saunus, Christian; Ast, Christian R; Morgenstern, Markus

2017-12-01

We present the design and calibration measurements of a scanning tunneling microscope setup in a 3 He ultrahigh-vacuum cryostat operating at 400 mK with a hold time of 10 days. With 2.70 m in height and 4.70 m free space needed for assembly, the cryostat fits in a one-story lab building. The microscope features optical access, an xy table, in situ tip and sample exchange, and enough contacts to facilitate atomic force microscopy in tuning fork operation and simultaneous magneto-transport measurements on the sample. Hence, it enables scanning tunneling spectroscopy on microstructured samples which are tuned into preselected transport regimes. A superconducting magnet provides a perpendicular field of up to 14 T. The vertical noise of the scanning tunneling microscope amounts to 1 pm rms within a 700 Hz bandwidth. Tunneling spectroscopy using one superconducting electrode revealed an energy resolution of 120 μeV. Data on tip-sample Josephson contacts yield an even smaller feature size of 60 μeV, implying that the system operates close to the physical noise limit.

Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens

PubMed Central

Grewe, Benjamin F.; Voigt, Fabian F.; van ’t Hoff, Marcel; Helmchen, Fritjof

2011-01-01

Functional two-photon Ca2+-imaging is a versatile tool to study the dynamics of neuronal populations in brain slices and living animals. However, population imaging is typically restricted to a single two-dimensional image plane. By introducing an electrically tunable lens into the excitation path of a two-photon microscope we were able to realize fast axial focus shifts within 15 ms. The maximum axial scan range was 0.7 mm employing a 40x NA0.8 water immersion objective, plenty for typically required ranges of 0.2–0.3 mm. By combining the axial scanning method with 2D acousto-optic frame scanning and random-access scanning, we measured neuronal population activity of about 40 neurons across two imaging planes separated by 40 μm and achieved scan rates up to 20–30 Hz. The method presented is easily applicable and allows upgrading of existing two-photon microscopes for fast 3D scanning. PMID:21750778

0.4 Microns Spatial Resolution with 1 GHz (lambda = 30 cm) Evanescent Microwave Probe

NASA Technical Reports Server (NTRS)

Tabib-Azar, M.; Su, D.-P.; Pohar, A.; LeClair, S. R.; Ponchak, George E.

1999-01-01

In this article we describe evanescent field imaging of material nonuniformities with a record resolution of 0.4 microns at 1 GHz (lambda(sub g)/750000), using a resonant stripline scanning microwave probe. A chemically etched tip is used as a point-like evanescent field emitter and a probe-sample distance modulation is employed to improve the signal-to-noise ratio. Images obtained by evanescent microwave probe, by optical microscope, and by scanning tunneling microscope are presented for comparison. Probe was calibrated to perform quantitative conductivity measurements. The principal factors affecting the ultimate resolution of evanescent microwave probe are also discussed.

Astigmatism correction in x-ray scanning photoemission microscope with use of elliptical zone plate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ade, H.; Ko, C.; Anderson, E.

1992-03-02

We report the impact of an elliptical, high resolution zone plate on the performance of an initially astigmatic soft x-ray scanning photoemission microscope. A zone plate with carefully calibrated eccentricity has been used to eliminate astigmatism arising from transport optics, and an improvement of about a factor of 3 in spatial resolution was achieved. The resolution is still dominated by the source size and chromatic aberrations rather than by diffraction and coma, and a further gain of about a factor of 2 in resolution is possible. Sub 100 nm photoemission microscopy with primary photoelectrons is now within reach.

Nano-scale luminescence characterization of individual InGaN/GaN quantum wells stacked in a microcavity using scanning transmission electron microscope cathodoluminescence

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schmidt, Gordon, E-mail: Gordon.Schmidt@ovgu.de; Müller, Marcus; Veit, Peter

2014-07-21

Using cathodoluminescence spectroscopy directly performed in a scanning transmission electron microscope at liquid helium temperatures, the optical and structural properties of a 62 InGaN/GaN multiple quantum well embedded in an AlInN/GaN based microcavity are investigated at the nanometer scale. We are able to spatially resolve a spectral redshift between the individual quantum wells towards the surface. Cathodoluminescence spectral linescans allow directly visualizing the critical layer thickness in the quantum well stack resulting in the onset of plastic relaxation of the strained InGaN/GaN system.

A novel graphene oxide-polyimide as optical waveguide material: Synthesis and thermo-optic switch properties

NASA Astrophysics Data System (ADS)

Cao, Tianlin; Zhao, Fanyu; Da, Zulin; Qiu, Fengxian; Yang, Dongya; Guan, Yijun; Cao, Guorong; Zhao, Zerun; Li, Jiaxin; Guo, Xiaotong

2016-10-01

In this work, a novel graphene oxide-polyimide (GOPI) as optical waveguide material was prepared. The structure, mechanical, thermal property and morphology of the GOPI was characterized by using fourier transform infrared, UV-visible spectroscopy, near-infrared spectrum, thermogravimetric analysis, differential scanning calorimetry, scanning electron microscope and transmission electron microscopy. The thermo-optic coefficients (dn/dT) are -9.16 × 10-4 (532 nm), -7.56 × 10-4 (650 nm) and -4.82 × 10-4 (850 nm) °C-1, respectively. Based on the thermo-optic effect of prepared GOPI as waveguide material, a Y-branch with branching angle of 0.143° and Mach-Zehnder thermo-optic switches were designed. Using finite difference beam propagation method (FD-BPM) method, the simulation results such as power consumptions and response times of two different thermo-optic switches were obtained.

Near-Field Scanning Optical Microscopy and Raman Microscopy.

NASA Astrophysics Data System (ADS)

Harootunian, Alec Tate

1987-09-01

Both a one dimensional near-field scanning optical microscope and Raman microprobe were constructed. In near -field scanning optical microscopy (NSOM) a subwavelength aperture is scanned in the near-field of the object. Radiation transmitted through the aperture is collected to form an image as the aperture scans over the object. The resolution of an NSOM system is essentially wavelength independent and is limited by the diameter of the aperture used to scan the object. NSOM was developed in an effort to provide a nondestructive in situ high spatial resolution probe while still utilizing photons at optical wavelengths. The Raman microprobe constructed provided vibrational Raman information with spatial resolution equivalent that of a conventional diffraction limited microscope. Both transmission studies and near-field diffration studies of subwavelength apertures were performed. Diffraction theories for a small aperture in an infinitely thin conducting screen, a slit in a thick conducting screen, and an aperture in a black screen were examined. All three theories indicate collimation of radiation to the size to the size of the subwavelength aperture or slit in the near-field. Theoretical calculations and experimental results indicate that light transmitted through subwavelength apertures is readily detectable. Light of wavelength 4579 (ANGSTROM) was transmitted through apertures with diameters as small as 300 (ANGSTROM). These studies indicate the feasibility of constructing an NSOM system. One dimensional transmission and fluorescence NSOM systems were constructed. Apertures in the tips of metallized glass pipettes width inner diameters of less than 1000 (ANGSTROM) were used as a light source in the NSOM system. A tunneling current was used to maintain the aperture position in the near-field. Fluorescence NSOM was demonstrated for the first time. Microspectroscopic and Raman microscopic studies of turtle cone oil droplets were performed. Both the Raman vibrational frequencies and the Raman excitation data indicate that the carotenoids are unaggregated. The carotenoid astaxanthin was identified in the orange and red droplets by Raman microscopy. Future applications for both Raman microscopy and near-field microscopy were proposed. Four methods of near-field distance regulation were also examined. Finally, theoretical exposure curves for near-field lithography were calculated. Both the near-field lithographic results and the near field diffraction studies indicate essentially wavelength independent resolution. (Abstract shortened with permission of author.).

LabVIEW control software for scanning micro-beam X-ray fluorescence spectrometer.

PubMed

Wrobel, Pawel; Czyzycki, Mateusz; Furman, Leszek; Kolasinski, Krzysztof; Lankosz, Marek; Mrenca, Alina; Samek, Lucyna; Wegrzynek, Dariusz

2012-05-15

Confocal micro-beam X-ray fluorescence microscope was constructed. The system was assembled from commercially available components - a low power X-ray tube source, polycapillary X-ray optics and silicon drift detector - controlled by an in-house developed LabVIEW software. A video camera coupled to optical microscope was utilized to display the area excited by X-ray beam. The camera image calibration and scan area definition software were also based entirely on LabVIEW code. Presently, the main area of application of the newly constructed spectrometer is 2-dimensional mapping of element distribution in environmental, biological and geological samples with micrometer spatial resolution. The hardware and the developed software can already handle volumetric 3-D confocal scans. In this work, a front panel graphical user interface as well as communication protocols between hardware components were described. Two applications of the spectrometer, to homogeneity testing of titanium layers and to imaging of various types of grains in air particulate matter collected on membrane filters, were presented. Copyright © 2012 Elsevier B.V. All rights reserved.

The original method for imaging of biological tissues in optical coherence tomography with usage of hyperchromatic lens

NASA Astrophysics Data System (ADS)

Egorov, D. I.

2017-06-01

Our study focuses on an analysis of the original method of investigation biological tissues in the spectral OCT (optical coherence tomography) with usage hyperchromatic lenses. Using hyperchromatic lens, i.e. the lens with uncorrected longitudinal color allows scanning in the depth of the object by changing the wavelength of the emitter. In this case, the depth of the scan will be determined not by the microlens depth of field, but the value of axial color. In our study, we demonstrated the advantages of this method of research on biological tissues existing. Spectral OCT schemes with the hyperchromatic lens could increase the depth of spectral scanning, eliminate the use of multi-channel systems with a set of microscope objectives, reduce the time of measurement. In our paper, we show the developed method of calculation of hyperchromatic lenses and hybrid hyperchromatic lens consisting of a diffractive and refractive component in spectral OCT systems. We also demonstrate the results of aberration calculation designed microscope lenses. We show examples of developed hyperchromatic lenses with the diffractive element and without it.

A 3D Polymer Based Printed Two-Dimensional Laser Scanner

NASA Astrophysics Data System (ADS)

Oyman, H. A.; Gokdel, Y. D.; Ferhanoglu, O.; Yalcinkaya, A. D.

2016-10-01

A two-dimensional (2D) polymer based scanning mirror with magnetic actuation is developed for imaging applications. Proposed device consists of a circular suspension holding a rectangular mirror and can generate a 2D scan pattern. Three dimensional (3D) printing technology which is used for implementation of the device, offers added flexibility in controlling the cross-sectional profile as well as the stress distribution compared to the traditional planar process technologies. The mirror device is developed to meet a portable, miniaturized confocal microscope application in mind, delivering 4.5 and 4.8 degrees of optical scan angles at 111 and 267 Hz, respectively. As a result of this mechanical performance, the resulting microscope incorporating the mirror is estimated to accomplish a field of view (FOV) of 350 µm × 350 µm.

Hyperspectral imaging with laser-scanning sum-frequency generation microscopy

PubMed Central

Hanninen, Adam; Shu, Ming Wai; Potma, Eric O.

2017-01-01

Vibrationally sensitive sum-frequency generation (SFG) microscopy is a chemically selective imaging technique sensitive to non-centrosymmetric molecular arrangements in biological samples. The routine use of SFG microscopy has been hampered by the difficulty of integrating the required mid-infrared excitation light into a conventional, laser-scanning nonlinear optical (NLO) microscope. In this work, we describe minor modifications to a regular laser-scanning microscope to accommodate SFG microscopy as an imaging modality. We achieve vibrationally sensitive SFG imaging of biological samples with sub-μm resolution at image acquisition rates of 1 frame/s, almost two orders of magnitude faster than attained with previous point-scanning SFG microscopes. Using the fast scanning capability, we demonstrate hyperspectral SFG imaging in the CH-stretching vibrational range and point out its use in the study of molecular orientation and arrangement in biologically relevant samples. We also show multimodal imaging by combining SFG microscopy with second-harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) on the same imaging platfrom. This development underlines that SFG microscopy is a unique modality with a spatial resolution and image acquisition time comparable to that of other NLO imaging techniques, making point-scanning SFG microscopy a valuable member of the NLO imaging family. PMID:28966861

Optimal resolution in Fresnel incoherent correlation holographic fluorescence microscopy

PubMed Central

Brooker, Gary; Siegel, Nisan; Wang, Victor; Rosen, Joseph

2011-01-01

Fresnel Incoherent Correlation Holography (FINCH) enables holograms and 3D images to be created from incoherent light with just a camera and spatial light modulator (SLM). We previously described its application to microscopic incoherent fluorescence wherein one complex hologram contains all the 3D information in the microscope field, obviating the need for scanning or serial sectioning. We now report experiments which have led to the optimal optical, electro-optic, and computational conditions necessary to produce holograms which yield high quality 3D images from fluorescent microscopic specimens. An important improvement from our previous FINCH configurations capitalizes on the polarization sensitivity of the SLM so that the same SLM pixels which create the spherical wave simulating the microscope tube lens, also pass the plane waves from the infinity corrected microscope objective, so that interference between the two wave types at the camera creates a hologram. This advance dramatically improves the resolution of the FINCH system. Results from imaging a fluorescent USAF pattern and a pollen grain slide reveal resolution which approaches the Rayleigh limit by this simple method for 3D fluorescent microscopic imaging. PMID:21445140

HelioScan: a software framework for controlling in vivo microscopy setups with high hardware flexibility, functional diversity and extendibility.

PubMed

Langer, Dominik; van 't Hoff, Marcel; Keller, Andreas J; Nagaraja, Chetan; Pfäffli, Oliver A; Göldi, Maurice; Kasper, Hansjörg; Helmchen, Fritjof

2013-04-30

Intravital microscopy such as in vivo imaging of brain dynamics is often performed with custom-built microscope setups controlled by custom-written software to meet specific requirements. Continuous technological advancement in the field has created a need for new control software that is flexible enough to support the biological researcher with innovative imaging techniques and provide the developer with a solid platform for quickly and easily implementing new extensions. Here, we introduce HelioScan, a software package written in LabVIEW, as a platform serving this dual role. HelioScan is designed as a collection of components that can be flexibly assembled into microscope control software tailored to the particular hardware and functionality requirements. Moreover, HelioScan provides a software framework, within which new functionality can be implemented in a quick and structured manner. A specific HelioScan application assembles at run-time from individual software components, based on user-definable configuration files. Due to its component-based architecture, HelioScan can exploit synergies of multiple developers working in parallel on different components in a community effort. We exemplify the capabilities and versatility of HelioScan by demonstrating several in vivo brain imaging modes, including camera-based intrinsic optical signal imaging for functional mapping of cortical areas, standard two-photon laser-scanning microscopy using galvanometric mirrors, and high-speed in vivo two-photon calcium imaging using either acousto-optic deflectors or a resonant scanner. We recommend HelioScan as a convenient software framework for the in vivo imaging community. Copyright © 2013 Elsevier B.V. All rights reserved.

Method and apparatus for chemical and topographical microanalysis

NASA Technical Reports Server (NTRS)

Kossakovski, Dmitri A. (Inventor); Baldeschwieler, John D. (Inventor); Beauchamp, Jesse L. (Inventor)

2002-01-01

A scanning probe microscope is combined with a laser induced breakdown spectrometer to provide spatially resolved chemical analysis of the surface correlated with the surface topography. Topographical analysis is achieved by scanning a sharp probe across the sample at constant distance from the surface. Chemical analysis is achieved by the means of laser induced breakdown spectroscopy by delivering pulsed laser radiation to the sample surface through the same sharp probe, and consequent collection and analysis of emission spectra from plasma generated on the sample by the laser radiation. The method comprises performing microtopographical analysis of the sample with a scanning probe, selecting a scanned topological site on the sample, generating a plasma plume at the selected scanned topological site, and measuring a spectrum of optical emission from the plasma at the selected scanned topological site. The apparatus comprises a scanning probe, a pulsed laser optically coupled to the probe, an optical spectrometer, and a controller coupled to the scanner, laser and spectrometer for controlling the operation of the scanner, laser and spectrometer. The probe and scanner are used for topographical profiling the sample. The probe is also used for laser radiation delivery to the sample for generating a plasma plume from the sample. Optical emission from the plasma plume is collected and delivered to the optical spectrometer so that analysis of emission spectrum by the optical spectrometer allows for identification of chemical composition of the sample at user selected sites.

VISUALIZATION FROM INTRAOPERATIVE SWEPT-SOURCE MICROSCOPE-INTEGRATED OPTICAL COHERENCE TOMOGRAPHY IN VITRECTOMY FOR COMPLICATIONS OF PROLIFERATIVE DIABETIC RETINOPATHY.

PubMed

Gabr, Hesham; Chen, Xi; Zevallos-Carrasco, Oscar M; Viehland, Christian; Dandrige, Alexandria; Sarin, Neeru; Mahmoud, Tamer H; Vajzovic, Lejla; Izatt, Joseph A; Toth, Cynthia A

2018-01-10

To evaluate the use of live volumetric (4D) intraoperative swept-source microscope-integrated optical coherence tomography in vitrectomy for proliferative diabetic retinopathy complications. In this prospective study, we analyzed a subgroup of patients with proliferative diabetic retinopathy complications who required vitrectomy and who were imaged by the research swept-source microscope-integrated optical coherence tomography system. In near real time, images were displayed in stereo heads-up display facilitating intraoperative surgeon feedback. Postoperative review included scoring image quality, identifying different diabetic retinopathy-associated pathologies and reviewing the intraoperatively documented surgeon feedback. Twenty eyes were included. Indications for vitrectomy were tractional retinal detachment (16 eyes), combined tractional-rhegmatogenous retinal detachment (2 eyes), and vitreous hemorrhage (2 eyes). Useful, good-quality 2D (B-scans) and 4D images were obtained in 16/20 eyes (80%). In these eyes, multiple diabetic retinopathy complications could be imaged. Swept-source microscope-integrated optical coherence tomography provided surgical guidance, e.g., in identifying dissection planes under fibrovascular membranes, and in determining residual membranes and traction that would benefit from additional peeling. In 4/20 eyes (20%), acceptable images were captured, but they were not useful due to high tractional retinal detachment elevation which was challenging for imaging. Swept-source microscope-integrated optical coherence tomography can provide important guidance during surgery for proliferative diabetic retinopathy complications through intraoperative identification of different complications and facilitation of intraoperative decision making.

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection.

PubMed

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.

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection

NASA Astrophysics Data System (ADS)

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.

Scanning tunneling microscopy and atomic force microscopy: application to biology and technology.

PubMed

Hansma, P K; Elings, V B; Marti, O; Bracker, C E

1988-10-14

The scanning tunneling microscope (STM) and the atomic force microscope (AFM) are scanning probe microscopes capable of resolving surface detail down to the atomic level. The potential of these microscopes for revealing subtle details of structure is illustrated by atomic resolution images including graphite, an organic conductor, an insulating layered compound, and individual adsorbed oxygen atoms on a semiconductor. Application of the STM for imaging biological materials directly has been hampered by the poor electron conductivity of most biological samples. The use of thin conductive metal coatings and replicas has made it possible to image some biological samples, as indicated by recently obtained images of a recA-DNA complex, a phospholipid bilayer, and an enzyme crystal. The potential of the AFM, which does not require a conductive sample, is shown with molecular resolution images of a nonconducting organic monolayer and an amino acid crystal that reveals individual methyl groups on the ends of the amino acids. Applications of these new microscopes to technology are demonstrated with images of an optical disk stamper, a diffraction grating, a thin-film magnetic recording head, and a diamond cutting tool. The STM has even been used to improve the quality of diffraction gratings and magnetic recording heads.

Large area scanning probe microscope in ultra-high vacuum demonstrated for electrostatic force measurements on high-voltage devices.

PubMed

Gysin, Urs; Glatzel, Thilo; Schmölzer, Thomas; Schöner, Adolf; Reshanov, Sergey; Bartolf, Holger; Meyer, Ernst

2015-01-01

The resolution in electrostatic force microscopy (EFM), a descendant of atomic force microscopy (AFM), has reached nanometre dimensions, necessary to investigate integrated circuits in modern electronic devices. However, the characterization of conducting or semiconducting power devices with EFM methods requires an accurate and reliable technique from the nanometre up to the micrometre scale. For high force sensitivity it is indispensable to operate the microscope under high to ultra-high vacuum (UHV) conditions to suppress viscous damping of the sensor. Furthermore, UHV environment allows for the analysis of clean surfaces under controlled environmental conditions. Because of these requirements we built a large area scanning probe microscope operating under UHV conditions at room temperature allowing to perform various electrical measurements, such as Kelvin probe force microscopy, scanning capacitance force microscopy, scanning spreading resistance microscopy, and also electrostatic force microscopy at higher harmonics. The instrument incorporates beside a standard beam deflection detection system a closed loop scanner with a scan range of 100 μm in lateral and 25 μm in vertical direction as well as an additional fibre optics. This enables the illumination of the tip-sample interface for optically excited measurements such as local surface photo voltage detection. We present Kelvin probe force microscopy (KPFM) measurements before and after sputtering of a copper alloy with chromium grains used as electrical contact surface in ultra-high power switches. In addition, we discuss KPFM measurements on cross sections of cleaved silicon carbide structures: a calibration layer sample and a power rectifier. To demonstrate the benefit of surface photo voltage measurements, we analysed the contact potential difference of a silicon carbide p/n-junction under illumination.

Microstructures and electrochemical behaviors of the friction stir welding dissimilar weld.

PubMed

Shen, Changbin; Zhang, Jiayan; Ge, Jiping

2011-06-01

By using optical microscope, the microstructures of 5083/6082 friction stir welding (FSW) weld and parent materials were analyzed. Meanwhile, at ambient temperature and in 0.2 mol/L NaHS03 and 0.6 mol/L NaCl solutionby gravimetric test, potentiodynamic polarization curve test, electrochemical impedance spectra (EIS) and scanning electron microscope (SEM) observation, the electrochemical behavior of 5083/6082 friction stir welding weld and parent materials were comparatively investigated by gravimetric test, potentiodynamic polarization curve test, electrochemical impedance spectra (EIS) and scanning electron microscope (SEM) observation. The results indicated that at given processing parameters, the anti-corrosion property of the dissimilar weld was superior to those of the 5083 and 6082 parent materials. Copyright © 2011 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

Failure analysis of an aluminum alloy material framework component induced by casting defects

NASA Astrophysics Data System (ADS)

Li, Bo; Hu, Weiye

2017-09-01

Failure analysis on a fractured radome framework component was carried out through visual observations, metallographic examination using optical microscope, fractog-raphy inspections using scanning electron microscope and chemical composition analysis. The failed frame was made of casting Al-Si7-Mg0.4 aluminum alloy. It had suffered a former vi-bration performance tests. It was indicated that the fractures were attributed to fatigue cracks which were induced by casting porosities at the outer surfaces of frame. Failure analysis was carefully conducted for the semi-penetrating crack appearing on the framework. According to the fractography inspected by scanning electron microscope, it was indicated that numerous casting porosities at the outer surface of the framework played the role of multiple fracture sources due to some applied stresses. Optical microstructure observations suggested that the dendrite-shaped casting porosities largely contributed to the crack-initiation. The groove-shaped structure at roots of spatial convex-bodies on the edge of casting porosities supplied the preferred paths of the crack-propagation. Besides, the brittle silicon eutectic particles distrib-uting along grain boundaries induced the intergranular fracture mode in the region of the over-load final fracture surface.

Eyecup scope—optical recordings of light stimulus-evoked fluorescence signals in the retina

PubMed Central

Hausselt, Susanne E.; Breuninger, Tobias; Castell, Xavier; Denk, Winfried; Margolis, David J.; Detwiler, Peter B.

2009-01-01

Dendritic signals play an essential role in processing visual information in the retina. To study them in neurites too small for electrical recording, we developed an instrument that combines a multi-photon (MP) microscope with a through-the-objective high-resolution visual stimulator. An upright microscope was designed that uses the objective lens for both MP imaging and delivery of visual stimuli to functionally intact retinal explants or eyecup preparations. The stimulator consists of a miniature liquid-crystal-on-silicon display coupled into the optical path of an infrared-excitation laser-scanning microscope. A pair of custom-made dichroic filters allows light from the excitation laser and three spectral bands (‘colors’) from the stimulator to reach the retina, leaving two intermediate bands for fluorescence imaging. Special optics allow displacement of the stimulator focus relative to the imaging focus. Spatially resolved changes in calcium-indicator fluorescence in response to visual stimuli were recorded in dendrites of different types of mammalian retinal neurons. PMID:19023590

Waveguide bends from nanometric silica wires

NASA Astrophysics Data System (ADS)

Tong, Limin; Lou, Jingyi; Mazur, Eric

2005-02-01

We propose to use bent silica wires with nanometric diameters to guide light as optical waveguide bend. We bend silica wires with scanning tunneling microscope probes under an optical microscope, and wire bends with bending radius smaller than 5 μm are obtained. Light from a He-Ne laser is launched into and guided through the wire bends, measured bending loss of a single bend is on the order of 1 dB. Brief introductions to the optical wave guiding and elastic bending properties of silica wires are also provided. Comparing with waveguide bends based on photonic bandgap structures, the waveguide bends from silica nanometric wires show advantages of simple structure, small overall size, easy fabrication and wide useful spectral range, which make them potentially useful in the miniaturization of photonic devices.

Ultrafast Graphene Photonics and Optoelectronics

DTIC Science & Technology

2017-04-14

SUBJECT TERMS Graphene, Ultrafast Optical Processin, Terahertz Electronics ; 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT SAR 18...Rep, (2016)) Fig. 4. (a) Images of scanning electron microscope for 1D and 2D gratings. (b) Ratio of the real part of the transmitted field

Reflectance confocal microscope for imaging oral tissues in vivo, potentially with line scanning as a low-cost approach for clinical use

NASA Astrophysics Data System (ADS)

Peterson, Gary; Abeytunge, Sanjeewa; Eastman, Zachary; Rajadhyaksha, Milind

2012-02-01

Reflectance confocal microscopy with a line scanning approach potentially offers a smaller, simpler and less expensive approach than traditional methods of point scanning for imaging in living tissues. With one moving mechanical element (galvanometric scanner), a linear array detector and off-the-shelf optics, we designed a compact (102x102x76mm) line scanning confocal reflectance microscope (LSCRM) for imaging human tissues in vivo in a clinical setting. Custom-designed electronics, based on field programmable gate array (FPGA) logic has been developed. With 405 nm illumination and a custom objective lens of numerical aperture 0.5, lateral resolution was measured to be 0.8 um (calculated 0.64 um). The calculated optical sectioning is 3.2 um. Preliminary imaging shows nuclear and cellular detail in human skin and oral epithelium in vivo. Blood flow is also visualized in the deeper connective tissue (lamina propria) in oral mucosa. Since a line is confocal only in one dimension (parallel) but not in the other, the detection is more sensitive to multiply scattered out of focus background noise than in the traditional point scanning configuration. Based on the results of our translational studies thus far, a simpler, smaller and lower-cost approach based on a LSCRM appears to be promising for clinical imaging.

The microscopic (optical and SEM) examination of putrefaction fluid deposits (PFD). Potential interest in forensic anthropology.

PubMed

Charlier, P; Georges, P; Bouchet, F; Huynh-Charlier, I; Carlier, R; Mazel, V; Richardin, P; Brun, L; Blondiaux, J; Lorin de la Grandmaison, G

2008-10-01

This article describes the potential interest in physical and forensic anthropology of the microscopic analysis of residues of putrefaction fluid, a calcified deposit frequently found associated with bone rests. Its sampling and analysis seem straightforward and relatively reproducible. Samples came from archeological material (Monterenzio Vecchia, an Etruscan necropolis from the north of Italy dated between the fifth and third century B.C.; body rests of Agnès Sorel, royal mistress died in 1450 A.D.; skull and grave of French King Louis the XI and Charlotte of Savoy dated from 1483 A.D.). All samples were studied by direct optical microscope and scanning electron microscopy. Many cytological, histological, and elemental analysis were possible, producing precious data for the identification of these remains and, in some cases, the cause of death.

Wide field-of-view, multi-region two-photon imaging of neuronal activity in the mammalian brain

PubMed Central

Stirman, Jeffrey N.; Smith, Ikuko T.; Kudenov, Michael W.; Smith, Spencer L.

2016-01-01

Two-photon calcium imaging provides an optical readout of neuronal activity in populations of neurons with subcellular resolution. However, conventional two-photon imaging systems are limited in their field of view to ~1 mm2, precluding the visualization of multiple cortical areas simultaneously. Here, we demonstrate a two-photon microscope with an expanded field of view (>9.5 mm2) for rapidly reconfigurable simultaneous scanning of widely separated populations of neurons. We custom designed and assembled an optimized scan engine, objective, and two independently positionable, temporally multiplexed excitation pathways. We used this new microscope to measure activity correlations between two cortical visual areas in mice during visual processing. PMID:27347754

A Low Temperature Scanning Force Microscope with a Vertical Cantilever and Interferometric Detection Scheme

NASA Astrophysics Data System (ADS)

Kim, Jeehoon; Williams, T. L.; Chu, Sang Lin; Korre, Hasan; Chalfin, Max; Hoffman, J. E.

2008-03-01

We have developed a fiber-optic interferometry system with a vertical cantilever for scanning force microscopy. A lens, mounted on a Pan-type walker, was used to collect the interference signal in the cavity between the cantilever and the single mode fiber. This vertical geometry has several advantages: (1) it is directly sensitive to lateral forces; (2) low spring constant vertical cantilevers may allow increased force sensitivity by solving the ``snap-in'' problem that occurs with soft horizontal cantilevers. We have sharpened vertical cantilevers by focused ion beam (FIB), achieving a tip radius of 20 nm. We will show test results of a magnetic force microscope (MFM) with this vertical cantilever system.

Corrugated metal-coated tapered tip for scanning near-field optical microscope.

PubMed

Antosiewicz, Tomasz J; Szoplik, Tomasz

2007-08-20

This paper addresses an important issue of light throughput of a metal-coated tapered tip for scanning near-field microscope (SNOM). Corrugations of the interface between the fiber core and metal coating in the form of parallel grooves of different profiles etched in the core considerably increase the energy throughput. In 2D FDTD simulations in the Cartesian coordinates we calculate near-field light emitted from such tips. For a certain wavelength range total intensity of forward emission from the corrugated tip is 10 times stronger than that from a classical tapered tip. When realized in practice the idea of corrugated tip may lead up to twice better resolution of SNOM.

Scientific Applications of Optical Instruments to Materials Research

NASA Technical Reports Server (NTRS)

Witherow, William K.

1997-01-01

Microgravity is a unique environment for materials and biotechnology processing. Microgravity minimizes or eliminates some of the effects that occur in one g. This can lead to the production of new materials or crystal structures. It is important to understand the processes that create these new materials. Thus, experiments are designed so that optical data collection can take place during the formation of the material. This presentation will discuss scientific application of optical instruments at MSFC. These instruments include a near-field scanning optical microscope, a miniaturized holographic system, and a phase-shifting interferometer.

A light field microscope imaging spectrometer based on the microlens array

NASA Astrophysics Data System (ADS)

Yao, Yu-jia; Xu, Feng; Xia, Yin-xiang

2017-10-01

A new light field spectrometry microscope imaging system, which was composed by microscope objective, microlens array and spectrometry system was designed in this paper. 5-D information (4-D light field and 1-D spectrometer) of the sample could be captured by the snapshot system in only one exposure, avoiding the motion blur and aberration caused by the scanning imaging process of the traditional imaging spectrometry. Microscope objective had been used as the former group while microlens array used as the posterior group. The optical design of the system was simulated by Zemax, the parameter matching condition between microscope objective and microlens array was discussed significantly during the simulation process. The result simulated in the image plane was analyzed and discussed.

Dual-modal three-dimensional imaging of single cells with isometric high resolution using an optical projection tomography microscope

NASA Astrophysics Data System (ADS)

Miao, Qin; Rahn, J. Richard; Tourovskaia, Anna; Meyer, Michael G.; Neumann, Thomas; Nelson, Alan C.; Seibel, Eric J.

2009-11-01

The practice of clinical cytology relies on bright-field microscopy using absorption dyes like hematoxylin and eosin in the transmission mode, while the practice of research microscopy relies on fluorescence microscopy in the epi-illumination mode. The optical projection tomography microscope is an optical microscope that can generate 3-D images of single cells with isometric high resolution both in absorption and fluorescence mode. Although the depth of field of the microscope objective is in the submicron range, it can be 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. Cells suspended in optical gel flow through a custom-designed microcapillary. Multiple pseudoprojection images are taken by rotating the microcapillary. After these pseudoprojection images are further aligned, computed tomography methods are applied to create 3-D reconstruction. 3-D reconstructed images of single cells are shown in both absorption and fluorescence mode. Fluorescence spatial resolution is measured at 0.35 μm in both axial and lateral dimensions. Since fluorescence and absorption images are taken in two different rotations, mechanical error may cause misalignment of 3-D images. This mechanical error is estimated to be within the resolution of the system.

Scanning Tunneling Optical Resonance Microscopy Developed

NASA Technical Reports Server (NTRS)

Bailey, Sheila G.; Raffaelle, Ryne P.; Lau, Janis E.; Jenkins, Phillip P.; Castro, Stephanie L.; Tin, Padetha; Wilt, David M.; Pal, Anna Maria; Fahey, Stephen D.

2004-01-01

The ability to determine the in situ optoelectronic properties of semiconductor materials has become especially important as the size of device architectures has decreased and the development of complex microsystems has increased. Scanning Tunneling Optical Resonance Microscopy, or STORM, can interrogate the optical bandgap as a function of its position within a semiconductor micro-structure. This technique uses a tunable solidstate titanium-sapphire laser whose output is "chopped" using a spatial light modulator and is coupled by a fiber-optic connector to a scanning tunneling microscope in order to illuminate the tip-sample junction. The photoenhanced portion of the tunneling current is spectroscopically measured using a lock-in technique. The capabilities of this technique were verified using semiconductor microstructure calibration standards that were grown by organometallic vapor-phase epitaxy. Bandgaps characterized by STORM measurements were found to be in good agreement with the bulk values determined by transmission spectroscopy and photoluminescence and with the theoretical values that were based on x-ray diffraction results.

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

NASA Astrophysics Data System (ADS)

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.

Multimodal ophthalmic imaging using spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography

NASA Astrophysics Data System (ADS)

El-Haddad, Mohamed T.; Malone, Joseph D.; Li, Jianwei D.; Bozic, Ivan; Arquitola, Amber M.; Joos, Karen M.; Patel, Shriji N.; Tao, Yuankai K.

2017-08-01

Ophthalmic surgery involves manipulation of delicate, layered tissue structures on milli- to micrometer scales. Traditional surgical microscopes provide an inherently two-dimensional view of the surgical field with limited depth perception which precludes accurate depth-resolved visualization of these tissue layers, and limits the development of novel surgical techniques. We demonstrate multimodal swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography (SS-SESLO-OCT) to address current limitations of image-guided ophthalmic microsurgery. SS-SESLO-OCT provides inherently co-registered en face and cross-sectional field-of-views (FOVs) at a line rate of 400 kHz and >2 GPix/s throughput. We show in vivo imaging of the anterior segment and retinal fundus of a healthy volunteer, and preliminary results of multi-volumetric mosaicking for ultrawide-field retinal imaging with 90° FOV. Additionally, a scan-head was rapid-prototyped with a modular architecture which enabled integration of SS-SESLO-OCT with traditional surgical microscope and slit-lamp imaging optics. Ex vivo surgical maneuvers were simulated in cadaveric porcine eyes. The system throughput enabled volumetric acquisition at 10 volumes-per-second (vps) and allowed visualization of surgical dynamics in corneal sweeps, compressions, and dissections, and retinal sweeps, compressions, and elevations. SESLO en face images enabled simple real-time co-registration with the surgical microscope FOV, and OCT cross-sections provided depth-resolved visualization of instrument-tissue interactions. Finally, we demonstrate novel augmented-reality integration with the surgical view using segmentation overlays to aid surgical guidance. SS-SESLO-OCT may benefit clinical diagnostics by enabling aiming, registration, and mosaicking; and intraoperative imaging by allowing for real-time surgical feedback, instrument tracking, and overlays of computationally extracted biomarkers of disease.

Experimental investigation on the caries characteristic of dental tissues by photothermal radiometry scanning imaging

NASA Astrophysics Data System (ADS)

Wang, Fei; Liu, Jun-yan; Wang, Xiao-chun; Wang, Yang

2018-03-01

In this paper, a one-dimensional (1D) thermal-wave model coupled diffuse-photon-density-wave for three-layer dental tissues using modulated laser stimulation was employed to illustrate the relationship between dental caries characteristic (i.e. caries layer thickness, optical absorption coefficient and optical scattering coefficient) and photothermal radiometry (PTR) signal. Experimental investigation of artificial caries was carried out using PTR scanning imaging. The PTR amplitude and phase delay were increased with dental demineralized treatment. The local caries characteristic parameters were obtained by the best-fitting method based on the 1D thermal-wave model. The PTR scanning imaging measurements illustrated that the optical absorption coefficient and scattering coefficient of caries region were much higher than those of the healthy enamel area. The demineralization thickness of caries region was measured by PTR scanning imaging and its average value shows in good agreement with the digital microscope. Experimental results show that PTR scanning imaging has the merits of high contrast for local inhomogeneity of dental caries; furthermore, this method is an allowance to provide a flexibility for non-contact quantitative evaluation of dental caries.

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

PubMed

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.

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

NASA Astrophysics Data System (ADS)

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

2017-09-01

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

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection

PubMed Central

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. PMID:20210471

Note: A three-dimensional calibration device for the confocal microscope.

PubMed

Jensen, K E; Weitz, D A; Spaepen, F

2013-01-01

Modern confocal microscopes enable high-precision measurement in three dimensions by collecting stacks of 2D (x-y) images that can be assembled digitally into a 3D image. It is difficult, however, to ensure position accuracy, particularly along the optical (z) axis where scanning is performed by a different physical mechanism than in x-y. We describe a simple device to calibrate simultaneously the x, y, and z pixel-to-micrometer conversion factors for a confocal microscope. By taking a known 2D pattern and positioning it at a precise angle with respect to the microscope axes, we created a 3D reference standard. The device is straightforward to construct and easy to use.

Analysis of particulate contamination on tape lift samples from the VETA optical surfaces

NASA Technical Reports Server (NTRS)

Germani, Mark S.

1992-01-01

Particulate contamination analysis was carried out on samples taken from the Verification Engineering Test Article (VETA) x-ray detection system. A total of eighteen tape lift samples were taken from the VETA optical surfaces. Initially, the samples were tested using a scanning electron microscope. Additionally, particle composition was determined by energy dispersive x-ray spectrometry. Results are presented in terms of particle loading per sample.

Imaging Single Cells in the Living Retina

PubMed Central

Williams, David R.

2011-01-01

A quarter century ago, we were limited to a macroscopic view of the retina inside the living eye. Since then, new imaging technologies, including confocal scanning laser ophthalmoscopy, optical coherence tomography, and adaptive optics fundus imaging, transformed the eye into a microscope in which individual cells can now be resolved noninvasively. These technologies have enabled a wide range of studies of the retina that were previously impossible. PMID:21596053

Dual-mode optical microscope based on single-pixel imaging

NASA Astrophysics Data System (ADS)

Rodríguez, A. D.; Clemente, P.; Tajahuerce, E.; Lancis, J.

2016-07-01

We demonstrate an inverted microscope that can image specimens in both reflection and transmission modes simultaneously with a single light source. The microscope utilizes a digital micromirror device (DMD) for patterned illumination altogether with two single-pixel photosensors for efficient light detection. The system, a scan-less device with no moving parts, works by sequential projection of a set of binary intensity patterns onto the sample that are codified onto a modified commercial DMD. Data to be displayed are geometrically transformed before written into a memory cell to cancel optical artifacts coming from the diamond-like shaped structure of the micromirror array. The 24-bit color depth of the display is fully exploited to increase the frame rate by a factor of 24, which makes the technique practicable for real samples. Our commercial DMD-based LED-illumination is cost effective and can be easily coupled as an add-on module for already existing inverted microscopes. The reflection and transmission information provided by our dual microscope complement each other and can be useful for imaging non-uniform samples and to prevent self-shadowing effects.

Holographic pixel super-resolution in portable lensless on-chip microscopy using a fiber-optic array.

PubMed

Bishara, Waheb; Sikora, Uzair; Mudanyali, Onur; Su, Ting-Wei; Yaglidere, Oguzhan; Luckhart, Shirley; Ozcan, Aydogan

2011-04-07

We report a portable lensless on-chip microscope that can achieve <1 µm resolution over a wide field-of-view of ∼ 24 mm(2) without the use of any mechanical scanning. This compact on-chip microscope weighs ∼ 95 g and is based on partially coherent digital in-line holography. Multiple fiber-optic waveguides are butt-coupled to light emitting diodes, which are controlled by a low-cost micro-controller to sequentially illuminate the sample. The resulting lensfree holograms are then captured by a digital sensor-array and are rapidly processed using a pixel super-resolution algorithm to generate much higher resolution holographic images (both phase and amplitude) of the objects. This wide-field and high-resolution on-chip microscope, being compact and light-weight, would be important for global health problems such as diagnosis of infectious diseases in remote locations. Toward this end, we validate the performance of this field-portable microscope by imaging human malaria parasites (Plasmodium falciparum) in thin blood smears. Our results constitute the first-time that a lensfree on-chip microscope has successfully imaged malaria parasites.

Method for identifying biochemical and chemical reactions and micromechanical processes using nanomechanical and electronic signal identification

DOEpatents

Holzrichter, J.F.; Siekhaus, W.J.

1997-04-15

A scanning probe microscope, such as an atomic force microscope (AFM) or a scanning tunneling microscope (STM), is operated in a stationary mode on a site where an activity of interest occurs to measure and identify characteristic time-varying micromotions caused by biological, chemical, mechanical, electrical, optical, or physical processes. The tip and cantilever assembly of an AFM is used as a micromechanical detector of characteristic micromotions transmitted either directly by a site of interest or indirectly through the surrounding medium. Alternatively, the exponential dependence of the tunneling current on the size of the gap in the STM is used to detect micromechanical movement. The stationary mode of operation can be used to observe dynamic biological processes in real time and in a natural environment, such as polymerase processing of DNA for determining the sequence of a DNA molecule. 6 figs.

Method for identifying biochemical and chemical reactions and micromechanical processes using nanomechanical and electronic signal identification

DOEpatents

Holzrichter, John F.; Siekhaus, Wigbert J.

1997-01-01

A scanning probe microscope, such as an atomic force microscope (AFM) or a scanning tunneling microscope (STM), is operated in a stationary mode on a site where an activity of interest occurs to measure and identify characteristic time-varying micromotions caused by biological, chemical, mechanical, electrical, optical, or physical processes. The tip and cantilever assembly of an AFM is used as a micromechanical detector of characteristic micromotions transmitted either directly by a site of interest or indirectly through the surrounding medium. Alternatively, the exponential dependence of the tunneling current on the size of the gap in the STM is used to detect micromechanical movement. The stationary mode of operation can be used to observe dynamic biological processes in real time and in a natural environment, such as polymerase processing of DNA for determining the sequence of a DNA molecule.

Z-scan studies of the nonlinear optical properties of gold nanoparticles prepared by electron beam deposition.

PubMed

Mezher, M H; Nady, A; Penny, R; Chong, W Y; Zakaria, R

2015-11-20

This paper details the fabrication process for placing single-layer gold (Au) nanoparticles on a planar substrate, and investigation of the resulting optical properties that can be exploited for nonlinear optics applications. Preparation of Au nanoparticles on the substrate involved electron beam deposition and subsequent thermal dewetting. The obtained thin films of Au had a variation in thicknesses related to the controllable deposition time during the electron beam deposition process. These samples were then subjected to thermal annealing at 600°C to produce a randomly distributed layer of Au nanoparticles. Observation from field-effect scanning electron microscope (FESEM) images indicated the size of Au nanoparticles ranges from ∼13 to ∼48  nm. Details of the optical properties related to peak absorption of localized surface plasmon resonance (LSPR) of the nanoparticle were revealed by use of UV-Vis spectroscopy. The Z-scan technique was used to measure the nonlinear effects on the fabricated Au nanoparticle layers where it strongly relates LSPR and nonlinear optical properties.

Structure and optical properties of TiO2 thin films deposited by ALD method

NASA Astrophysics Data System (ADS)

Szindler, Marek; Szindler, Magdalena M.; Boryło, Paulina; Jung, Tymoteusz

2017-12-01

This paper presents the results of study on titanium dioxide thin films prepared by atomic layer deposition method on a silicon substrate. The changes of surface morphology have been observed in topographic images performed with the atomic force microscope (AFM) and scanning electron microscope (SEM). Obtained roughness parameters have been calculated with XEI Park Systems software. Qualitative studies of chemical composition were also performed using the energy dispersive spectrometer (EDS). The structure of titanium dioxide was investigated by X-ray crystallography. A variety of crystalline TiO2 was also confirmed by using the Raman spectrometer. The optical reflection spectra have been measured with UV-Vis spectrophotometry.

Scanning thin-sheet laser imaging microscopy (sTSLIM) with structured illumination and HiLo background rejection.

PubMed

Schröter, Tobias J; Johnson, Shane B; John, Kerstin; Santi, Peter A

2012-01-01

We report replacement of one side of a static illumination, dual sided, thin-sheet laser imaging microscope (TSLIM) with an intensity modulated laser scanner in order to implement structured illumination (SI) and HiLo image demodulation techniques for background rejection. The new system is equipped with one static and one scanned light-sheet and is called a scanning thin-sheet laser imaging microscope (sTSLIM). It is an optimized version of a light-sheet fluorescent microscope that is designed to image large specimens (<15 mm in diameter). In this paper we describe the hardware and software modifications to TSLIM that allow for static and uniform light-sheet illumination with SI and HiLo image demodulation. The static light-sheet has a thickness of 3.2 µm; whereas, the scanned side has a light-sheet thickness of 4.2 µm. The scanned side images specimens with subcellular resolution (<1 µm lateral and <4 µm axial resolution) with a size up to 15 mm. SI and HiLo produce superior contrast compared to both the uniform static and scanned light-sheets. HiLo contrast was greater than SI and is faster and more robust than SI because as it produces images in two-thirds of the time and exhibits fewer intensity streaking artifacts. 2011 Optical Society of America

Optical, structural and nonlinear optical properties of laser ablation synthesized Ag nanoparticles and photopolymer nanocomposites based on them

NASA Astrophysics Data System (ADS)

Zulina, Natalia A.; Pavlovetc, Ilia M.; Baranov, Mikhail A.; Denisyuk, Igor Yu.

2017-03-01

In this work Ag nanoparticles (NPs) stable colloid solution were prepared by laser ablation of chemically pure silver rod in liquid monomer isodecyl acrylate (IDA). Sizes of obtained nanoparticles were determined by scanning electron microscope and vary from 27 to 120 nm. Nanocomposites films were prepared from obtained stable colloid solution of AgNPs by photocuring. For aliphatic polymer IDA long molecules cross-linking Diurethane dimethacrylate, 1,6-Hexandiol diacrylate and Tetra (ethylene glycol) diacrylate were used. Prepared nanomaterials exhibit strong third-order nonlinear optical responses, which was estimated by using z-scan technique. The third-order nonlinear optical coefficients of the studied nanocomposites were found to be up to Reχ(3)=1.31×10-5 (esu) and Imχ(3)=7.64×10-5 (esu).

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

PubMed

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.

Resolution and throughput optimized intraoperative spectrally encoded coherence tomography and reflectometry (iSECTR) for multimodal imaging during ophthalmic microsurgery

NASA Astrophysics Data System (ADS)

Malone, Joseph D.; El-Haddad, Mohamed T.; Leeburg, Kelsey C.; Terrones, Benjamin D.; Tao, Yuankai K.

2018-02-01

Limited visualization of semi-transparent structures in the eye remains a critical barrier to improving clinical outcomes and developing novel surgical techniques. While increases in imaging speed has enabled intraoperative optical coherence tomography (iOCT) imaging of surgical dynamics, several critical barriers to clinical adoption remain. Specifically, these include (1) static field-of-views (FOVs) requiring manual instrument-tracking; (2) high frame-rates require sparse sampling, which limits FOV; and (3) small iOCT FOV also limits the ability to co-register data with surgical microscopy. We previously addressed these limitations in image-guided ophthalmic microsurgery by developing microscope-integrated multimodal intraoperative swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography. Complementary en face images enabled orientation and coregistration with the widefield surgical microscope view while OCT imaging enabled depth-resolved visualization of surgical instrument positions relative to anatomic structures-of-interest. In addition, we demonstrated novel integrated segmentation overlays for augmented-reality surgical guidance. Unfortunately, our previous system lacked the resolution and optical throughput for in vivo retinal imaging and necessitated removal of cornea and lens. These limitations were predominately a result of optical aberrations from imaging through a shared surgical microscope objective lens, which was modeled as a paraxial surface. Here, we present an optimized intraoperative spectrally encoded coherence tomography and reflectometry (iSECTR) system. We use a novel lens characterization method to develop an accurate model of surgical microscope objective performance and balance out inherent aberrations using iSECTR relay optics. Using this system, we demonstrate in vivo multimodal ophthalmic imaging through a surgical microscope

Cathodoluminescence in the scanning transmission electron microscope.

PubMed

Kociak, M; Zagonel, L F

2017-05-01

Cathodoluminescence (CL) is a powerful tool for the investigation of optical properties of materials. In recent years, its combination with scanning transmission electron microscopy (STEM) has demonstrated great success in unveiling new physics in the field of plasmonics and quantum emitters. Most of these results were not imaginable even twenty years ago, due to conceptual and technical limitations. The purpose of this review is to present the recent advances that broke these limitations, and the new possibilities offered by the modern STEM-CL technique. We first introduce the different STEM-CL operating modes and the technical specificities in STEM-CL instrumentation. Two main classes of optical excitations, namely the coherent one (typically plasmons) and the incoherent one (typically light emission from quantum emitters) are investigated with STEM-CL. For these two main classes, we describe both the physics of light production under electron beam irradiation and the physical basis for interpreting STEM-CL experiments. We then compare STEM-CL with its better known sister techniques: scanning electron microscope CL, photoluminescence, and electron energy-loss spectroscopy. We finish by comprehensively reviewing recent STEM-CL applications. Copyright © 2016 Elsevier B.V. All rights reserved.

Cathodoluminescence in the scanning transmission electron microscope.

PubMed

Kociak, M; Zagonel, L F

2016-12-19

Cathodoluminescence (CL) is a powerful tool for the investigation of optical properties of materials. In recent years, its combination with scanning transmission electron microscopy (STEM) has demonstrated great success in unveiling new physics in the field of plasmonics and quantum emitters. Most of these results were not imaginable even twenty years ago, due to conceptual and technical limitations. The purpose of this review is to present the recent advances that broke these limitations, and the new possibilities offered by the modern STEM-CL technique. We first introduce the different STEM-CL operating modes and the technical specificities in STEM-CL instrumentation. Two main classes of optical excitations, namely the coherent one (typically plasmons) and the incoherent one (typically light emission from quantum emitters) are investigated with STEM-CL. For these two main classes, we describe both the physics of light production under electron beam irradiation and the physical basis for interpreting STEM-CL experiments. We then compare STEM-CL with its better known sister techniques: scanning electron microscope CL, photoluminescence, and electron energy-loss spectroscopy. We finish by comprehensively reviewing recent STEM-CL applications. Copyright © 2016 Elsevier B.V. All rights reserved.

Biological applications of near-field scanning optical microscopy

NASA Astrophysics Data System (ADS)

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.

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

NASA Astrophysics Data System (ADS)

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

2001-10-01

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

Experimental Plan for the Development of Equivalent Crack Size Distributions and a Monte Carlo Model of Fatigue in Low and High-kt Specimens of Corroded AA7050-T7451

DTIC Science & Technology

2012-03-01

18 3.3.7 Fractography ... Fractography : The fracture surfaces of the fractured fatigue specimens will be examined optically and in the scanning electron microscope to measure the size...scanned and added to the report containing the results from these fatigue tests. 3.3.7 Fractography Once a specimen has been tested, and assuming

Transfer function characteristics of super resolving systems

NASA Technical Reports Server (NTRS)

Milster, Tom D.; Curtis, Craig H.

1992-01-01

Signal quality in an optical storage device greatly depends on the optical system transfer function used to write and read data patterns. The problem is similar to analysis of scanning optical microscopes. Hopkins and Braat have analyzed write-once-read-many (WORM) optical data storage devices. Herein, transfer function analysis of magnetooptic (MO) data storage devices is discussed with respect to improving transfer-function characteristics. Several authors have described improving the transfer function as super resolution. However, none have thoroughly analyzed the MO optical system and effects of the medium. Both the optical system transfer function and effects of the medium of this development are discussed.

Crystal Structure, Magnetic and Optical Properties of Mn-Doped BiFeO₃ by Hydrothermal Synthesis.

PubMed

Zhang, Ning; Wei, Qinhua; Qin, Laishun; Chen, Da; Chen, Zhi; Niu, Feng; Wang, Jiangying; Huanag, Yuexiang

2017-01-01

In this paper, Mn doped BiFeO₃ were firstly synthesized by hydrothermal process. The influence of Mn doping on structural, optical and magnetic properties of BiFeO₃ was studied. The different amounts of Mn doping in BiFeO₃ were characterized by X-ray diffraction, Scanning Electron Microscope, Energy Dispersive X-ray Spectroscope, UV-Vis diffuse reflectance spectroscopy and magnetic measurements. The X-ray diffraction (XRD) patterns confirmed the formation of pure phase rhombohedral structure in BiFe(1−x) Mn (x) O₃ (x = 0.01, 0.03, 0.05, 0.07) samples. The morphologies and chemical compositions of as-prepared samples could be observed by Scanning Electron Microscope (SEM) and Energy Dispersive X-ray Spectroscope (EDS). A relative large saturated magnetization (Ms) of 0.53 emu/g for x = 0.07 sample was obtained at room temperature, which is considered to be Mn ions doping. UV-Vis diffuse reflectance spectroscopy showed strong absorption of light in the range of 200–1000 nm, indicating the optical band gap in the visible region for these samples. This implied that BiFe(1−x) Mn(x)O₃ may be a potential photocatalyst for utilizing solar energy.

Optical sectioning and 3D reconstructions as an alternative to scanning electron microscopy for analysis of cell shape.

PubMed

Landis, Jacob B; Ventura, Kayla L; Soltis, Douglas E; Soltis, Pamela S; Oppenheimer, David G

2015-04-01

Visualizing flower epidermal cells is often desirable for investigating the interaction between flowers and their pollinators, in addition to the broader range of ecological interactions in which flowers are involved. We developed a protocol for visualizing petal epidermal cells without the limitations of the commonly used method of scanning electron microscopy (SEM). Flower material was collected and fixed in glutaraldehyde, followed by dehydration in an ethanol series. Flowers were dissected to collect petals, and subjected to a Histo-Clear series to remove the cuticle. Material was then stained with aniline blue, mounted on microscope slides, and imaged using a compound fluorescence microscope to obtain optical sections that were reconstructed into a 3D image. This optical sectioning method yielded high-quality images of the petal epidermal cells with virtually no damage to cells. Flowers were processed in larger batches than are possible using common SEM methods. Also, flower size was not a limiting factor as often observed in SEM studies. Flowers up to 5 cm in length were processed and mounted for visualization. This method requires no special equipment for sample preparation prior to imaging and should be seen as an alternative method to SEM.

Dynamic performance of MEMS deformable mirrors for use in an active/adaptive two-photon microscope

NASA Astrophysics Data System (ADS)

Zhang, Christian C.; Foster, Warren B.; Downey, Ryan D.; Arrasmith, Christopher L.; Dickensheets, David L.

2016-03-01

Active optics can facilitate two-photon microscopic imaging deep in tissue. We are investigating fast focus control mirrors used in concert with an aberration correction mirror to control the axial position of focus and system aberrations dynamically during scanning. With an adaptive training step, sample-induced aberrations may be compensated as well. If sufficiently fast and precise, active optics may be able to compensate under-corrected imaging optics as well as sample aberrations to maintain diffraction-limited performance throughout the field of view. Toward this end we have measured a Boston Micromachines Corporation Multi-DM 140 element deformable mirror, and a Revibro Optics electrostatic 4-zone focus control mirror to characterize dynamic performance. Tests for the Multi-DM included both step response and sinusoidal frequency sweeps of specific Zernike modes. For the step response we measured 10%-90% rise times for the target Zernike amplitude, and wavefront rms error settling times. Frequency sweeps identified the 3dB bandwidth of the mirror when attempting to follow a sinusoidal amplitude trajectory for a specific Zernike mode. For five tested Zernike modes (defocus, spherical aberration, coma, astigmatism and trefoil) we find error settling times for mode amplitudes up to 400nm to be less than 52 us, and 3 dB frequencies range from 6.5 kHz to 10 kHz. The Revibro Optics mirror was tested for step response only, with error settling time of 80 μs for a large 3 um defocus step, and settling time of only 18 μs for a 400nm spherical aberration step. These response speeds are sufficient for intra-scan correction at scan rates typical of two-photon microscopy.

Corrosion resistance evaluation of Pd-free Ag-Au-Pt-Cu dental alloys.

PubMed

Fujita, Takeshi; Shiraishi, Takanobu; Takuma, Yasuko; Hisatsune, Kunihiro

2011-01-01

The corrosion resistance of nine experimental Pd-free Ag-Au-Pt-Cu dental alloys in a 0.9% NaCl solution was investigated using cyclic voltammetry (CV), optical microscopy, and scanning electron microscopy (SEM). CV measurements revealed that the breakdown potential (E(bd)) and zero current potential (E(zc)) increased with increasing Au/(Au+Ag) atomic ratio. Thus, the Au/(Au+Ag) atomic ratio, but not the Cu content, influenced the corrosion resistance of Ag-Au-Pt-Cu alloys. After the forward scan of CV, both optical and scanning electron microscope images showed that in all the experimental alloys, the matrix phase was corroded but not the second phase. From corrosion resistance viewpoint, the Ag-Au-Pt-Cu alloys seemed to be suitable for clinical application.

Full field vertical scanning in short coherence digital holographic microscope.

PubMed

Monemahghdoust, Zahra; Montfort, Frederic; Cuche, Etienne; Emery, Yves; Depeursinge, Christian; Moser, Christophe

2013-05-20

In Digital holography Microscopes (DHM) implemented in the so-called "off axis" configuration, the object and reference wave fronts are not co-planar but form an angle of a few degrees. This results into two main drawbacks. First, the contrast of the interference is not uniform spatially when the light source has low coherence. The interference contrast is optimal along a line, but decreases when moving away from it, resulting in a lower image quality. Second, the non-coplanarity between the coherence plane of both wavefronts impacts the coherence vertical scanning measurement mode: when the optical path difference between the signal and the reference beam is changed, the region of maximum interference contrast shifts laterally in the plane of the objective. This results in more complex calculations to extract the topography of the sample and requires scanning over a much larger vertical range, leading to a longer measurement time. We have previously shown that by placing a volume diffractive optical element (VDOE) in the reference arm, the wavefront can be made coplanar with the object wavefront and the image plane of the microscope objective, resulting in a uniform and optimal interferogram. In this paper, we demonstrate a vertical scanning speed improvement by an order of magnitude. Noise in the phase and intensity images caused by scattering and non-uniform diffraction in the VDOE is analyzed quantitatively. Five VDOEs were fabricated with an identical procedure. We observe that VDOEs introduce a small intensity non-uniformity in the reference beam which results in a 20% noise increase in the extracted phase image as compared to the noise in extracted phase image when the VDOE is removed. However, the VDOE has no impact on the temporal noise measured from extracted phase images.

Emerging fiber optic endomicroscopy technologies towards noninvasive real-time visualization of histology in situ

NASA Astrophysics Data System (ADS)

Xi, Jiefeng; Zhang, Yuying; Huo, Li; Chen, Yongping; Jabbour, Toufic; Li, Ming-Jun; Li, Xingde

2010-09-01

This paper reviews our recent developments of ultrathin fiber-optic endomicroscopy technologies for transforming high-resolution noninvasive optical imaging techniques to in vivo and clinical applications such as early disease detection and guidance of interventions. Specifically we describe an all-fiber-optic scanning endomicroscopy technology, which miniaturizes a conventional bench-top scanning laser microscope down to a flexible fiber-optic probe of a small footprint (i.e. ~2-2.5 mm in diameter), capable of performing two-photon fluorescence and second harmonic generation microscopy in real time. This technology aims to enable realtime visualization of histology in situ without the need for tissue removal. We will also present a balloon OCT endoscopy technology which permits high-resolution 3D imaging of the entire esophagus for detection of neoplasia, guidance of biopsy and assessment of therapeutic outcome. In addition we will discuss the development of functional polymeric fluorescent nanocapsules, which use only FAD approved materials and potentially enable fast track clinical translation of optical molecular imaging and targeted therapy.

Application of Environmental Scanning Electron Microscope-Nanomanipulation System on Spheroplast Yeast Cells Surface Observation.

PubMed

Rad, Maryam Alsadat; Ahmad, Mohd Ridzuan; Nakajima, Masahiro; Kojima, Seiji; Homma, Michio; Fukuda, Toshio

2017-01-01

The preparation and observations of spheroplast W303 cells are described with Environmental Scanning Electron Microscope (ESEM). The spheroplasting conversion was successfully confirmed qualitatively, by the evaluation of the morphological change between the normal W303 cells and the spheroplast W303 cells, and quantitatively, by determining the spheroplast conversion percentage based on the OD 800 absorbance data. From the optical microscope observations as expected, the normal cells had an oval shape whereas spheroplast cells resemble a spherical shape. This was also confirmed under four different mediums, that is, yeast peptone-dextrose (YPD), sterile water, sorbitol-EDTA-sodium citrate buffer (SCE), and sorbitol-Tris-Hcl-CaCl 2 (CaS). It was also observed that the SCE and CaS mediums had a higher number of spheroplast cells as compared to the YPD and sterile water mediums. The OD 800 absorbance data also showed that the whole W303 cells were fully converted to the spheroplast cells after about 15 minutes. The observations of the normal and the spheroplast W303 cells were then performed under an environmental scanning electron microscope (ESEM). The normal cells showed a smooth cell surface whereas the spheroplast cells had a bleb-like surface after the loss of its integrity when removing the cell wall.

A combined confocal and magnetic resonance microscope for biological studies

NASA Astrophysics Data System (ADS)

Majors, Paul D.; Minard, Kevin R.; Ackerman, Eric J.; Holtom, Gary R.; Hopkins, Derek F.; Parkinson, Christopher I.; Weber, Thomas J.; Wind, Robert A.

2002-12-01

Complementary data acquired with different microscopy techniques provide a basis for establishing a more comprehensive understanding of cell function in health and disease, particularly when results acquired with different methodologies can be correlated in time and space. In this article, a novel microscope is described for studying live cells simultaneously with both confocal scanning laser fluorescence optical microscopy and magnetic resonance microscopy. The various design considerations necessary for integrating these two complementary techniques are discussed, the layout and specifications of the instrument are given, and examples of confocal and magnetic resonance images of large frog cells and model tumor spheroids obtained with the compound microscope are presented.

Development of 1500mm Wide Wrought Magnesium Alloys by Twin Roll Casting Technique in Turkey

NASA Astrophysics Data System (ADS)

Duygulu, Ozgur; Ucuncuoglu, Selda; Oktay, Gizem; Temur, Deniz Sultan; Yucel, Onuralp; Kaya, Ali Arslan

Magnesium alloy AZ31, AZ61, AZ91, AM50 and AM60 sheets were produced by twin roll casting first time in Turkey. Sheets of 4.5-6.5mm thick and 1500mm width were successfully achieved. Microstructure of the sheet was analyzed by optical microscope, scanning electron microscope (SEM) and transmission electron microscope (TEM). Semi-quantitative analyses were performed by SEM-EDS. In addition, X-ray studies were performed for both characterization and texture purposes. Mechanical properties were investigated by tensile tests and also hardness measurements. Homogenization and annealing heat treatments were performed on the produced sheets.

Fiber-optic fluorescence imaging

PubMed Central

Flusberg, Benjamin A; Cocker, Eric D; Piyawattanametha, Wibool; Jung, Juergen C; Cheung, Eunice L M; Schnitzer, Mark J

2010-01-01

Optical fibers guide light between separate locations and enable new types of fluorescence imaging. Fiber-optic fluorescence imaging systems include portable handheld microscopes, flexible endoscopes well suited for imaging within hollow tissue cavities and microendoscopes that allow minimally invasive high-resolution imaging deep within tissue. A challenge in the creation of such devices is the design and integration of miniaturized optical and mechanical components. Until recently, fiber-based fluorescence imaging was mainly limited to epifluorescence and scanning confocal modalities. Two new classes of photonic crystal fiber facilitate ultrashort pulse delivery for fiber-optic two-photon fluorescence imaging. An upcoming generation of fluorescence imaging devices will be based on microfabricated device components. PMID:16299479

Controlled core removal from a D-shaped optical fiber.

PubMed

Markos, Douglas J; Ipson, Benjamin L; Smith, Kevin H; Schultz, Stephen M; Selfridge, Richard H; Monte, Thomas D; Dyott, Richard B; Miller, Gregory

2003-12-20

The partial removal of a section of the core from a continuous D-shaped optical fiber is presented. In the core removal process, selective chemical etching is used with hydrofluoric (HF) acid. A 25% HF acid solution removes the cladding material above the core, and a 5% HF acid solution removes the core. A red laser with a wavelength of 670 nm is transmitted through the optical fiber during the etching. The power transmitted through the optical fiber is correlated to the etch depth by scanning electron microscope imaging. The developed process provides a repeatable method to produce an optical fiber with a specific etch depth.

Photon theory hypothesis about photon tunneling microscope's subwavelength resolution

NASA Astrophysics Data System (ADS)

Zhu, Yanbin; Ma, Junfu

1995-09-01

The foundation for the invention of the photon scanning tunneling microscope (PSTM) are the near field scanning optical microscope, the optical fiber technique, the total internal reflection, high sensitive opto-electronic detecting technique and computer technique etc. Recent research results show the subwavelength resolution of 1 - 3 nm is obtained. How to explain the PSTM has got such high subwavelength resolution? What value is the PSTM's limiting of subwavelength resolution? For resolving these problems this paper presented a photon theory hypothesis about PSTM that is based on the following two basic laws: (1) Photon is not only a carrier bringing energy and optical information, but also is a particle occupied fixed space size. (2) When a photon happened reflection, refraction, scattering, etc., only changed its energy and optical information carried, its particle size doesn't change. g (DOT) pphoton equals constant. Using these two basic laws to PSTM, the `evanescent field' is practically a weak photon distribution field and the detecting fiber tip diameter is practically a `gate' which size controlled the photon numbers into fiber tip. Passing through some calculation and inference, the following three conclusions can be given: (1) Under the PSTM's detection system sensitivity is high enough, the diameter D of detecting fiber tip and the near field detecting distance Z are the two most important factors to decide the subwavelength resolution of PSTM. (2) The limiting of PSTM's resolution will be given upon the conditions of D equals pphoton and Z equals pphoton, where pphoton is one photon size. (2) The final resolution limit R of PSTM will be lim R equals pphoton, D yields pphoton, Z yields pphoton.

Plasmonic Imaging of Electrochemical Reactions of Single Nanoparticles.

PubMed

Fang, Yimin; Wang, Hui; Yu, Hui; Liu, Xianwei; Wang, Wei; Chen, Hong-Yuan; Tao, N J

2016-11-15

Electrochemical reactions are involved in many natural phenomena, and are responsible for various applications, including energy conversion and storage, material processing and protection, and chemical detection and analysis. An electrochemical reaction is accompanied by electron transfer between a chemical species and an electrode. For this reason, it has been studied by measuring current, charge, or related electrical quantities. This approach has led to the development of various electrochemical methods, which have played an essential role in the understanding and applications of electrochemistry. While powerful, most of the traditional methods lack spatial and temporal resolutions desired for studying heterogeneous electrochemical reactions on electrode surfaces and in nanoscale materials. To overcome the limitations, scanning probe microscopes have been invented to map local electrochemical reactions with nanometer resolution. Examples include the scanning electrochemical microscope and scanning electrochemical cell microscope, which directly image local electrochemical reaction current using a scanning electrode or pipet. The use of a scanning probe in these microscopes provides high spatial resolution, but at the expense of temporal resolution and throughput. This Account discusses an alternative approach to study electrochemical reactions. Instead of measuring electron transfer electrically, it detects the accompanying changes in the reactant and product concentrations on the electrode surface optically via surface plasmon resonance (SPR). SPR is highly surface sensitive, and it provides quantitative information on the surface concentrations of reactants and products vs time and electrode potential, from which local reaction kinetics can be analyzed and quantified. The plasmonic approach allows imaging of local electrochemical reactions with high temporal resolution and sensitivity, making it attractive for studying electrochemical reactions in biological systems and nanoscale materials with high throughput. The plasmonic approach has two imaging modes: electrochemical current imaging and interfacial impedance imaging. The former images local electrochemical current associated with electrochemical reactions (faradic current), and the latter maps local interfacial impedance, including nonfaradic contributions (e.g., double layer charging). The plasmonic imaging technique can perform voltammetry (cyclic or square wave) in an analogous manner to the traditional electrochemical methods. It can also be integrated with bright field, dark field, and fluorescence imaging capabilities in one optical setup to provide additional capabilities. To date the plasmonic imaging technique has found various applications, including mapping of heterogeneous surface reactions, analysis of trace substances, detection of catalytic reactions, and measurement of graphene quantum capacitance. The plasmonic and other emerging optical imaging techniques (e.g., dark field and fluorescence microscopy), together with the scanning probe-based electrochemical imaging and single nanoparticle analysis techniques, provide new capabilities for one to study single nanoparticle electrochemistry with unprecedented spatial and temporal resolutions. In this Account, we focus on imaging of electrochemical reactions at single nanoparticles.

Harmonic demodulation and minimum enhancement factors in field-enhanced near-field optical microscopy.

PubMed

Scarpettini, A F; Bragas, A V

2015-01-01

Field-enhanced scanning optical microscopy relies on the design and fabrication of plasmonic probes which had to provide optical and chemical contrast at the nanoscale. In order to do so, the scattering containing the near-field information recorded in a field-enhanced scanning optical microscopy experiment, has to surpass the background light, always present due to multiple interferences between the macroscopic probe and sample. In this work, we show that when the probe-sample distance is modulated with very low amplitude, the higher the harmonic demodulation is, the better the ratio between the near-field signal and the interferometric background results. The choice of working at a given n harmonic is dictated by the experiment when the signal at the n + 1 harmonic goes below the experimental noise. We demonstrate that the optical contrast comes from the nth derivative of the near-field scattering, amplified by the interferometric background. By modelling the far and near field we calculate the probe-sample approach curves, which fit very well the experimental ones. After taking a great amount of experimental data for different probes and samples, we conclude with a table of the minimum enhancement factors needed to have optical contrast with field-enhanced scanning optical microscopy. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

Rapid microwave assisted synthesis of YIn1-xMnxO3 blue pigments: Synthesis, microstructure and optical properties

NASA Astrophysics Data System (ADS)

Zhou, Yuncheng; Jiang, Peng; Kuang, Jianlei; Yang, Xueshan; Cao, Wenbin

2018-07-01

The YIn1-xMnxO3 (0.1 ≤x ≤ 0.5) blue pigment samples are successfully prepared through a sol-gel process followed by microwave assisted sintering process. All the samples are shown single phases in the X-ray diffraction results. In the morphology study from scanning electronic microscope, the samples are composed of loosely connected small particles. The oxidation state of Mn is confirmed to be 3 + from the results of X-ray photonelectronic scan. The optical properties are characterized by UV-Visible spectrum and UV-visible-NIR spectrum. The samples exhibit intense blue color and they show small absorption in infrared region.

Microstructure-related properties of magnesium fluoride films at 193nm by oblique-angle deposition.

PubMed

Guo, Chun; Kong, Mingdong; Lin, Dawei; Liu, Cunding; Li, Bincheng

2013-01-14

Magnesium fluoride (MgF2) films deposited by resistive heating evaporation with oblique-angle deposition have been investigated in details. The optical and micro-structural properties of single-layer MgF2 films were characterized by UV-VIS and FTIR spectrophotometers, scanning electron microscope (SEM), atomic force microscope (AFM), and x-ray diffraction (XRD), respectively. The dependences of the optical and micro-structural parameters of the thin films on the deposition angle were analyzed. It was found that the MgF2 film in a columnar microstructure was negatively inhomogeneous of refractive index and polycrystalline. As the deposition angle increased, the optical loss, extinction coefficient, root-mean-square (rms) roughness, dislocation density and columnar angle of the MgF2 films increased, while the refractive index, packing density and grain size decreased. Furthermore, IR absorption of the MgF2 films depended on the columnar structured growth.

Development and applications of optical interferometric micrometrology in the angstrom and subangstrom range

NASA Technical Reports Server (NTRS)

Lauer, James L.; Abel, Phillip B.

1988-01-01

The recent development of the scanning electron tunneling microscope and the atomic force microscope requires absolute standards for measurements in the angstrom and subangstrom range. Optical interferometry with lasers and multiple mode laser resonances can provide absolute measurements as the laser wavelengths are very accurately known. A key feature of such measurements is the use of piezoelectric crystals as translators of the highest accuracy for very small disturbances. However, the dimensional changes of these crystals resulting from electrical potential changes depend on many variables, among them the method of mounting, so that accurate calibrations are necessary. Starting from advances in optical metrology made by physicists trying to find gravity waves, advances which led to measurements down to 10 to the -5 A, the author designed and built a much simpler system for the angstrom range. The major limiting factors were mechanical vibrations, air currents, thermal changes and laser instabilities.

Optical toolkits for in vivo deep tissue laser scanning microscopy: a primer

NASA Astrophysics Data System (ADS)

Lee, Woei Ming; McMenamin, Thomas; Li, Yongxiao

2018-06-01

Life at the microscale is animated and multifaceted. The impact of dynamic in vivo microscopy in small animals has opened up opportunities to peer into a multitude of biological processes at the cellular scale in their native microenvironments. Laser scanning microscopy (LSM) coupled with targeted fluorescent proteins has become an indispensable tool to enable dynamic imaging in vivo at high temporal and spatial resolutions. In the last few decades, the technique has been translated from imaging cells in thin samples to mapping cells in the thick biological tissue of living organisms. Here, we sought to provide a concise overview of the design considerations of a LSM that enables cellular and subcellular imaging in deep tissue. Individual components under review include: long working distance microscope objectives, laser scanning technologies, adaptive optics devices, beam shaping technologies and photon detectors, with an emphasis on more recent advances. The review will conclude with the latest innovations in automated optical microscopy, which would impact tracking and quantification of heterogeneous populations of cells in vivo.

Method to deterministically study photonic nanostructures in different experimental instruments.

PubMed

Husken, B H; Woldering, L A; Blum, C; Vos, W L

2009-01-01

We describe an experimental method to recover a single, deterministically fabricated nanostructure in various experimental instruments without the use of artificially fabricated markers, with the aim to study photonic structures. Therefore, a detailed map of the spatial surroundings of the nanostructure is made during the fabrication of the structure. These maps are made using a series of micrographs with successively decreasing magnifications. The graphs reveal intrinsic and characteristic geometric features that can subsequently be used in different setups to act as markers. As an illustration, we probe surface cavities with radii of 65 nm on a silica opal photonic crystal with various setups: a focused ion beam workstation; a scanning electron microscope (SEM); a wide field optical microscope and a confocal microscope. We use cross-correlation techniques to recover a small area imaged with the SEM in a large area photographed with the optical microscope, which provides a possible avenue to automatic searching. We show how both structural and optical reflectivity data can be obtained from one and the same nanostructure. Since our approach does not use artificial grids or markers, it is of particular interest for samples whose structure is not known a priori, like samples created solely by self-assembly. In addition, our method is not restricted to conducting samples.

Video-rate in vivo fluorescence imaging with a line-scanned dual-axis confocal microscope.

PubMed

Chen, Ye; Wang, Danni; Khan, Altaz; Wang, Yu; Borwege, Sabine; Sanai, Nader; Liu, Jonathan T C

2015-10-01

Video-rate optical-sectioning microscopy of living organisms would allow for the investigation of dynamic biological processes and would also reduce motion artifacts, especially for in vivo imaging applications. Previous feasibility studies, with a slow stage-scanned line-scanned dual-axis confocal (LS-DAC) microscope, have demonstrated that LS-DAC microscopy is capable of imaging tissues with subcellular resolution and high contrast at moderate depths of up to several hundred microns. However, the sensitivity and performance of a video-rate LS-DAC imaging system, with low-numerical aperture optics, have yet to be demonstrated. Here, we report on the construction and validation of a video-rate LS-DAC system that possesses sufficient sensitivity to visualize fluorescent contrast agents that are topically applied or systemically delivered in animal and human tissues. We present images of murine oral mucosa that are topically stained with methylene blue, and images of protoporphyrin IX-expressing brain tumor from glioma patients that have been administered 5-aminolevulinic acid prior to surgery. In addition, we demonstrate in vivo fluorescence imaging of red blood cells trafficking within the capillaries of a mouse ear, at frame rates of up to 30 fps. These results can serve as a benchmark for miniature in vivo microscopy devices under development.

Video-rate in vivo fluorescence imaging with a line-scanned dual-axis confocal microscope

NASA Astrophysics Data System (ADS)

Chen, Ye; Wang, Danni; Khan, Altaz; Wang, Yu; Borwege, Sabine; Sanai, Nader; Liu, Jonathan T. C.

2015-10-01

Video-rate optical-sectioning microscopy of living organisms would allow for the investigation of dynamic biological processes and would also reduce motion artifacts, especially for in vivo imaging applications. Previous feasibility studies, with a slow stage-scanned line-scanned dual-axis confocal (LS-DAC) microscope, have demonstrated that LS-DAC microscopy is capable of imaging tissues with subcellular resolution and high contrast at moderate depths of up to several hundred microns. However, the sensitivity and performance of a video-rate LS-DAC imaging system, with low-numerical aperture optics, have yet to be demonstrated. Here, we report on the construction and validation of a video-rate LS-DAC system that possesses sufficient sensitivity to visualize fluorescent contrast agents that are topically applied or systemically delivered in animal and human tissues. We present images of murine oral mucosa that are topically stained with methylene blue, and images of protoporphyrin IX-expressing brain tumor from glioma patients that have been administered 5-aminolevulinic acid prior to surgery. In addition, we demonstrate in vivo fluorescence imaging of red blood cells trafficking within the capillaries of a mouse ear, at frame rates of up to 30 fps. These results can serve as a benchmark for miniature in vivo microscopy devices under development.

Swept source optical coherence microscopy using a 1310 nm VCSEL light source

PubMed Central

Ahsen, Osman O.; Tao, Yuankai K.; Potsaid, Benjamin M.; Sheikine, Yuri; Jiang, James; Grulkowski, Ireneusz; Tsai, Tsung-Han; Jayaraman, Vijaysekhar; Kraus, Martin F.; Connolly, James L.; Hornegger, Joachim; Cable, Alex; Fujimoto, James G.

2013-01-01

We demonstrate high speed, swept source optical coherence microscopy (OCM) using a MEMS tunable vertical cavity surface-emitting laser (VCSEL) light source. The light source had a sweep rate of 280 kHz, providing a bidirectional axial scan rate of 560 kHz. The sweep bandwidth was 117 nm centered at 1310 nm, corresponding to an axial resolution of 13.1 µm in air, corresponding to 8.1 µm (9.6 µm spectrally shaped) in tissue. Dispersion mismatch from different objectives was compensated numerically, enabling magnification and field of view to be easily changed. OCM images were acquired with transverse resolutions between 0.86 µm - 3.42 µm using interchangeable 40X, 20X and 10X objectives with ~600 µm x 600 µm, ~1 mm x 1 mm and ~2 mm x 2 mm field-of-view (FOV), respectively. Parasitic variations in path length with beam scanning were corrected numerically. These features enable swept source OCM to be integrated with a wide range of existing scanning microscopes. Large FOV mosaics were generated by serially acquiring adjacent overlapping microscopic fields and combining them in post-processing. Fresh human colon, thyroid and kidney specimens were imaged ex vivo and compared to matching histology sections, demonstrating the ability of OCM to image tissue specimens. PMID:23938673

Random-access scanning microscopy for 3D imaging in awake behaving animals

PubMed Central

Nadella, K. M. Naga Srinivas; Roš, Hana; Baragli, Chiara; Griffiths, Victoria A.; Konstantinou, George; Koimtzis, Theo; Evans, Geoffrey J.; Kirkby, Paul A.; Silver, R. Angus

2018-01-01

Understanding how neural circuits process information requires rapid measurements from identified neurons distributed in 3D space. Here we describe an acousto-optic lens two-photon microscope that performs high-speed focussing and line-scanning within a volume spanning hundreds of micrometres. We demonstrate its random access functionality by selectively imaging cerebellar interneurons sparsely distributed in 3D and by simultaneously recording from the soma, proximal and distal dendrites of neocortical pyramidal cells in behaving mice. PMID:27749836

Laser scanning confocal microscopy: history, applications, and related optical sectioning techniques.

PubMed

Paddock, Stephen W; Eliceiri, Kevin W

2014-01-01

Confocal microscopy is an established light microscopical technique for imaging fluorescently labeled specimens with significant three-dimensional structure. Applications of confocal microscopy in the biomedical sciences include the imaging of the spatial distribution of macromolecules in either fixed or living cells, the automated collection of 3D data, the imaging of multiple labeled specimens and the measurement of physiological events in living cells. The laser scanning confocal microscope continues to be chosen for most routine work although a number of instruments have been developed for more specific applications. Significant improvements have been made to all areas of the confocal approach, not only to the instruments themselves, but also to the protocols of specimen preparation, to the analysis, the display, the reproduction, sharing and management of confocal images using bioinformatics techniques.

Crystallographic Study of Itokawa Particle, RA-QD02-0127 by Using Energy-Scanning X-Ray Diffraction Method with Synchrotron Radiation

NASA Technical Reports Server (NTRS)

Hagiya, K.; Ohsumi, K.; Komatsu, M.; Mikouchi, T.; Zolensky, M. E.; Hirata, A.; Yamaguchi, S.; Kurokawa, A.

2016-01-01

The petrographic study of Itokawa particle, RA-QD02-0127 has been performed by SEM-EDS and optical microscope observations. The purpose of this study is to understand better the metamorphic and impact shock history of asteroid Itokawa, and other S-class asteroids.

Scanning digital lithography providing high speed large area patterning with diffraction limited sub-micron resolution

NASA Astrophysics Data System (ADS)

Wen, Sy-Bor; Bhaskar, Arun; Zhang, Hongjie

2018-07-01

A scanning digital lithography system using computer controlled digital spatial light modulator, spatial filter, infinity correct optical microscope and high precision translation stage is proposed and examined. Through utilizing the spatial filter to limit orders of diffraction modes for light delivered from the spatial light modulator, we are able to achieve diffraction limited deep submicron spatial resolution with the scanning digital lithography system by using standard one inch level optical components with reasonable prices. Raster scanning of this scanning digital lithography system using a high speed high precision x-y translation stage and piezo mount to real time adjust the focal position of objective lens allows us to achieve large area sub-micron resolved patterning with high speed (compared with e-beam lithography). It is determined in this study that to achieve high quality stitching of lithography patterns with raster scanning, a high-resolution rotation stage will be required to ensure the x and y directions of the projected pattern are in the same x and y translation directions of the nanometer precision x-y translation stage.

Rheological Analysis of Live and Dead Microalgae Suspensions

NASA Astrophysics Data System (ADS)

Song, Young Seok; Kang, Chul; Jeong, Jiwon; Kim, Kyu-Oh; Lim, Eunju

2018-04-01

We investigate the rheological properties of microalgae suspensions that are currently being used in various applications. Two kinds of microalgae, chlorella and Synechococcus, were used for preparation of the suspensions, and their rheological characteristics were analyzed experimentally. In order to evaluate the viability of algae, we performed live and dead tests using trypan blue staining assays. Morphological analyses for the algae were conducted using a scanning electron microscope (SEM) and an optical microscope (OP). We examined the viscoelastic behavior of the live and the dead algae suspensions by performing dynamic oscillatory shear tests.

Miniature near-infrared dual-axes confocal microscope utilizing a two-dimensional microelectromechanical systems scanner

PubMed Central

Liu, Jonathan T. C.; Mandella, Michael J.; Ra, Hyejun; Wong, Larry K.; Solgaard, Olav; Kino, Gordon S.; Piyawattanametha, Wibool; Contag, Christopher H.; Wang, Thomas D.

2007-01-01

The first, to our knowledge, miniature dual-axes confocal microscope has been developed, with an outer diameter of 10 mm, for subsurface imaging of biological tissues with 5–7 μm resolution. Depth-resolved en face images are obtained at 30 frames per second, with a field of view of 800 × 100 μm, by employing a two-dimensional scanning microelectromechanical systems mirror. Reflectance and fluorescence images are obtained with a laser source at 785 nm, demonstrating the ability to perform real-time optical biopsy. PMID:17215937

Two-photon imaging in living brain slices.

PubMed

Mainen, Z F; Maletic-Savatic, M; Shi, S H; Hayashi, Y; Malinow, R; Svoboda, K

1999-06-01

Two-photon excitation laser scanning microscopy (TPLSM) has become the tool of choice for high-resolution fluorescence imaging in intact neural tissues. Compared with other optical techniques, TPLSM allows high-resolution imaging and efficient detection of fluorescence signal with minimal photobleaching and phototoxicity. The advantages of TPLSM are especially pronounced in highly scattering environments such as the brain slice. Here we describe our approaches to imaging various aspects of synaptic function in living brain slices. To combine several imaging modes together with patch-clamp electrophysiological recordings we found it advantageous to custom-build an upright microscope. Our design goals were primarily experimental convenience and efficient collection of fluorescence. We describe our TPLSM imaging system and its performance in detail. We present dynamic measurements of neuronal morphology of neurons expressing green fluorescent protein (GFP) and GFP fusion proteins as well as functional imaging of calcium dynamics in individual dendritic spines. Although our microscope is a custom instrument, its key advantages can be easily implemented as a modification of commercial laser scanning microscopes. Copyright 1999 Academic Press.

Developments in optical modeling methods for metrology

NASA Astrophysics Data System (ADS)

Davidson, Mark P.

1999-06-01

Despite the fact that in recent years the scanning electron microscope has come to dominate the linewidth measurement application for wafer manufacturing, there are still many applications for optical metrology and alignment. These include mask metrology, stepper alignment, and overlay metrology. Most advanced non-optical lithographic technologies are also considering using topics for alignment. In addition, there have been a number of in-situ technologies proposed which use optical measurements to control one aspect or another of the semiconductor process. So optics is definitely not dying out in the semiconductor industry. In this paper a description of recent advances in optical metrology and alignment modeling is presented. The theory of high numerical aperture image simulation for partially coherent illumination is discussed. The implications of telecentric optics on the image simulation is also presented. Reciprocity tests are proposed as an important measure of numerical accuracy. Diffraction efficiencies for chrome gratings on reticles are one good way to test Kirchoff's approximation as compared to rigorous calculations. We find significant differences between the predictions of Kirchoff's approximation and rigorous methods. The methods for simulating brightfield, confocal, and coherence probe microscope imags are outlined, as are methods for describing aberrations such as coma, spherical aberration, and illumination aperture decentering.

The influence of voltage applied between the electrodes on optical and morphological properties of the InGaN thin films grown by thermionic vacuum arc.

PubMed

Özen, Soner; Şenay, Volkan; Pat, Suat; Korkmaz, Şadan

2016-01-01

The aim of this research is to investigate the optical and morphological properties of the InGaN thin films deposited onto amorphous glass substrates in two separate experiments with two different voltages applied between the electrodes, i.e. 500 and 600 V by means of the thermionic vacuum arc technique. This technique is original for thin film deposition and it enables thin film production in a very short period of time. The optical and morphological properties of the films were investigated by using field emission scanning electron microscope, atomic force microscope, spectroscopic ellipsometer, reflectometer, spectrophotometer, and optical tensiometer. Optical properties were also supported by empirical relations. The deposition rates were calculated as 3 and 3.3 nm/sec for 500 and 600 V, respectively. The increase in the voltage also increased the refractive index, grain size, root mean square roughness and surface free energy. According to the results of the wetting experiments, InGaN samples were low-wettable, also known as hydrophobic. © Wiley Periodicals, Inc.

Creating optical near-field orbital angular momentum in a gold metasurface.

PubMed

Chen, Ching-Fu; Ku, Chen-Ta; Tai, Yi-Hsin; Wei, Pei-Kuen; Lin, Heh-Nan; Huang, Chen-Bin

2015-04-08

Nanocavities inscribed in a gold thin film are optimized and designed to form a metasurface. We demonstrate both numerically and experimentally the creation of surface plasmon (SP) vortex carrying orbital angular momentum in the metasurface under linearly polarized optical excitation that carries no optical angular momentum. Moreover, depending on the orientation of the exciting linearly polarized light, we show that the metasurface is capable of providing dynamic switching between SP vortex formation or SP subwavelength focusing. The resulting SP intensities are experimentally measured using a near-field scanning optical microscope and are found in excellent quantitative agreements as compared to the numerical results.

Handheld optical-resolution photoacoustic microscopy

NASA Astrophysics Data System (ADS)

Lin, Li; Zhang, Pengfei; Xu, Song; Shi, Junhui; Li, Lei; Yao, Junjie; Wang, Lidai; Zou, Jun; Wang, Lihong V.

2017-04-01

Optical-resolution photoacoustic microscopy (OR-PAM) offers label-free in vivo imaging with high spatial resolution by acoustically detecting optical absorption contrasts via the photoacoustic effect. We developed a compact handheld OR-PAM probe for fast photoacoustic imaging. Different from benchtop microscopes, the handheld probe provides flexibility in imaging various anatomical sites. Resembling a cup in size, the probe uses a two-axis water-immersible microelectromechanical system mirror to scan both the illuminating optical beam and resultant acoustic beam. The system performance was tested in vivo by imaging the capillary bed in a mouse ear and both the capillary bed and a mole on a human volunteer.

Characterization of Polycapillary Optics in a TES Microcalorimeter EDS System Installed on an SEM

NASA Astrophysics Data System (ADS)

Takano, A.; Maehata, K.; Iyomoto, N.; Yasuda, K.; Maeno, H.; Shiiyama, K.; Tanaka, K.

2016-08-01

Energy-dispersive spectroscopic measurements are performed using a superconducting transition-edge sensor (TES) microcalorimeter mounted on a scanning electron microscope (SEM) for advanced research at Kyushu University. Because the sensitive area of the TES microcalorimeter is about 0.02~mm2, polycapillary optics is used to collect the X-rays emitted by the SEM specimen on the TES microcalorimeter. The X-ray transmission efficiency of the polycapillary optics is obtained by analyzing the X-ray energy spectra measured by the TES microcalorimeter. The obtained transmission efficiency of the polycapillary optics is reproduced by the calculated results of the simulation.

Lensless on-chip imaging of cells provides a new tool for high-throughput cell-biology and medical diagnostics.

PubMed

Mudanyali, Onur; Erlinger, Anthony; Seo, Sungkyu; Su, Ting-Wei; Tseng, Derek; Ozcan, Aydogan

2009-12-14

Conventional optical microscopes image cells by use of objective lenses that work together with other lenses and optical components. While quite effective, this classical approach has certain limitations for miniaturization of the imaging platform to make it compatible with the advanced state of the art in microfluidics. In this report, we introduce experimental details of a lensless on-chip imaging concept termed LUCAS (Lensless Ultra-wide field-of-view Cell monitoring Array platform based on Shadow imaging) that does not require any microscope objectives or other bulky optical components to image a heterogeneous cell solution over an ultra-wide field of view that can span as large as approximately 18 cm(2). Moreover, unlike conventional microscopes, LUCAS can image a heterogeneous cell solution of interest over a depth-of-field of approximately 5 mm without the need for refocusing which corresponds to up to approximately 9 mL sample volume. This imaging platform records the shadows (i.e., lensless digital holograms) of each cell of interest within its field of view, and automated digital processing of these cell shadows can determine the type, the count and the relative positions of cells within the solution. Because it does not require any bulky optical components or mechanical scanning stages it offers a significantly miniaturized platform that at the same time reduces the cost, which is quite important for especially point of care diagnostic tools. Furthermore, the imaging throughput of this platform is orders of magnitude better than conventional optical microscopes, which could be exceedingly valuable for high-throughput cell-biology experiments.

Lensless On-chip Imaging of Cells Provides a New Tool for High-throughput Cell-Biology and Medical Diagnostics

PubMed Central

Mudanyali, Onur; Erlinger, Anthony; Seo, Sungkyu; Su, Ting-Wei; Tseng, Derek; Ozcan, Aydogan

2009-01-01

Conventional optical microscopes image cells by use of objective lenses that work together with other lenses and optical components. While quite effective, this classical approach has certain limitations for miniaturization of the imaging platform to make it compatible with the advanced state of the art in microfluidics. In this report, we introduce experimental details of a lensless on-chip imaging concept termed LUCAS (Lensless Ultra-wide field-of-view Cell monitoring Array platform based on Shadow imaging) that does not require any microscope objectives or other bulky optical components to image a heterogeneous cell solution over an ultra-wide field of view that can span as large as ~18 cm2. Moreover, unlike conventional microscopes, LUCAS can image a heterogeneous cell solution of interest over a depth-of-field of ~5 mm without the need for refocusing which corresponds to up to ~9 mL sample volume. This imaging platform records the shadows (i.e., lensless digital holograms) of each cell of interest within its field of view, and automated digital processing of these cell shadows can determine the type, the count and the relative positions of cells within the solution. Because it does not require any bulky optical components or mechanical scanning stages it offers a significantly miniaturized platform that at the same time reduces the cost, which is quite important for especially point of care diagnostic tools. Furthermore, the imaging throughput of this platform is orders of magnitude better than conventional optical microscopes, which could be exceedingly valuable for high-throughput cell-biology experiments. PMID:20010542

PVP capped CdS nanoparticles for UV-LED applications

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sivaram, H.; Selvakumar, D.; Jayavel, R., E-mail: rjvel@annauniv.edu

Polyvinlypyrrolidone (PVP) capped cadmium sulphide (CdS) nanoparticles are synthesized by wet chemical method. The powder X-ray diffraction (XRD) result indicates that the nanoparticles are crystallized in cubic phase. The optical properties are characterized by UV-Vis absorption. The morphology of CdS nanoparticles are studied using Scanning electron microscope (SEM). The thermal behavior of the as prepared nanoparticles has been examined by Thermo gravimetric analysis (TGA). The optical absorption study of pvp capped CdS reveal a red shift confirms the UV-LED applications.

Generation of a frequency comb and applications thereof

DOEpatents

Hagmann, Mark J; Yarotski, Dmitry A

2013-12-03

Apparatus for generating a microwave frequency comb (MFC) in the DC tunneling current of a scanning tunneling microscope (STM) by fast optical rectification, cause by nonlinearity of the DC current vs. voltage curve for the tunneling junction, of regularly-spaced, short pulses of optical radiation from a focused mode-locked, ultrafast laser, directed onto the tunneling junction, is described. Application of the MFC to high resolution dopant profiling in semiconductors is simulated. Application of the MFC to other measurements is described.

Three-dimensional non-destructive optical evaluation of laser-processing performance using optical coherence tomography.

PubMed

Kim, Youngseop; Choi, Eun Seo; Kwak, Wooseop; Shin, Yongjin; Jung, Woonggyu; Ahn, Yeh-Chan; Chen, Zhongping

2008-06-01

We demonstrate the use of optical coherence tomography (OCT) as a non-destructive diagnostic tool for evaluating laser-processing performance by imaging the features of a pit and a rim. A pit formed on a material at different laser-processing conditions is imaged using both a conventional scanning electron microscope (SEM) and OCT. Then using corresponding images, the geometrical characteristics of the pit are analyzed and compared. From the results, we could verify the feasibility and the potential of the application of OCT to the monitoring of the laser-processing performance.

Three-dimensional non-destructive optical evaluation of laser-processing performance using optical coherence tomography

PubMed Central

Kim, Youngseop; Choi, Eun Seo; Kwak, Wooseop; Shin, Yongjin; Jung, Woonggyu; Ahn, Yeh-Chan; Chen, Zhongping

2014-01-01

We demonstrate the use of optical coherence tomography (OCT) as a non-destructive diagnostic tool for evaluating laser-processing performance by imaging the features of a pit and a rim. A pit formed on a material at different laser-processing conditions is imaged using both a conventional scanning electron microscope (SEM) and OCT. Then using corresponding images, the geometrical characteristics of the pit are analyzed and compared. From the results, we could verify the feasibility and the potential of the application of OCT to the monitoring of the laser-processing performance. PMID:24932051

An atomic force microscope for the study of the effects of tip sample interactions on dimensional metrology

NASA Astrophysics Data System (ADS)

Yacoot, Andrew; Koenders, Ludger; Wolff, Helmut

2007-02-01

An atomic force microscope (AFM) has been developed for studying interactions between the AFM tip and the sample. Such interactions need to be taken into account when making quantitative measurements. The microscope reported here has both the conventional beam deflection system and a fibre optical interferometer for measuring the movement of the cantilever. Both can be simultaneously used so as to not only servo control the tip movements, but also detect residual movement of the cantilever. Additionally, a high-resolution homodyne differential optical interferometer is used to measure the vertical displacement between the cantilever holder and the sample, thereby providing traceability for vertical height measurements. The instrument is compatible with an x-ray interferometer, thereby facilitating high resolution one-dimensional scans in the X-direction whose metrology is based on the silicon d220 lattice spacing (0.192 nm). This paper concentrates on the first stage of the instrument's development and presents some preliminary results validating the instrument's performance and showing its potential.

Design of high-performance adaptive objective lens with large optical depth scanning range for ultrabroad near infrared microscopic imaging

PubMed Central

Lan, Gongpu; Mauger, Thomas F.; Li, Guoqiang

2015-01-01

We report on the theory and design of adaptive objective lens for ultra broadband near infrared light imaging with large dynamic optical depth scanning range by using an embedded tunable lens, which can find wide applications in deep tissue biomedical imaging systems, such as confocal microscope, optical coherence tomography (OCT), two-photon microscopy, etc., both in vivo and ex vivo. This design is based on, but not limited to, a home-made prototype of liquid-filled membrane lens with a clear aperture of 8mm and the thickness of 2.55mm ~3.18mm. It is beneficial to have an adaptive objective lens which allows an extended depth scanning range larger than the focal length zoom range, since this will keep the magnification of the whole system, numerical aperture (NA), field of view (FOV), and resolution more consistent. To achieve this goal, a systematic theory is presented, for the first time to our acknowledgment, by inserting the varifocal lens in between a front and a back solid lens group. The designed objective has a compact size (10mm-diameter and 15mm-length), ultrabroad working bandwidth (760nm - 920nm), a large depth scanning range (7.36mm in air) — 1.533 times of focal length zoom range (4.8mm in air), and a FOV around 1mm × 1mm. Diffraction-limited performance can be achieved within this ultrabroad bandwidth through all the scanning depth (the resolution is 2.22 μm - 2.81 μm, calculated at the wavelength of 800nm with the NA of 0.214 - 0.171). The chromatic focal shift value is within the depth of focus (field). The chromatic difference in distortion is nearly zero and the maximum distortion is less than 0.05%. PMID:26417508

Numerical dispersion compensation for Partial Coherence Interferometry and Optical Coherence Tomography.

PubMed

Fercher, A; Hitzenberger, C; Sticker, M; Zawadzki, R; Karamata, B; Lasser, T

2001-12-03

Dispersive samples introduce a wavelength dependent phase distortion to the probe beam. This leads to a noticeable loss of depth resolution in high resolution OCT using broadband light sources. The standard technique to avoid this consequence is to balance the dispersion of the sample byarrangingadispersive materialinthereference arm. However, the impact of dispersion is depth dependent. A corresponding depth dependent dispersion balancing technique is diffcult to implement. Here we present a numerical dispersion compensation technique for Partial Coherence Interferometry (PCI) and Optical Coherence Tomography (OCT) based on numerical correlation of the depth scan signal with a depth variant kernel. It can be used a posteriori and provides depth dependent dispersion compensation. Examples of dispersion compensated depth scan signals obtained from microscope cover glasses are presented.

Digital Hilbert transformation for separation measurement of thicknesses and refractive indices of layered objects by use of a wavelength-scanning heterodyne interference confocal microscope.

PubMed

Watanabe, Yuuki; Yamaguchi, Ichirou

2002-08-01

A wavelength-scanning heterodyne interference confocal microscope quickly accomplishes the simultaneous measurement of the thickness and the refractive index of a sample by detection of the amplitude and the phase of the interference signal during a sample scan. However, the measurement range of the optical path difference (OPD) that is obtained from the phase changes is limited by the time response of the phase-locked loop circuit in the FM demodulator. To overcome this limitation and to improve the accuracy of the separation measurement, we propose an OPD detection using digital signal processing with a Hilbert transform. The measurement range is extended approximately five times, and the resolution of the OPD is improved to 5.5 from 9 microm without the electrical noise of the FM demodulator circuit. By applying this method for simultaneous measurement of thickness and the refractive index, we can measure samples 20-30-microm thick with refractive indices between 1 and 1.5.

Digital Hilbert transformation for separation measurement of thicknesses and refractive indices of layered objects by use of a wavelength-scanning heterodyne interference confocal microscope

NASA Astrophysics Data System (ADS)

Watanabe, Yuuki; Yamaguchi, Ichirou

2002-08-01

A wavelength-scanning heterodyne interference confocal microscope quickly accomplishes the simultaneous measurement of the thickness and the refractive index of a sample by detection of the amplitude and the phase of the interference signal during a sample scan. However, the measurement range of the optical path difference (OPD) that is obtained from the phase changes is limited by the time response of the phase-locked loop circuit in the FM demodulator. To overcome this limitation and to improve the accuracy of the separation measurement, we propose an OPD detection using digital signal processing with a Hilbert transform. The measurement range is extended approximately five times, and the resolution of the OPD is improved to 5.5 from 9 mum without the electrical noise of the FM demodulator circuit. By applying this method for simultaneous measurement of thickness and the refractive index, we can measure samples 20-30-mum thick with refractive indices between 1 and 1.5.

Light-sheet microscopy by confocal line scanning of dual-Bessel beams

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Pengfei; Phipps, Mary Elizabeth; Goodwin, Peter Marvin

Here, we have developed a light-sheet microscope that uses confocal scanning of dual-Bessel beams for illumination. A digital micromirror device (DMD) is placed in the intermediate image plane of the objective used to collect fluorescence and is programmed with two lines of pixels in the “on” state such that the DMD functions as a spatial filter to reject the out-of-focus background generated by the side-lobes of the Bessel beams. The optical sectioning and out-of-focus background rejection capabilities of this microscope were demonstrated by imaging of fluorescently stained actin in human A431 cells. The dual-Bessel beam system enables twice as manymore » photons to be detected per imaging scan, which is useful for low light applications (e.g., single-molecule localization) or imaging at high speed with a superior signal to noise. While demonstrated for two Bessel beams, this approach is scalable to a larger number of beams.« less

Light-sheet microscopy by confocal line scanning of dual-Bessel beams

DOE PAGES

Zhang, Pengfei; Phipps, Mary Elizabeth; Goodwin, Peter Marvin; ...

2016-10-25

Here, we have developed a light-sheet microscope that uses confocal scanning of dual-Bessel beams for illumination. A digital micromirror device (DMD) is placed in the intermediate image plane of the objective used to collect fluorescence and is programmed with two lines of pixels in the “on” state such that the DMD functions as a spatial filter to reject the out-of-focus background generated by the side-lobes of the Bessel beams. The optical sectioning and out-of-focus background rejection capabilities of this microscope were demonstrated by imaging of fluorescently stained actin in human A431 cells. The dual-Bessel beam system enables twice as manymore » photons to be detected per imaging scan, which is useful for low light applications (e.g., single-molecule localization) or imaging at high speed with a superior signal to noise. While demonstrated for two Bessel beams, this approach is scalable to a larger number of beams.« less

A Miniaturized Variable Pressure Scanning Electron Microscope (MVP-SEM) for the Surface of Mars: An Instrument for the Planetary Science Community

NASA Technical Reports Server (NTRS)

Edmunson, J.; Gaskin, J. A.; Danilatos, G.; Doloboff, I. J.; Effinger, M. R.; Harvey, R. P.; Jerman, G. A.; Klein-Schoder, R.; Mackie, W.; Magera, B.;

Path-separated electron interferometry in a scanning transmission electron microscope

NASA Astrophysics Data System (ADS)

Yasin, Fehmi S.; Harvey, Tyler R.; Chess, Jordan J.; Pierce, Jordan S.; McMorran, Benjamin J.

2018-05-01

We report a path-separated electron interferometer within a scanning transmission electron microscope. In this setup, we use a nanofabricated grating as an amplitude-division beamsplitter to prepare multiple spatially separated, coherent electron probe beams. We achieve path separations of 30 nm. We pass the  +1 diffraction order probe through amorphous carbon while passing the 0th and  ‑1 orders through vacuum. The probes are then made to interfere via imaging optics, and we observe an interference pattern at the CCD detector with up to 39.7% fringe visibility. We show preliminary experimental results in which the interference pattern was recorded during a 1D scan of the diffracted probes across a test phase object. These results qualitatively agree with a modeled interference predicted by an independent measurement of the specimen thickness. This experimental design can potentially be applied to phase contrast imaging and fundamental physics experiments, such as an exploration of electron wave packet coherence length.

Ultra-Compact Multitip Scanning Probe Microscope with an Outer Diameter of 50 mm

NASA Astrophysics Data System (ADS)

Cherepanov, Vasily; Zubkov, Evgeny; Junker, Hubertus; Korte, Stefan; Blab, Marcus; Coenen, Peter; Voigtländer, Bert

We present a multitip scanning tunneling microscope (STM) where four independent STM units are integrated on a diameter of 50 mm. The coarse positioning of the tips is done under the control of an optical microscope or an SEM in vacuum. The heart of this STM is a new type of piezoelectric coarse approach called Koala Drive which can have a diameter greater than 2.5 mm and a length smaller than 10 mm. Alternating movements of springs move a central tube which holds the STM tip or AFM sensor. This new operating principle provides a smooth travel sequence and avoids shaking which is intrinsically present for nanopositioners based on inertial motion with saw tooth driving signals. Inserting the Koala Drive in a piezo tube for xyz-scanning integrates a complete STM inside a 4 mm outer diameter piezo tube of <10 mm length. The use of the Koala Drive makes the scanning probe microscopy design ultra-compact and accordingly leads to a high mechanical stability. The drive is UHV, low temperature, and magnetic field compatible. The compactness of the Koala Drive allows building a four-tip STM as small as a single-tip STM with a drift of <0.2 nm/min and lowest resonance frequencies of 2.5 (xy) and 5.5 kHz (z). We present examples of the performance of the multitip STM designed using the Koala Drive.

Fabrication of large area plasmonic nanoparticle grating structure on silver halide based transmission electron microscope film and its application as a surface enhanced Raman spectroscopy substrate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sudheer,, E-mail: sudheer@rrcat.gov.in; Tiwari, P.; Singh, M. N.

The plasmonic responses of silver nanoparticle grating structures of different periods made on silver halide based electron microscope film are investigated. Raster scan of the conventional scanning electron microscope (SEM) is used to carry out electron beam lithography for fabricating the plasmonic nanoparticle grating (PNG) structures. Morphological characterization of the PNG structures, carried out by the SEM and the atomic force microscope, indicates that the depth of the groove decreases with a decrease in the grating period. Elemental characterization performed by the energy dispersive spectroscopy and the x-ray diffraction shows the presence of nanoparticles of silver in the PNG grating.more » The optical characterization of the gratings shows that the localized surface plasmon resonance peak shifts from 366 to 378 nm and broadens with a decrease in grating period from 10 to 2.5 μm. The surface enhanced Raman spectroscopy of the Rhodamine-6G dye coated PNG structure shows the maximum enhancement by two orders of magnitude in comparison to the randomly distributed silver nanoparticles having similar size and shape as the PNG structure.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cooke, Gary A.; Pestovich, John A.; Huber, Heinz J.

This report presents the results for solid phase characterization (SPC) of solid samples removed from tank 241-C-108 (C-108) on August 12-13,2012, using the off-riser sampler. Samples were received at the 222-S Laboratory on August 13 and were described and photographed. The SPC analyses that were performed include scanning electron microscopy (SEM) using the ASPEX(R)l scanning electron microscope, X-ray diffraction (XRD) using the Rigaku(R) 2 MiniFlex X-ray diffractometer, and polarized light microscopy (PLM) using the Nikon(R) 3 Eclipse Pol optical microscope. The SEM is equipped with an energy dispersive X-ray spectrometer (EDS) to provide chemical information. Gary A. Cooke conducted themore » SEM analysis, John A. Pestovich performed the XRD analysis, and Dr. Heinz J. Huber performed the PLM examination. The results of these analyses are presented here.« less

Improved silicon nitride for advanced heat engines

NASA Technical Reports Server (NTRS)

Yeh, Harry C.; Fang, Ho T.

1991-01-01

The results of a four year program to improve the strength and reliability of injection-molded silicon nitride are summarized. Statistically designed processing experiments were performed to identify and optimize critical processing parameters and compositions. Process improvements were monitored by strength testing at room and elevated temperatures, and microstructural characterization by optical, scanning electron microscopes, and scanning transmission electron microscope. Processing modifications resulted in a 20 percent strength and 72 percent Weibull slope improvement of the baseline material. Additional sintering aids screening and optimization experiments succeeded in developing a new composition (GN-10) capable of 581.2 MPa at 1399 C. A SiC whisker toughened composite using this material as a matrix achieved a room temperature toughness of 6.9 MPa m(exp .5) by the Chevron notched bar technique. Exploratory experiments were conducted on injection molding of turbocharger rotors.

Nonlinear Polarimetric Microscopy for Biomedical Imaging

NASA Astrophysics Data System (ADS)

Samim, Masood

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

Computational modeling of optical projection tomographic microscopy using the finite difference time domain method.

PubMed

Coe, Ryan L; Seibel, Eric J

2012-12-01

We present a method for modeling image formation in optical projection tomographic microscopy (OPTM) using high numerical aperture (NA) condensers and objectives. Similar to techniques used in computed tomography, OPTM produces three-dimensional, reconstructed images of single cells from two-dimensional projections. The model is capable of simulating axial scanning of a microscope objective to produce projections, which are reconstructed using filtered backprojection. Simulation of optical scattering in transmission optical microscopy is designed to analyze all aspects of OPTM image formation, such as degree of specimen staining, refractive-index matching, and objective scanning. In this preliminary work, a set of simulations is performed to examine the effect of changing the condenser NA, objective scan range, and complex refractive index on the final reconstruction of a microshell with an outer radius of 1.5 μm and an inner radius of 0.9 μm. The model lays the groundwork for optimizing OPTM imaging parameters and triaging efforts to further improve the overall system design. As the model is expanded in the future, it will be used to simulate a more realistic cell, which could lead to even greater impact.

Effect of reduction time on third order optical nonlinearity of reduced graphene oxide

NASA Astrophysics Data System (ADS)

Sreeja, V. G.; Vinitha, G.; Reshmi, R.; Anila, E. I.; Jayaraj, M. K.

2017-04-01

We report the influence of reduction time on structural, linear and nonlinear optical properties of reduced graphene oxide (rGO) thin films synthesized by spin coating method. We observed that the structural, linear and nonlinear optical properties can be tuned with reduction time in GO is due to the increased structural ordering because of the restoration of sp2 carbon atoms with the time of reduction. The nonlinear absorption studies by open aperture Z-scan technique exhibited a saturable absorption. The nonlinear refraction studies showed the self de focusing nature of rGO by closed aperture Z scan technique. The nonlinear absorption coefficient and saturation intensity varies with the time for reduction of GO which is attributed to the depletion of valence band and the conduction band filling effect. Our results emphasize duration for reduction of GO dependent optical nonlinearity of rGO thin films to a great extent and explore its applications Q switched mode locking laser systems for generating ultra short laser pulses and in optical sensors. The rGO coated films were characterized by X-Ray diffraction method (XRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, UV-Vis absorption spectroscopy (UV-Vis), Photoluminescence (PL) and Scanning electron microscope (SEM) measurements.

Special Features of the Optical Absorption and Photoconductivity of Indium Monoselenide Upon Laser Excitation

NASA Astrophysics Data System (ADS)

Kyazym-Zade, A. G.; Salmanov, V. M.; Guseinov, A. G.; Mamedov, R. M.; Salmanova, A. A.; Akhmedova, F. Sh.

2018-02-01

The successive ionic layer adsorption and reaction (SILAR) method is used to prepare InSe thin films and InSe nanoparticles. Shapes and sizes of the obtained nanoparticles are investigated using a scanning electron microscope and an atomic force microscope. The main parameters of the examined structures, nanoparticle sizes (4-20 nm), and band gap ( E g = 1.60 eV) for nanoparticles with the least sizes are determined. Superfast (1.5·10-8 s) photocurrent relaxation and stimulated emission with line half-width of 8 Å have been observed upon exposure to laser radiation.

Multimodality hard-x-ray imaging of a chromosome with nanoscale spatial resolution

DOE PAGES

Yan, Hanfei; Nazaretski, Evgeny; Lauer, Kenneth R.; ...

2016-02-05

Here, we developed a scanning hard x-ray microscope using a new class of x-ray nano-focusing optic called a multilayer Laue lens and imaged a chromosome with nanoscale spatial resolution. The combination of the hard x-ray's superior penetration power, high sensitivity to elemental composition, high spatial-resolution and quantitative analysis creates a unique tool with capabilities that other microscopy techniques cannot provide. Using this microscope, we simultaneously obtained absorption-, phase-, and fluorescence-contrast images of Pt-stained human chromosome samples. The high spatial-resolution of the microscope and its multi-modality imaging capabilities enabled us to observe the internal ultra-structures of a thick chromosome without sectioningmore » it.« less

Multimodality hard-x-ray imaging of a chromosome with nanoscale spatial resolution

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yan, Hanfei; Nazaretski, Evgeny; Lauer, Kenneth R.

Here, we developed a scanning hard x-ray microscope using a new class of x-ray nano-focusing optic called a multilayer Laue lens and imaged a chromosome with nanoscale spatial resolution. The combination of the hard x-ray's superior penetration power, high sensitivity to elemental composition, high spatial-resolution and quantitative analysis creates a unique tool with capabilities that other microscopy techniques cannot provide. Using this microscope, we simultaneously obtained absorption-, phase-, and fluorescence-contrast images of Pt-stained human chromosome samples. The high spatial-resolution of the microscope and its multi-modality imaging capabilities enabled us to observe the internal ultra-structures of a thick chromosome without sectioningmore » it.« less

Microstructure analysis in the coupling region of fiber coupler with a novel electrical micro-heater

NASA Astrophysics Data System (ADS)

Shuai, Cijun; Gao, Chengde; Nie, Yi; Hu, Huanlong; Peng, Shuping

2011-12-01

Fused-tapered fiber coupler is widely used in optical-fiber communication, optical-fiber sensor and optical signal processing. Its optical performance is mainly determined by the glass properties in the coupling region. In this study, the effect of fused biconical taper (FBT) process on glass microstructure of fiber coupler was investigated by testing the microstructure of the cross-section of coupling region. The fiber coupler is fabricated with a novel home-designed electrical heater. Our experimental results show that the boundary between fiber core and fiber cladding become vague or indistinct after FBT under transmission electron microscopy (TEM) and Ge 2+ in fiber core diffuses into fiber cladding. Crystallizations are observed in coupling region under scanning electron microscope (SEM) and microscopic infrared (IR), and the micro crystallizations become smaller with the drawing speed increasing. The wave number of fiberglass increases after FBT and it is in proportion to the drawing speed. The analysis of the microstructure in the coupling region explored the mechanism of the improvement in the performance of fiber couplers which can be used for the guidance of fabrication process.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yashchuk, V.V.; Conley, R.; Anderson, E.H.

Verification of the reliability of metrology data from high quality X-ray optics requires that adequate methods for test and calibration of the instruments be developed. For such verification for optical surface profilometers in the spatial frequency domain, a modulation transfer function (MTF) calibration method based on binarypseudo-random (BPR) gratings and arrays has been suggested and and proven to be an effective calibration method for a number of interferometric microscopes, a phase shifting Fizeau interferometer, and a scatterometer. Here we describe the details of development of binarypseudo-random multilayer (BPRML) test samples suitable for characterization of scanning (SEM) and transmission (TEM) electronmore » microscopes. We discuss the results of TEM measurements with the BPRML test samples fabricated from a WiSi{sub 2}/Si multilayer coating with pseudo-randomly distributed layers. In particular, we demonstrate that significant information about the metrological reliability of the TEM measurements can be extracted even when the fundamental frequency of the BPRML sample is smaller than the Nyquist frequency of the measurements. The measurements demonstrate a number of problems related to the interpretation of the SEM and TEM data. Note that similar BPRML testsamples can be used to characterize X-ray microscopes. Corresponding work with X-ray microscopes is in progress.« less

OPTiM: Optical projection tomography integrated microscope using open-source hardware and software

PubMed Central

Andrews, Natalie; Davis, Samuel; Bugeon, Laurence; Dallman, Margaret D.; McGinty, James

2017-01-01

We describe the implementation of an OPT plate to perform optical projection tomography (OPT) on a commercial wide-field inverted microscope, using our open-source hardware and software. The OPT plate includes a tilt adjustment for alignment and a stepper motor for sample rotation as required by standard projection tomography. Depending on magnification requirements, three methods of performing OPT are detailed using this adaptor plate: a conventional direct OPT method requiring only the addition of a limiting aperture behind the objective lens; an external optical-relay method allowing conventional OPT to be performed at magnifications >4x; a remote focal scanning and region-of-interest method for improved spatial resolution OPT (up to ~1.6 μm). All three methods use the microscope’s existing incoherent light source (i.e. arc-lamp) and all of its inherent functionality is maintained for day-to-day use. OPT acquisitions are performed on in vivo zebrafish embryos to demonstrate the implementations’ viability. PMID:28700724

Molecular imaging of photodynamic therapy

NASA Astrophysics Data System (ADS)

Chang, Sung K.; Errabelli, Divya; Rizvi, Imran; Solban, Nicolas; O'Riordan, Katherine; Hasan, Tayyaba

2006-02-01

Recent advances in light sources, detectors and other optical imaging technologies coupled with the development of novel optical contrast agents have enabled real-time, high resolution, in vivo monitoring of molecular targets. Noninvasive monitoring of molecular targets is particularly relevant to photodynamic therapy (PDT), including the delivery of photosensitizer in the treatment site and monitoring of molecular and physiological changes following treatment. Our lab has developed optical imaging technologies to investigate these various aspects of photodynamic therapy (PDT). We used a laser scanning confocal microscope to monitor the pharmacokinetics of various photosensitizers in in vitro as well as ex vivo samples, and developed an intravital fluorescence microscope to monitor photosensitizer delivery in vivo in small animals. A molecular specific contrast agent that targets the vascular endothelial growth factor (VEGF) was developed to monitor the changes in the protein expression following PDT. We were then able to study the physiological changes due to post-treatment VEGF upregulation by quantifying vascular permeability with in vivo imaging.

Piezoresistor-equipped fluorescence-based cantilever probe for near-field scanning.

PubMed

Kan, Tetsuo; Matsumoto, Kiyoshi; Shimoyama, Isao

2007-08-01

Scanning near-field optical microscopes (SNOMs) with fluorescence-based probes are promising tools for evaluating the optical characteristics of nanoaperture devices used for biological investigations, and this article reports on the development of a microfabricated fluorescence-based SNOM probe with a piezoresistor. The piezoresistor was built into a two-legged root of a 160-microm-long cantilever. To improve the displacement sensitivity of the cantilever, the piezoresistor's doped area was shallowly formed on the cantilever surface. A fluorescent bead, 500 nm in diameter, was attached to the bottom of the cantilever end as a light-intensity-sensitive material in the visible-light range. The surface of the scanned sample was simply detected by the probe's end being displaced by contact with the sample. Measuring displacements piezoresistively is advantageous because it eliminates the noise arising from the use of the optical-lever method and is free of any disturbance in the absorption or the emission spectrum of the fluorescent material at the probe tip. The displacement sensitivity was estimated to be 6.1 x 10(-6) nm(-1), and the minimum measurable displacement was small enough for near-field measurement. This probe enabled clear scanning images of the light field near a 300 x 300 nm(2) aperture to be obtained in the near-field region where the tip-sample distance is much shorter than the light wavelength. This scanning result indicates that the piezoresistive way of tip-sample distance regulation is effective for characterizing nanoaperture optical devices.

EDITORIAL: Nanotechnology in motion Nanotechnology in motion

NASA Astrophysics Data System (ADS)

Demming, Anna

2012-02-01

Microscopes provide tools of inimitable value for probing the building blocks of the world around us. The identity of the inventor of the first microscope remains under debate, but a name unequivocally linked with early developments in microscopy is Robert Hooke. His Micrographia published in 1665, was the first ever bestseller in science and brought topics in microscopy to the broader public eye with pages of detailed micrographs, most famously the fly's eye and plant cells. Since the first microscopes in the late 16th century, ingenious alternatives to the original optical microscopes have been developed to create images of the world at ever smaller dimensions. Innovations include scanning probe techniques such as the atomic force microscope [1]. As Toshio Ando describes in a review in this issue [2], these devices have also entered a new era in the past decade with the development of high-speed atomic force microscopy. Now, we can not only see the nanoscale components that make up the world around us, but we can watch them at work. One of the first innovations in optical microscopy was the use of dyes. This principle first came into practice with the use of ultraviolet light to reveal previously indistinguishable features. As explained by a researcher in the early 1930s, 'It is obvious that if the dyes used for selective staining in ordinary microscopical work are supplemented by substances which cause a particular detail of the structure to fluoresce with a specific colour in ultraviolet light, then many strings will be added to the bow of the practical microscopist' [3]. More recently, emphasis on the role of plasmons—collective oscillations of electrons in nanoscale metal structures—has received considerable research attention. Plasmons enhance the local electromagnetic field and can lead to increased fluorescence rates from nearby fluorophores depending on the efficiency of the counteracting process, non-radiative transfer [4]. The 1930s also saw the development of the electron microscope, which aimed to exceed the resolving power of diffraction-limited optical microscopes. Since the diffraction limit is proportional to the incident wavelength, the shorter wavelength electron beam allows smaller features to be resolved than optical light. Ernst Ruska shared the Nobel Prize for Physics in 1986 for his work in developing the transmission electron microscope [5]. The technique continues to provide an invaluable tool in nanotechnology studies, as demonstrated recently by a collaboration of researchers in the US, Singapore and Korea used electron and atomic force microscopy in their investigation of the deposition of gold nanoparticles on graphene and the enhanced conductivity of the doped film [6]. The other half of the 1986 Nobel Prize was awarded jointly to Gerd Binnig and Heinrich Rohrer 'for their design of the scanning tunnelling microscope'. The scanning tunnelling microscope offered the first glimpses of atomic scale features, galvanizing research in nanoscale science and technology into a burst of fruitful activity that persists to this day. Instead of using the diffraction and scattering of beams to 'see' nanoscale structures, the atomic force microscope developed by Binnig, Quate and Gerber in the 1980s [1] determines the surface topology 'by touch'. The device uses nanoscale changes in the forces exerted on a tip as it scans the sample surface to generate an image. As might be expected, innovations on the original atomic force microscope have now been developed achieving ever greater sensitivities for imaging soft matter without destroying it. Recent work by collaborators at the University of Bristol and the University of Glasgow used a cigar-shaped nanoparticle held in optical tweezers as the scanning tip. The technique is not diffraction limited, imparts less force on samples than contact scanning probe microscopy techniques, and allows highly curved and strongly scattering samples to be imaged [7]. In this issue, Toshio Ando from the University of Kanazawa provides an overview of developments that have allowed atomic force microscopy to move from rates of the order of one frame a minute to over a thousand frames per second in constant height mode, as reported by Mervyn Miles and colleagues at Bristol University and University College London [8]. Among the pioneers in the field, Ando's group demonstrated the ability to record the Brownian motion of myosin V molecules on mica with image capture rates of 100 x 100 pixels in 80 ms over a decade ago [9]. The developments unleash the potential of atomic force microscopy to observe the dynamics of biological and materials systems. If seeing is believing, the ability to present real motion pictures of the nanoworld cannot fail to capture the public imagination and stimulate burgeoning new avenues of scientific endeavour. Nearly 350 years on from the publication Micrographia, images in microscopy have moved from the page to the movies. References [1] Binnig G, Quate C F, and Gerber Ch 1986 Phys. Rev. Lett. 56 930-3 [2] Ando T 2012 Nanotechnology 23 062001 [3] J G 1934 Nature 134 635-6 [4] Bharadwaj P, Anger P and Novotny L 2007 Nanotechnology 18 044017 [5] The Nobel Prize in Physics 1986 Nobelprize.org [6] Kim K K, Reina A, Shi Y, Park H, Li L-J, Lee Y H and Kong J 2010 Nanotechnology 21 285205 [7] Phillips D B, Grieve J A, Olof S N, Kocher S J, Bowman R, Padgett M J, Miles M J and Carberry D M 2011 Nanotechnology 22 285503 [8] Picco L M, Bozec L, Ulcinas A, Engledew D J, Antognozzi M, Horton M A and Miles M J 2007 Nanotechnology 18 044030 [9] Ando T, Kodera N, Takai E, Maruyama D, Saito K and Toda A 2001 Proc. Natl. Acad. Sci. 98 12468

Note: Laser beam scanning using a ferroelectric liquid crystal spatial light modulator

NASA Astrophysics Data System (ADS)

Das, Abhijit; Boruah, Bosanta R.

2014-04-01

In this work we describe laser beam scanning using a ferroelectric liquid crystal spatial light modulator. Commercially available ferroelectric liquid crystal spatial light modulators are capable of displaying 85 colored images in 1 s using a time dithering technique. Each colored image, in fact, comprises 24 single bit (black and white) images displayed sequentially. We have used each single bit image to write a binary phase hologram. For a collimated laser beam incident on the hologram, one of the diffracted beams can be made to travel along a user defined direction. We have constructed a beam scanner employing the above arrangement and demonstrated its use to scan a single laser beam in a laser scanning optical sectioning microscope setup.

Combined low-temperature scanning tunneling/atomic force microscope for atomic resolution imaging and site-specific force spectroscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schwarz, Udo; Albers, Boris J.; Liebmann, Marcus

2008-02-27

The authors present the design and first results of a low-temperature, ultrahigh vacuum scanning probe microscope enabling atomic resolution imaging in both scanning tunneling microscopy (STM) and noncontact atomic force microscopy (NC-AFM) modes. A tuning-fork-based sensor provides flexibility in selecting probe tip materials, which can be either metallic or nonmetallic. When choosing a conducting tip and sample, simultaneous STM/NC-AFM data acquisition is possible. Noticeable characteristics that distinguish this setup from similar systems providing simultaneous STM/NC-AFM capabilities are its combination of relative compactness (on-top bath cryostat needs no pit), in situ exchange of tip and sample at low temperatures, short turnaroundmore » times, modest helium consumption, and unrestricted access from dedicated flanges. The latter permits not only the optical surveillance of the tip during approach but also the direct deposition of molecules or atoms on either tip or sample while they remain cold. Atomic corrugations as low as 1 pm could successfully be resolved. In addition, lateral drifts rates of below 15 pm/h allow long-term data acquisition series and the recording of site-specific spectroscopy maps. Results obtained on Cu(111) and graphite illustrate the microscope's performance.« less

EDITORIAL: Scanning probe microscopy: a visionary development Scanning probe microscopy: a visionary development

NASA Astrophysics Data System (ADS)

Demming, Anna

2013-07-01

The development of scanning probe microscopy repositioned modern physics. When Rohrer and Binnig first used electronic tunnelling effects to image atoms and quantum states they did more than pin down theoretical hypotheses to real-world observables; the scanning tunnelling microscope fed imaginations, prompting researchers to consider new directions and possibilities [1]. As Rohrer once commented, 'We could show that you can easily manipulate or position something small in space with an accuracy of 10 pm.... When you can do that, you simply have ideas of what you can do' [2]. The development heralded a cavalry of scanning probe techniques—such as atomic force microscopy (AFM) [3-5], scanning near-field optical microscopy (SNOM) [6-8] and Kelvin probe force microscopy (KPFM) [9, 10]—that still continue to bring nanomaterials and nanoscale phenomena into fresh focus. Not long after the development of scanning tunnelling microscopy, Binnig, Quate and Gerber collaborating in California in the US published work on a new type of microscope also capable of atomic level resolution [3]. The original concept behind scanning tunnelling microscopy uses electrical conductance, which places substantial limitations on the systems that it can image. Binnig, Quate and Gerber developed the AFM to 'feel' the topology of surfaces like the needle of an old fashioned vinyl player. In this way insulators could be imaged as well. The development of a force modulation mode AFM extended the tool's reach to soft materials making images of biological samples accessible with the technique [4]. There have now been a number of demonstrations of image capture at rates that allow dynamics at the nanoscale to be tracked in real time, opening further possibilities in applications of the AFM as described in a recent review by Toshio Ando at Kanazawa University [5]. Researchers also found a way to retrieve optical information at 'super-resolution' [6, 7]. Optical microscopy provides spectral details that harbour a wealth of additional information about the sample and its environment, like switching from black and white to technicolour. With the invention of SNOM these details were no longer restricted by the diffraction limit to a resolution of half the wavelength of the incident light. The principle behind SNOM remains very similar to STM but instead of measuring an electronic current, information is captured from the non-propagating optical near field, where the diffraction limit does not apply. SNOM continues to be an invaluable imaging technique as demonstrated recently by researchers in Spain and Korea, who used it to measure near-infrared-to-visible upconversion and cathodoluminescence emission properties of Ln3+ in nanocrystalline Ln-doped Lu2O3 materials with 1D morphology [8]. Their work holds promise for controlled incorporation of such optically active nanostructures in future photonic structures and applications. The cantilever-probe system provides a number of highly sensitive interactions that can be exploited to extract details of a sample system. The potential offset between the probe and surface manifests itself in a force and this too has been used in KPFM [9]. The finite tip size has a profound effect on the measured image in scanning probe-microscopes in general. In KPFM, as Rosenwaks and colleagues in Israel, US and Germany point out in this issue [10] the influence of the tip and cantilever on measurements is particularly significant because of the long range nature of the electrostatic forces involved. Measurements at any one point provide a weighted average of the contact potential difference of the sample and to obtain a quantitative image this averaging must be taken into account. Rosenwaks and colleagues tackle this challenge in the work reported in this issue, presenting an algorithm for reconstructing a sample surface potential from its KPFM image. Their study also reveals that the averaging effects are far more significant for amplitude modulated KPFM measurements compared with the frequency modulated mode. Rohrer and Binnig shared the Nobel Prize for Physics 'for their design of the scanning tunnelling microscope' [11]. They are widely recognized among the founding fathers of nanoscience. In an interview in 2005 Rohrer once commented on the benefits of changing fields even if it leaves you feeling a little 'lost and lonely' at first. In fact he attributed his ability to contribute his Nobel Prize winning work to science at a comparatively senior age to the fact that he had changed fields. 'You cannot be the star from the beginning, but I think what is important is that you might bring in a different way of thinking. You have a certain lightness to approach something that is the expert opinion' [2]. In nanotechnology where such a formidable range of disciplines seem to feed into the research such words may be particularly encouraging. Rohrer passed away on 16 May 2013, but the awesome legacy of his life's work continues. With the scanning tunnelling microscope the lofty eccentricities of quantum mechanical theory literally came into view, quite an inspiration. References [1] Binning G, Rohrer H, Gerber Ch and Weibel E 1982 Surface studies by scanning tunneling microscopy Phys. Rev. Lett. 49 57-61 [2] Weiss P S 2007 A conversation with Dr. Heinrich Rohrer: STM Co-inventor and one of the founding fathers of nanoscience ACS Nano 1 3-5 [3] Binnig G, Quate C F and Gerber Ch 1986 Atomic force microscope Phys. Rev. Lett. 56 930-3 [4] Maivald P, Butt H J, Gould S A C, Prater C B, Drake B, Gurley J A, Elings V B and Hansma P K 1991 Using force modulation to image surface elasticities with the atomic force microscope Nanotechnology 2 103-6 [5] Ando T 2012 High-speed atomic force microscopy coming of age Nanotechnology 23 062001 [6] Betzig E, Isaacson M, Barshatzky H, Lewis A and Lin K 1988 Super-resolution imaging with near-field scanning optical microscopy (NSOM) Ultramicroscopy 25 155-63 [7] Thio T, Lezec H J, Ebbesen T W, Pellerin K M, Lewen G D, Nahata A and Linke R A 2002 Giant optical transmission of sub-wavelength apertures: physics and applications Nanotechnology 13 429-32 [8] Barrera E W, Pujol M C, Díaz F, Choi S B, Rotermund F, Park K H, Jeong M S and Cascales C 2011 Emission properties of hydrothermal Yb3+, Er3+ and Yb3+, Tm3+-codoped Lu2O3 nanorods: upconversion, cathodoluminescence and assessment of waveguide behaviour Nanotechnology 22 075205 [9] Nonnenmacher M, O'Boyle M P and Wickramasinghe H K 1991 Kelvin probe force microscopy Appl. Phys. Lett. 58 2921-3 [10] Cohen G, Halpern E, Nanayakkara S U, Luther J M, Held C, Bennewitz R, Boag A and Rosenwaks Y 2013 Reconstruction of surface potential from Kelvin probe force microscopy images Nanotechnology 24 295702 [11] 1986 The Nobel Prize in Physics www.nobelprize.org/nobel prizes/physics/laureates/1986/ index.html

Near-Field Enhanced Photochemistry of Single Molecules in a Scanning Tunneling Microscope Junction.

PubMed

Böckmann, Hannes; Gawinkowski, Sylwester; Waluk, Jacek; Raschke, Markus B; Wolf, Martin; Kumagai, Takashi

2018-01-10

Optical near-field excitation of metallic nanostructures can be used to enhance photochemical reactions. The enhancement under visible light illumination is of particular interest because it can facilitate the use of sunlight to promote photocatalytic chemical and energy conversion. However, few studies have yet addressed optical near-field induced chemistry, in particular at the single-molecule level. In this Letter, we report the near-field enhanced tautomerization of porphycene on a Cu(111) surface in a scanning tunneling microscope (STM) junction. The light-induced tautomerization is mediated by photogenerated carriers in the Cu substrate. It is revealed that the reaction cross section is significantly enhanced in the presence of a Au tip compared to the far-field induced process. The strong enhancement occurs in the red and near-infrared spectral range for Au tips, whereas a W tip shows a much weaker enhancement, suggesting that excitation of the localized plasmon resonance contributes to the process. Additionally, using the precise tip-surface distance control of the STM, the near-field enhanced tautomerization is examined in and out of the tunneling regime. Our results suggest that the enhancement is attributed to the increased carrier generation rate via decay of the excited near-field in the STM junction. Additionally, optically excited tunneling electrons also contribute to the process in the tunneling regime.

Magnetic microscopic imaging with an optically pumped magnetometer and flux guides

DOE PAGES

Kim, Young Jin; Savukov, Igor Mykhaylovich; Huang, Jen -Huang; ...

2017-01-23

Here, by combining an optically pumped magnetometer (OPM) with flux guides (FGs) and by installing a sample platform on automated translation stages, we have implemented an ultra-sensitive FG-OPM scanning magnetic imaging system that is capable of detecting magnetic fields of ~20 pT with spatial resolution better than 300 μm (expected to reach ~10 pT sensitivity and ~100 μm spatial resolution with optimized FGs). As a demonstration of one possible application of the FG-OPM device, we conducted magnetic imaging of micron-size magnetic particles. Magnetic imaging of such particles, including nano-particles and clusters, is very important for many fields, especially for medicalmore » cancer diagnostics and biophysics applications. For rapid, precise magnetic imaging, we constructed an automatic scanning system, which holds and moves a target sample containing magnetic particles at a given stand-off distance from the FG tips. We show that the device was able to produce clear microscopic magnetic images of 10 μm-size magnetic particles. In addition, we also numerically investigated how the magnetic flux from a target sample at a given stand-off distance is transmitted to the OPM vapor cell.« less

Fabrication of 2D and 3D photonic structures using laser lithography

NASA Astrophysics Data System (ADS)

Gaso, P.; Jandura, D.; Pudis, D.

2016-12-01

In this paper we demonstrate possibilities of three-dimensional (3D) printing technology based on two photon polymerization. We used three-dimensional dip-in direct-laser-writing (DLW) optical lithography to fabricate 2D and 3D optical structures for optoelectronics and for optical sensing applications. DLW lithography allows us use a non conventional way how to couple light into the waveguide structure. We prepared ring resonator and we investigated its transmission spectral characteristic. We present 3D inverse opal structure from its design to printing and scanning electron microscope (SEM) imaging. Finally, SEM images of some prepared photonic crystal structures were performed.

Optical Tweezers for Sample Fixing in Micro-Diffraction Experiments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Amenitsch, H.; Rappolt, M.; Sartori, B.

2007-01-19

In order to manipulate, characterize and measure the micro-diffraction of individual structural elements down to single phospholipid liposomes we have been using optical tweezers (OT) combined with an imaging microscope. We were able to install the OT system at the microfocus beamline ID13 at the ESRF and trap clusters of about 50 multi-lamellar liposomes (< 10 {mu}m large cluster). Further we have performed a scanning diffraction experiment with a 1 micrometer beam to demonstrate the fixing capabilities and to confirm the size of the liposome cluster by X-ray diffraction.

Experimental and theoretical analysis for improved microscope design of optical projection tomographic microscopy.

PubMed

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.

On-chip beam positioning sensor via frequency locked cascaded ring resonators

NASA Astrophysics Data System (ADS)

Naiman, Alex; Stern, Liron; Levy, Uriel

2018-05-01

We demonstrate an approach for on-chip beam positioning with a position accuracy of up to 100 nm. This approach is based on tracking the resonance of two adjacent microring resonators that are implemented on a silicon on insulator chip. We demonstrate the functionality of our approach by illuminating the chip through a Near Field Scanning Optical Microscope tip and monitoring the shift of the microring resonances due to the thermo-optic effect. We also discuss the contribution of different effects such as free carrier absorption and dispersion to the resonance shift.

Performance of a three-dimensional-printed microscanner in a laser scanning microscopy application

NASA Astrophysics Data System (ADS)

Oyman, Hilmi Artun; Gokdel, Yigit Daghan; Ferhanoglu, Onur; Yalcinkaya, Arda Deniz

2018-04-01

A magnetically actuated microscanner is used in a laser scanning microscopy application. Stress distribution along the circular-profiled flexure is compared with a rectangular counterpart in finite-element environment. Magnetic actuation mechanism of the scanning unit is explained in detail. Moreover, reliability of the scanner is tested for 3×106 cycle. The scanning device is designed to meet a confocal microscopy application providing 100 μm×100 μm field of view and <3-μm lateral resolution. The resonance frequencies of the device were analytically modeled, where we obtained 130- and 268-Hz resonance values for the out-of-plane and torsion modes, respectively. The scanning device provided an optical scan angle about 2.5 deg for 170-mA drive current, enabling the desired field of view for our custom built confocal microscope setup. Finally, imaging experiments were conducted on a resolution target, showcasing the desired scan area and resolution.

Investigation of photoconductivity of individual InAs/GaAs(001) quantum dots by Scanning Near-field Optical Microscopy

NASA Astrophysics Data System (ADS)

Filatov, D. O.; Kazantseva, I. A.; Baidus', N. V.; Gorshkov, A. P.; Mishkin, V. P.

2017-10-01

The spatial distribution of the photocurrent in the input window plane of a GaAs-based p-i-n photodiode with embedded self-assembled InAs quantum dots (QDs) has been studied with the photoexcitation through a Scanning Near-field Optical Microscope (SNOM) probe at the emission wavelength greater than the intrinsic absorption edge of the host material (GaAs). The inhomogeneities related to the interband absorption in the individual InAs/GaAs(001) QDs have been observed in the photocurrent SNOM images. Thus, the possibility of imaging the individual InAs/GaAs(001) QDs in the photocurrent SNOM images with the lateral spatial resolution ˜ 100 nm (of the same order of magnitude as the SNOM probe aperture size) has been demonstrated.

Miscibility, Crystallization, and Rheological Behavior of Solution Casting Poly(3-hydroxybutyrate)/poly(ethylene succinate) Blends Probed by Differential Scanning Calorimetry, Rheology, and Optical Microscope Techniques

NASA Astrophysics Data System (ADS)

Sun, Wei-hua; Qiao, Xiao-ping; Cao, Qi-kun; Liu, Jie-ping

2010-02-01

The miscibility and crystallization of solution casting biodegradable poly(3-hydroxybutyrate)/poly(ethylene succinate) (PHB/PES) blends was investigated by differential scanning calorimetry, rheology, and optical microscopy. The blends showed two glass transition temperatures and a depression of melting temperature of PHB with compositions in phase diagram, which indicated that the blend was partially miscible. The morphology observation supported this result. It was found that the PHB and PES can crystallize simultaneously or upon stepwise depending on the crystallization temperatures and compositions. The spherulite growth rate of PHB increased with increasing of PES content. The influence of compositions on the spherulitic growth rate for the partially miscible polymer blends was discussed.

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

PubMed

Sekine, Ryojun; Aoki, Hiroyuki; Ito, Shinzaburo

2009-10-01

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

Near-field microscopy of waveguide architectures of InGaN/GaN diode lasers

NASA Astrophysics Data System (ADS)

Friede, Sebastian; Tomm, Jens W.; Kühn, Sergei; Hoffmann, Veit; Wenzel, Hans; Weyers, Markus

2016-11-01

Waveguide (WG) architectures of 420 nm emitting InGaN/GaN diode lasers are analyzed by photoluminescence and photocurrent spectroscopy using a nearfield scanning optical microscope that scans along their front facets. The components of the ‘optical active cavity’, quantum wells, WGs, and cladding layers are individually inspected with a spatial resolution of ∼100 nm. Separate analysis of the p- and n-sections of the WG was achieved, and reveals defect levels in the p-part. Moreover, it is demonstrated that the homogeneity of the n-WG section directly affects the quantum wells that are grown on top of this layer. Substantially increased carrier capture efficiencies into InGaN/GaN-WGs compared to GaN-WGs are demonstrated.

Optical diffraction tomography with fully and partially coherent illumination in high numerical aperture label-free microscopy [Invited].

PubMed

Soto, Juan M; Rodrigo, José A; Alieva, Tatiana

2018-01-01

Quantitative label-free imaging is an important tool for the study of living microorganisms that, during the last decade, has attracted wide attention from the optical community. Optical diffraction tomography (ODT) is probably the most relevant technique for quantitative label-free 3D imaging applied in wide-field microscopy in the visible range. The ODT is usually performed using spatially coherent light illumination and specially designed holographic microscopes. Nevertheless, the ODT is also compatible with partially coherent illumination and can be realized in conventional wide-field microscopes by applying refocusing techniques, as it has been recently demonstrated. Here, we compare these two ODT modalities, underlining their pros and cons and discussing the optical setups for their implementation. In particular, we pay special attention to a system that is compatible with a conventional wide-field microscope that can be used for both ODT modalities. It consists of two easily attachable modules: the first for sample illumination engineering based on digital light processing technology; the other for focus scanning by using an electrically driven tunable lens. This hardware allows for a programmable selection of the wavelength and the illumination design, and provides fast data acquisition as well. Its performance is experimentally demonstrated in the case of ODT with partially coherent illumination providing speckle-free 3D quantitative imaging.

Gabor domain optical coherence microscopy

NASA Astrophysics Data System (ADS)

Murali, Supraja

Time domain Optical Coherence Tomography (TD-OCT), first reported in 1991, makes use of the low temporal coherence properties of a NIR broadband laser to create depth sectioning of up to 2mm under the surface using optical interferometry and point to point scanning. Prior and ongoing work in OCT in the research community has concentrated on improving axial resolution through the development of broadband sources and speed of image acquisition through new techniques such as Spectral domain OCT (SD-OCT). In SD-OCT, an entire depth scan is acquired at once with a low numerical aperture (NA) objective lens focused at a fixed point within the sample. In this imaging geometry, a longer depth of focus is achieved at the expense of lateral resolution, which is typically limited to 10 to 20 mum. Optical Coherence Microscopy (OCM), introduced in 1994, combined the advantages of high axial resolution obtained in OCT with high lateral resolution obtained by increasing the NA of the microscope placed in the sample arm. However, OCM presented trade-offs caused by the inverse quadratic relationship between the NA and the DOF of the optics used. For applications requiring high lateral resolution, such as cancer diagnostics, several solutions have been proposed including the periodic manual re-focusing of the objective lens in the time domain as well as the spectral domain C-mode configuration in order to overcome the loss in lateral resolution outside the DOF. In this research, we report for the first time, high speed, sub-cellular imaging (lateral resolution of 2 mum) in OCM using a Gabor domain image processing algorithm with a custom designed and fabricated dynamic focus microscope interfaced to a Ti:Sa femtosecond laser centered at 800 nm within an SD-OCM configuration. It is envisioned that this technology will provide a non-invasive replacement for the current practice of multiple biopsies for skin cancer diagnosis. The research reported here presents three important advances to this technology all of which have been demonstrated in full functional hardware conceived and built during the course of this research. First, it has been demonstrated that the coherence gate created by the femtosecond laser can be coupled into a scanning optical microscope using optical design methods to include liquid lens technology that enables scanning below the surface of skin with no moving parts and at high resolution throughout a 2x2x2 mm imaging cube. Second, the integration the variable-focus liquid lens technology within a fixed-optics microscope custom optical design helped increase the working NA by an order of magnitude over the limitation imposed by the liquid lens alone. Thus, this design has enabled homogenous axial and lateral resolution at the micron-level (i.e., 2 mum) while imaging in the spectral domain, and still maintaining in vivo speeds. The latest images in biological specimens clearly demonstrate sub-cellular resolution in all dimensions throughout the imaging volume. Third, this new modality for data collection has been integrated with an automated Gabor domain image registration and fusion algorithm to provide full resolution images across the data cube in real-time. We refer to this overall OCM method as Gabor domain OCM (GD-OCM). These advantages place GD-OCM in a unique position with respect to the diagnosis of cancer, because when fully developed, it promises to enable fast and accurate screening for early symptoms that could lead to prevention. The next step for this technology is to apply it directly, in a clinical environment. This step is underway and is expected to be reported by the next generation of researchers within this group.

Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy.

PubMed

Schulz, Olaf; Pieper, Christoph; Clever, Michaela; Pfaff, Janine; Ruhlandt, Aike; Kehlenbach, Ralph H; Wouters, Fred S; Großhans, Jörg; Bunt, Gertrude; Enderlein, Jörg

2013-12-24

We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions.

Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy

PubMed Central

Schulz, Olaf; Pieper, Christoph; Clever, Michaela; Pfaff, Janine; Ruhlandt, Aike; Kehlenbach, Ralph H.; Wouters, Fred S.; Großhans, Jörg; Bunt, Gertrude; Enderlein, Jörg

2013-01-01

We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions. PMID:24324140

Probing plasmons in three dimensions by combining complementary spectroscopies in a scanning transmission electron microscope

DOE PAGES

Hachtel, Jordan A.; Marvinney, Claire; Mouti, Anas; ...

2016-03-02

The nanoscale optical response of surface plasmons in three-dimensional metallic nanostructures plays an important role in many nanotechnology applications, where precise spatial and spectral characteristics of plasmonic elements control device performance. Electron energy loss spectroscopy (EELS) and cathodoluminescence (CL) within a scanning transmission electron microscope have proven to be valuable tools for studying plasmonics at the nanoscale. Each technique has been used separately, producing three-dimensional reconstructions through tomography, often aided by simulations for complete characterization. Here we demonstrate that the complementary nature of the two techniques, namely that EELS probes beam-induced electronic excitations while CL probes radiative decay, allows usmore » to directly obtain a spatially- and spectrally-resolved picture of the plasmonic characteristics of nanostructures in three dimensions. Furthermore, the approach enables nanoparticle-by-nanoparticle plasmonic analysis in three dimensions to aid in the design of diverse nanoplasmonic applications.« less

Construction and Application of a Terahertz Scanning Near-Field Microscope for Study of Correlated Electron Materials at Cryogenic Temperatures and Nanometer Length Scales

NASA Astrophysics Data System (ADS)

Stinson, Harry Theodore, III

This dissertation describes the design and construction of the world's first cryogenic apertureless near-field microscope designed for terahertz sources and detectors. I first provide motivation for the creation of this instrument in the context of spectroscopy of correlated electron materials, and background information on the two techniques that the instrument combines, scanning near-field optical microscopy and terahertz time-domain spectroscopy. I then detail key components of the instrument design, including proof-of-principle results obtained at room and cryogenic temperatures. Following this, I discuss an imaging experiment performed with this instrument on vanadium dioxide, an insulator-metal transition material, which sheds new light on the nature of the phase transition and provides support for a new model Hamiltonian for the system. Finally, I discuss a theoretical proposal for the study of cuprate superconductors using this instrument.

Handheld nonlinear microscope system comprising a 2 MHz repetition rate, mode-locked Yb-fiber laser for in vivo biomedical imaging

PubMed Central

Krolopp, Ádám; Csákányi, Attila; Haluszka, Dóra; Csáti, Dániel; Vass, Lajos; Kolonics, Attila; Wikonkál, Norbert; Szipőcs, Róbert

2016-01-01

A novel, Yb-fiber laser based, handheld 2PEF/SHG microscope imaging system is introduced. It is suitable for in vivo imaging of murine skin at an average power level as low as 5 mW at 200 kHz sampling rate. Amplified and compressed laser pulses having a spectral bandwidth of 8 to 12 nm at around 1030 nm excite the biological samples at a ~1.89 MHz repetition rate, which explains how the high quality two-photon excitation fluorescence (2PEF) and second harmonic generation (SHG) images are obtained at the average power level of a laser pointer. The scanning, imaging and detection head, which comprises a conventional microscope objective for beam focusing, has a physical length of ~180 mm owing to the custom designed imaging telescope system between the laser scanner mirrors and the entrance aperture of the microscope objective. Operation of the all-fiber, all-normal dispersion Yb-fiber ring laser oscillator is electronically controlled by a two-channel polarization controller for Q-switching free mode-locked operation. The whole nonlinear microscope imaging system has the main advantages of the low price of the fs laser applied, fiber optics flexibility, a relatively small, light-weight scanning and detection head, and a very low risk of thermal or photochemical damage of the skin samples. PMID:27699118

Handheld nonlinear microscope system comprising a 2 MHz repetition rate, mode-locked Yb-fiber laser for in vivo biomedical imaging.

PubMed

Krolopp, Ádám; Csákányi, Attila; Haluszka, Dóra; Csáti, Dániel; Vass, Lajos; Kolonics, Attila; Wikonkál, Norbert; Szipőcs, Róbert

2016-09-01

A novel, Yb-fiber laser based, handheld 2PEF/SHG microscope imaging system is introduced. It is suitable for in vivo imaging of murine skin at an average power level as low as 5 mW at 200 kHz sampling rate. Amplified and compressed laser pulses having a spectral bandwidth of 8 to 12 nm at around 1030 nm excite the biological samples at a ~1.89 MHz repetition rate, which explains how the high quality two-photon excitation fluorescence (2PEF) and second harmonic generation (SHG) images are obtained at the average power level of a laser pointer. The scanning, imaging and detection head, which comprises a conventional microscope objective for beam focusing, has a physical length of ~180 mm owing to the custom designed imaging telescope system between the laser scanner mirrors and the entrance aperture of the microscope objective. Operation of the all-fiber, all-normal dispersion Yb-fiber ring laser oscillator is electronically controlled by a two-channel polarization controller for Q-switching free mode-locked operation. The whole nonlinear microscope imaging system has the main advantages of the low price of the fs laser applied, fiber optics flexibility, a relatively small, light-weight scanning and detection head, and a very low risk of thermal or photochemical damage of the skin samples.

Intraocular Gnathostoma spinigerum. Clinicopathologic study of two cases with review of literature.

PubMed

Biswas, J; Gopal, L; Sharma, T; Badrinath, S S

1994-01-01

Live intraocular nematode is a rare occurrence that is mostly reported in Southeast Asian countries. Common nematodes that are seen live in the eye are microfilaria, Gnathostoma, and Angiostrongylus. Approximately 12 cases of intraocular gnathostomiasis have been reported in the literature. Two cases of intraocular gnathostoma, removed by vitrectomy in the first case and by paracentesis in the second case, are reported. Morphologic study of the parasites in wet preparation was performed under dissecting microscope and fixed in Karnovosky's fixative. Light microscopic and scanning electron microscopic studies were also performed. The first patient had anterior uveitis, multiple iris holes, and dense vitreous haze with fibrous proliferation over the optic disc. On resolution of the vitreous haze, a live worm was seen in the vitreous cavity. The second patient had anterior uveitis with secondary glaucoma, multiple iris holes, mild vitritis, and focal subretinal haemorrhage with subretinal tracts. Four days later a live worm was seen in the anterior chamber and removed. Microscopic study of the parasites from both patients revealed typical head bulb with four circumferential rows of hooklets, and fine cuticular spines were seen on the surface of the body. Iris holes, uveitis, and subretinal haemorrhage with subretinal tract can be characteristic features of intraocular gnathostomiasis. Identification of this parasite can be made by typical features, which can be identified on light and scanning electron microscopic study.

Scanning electron microscopic characteristics of commercially available 1- and 3-piece intraocular lenses.

PubMed

Brockmann, Tobias; Brockmann, Claudia; Nietzsche, Sandor; Bertelmann, Eckart; Strobel, Juergen; Dawczynski, Jens

2013-12-01

To evaluate commercially available 1- and 3-piece intraocular lenses (IOLs) with scanning electron microscopy (SEM). Department of Ophthalmology and Electron Microscopy Center, University Hospital Jena, Jena, Germany. Experimental study. Seven +23.0 diopter IOLs of different design and material and from different manufacturers were chosen for a detailed assessment. Scanning electron microscopy was used at standardized magnifications to assess typical IOL characteristics. The particular focus was the optic edge, the optic surface, the haptic–optic junction, and the haptic. All square-edged IOLs had a curvature radius of less than 10 μm, while the mean optic edge thickness ranged between 216 μm and 382 μm. A 360-degree square-edged boundary was present in all 3-piece IOLs and in a single 1-piece model. Relevant production remnants on the optic edge were observed in 1 case. Regarding the haptic, 3-piece IOLs had uniformly shaped fibers with a mean thickness of 177 μm ± 51 (SD) (range 116 to 220 μm). Chemical adhesives were used to attach the haptic in 1 case, where alterations of the IOL material were observed. In another case, the haptic fiber was press-fitted into the optic, which resulted in bulging of the optic profile. Inspection of surface characteristics showed wavelike patterns in 2 IOLs. Taking clinical relevance into account, all IOLs were of high manufacturing quality. Certain attention was paid in creating a sharp optic edge. Surface irregularities of 2 IOLs were attributed to the manufacturing technique. Methods for implementing the haptic–optic junction were diverse.

Video-Rate Confocal Microscopy for Single-Molecule Imaging in Live Cells and Superresolution Fluorescence Imaging

PubMed Central

Lee, Jinwoo; Miyanaga, Yukihiro; Ueda, Masahiro; Hohng, Sungchul

2012-01-01

There is no confocal microscope optimized for single-molecule imaging in live cells and superresolution fluorescence imaging. By combining the swiftness of the line-scanning method and the high sensitivity of wide-field detection, we have developed a, to our knowledge, novel confocal fluorescence microscope with a good optical-sectioning capability (1.0 μm), fast frame rates (<33 fps), and superior fluorescence detection efficiency. Full compatibility of the microscope with conventional cell-imaging techniques allowed us to do single-molecule imaging with a great ease at arbitrary depths of live cells. With the new microscope, we monitored diffusion motion of fluorescently labeled cAMP receptors of Dictyostelium discoideum at both the basal and apical surfaces and obtained superresolution fluorescence images of microtubules of COS-7 cells at depths in the range 0–85 μm from the surface of a coverglass. PMID:23083712

An integrated single- and two-photon non-diffracting light-sheet microscope

NASA Astrophysics Data System (ADS)

Lau, Sze Cheung; Chiu, Hoi Chun; Zhao, Luwei; Zhao, Teng; Loy, M. M. T.; Du, Shengwang

2018-04-01

We describe a fluorescence optical microscope with both single-photon and two-photon non-diffracting light-sheet excitations for large volume imaging. With a special design to accommodate two different wavelength ranges (visible: 400-700 nm and near infrared: 800-1200 nm), we combine the line-Bessel sheet (LBS, for single-photon excitation) and the scanning Bessel beam (SBB, for two-photon excitation) light sheet together in a single microscope setup. For a transparent thin sample where the scattering can be ignored, the LBS single-photon excitation is the optimal imaging solution. When the light scattering becomes significant for a deep-cell or deep-tissue imaging, we use SBB light-sheet two-photon excitation with a longer wavelength. We achieved nearly identical lateral/axial resolution of about 350/270 nm for both imagings. This integrated light-sheet microscope may have a wide application for live-cell and live-tissue three-dimensional high-speed imaging.

Enhanced Thermo-Optical Switching of Paraffin-Wax Composite Spots under Laser Heating

PubMed Central

Said, Asmaa; Salah, Abeer; Abdel Fattah, Gamal

2017-01-01

Thermo-optical switches are of particular significance in communications networks where increasingly high switching speeds are required. Phase change materials (PCMs), in particular those based on paraffin wax, provide wealth of exciting applications with unusual thermally-induced switching properties, only limited by paraffin’s rather low thermal conductivity. In this paper, the use of different carbon fillers as thermal conductivity enhancers for paraffin has been investigated, and a novel structure based on spot of paraffin wax as a thermo-optic switch is presented. Thermo-optical switching parameters are enhanced with the addition of graphite and graphene, due to the extreme thermal conductivity of the carbon fillers. Differential Scanning Calorimetry (DSC) and Scanning electron microscope (SEM) are performed on paraffin wax composites, and specific heat capacities are calculated based on DSC measurements. Thermo-optical switching based on transmission is measured as a function of the host concentration under conventional electric heating and laser heating of paraffin-carbon fillers composites. Further enhancements in thermo-optical switching parameters are studied under Nd:YAG laser heating. This novel structure can be used in future networks with huge bandwidth requirements and electric noise free remote aerial laser switching applications. PMID:28772884

Enhanced Thermo-Optical Switching of Paraffin-Wax Composite Spots under Laser Heating.

PubMed

Said, Asmaa; Salah, Abeer; Fattah, Gamal Abdel

2017-05-12

Thermo-optical switches are of particular significance in communications networks where increasingly high switching speeds are required. Phase change materials (PCMs), in particular those based on paraffin wax, provide wealth of exciting applications with unusual thermally-induced switching properties, only limited by paraffin's rather low thermal conductivity. In this paper, the use of different carbon fillers as thermal conductivity enhancers for paraffin has been investigated, and a novel structure based on spot of paraffin wax as a thermo-optic switch is presented. Thermo-optical switching parameters are enhanced with the addition of graphite and graphene, due to the extreme thermal conductivity of the carbon fillers. Differential Scanning Calorimetry (DSC) and Scanning electron microscope (SEM) are performed on paraffin wax composites, and specific heat capacities are calculated based on DSC measurements. Thermo-optical switching based on transmission is measured as a function of the host concentration under conventional electric heating and laser heating of paraffin-carbon fillers composites. Further enhancements in thermo-optical switching parameters are studied under Nd:YAG laser heating. This novel structure can be used in future networks with huge bandwidth requirements and electric noise free remote aerial laser switching applications.

Comparative study of image contrast in scanning electron microscope and helium ion microscope.

PubMed

O'Connell, R; Chen, Y; Zhang, H; Zhou, Y; Fox, D; Maguire, P; Wang, J J; Rodenburg, C

2017-12-01

Images of Ga + -implanted amorphous silicon layers in a 110 n-type silicon substrate have been collected by a range of detectors in a scanning electron microscope and a helium ion microscope. The effects of the implantation dose and imaging parameters (beam energy, dwell time, etc.) on the image contrast were investigated. We demonstrate a similar relationship for both the helium ion microscope Everhart-Thornley and scanning electron microscope Inlens detectors between the contrast of the images and the Ga + density and imaging parameters. These results also show that dynamic charging effects have a significant impact on the quantification of the helium ion microscope and scanning electron microscope contrast. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Metal-filled carbon nanotube based optical nanoantennas: bubbling, reshaping, and in situ characterization.

PubMed

Fan, Zheng; Tao, Xinyong; Cui, Xudong; Fan, Xudong; Zhang, Xiaobin; Dong, Lixin

2012-09-21

Controlled fabrication of metal nanospheres on nanotube tips for optical antennas is investigated experimentally. Resembling soap bubble blowing using a straw, the fabrication process is based on nanofluidic mass delivery at the attogram scale using metal-filled carbon nanotubes (m@CNTs). Two methods have been investigated including electron-beam-induced bubbling (EBIB) and electromigration-based bubbling (EMBB). EBIB involves the bombardment of an m@CNT with a high energy electron beam of a transmission electron microscope (TEM), with which the encapsulated metal is melted and flowed out from the nanotube, generating a metallic particle on a nanotube tip. In the case where the encapsulated materials inside the CNT have a higher melting point than what the beam energy can reach, EMBB is an optional process to apply. Experiments show that, under a low bias (2.0-2.5 V), nanoparticles can be formed on the nanotube tips. The final shape and crystallinity of the nanoparticles are determined by the cooling rate. Instant cooling occurs with a relatively large heat sink and causes the instant shaping of the solid deposit, which is typically similar to the shape of the molten state. With a smaller heat sink as a probe, it is possible to keep the deposit in a molten state. Instant cooling by separating the deposit from the probe can result in a perfect sphere. Surface and volume plasmons characterized with electron energy loss spectroscopy (EELS) prove that resonance occurs between a pair of as-fabricated spheres on the tip structures. Such spheres on pillars can serve as nano-optical antennas and will enable devices such as scanning near-field optical microscope (SNOM) probes, scanning anodes for field emitters, and single molecule detectors, which can find applications in bio-sensing, molecular detection, and high-resolution optical microscopy.

Characterization of a Hybrid Optical Microscopy/Laser Ablation Liquid Vortex Capture/Electrospray Ionization System for Mass Spectrometry Imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cahill, John F.; Kertesz, Vilmos; Van Berkel, Gary J.

Herein, a commercial optical microscope, laser microdissection instrument was coupled with an electrospray ionization mass spectrometer via a low profile liquid vortex capture probe to yield a hybrid optical microscopy/mass spectrometry imaging system. The instrument has bright-field and fluorescence microscopy capabilities in addition to a highly focused UV laser beam that is utilized for laser ablation of samples. With this system, material laser ablated from a sample using the microscope was caught by a liquid vortex capture probe and transported in solution for analysis by electrospray ionization mass spectrometry. Both lane scanning and spot sampling mass spectral imaging modes weremore » used. The smallest area the system was able to ablate was ~0.544 μm × ~0.544 μm, achieved by oversampling of the smallest laser ablation spot size that could be obtained (~1.9 μm). With use of a model photoresist surface, known features as small as ~1.5 μm were resolved. The capabilities of the system with real world samples were demonstrated first with a blended polymer thin film containing poly(2-vinylpyridine) and poly(N-vinylcarbazole). Using spot sampling imaging, sub-micrometer sized features (0.62, 0.86, and 0.98 μm) visible by optical microscopy were clearly distinguished in the mass spectral images. A second real world example showed the imaging of trace amounts of cocaine in mouse brain thin tissue sections. Lastly, with use of a lane scanning mode with ~6 μm × ~6 μm data pixels, features in the tissue as small as 15 μm in size could be distinguished in both the mass spectral and optical images.« less

Characterization of a Hybrid Optical Microscopy/Laser Ablation Liquid Vortex Capture/Electrospray Ionization System for Mass Spectrometry Imaging

DOE PAGES

Cahill, John F.; Kertesz, Vilmos; Van Berkel, Gary J.

2015-10-22

Herein, a commercial optical microscope, laser microdissection instrument was coupled with an electrospray ionization mass spectrometer via a low profile liquid vortex capture probe to yield a hybrid optical microscopy/mass spectrometry imaging system. The instrument has bright-field and fluorescence microscopy capabilities in addition to a highly focused UV laser beam that is utilized for laser ablation of samples. With this system, material laser ablated from a sample using the microscope was caught by a liquid vortex capture probe and transported in solution for analysis by electrospray ionization mass spectrometry. Both lane scanning and spot sampling mass spectral imaging modes weremore » used. The smallest area the system was able to ablate was ~0.544 μm × ~0.544 μm, achieved by oversampling of the smallest laser ablation spot size that could be obtained (~1.9 μm). With use of a model photoresist surface, known features as small as ~1.5 μm were resolved. The capabilities of the system with real world samples were demonstrated first with a blended polymer thin film containing poly(2-vinylpyridine) and poly(N-vinylcarbazole). Using spot sampling imaging, sub-micrometer sized features (0.62, 0.86, and 0.98 μm) visible by optical microscopy were clearly distinguished in the mass spectral images. A second real world example showed the imaging of trace amounts of cocaine in mouse brain thin tissue sections. Lastly, with use of a lane scanning mode with ~6 μm × ~6 μm data pixels, features in the tissue as small as 15 μm in size could be distinguished in both the mass spectral and optical images.« less

A handheld MEMS-based line-scanned dual-axis confocal microscope for early cancer detection and surgical guidance (Conference Presentation)

NASA Astrophysics Data System (ADS)

Chen, Ye; Yin, Chengbo; Wei, Linpeng; Glaser, Adam K.; Abeytunge, Sanjee; Peterson, Gary; Mandella, Michael J.; Sanai, Nader; Rajadhyaksha, Milind; Liu, Jonathan T.

2017-02-01

Considerable efforts have been recently undertaken to develop miniature optical-sectioning microscopes for in vivo microendoscopy and point-of-care pathology. These devices enable in vivo interrogation of disease as a real-time and noninvasive alternative to gold-standard histopathology, and therefore could have a transformative impact for the early detection of cancer as well as for guiding tumor-resection procedures. Regardless of the specific modality, various trade-offs in size, speed, field of view, resolution, contrast, and sensitivity are necessary to optimize a device for a particular application. Here, a miniature MEMS-based line-scanned dual-axis confocal (LS-DAC) microscope, with a 12-mm diameter distal tip, has been developed for point-of-care pathology. The dual-axis architecture has demonstrated superior rejection of out-of-focus and multiply scattered photons compared to a conventional single-axis confocal configuration. The use of line scanning enables fast frame rates (≥15 frames/sec), which mitigates motion artifacts of a handheld device during clinical use. We have developed a method to actively align the illumination and collection beams in this miniature LS-DAC microscope through the use of a pair of rotatable alignment mirrors. Incorporation of a custom objective lens, with a small form factor for in vivo application, enables the device to achieve an axial and lateral resolution of 2.0 and 1.1 microns, respectively. Validation measurements with reflective targets, as well as in vivo and ex vivo images of tissues, demonstrate that this high-speed LS-DAC microscope can achieve high-contrast imaging of fluorescently labeled tissues with sufficient sensitivity for applications such as oral cancer detection and guiding brain-tumor resections.

Nanoscale Mineralogy and Composition of Experimental Regolith Agglutinates Produced under Asteroidal Impact Conditions

NASA Technical Reports Server (NTRS)

Christoffersen, Roy; Cintala, M. J.; Keller, L. P.; See, T. H.; Horz, F.

2013-01-01

On the Moon, the energetics of smaller impactors and the physical/chemical characteristics of the granular regolith target combine to form a key product of lunar space weathering: chemically reduced shock melts containing optically-active nanophase Fe metal grains (npFe0) [1]. In addition to forming the optically dark glassy matrix phase in lunar agglutinitic soil particles [1], these shock melts are becoming increasingly recognized for their contribution to optically active patina coatings on a wide range of exposed rock and grain surfaces in the lunar regolith [2]. In applying the lessons of lunar space weathering to asteroids, the potential similarities and differences in regolith-hosted shock melts on the Moon compared to those on asteroids has become a topic of increasing interest [3,4]. In a series of impact experiments performed at velocities applicable to the asteroid belt [5], Horz et al. [6] and See and Horz [7] have previously shown that repeated impacts into a gabbroic regolith analog target can produce melt-welded grain aggregates morphologically very similar to lunar agglutinates [6,7]. Although these agglutinate-like particles were extensively analyzed by electron microprobe and scanning electron microscopy (SEM) as part of the original study [7], a microstructural and compositional comparison of these aggregates to lunar soil agglutinates at sub-micron scales has yet to be made. To close this gap, we characterized a representative set of these aggregates using a JEOL 7600 field-emission scanning electron microscope (FE-SEM), and JEOL 2500SE field-emission scanning transmission electron microscope (FE-STEM) both optimized for energy dispersive X-ray spectroscopy (EDX) compositional spectrum imaging at respective analytical spatial resolutions of 0.5 to 1 micron, and 2 to 4 nm.

Effect of 60Co γ-irradiation on structural and optical properties of thin films of Ga10Se80Hg10

NASA Astrophysics Data System (ADS)

Ahmad, Shabir; Asokan, K.; Shahid Khan, Mohd.; Zulfequar, M.

2015-08-01

Thin films of Ga10Se80Hg10 have been deposited onto a chemically cleaned Al2O3 substrates by thermal evaporation technique under vacuum. The investigated thin films are irradiated by 60Co γ-rays in the dose range of 50-150 kGy. X-ray diffraction patterns of the investigated thin films confirm the preferred crystallite growth occurs in the tetragonal phase structure. It also shows, the average crystallite size increases after γ-exposure, which indicates the crystallinity of the material increases after γ-irradiation. These results were further supported by surface morphological analysis carried out by scanning electron microscope and atomic force microscope which also shows the crystallinity of the material increases with increasing the γ-irradiation dose. The optical transmission spectra of the thin films at normal incidence were investigated in the spectral range from 190 to 1100 nm. Using the transmission spectra, the optical constants like refractive index (n) and extinction coefficient (k) were calculated based on Swanepoel's method. The optical band gap (Eg) was also estimated using Tauc's extrapolation procedure. The optical analysis shows: the value of optical band gap of investigated thin films decreases and the corresponding absorption coefficient increases continuously with increasing dose of γ-irradiation.

The microscopic (optical and SEM) examination of dental calculus deposits (DCD). Potential interest in forensic anthropology of a bio-archaeological method.

PubMed

Charlier, Philippe; Huynh-Charlier, Isabelle; Munoz, Olivia; Billard, Michel; Brun, Luc; de la Grandmaison, Geoffroy Lorin

2010-07-01

This article describes the potential interest in forensic anthropology of the microscopic analysis of dental calculus deposits (DCD), a calcified residue frequently found on the surface of teeth. Its sampling and analysis seem straightforward and relatively reproducible. Samples came from archaeological material (KHB-1 Ra's al-Khabbah and RH-5 Ra's al-Hamra, two Prehistoric graveyards located in the Sultanate of Oman, dated between the 5th and 4th millennium B.C.; Montenzio Vecchia, an Etruscan-Celtic necropolis from the north of Italy, dated between the 5th and 3rd century B.C.; body rests of Agnès Sorel, French royal mistress died in 1450 A.D.; skeleton of Pierre Hazard, French royal notary from the 15th century A.D.). Samples were studies by direct optical microscope (OM) or scanning electron microscopy (SEM). Many cytological, histological and elemental analyses were possible, producing precious data for the identification of these remains, the reconstitution of their alimentation and occupational habits, and propositions for manner of death. Copyright 2010 Elsevier Ireland Ltd. All rights reserved.

In vivo study of rat cortical hemodynamics using a stereotaxic-apparatus-compatible photoacoustic microscope.

PubMed

Guo, Heng; Chen, Qian; Qi, Weizhi; Chen, Xingxing; Xi, Lei

2018-04-19

Brain imaging is an important technique in cognitive neuroscience. In this article, we designed a stereotaxic-apparatus-compatible photoacoustic microscope for the studies of rat cortical hemodynamics. Compared with existing optical resolution photoacoustic microscopy (ORPAM) systems, the probe owns feature of fast, light and miniature. In this microscope, we integrated a miniaturized ultrasound transducer with a center frequency of 10 MHz to detect photoacoustic signals and a 2-dimensional (2D) microelectromechanical system (MEMS) scanner to achieve raster scanning of the optical focus. Based on phantom evaluation, this imaging probe has a high lateral resolution of 3.8 μm and an effective imaging domain of 2 × 2 mm 2 . Different from conventional ORPAMs, combining with standard stereotaxic apparatus enables broad studies of rodent brains without any motion artifact. To show its capability, we successfully captured red blood cell flow in the capillary, monitored the vascular changes during bleeding and blood infusion and visualized cortical hemodynamics induced by middle cerebral artery occlusion. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

An estimate of biofilm properties using an acoustic microscope.

PubMed

Good, Morris S; Wend, Christopher F; Bond, Leonard J; Mclean, Jeffrey S; Panetta, Paul D; Ahmed, Salahuddin; Crawford, Susan L; Daly, Don S

2006-09-01

Noninvasive measurements over a biofilm, a three-dimensional (3-D) community of microorganisms immobilized at a substratum, were made using an acoustic microscope operating at frequencies up to 70 MHz. The microscope scanned a 2.5-mm by 2.5-mm region of a living biofilm having a nominal thickness of 100 microm. Spatial variation of surface heterogeneity, thickness, interior structure, and biomass were estimated. Thickness was estimated as the product of the speed of sound of the medium and the interim between the highest signal peak and that of the substratum plane without biofilm. The thickest portions of biofilm were 145 microm; however, slender structures attributed as streamers extended above, with one obtaining a 274-microm height above the substratum. Three-dimensional iso-contours of amplitude were used to estimate the internal structure of the biofilm. Backscatter amplitude was examined at five zones of increasing height from the substratum to examine biomass distribution. Ultrasound-based estimates of thickness were corroborated with optical microscopy. The experimental acoustic and optical systems, methods used to estimate biofilm properties, and potential applications for the resulting data are discussed.

A Student-Built Scanning Tunneling Microscope

ERIC Educational Resources Information Center

Ekkens, Tom

2015-01-01

Many introductory and nanotechnology textbooks discuss the operation of various microscopes including atomic force (AFM), scanning tunneling (STM), and scanning electron microscopes (SEM). In a nanotechnology laboratory class, students frequently utilize microscopes to obtain data without a thought about the detailed operation of the tool itself.…

Microscopic observations of self-healing products in calcareous fly ash mortars.

PubMed

Jóźwiak-Niedźwiedzka, Daria

2015-01-01

The results of microstructural characterization of mortars containing fly ash class C (High Calcium Fly Ash) from combustion of lignite are presented. The evaluation of the microstructure was performed using scanning electron microscope, optical, and confocal microscope. The tested beams were bent till the crack and microcracks opening, which were healed during the different curing time. The results showed that the replacement of cement with fly ash class C influenced the process of crack healing. The addition of HCFA, at both 30% and 60%, speeds up the self-healing process in cracks and particularly in micro-cracks. In the research, the completely filling up of the cracks by new phases has not been observed, only the beginning of such process has been noticed. © 2014 Wiley Periodicals, Inc.

Controlled waveguide coupling for photon emission from colloidal PbS quantum dot using tunable microcavity made of optical polymer and silicon

NASA Astrophysics Data System (ADS)

Nozaka, Takahiro; Mukai, Kohki

2016-04-01

A tunable microcavity device composed of optical polymer and Si with a colloidal quantum dot (QD) is proposed as a single-photon source for planar optical circuit. Cavity size is controlled by electrostatic micromachine behavior with the air bridge structure to tune timing of photon injection into optical waveguide from QD. Three-dimensional positioning of a QD in the cavity structure is available using a nanohole on Si processed by scanning probe microscope lithography. We fabricated the prototype microcavity with PbS-QD-mixed polymenthyl methacrylate on a SOI (semiconductor-on-insulator) substrate to show the tunability of cavity size as the shift of emission peak wavelength of QD ensemble.

Effect of pulse temporal shape on optical trapping and impulse transfer using ultrashort pulsed lasers.

PubMed

Shane, Janelle C; Mazilu, Michael; Lee, Woei Ming; Dholakia, Kishan

2010-03-29

We investigate the effects of pulse duration on optical trapping with high repetition rate ultrashort pulsed lasers, through Lorentz-Mie theory, numerical simulation, and experiment. Optical trapping experiments use a 12 femtosecond duration infrared pulsed laser, with the trapping microscope's temporal dispersive effects measured and corrected using the Multiphoton Intrapulse Interference Phase Scan method. We apply pulse shaping to reproducibly stretch pulse duration by 1.5 orders of magnitude and find no material-independent effects of pulse temporal profile on optical trapping of 780nm silica particles, in agreement with our theory and simulation. Using pulse shaping, we control two-photon fluorescence in trapped fluorescent particles, opening the door to other coherent control applications with trapped particles.

Influence of Ultrasonic Surface Rolling on Microstructure and Wear Behavior of Selective Laser Melted Ti-6Al-4V Alloy

PubMed Central

Wang, Zhen; Xiao, Zhiyu; Huang, Chuanshou; Wen, Liping; Zhang, Weiwen

2017-01-01

The present article studied the effect of ultrasonic surface rolling process (USRP) on the microstructure and wear behavior of a selective laser melted Ti-6Al-4V alloy. Surface characteristics were investigated using optical microscope, nano-indentation, scanning electron microscope, transmission electron microscope and laser scanning confocal microscope. Results indicated that the thickness of pore-free surfaces increased to 100~200 μm with the increasing ultrasonic surface rolling numbers. Severe work hardening occurred in the densified layer, resulting in the formation of refined grains, dislocation walls and deformation twins. After 1000 N 6 passes, about 15.5% and 14.1% increment in surficial Nano-hardness and Vickers-hardness was obtained, respectively. The hardness decreased gradually from the top surface to the substrate. Wear tests revealed that the friction coefficient declined from 0.74 (polished surface) to 0.64 (USRP treated surface) and the wear volume reduced from 0.205 mm−3 to 0.195 mm−3. The difference in wear volume between USRP treated and polished samples increased with sliding time. The enhanced wear resistance was concluded to be associated with the improvement of hardness and shear resistance and also the inhibition of delamination initiation. PMID:29048344

Brush Plating of Nickel-Tungsten Alloy for Engineering Application

DTIC Science & Technology

2012-08-01

ASETS Defense ‘12 1 Brush Plating of Nickel-Tungsten Alloy for Engineering Application Zhimin Zhong & Sid Clouser Report Documentation Page Form...COVERED 00-00-2012 to 00-00-2012 4. TITLE AND SUBTITLE Brush Plating of Nickel-Tungsten Alloy for Engineering Application 5a. CONTRACT NUMBER...6 Surface morphology Visual appearance, scanning electron and optical microscope images. Smooth, fine grained, micro- cracked surface morphology

Nanoroughened plasmonic films for enhanced biosensing detection

NASA Astrophysics Data System (ADS)

LeMoal, Eric; Lévêque-Fort, Sandrine; Potier, Marie-Claude; Fort, Emmanuel

2009-06-01

Although fluorescence is the prevailing labeling technique in biosensing applications, sensitivity improvement is still a striving challenge. We show that coating standard microscope slides with nanoroughened silver films provides a high fluorescence signal enhancement due to plasmonic interactions. As a proof of concept, we applied these films with tailored plasmonic properties to DNA microarrays. Using common optical scanning devices, we achieved signal amplifications of more than 40-fold.

Easy performance of 6-color confocal immunofluorescence with 4-laser line microscopes.

PubMed

Eissing, Nathalie; Heger, Lukas; Baranska, Anna; Cesnjevar, Robert; Büttner-Herold, Maike; Söder, Stephan; Hartmann, Arndt; Heidkamp, Gordon F; Dudziak, Diana

2014-09-01

Confocal laser scanning microscopy is an advanced technique for imaging tissue samples in vitro and in vivo at high optical resolution. The development of new fluorochrome variants do not only make it possible to perform multicolor flow cytometry of single cells, but in combination with high resolution laser scanning systems also to investigate the distribution of cells in lymphoid tissues by confocal immunofluorescence analyses, thus allowing the distinction of various cell populations directly in the tissue. Here, we provide a protocol for the visualization of at least six differently fluorochrome-labeled antibodies at the same time using a conventional confocal laser scanning microscope with four laser lines (405 nm, 488 nm, 555 nm, and 639 nm laser wavelength) in both murine and human tissue samples. We further demonstrate that compensation correction algorithms are not necessary to reduce spillover of fluorochromes into other channels when the used fluorochromes are combined according to their specific emission bands and the varying Stokes shift for co-excited fluorochromes with the same laser line. Copyright © 2014 Elsevier B.V. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sharma, Rabindar K., E-mail: rkrksharma6@gmail.com; Reddy, G. B., E-mail: rkrksharma6@gmail.com

In this report, we synthesize vertically aligned molybdenum trioxide (α−MoO{sub 3}) nanoflakes (NFs) with high aspect ratio (height/thickness >15) on the cobalt coated glass substrates by the plasma assisted sublimation process, employing Mo metal strip as a sublimation source. The effect of substrate temperature, nature of substrate as well as the geometry of the sublimation source (Mo-strip) have been investigated on the morphological, structural and optical properties of the grown NFs, keeping plasma parameters as fixed. The surface morphology, crystalline structure and optical properties of MoO{sub 3} NFs have been studied systematically by using scanning electron microscope (SEM), transmission electronmore » microscope (TEM) with selected area electron diffraction (SAED), X-ray diffractometer, and IR- spectroscopy. The experimental observations endorse that the characteristics of MoO{sub 3} NFs are strongly depend on substrate temperature, substrate nature as well as geometry of Mo-strip. All the observed results are well in consonance with each other.« less

In vitro assessments on bacterial adhesion and corrosion performance of TiN coating on Ti6Al4V titanium alloy synthesized by multi-arc ion plating

NASA Astrophysics Data System (ADS)

Lin, Naiming; Huang, Xiaobo; Zhang, Xiangyu; Fan, Ailan; Qin, Lin; Tang, Bin

2012-07-01

TiN coating was synthesized on Ti6Al4V titanium alloy surface by multi-arc ion plating (MIP) technique. Surface morphology, cross sectional microstructure, elemental distributions and phase compositions of the obtained coating were analyzed by means of scanning electron microscope (SEM), optical microscope (OM), glow discharge optical emission spectroscope (GDOES) and X-ray diffraction (XRD). Bacterial adhesion and corrosion performance of Ti6Al4V and the TiN coating were assessed via in vitro bacterial adhesion tests and corrosion experiments, respectively. The results indicated that continuous and compact coating which was built up by pure TiN with a typical columnar crystal structure has reached a thickness of 1.5 μm. This TiN coating could significantly reduce the bacterial adhesion and enhance the corrosion resistance of Ti6Al4V substrate.

Tapered fiber coupling of single photons emitted by a deterministically positioned single nitrogen vacancy center

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liebermeister, Lars, E-mail: lars.liebermeister@physik.uni-muenchen.de; Petersen, Fabian; Münchow, Asmus v.

2014-01-20

A diamond nano-crystal hosting a single nitrogen vacancy (NV) center is optically selected with a confocal scanning microscope and positioned deterministically onto the subwavelength-diameter waist of a tapered optical fiber (TOF) with the help of an atomic force microscope. Based on this nano-manipulation technique, we experimentally demonstrate the evanescent coupling of single fluorescence photons emitted by a single NV-center to the guided mode of the TOF. By comparing photon count rates of the fiber-guided and the free-space modes and with the help of numerical finite-difference time domain simulations, we determine a lower and upper bound for the coupling efficiency ofmore » (9.5 ± 0.6)% and (10.4 ± 0.7)%, respectively. Our results are a promising starting point for future integration of single photon sources into photonic quantum networks and applications in quantum information science.« less

Fast widefield techniques for fluorescence and phase endomicroscopy

NASA Astrophysics Data System (ADS)

Ford, Tim N.

Endomicroscopy is a recent development in biomedical optics which gives researchers and physicians microscope-resolution views of intact tissue to complement macroscopic visualization during endoscopy screening. This thesis presents HiLo endomicroscopy and oblique back-illumination endomicroscopy, fast wide-field imaging techniques with fluorescence and phase contrast, respectively. Fluorescence imaging in thick tissue is often hampered by strong out-of-focus background signal. Laser scanning confocal endomicroscopy has been developed for optically-sectioned imaging free from background, but reliance on mechanical scanning fundamentally limits the frame rate and represents significant complexity and expense. HiLo is a fast, simple, widefield fluorescence imaging technique which rejects out-of-focus background signal without the need for scanning. It works by acquiring two images of the sample under uniform and structured illumination and synthesizing an optically sectioned result with real-time image processing. Oblique back-illumination microscopy (OBM) is a label-free technique which allows, for the first time, phase gradient imaging of sub-surface morphology in thick scattering tissue with a reflection geometry. OBM works by back-illuminating the sample with the oblique diffuse reflectance from light delivered via off-axis optical fibers. The use of two diametrically opposed illumination fibers allows simultaneous and independent measurement of phase gradients and absorption contrast. Video-rate single-exposure operation using wavelength multiplexing is demonstrated.

Living Matter Observations with a Novel Hyperspectral Supercontinuum Confocal Microscope for VIS to Near-IR Reflectance Spectroscopy

PubMed Central

Bertani, Francesca R.; Ferrari, Luisa; Mussi, Valentina; Botti, Elisabetta; Costanzo, Antonio; Selci, Stefano

2013-01-01

A broad range hyper-spectroscopic microscope fed by a supercontinuum laser source and equipped with an almost achromatic optical layout is illustrated with detailed explanations of the design, implementation and data. The real novelty of this instrument, a confocal spectroscopic microscope capable of recording high resolution reflectance data in the VIS-IR spectral range from about 500 nm to 2.5 μm wavelengths, is the possibility of acquiring spectral data at every physical point as defined by lateral coordinates, X and Y, as well as at a depth coordinate, Z, as obtained by the confocal optical sectioning advantage. With this apparatus we collect each single scanning point as a whole spectrum by combining two linear spectral detector arrays, one CCD for the visible range, and one InGaAs infrared array, simultaneously available at the sensor output channel of the home made instrument. This microscope has been developed for biomedical analysis of human skin and other similar applications. Results are shown illustrating the technical performances of the instrument and the capability in extracting information about the composition and the structure of different parts or compartments in biological samples as well as in solid statematter. A complete spectroscopic fingerprinting of samples at microscopic level is shown possible by using statistical analysis on raw data or analytical reflectance models based on Abelés matrix transfer methods. PMID:24233077

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kang, Hyeonggon; Attota, Ravikiran, E-mail: ravikiran.attota@nist.gov; Tondare, Vipin

We present a method that uses conventional optical microscopes to determine the number of nanoparticles in a cluster, which is typically not possible using traditional image-based optical methods due to the diffraction limit. The method, called through-focus scanning optical microscopy (TSOM), uses a series of optical images taken at varying focus levels to achieve this. The optical images cannot directly resolve the individual nanoparticles, but contain information related to the number of particles. The TSOM method makes use of this information to determine the number of nanoparticles in a cluster. Initial good agreement between the simulations and the measurements ismore » also presented. The TSOM method can be applied to fluorescent and non-fluorescent as well as metallic and non-metallic nano-scale materials, including soft materials, making it attractive for tag-less, high-speed, optical analysis of nanoparticles down to 45 nm diameter.« less

Microstructural study of the nickel-base alloy WAZ-20 using qualitative and quantitative electron optical techniques

NASA Technical Reports Server (NTRS)

Young, S. G.

1973-01-01

The NASA nickel-base alloy WAZ-20 was analyzed by advanced metallographic techniques to qualitatively and quantitatively characterize its phases and stability. The as-cast alloy contained primary gamma-prime, a coarse gamma-gamma prime eutectic, a gamma-fine gamma prime matrix, and MC carbides. A specimen aged at 870 C for 1000 hours contained these same constituents and a few widely scattered high W particles. No detrimental phases (such as sigma or mu) were observed. Scanning electron microscope, light metallography, and replica electron microscope methods are compared. The value of quantitative electron microprobe techniques such as spot and area analysis is demonstrated.

Refractive index profiles of Ge-doped optical fibers with nanometer spatial resolution using atomic force microscopy.

PubMed

Pace, P; Huntington, Shane; Lyytikäinen, K; Roberts, A; Love, J

2004-04-05

We show a quantitative connection between Refractive Index Profiles (RIP) and measurements made by an Atomic Force Microscope (AFM). Germanium doped fibers were chemically etched in hydrofluoric acid solution (HF) and the wet etching characteristics of germanium were studied using an AFM. The AFM profiles were compared to both a concentration profile of the preform determined using a Scanning Electron Microscope (SEM) and a RIP of the fiber measured using a commercial profiling instrument, and were found to be in excellent agreement. It is now possible to calculate the RIP of a germanium doped fiber directly from an AFM profile.

A Novel Preparation Method of SiC Reinforced Aluminum Composite Through Vertical Rotatory Furnace

NASA Astrophysics Data System (ADS)

Nassar, Amal Ebrahim; Nassar, Eman Ebrahim; Younis, Mona Ahmed

2018-04-01

The aluminum composite was prepared successfully by stirring using internal blade installed inside a vertical rotatory furnace. Pure aluminum was used as matrix and silicon carbide particles with 10 weight percentage as reinforcement. To evaluate the efficiency of the suggested stirrer, the microstructure of the samples was analyzed using scanning electron microscope, image analyzer software available with optical microscope and energy dispersive X-ray spectroscopy analysis. Furthermore, mechanical properties were studied by measuring ultimate and yield strength, wear resistance, hardness and porosity. It was found that the particle distribution was enhanced and consequently improved the mechanical properties of the composite.

Beyond experimental noise: Analyzing single-molecule data of heterogeneous systems. Comment on "Extracting physics of life at the molecular level: A review of single-molecule data analyses" by W. Colomb and S.K. Sarkar

NASA Astrophysics Data System (ADS)

Meroz, Yasmine

2015-06-01

In the 1980s the world witnessed the advent of single-molecule experiments. The first atomic resolution characterization of a surface was reported by scanning tunneling microscope (STM) in 1982 [1], followed by atomic force microscope (AFM) in 1986 [2]. The first optical detection and spectroscopy of a single molecule in a solid took place in 1989 [3,4], in a time where essentially all chemical experiments were made on bulk, i.e. averaging over millions of copies of the same molecule.

A new scanning system for alpha decay events as calibration sources for range-energy relation in nuclear emulsion

NASA Astrophysics Data System (ADS)

Yoshida, J.; Kinbara, S.; Mishina, A.; Nakazawa, K.; Soe, M. K.; Theint, A. M. M.; Tint, K. T.

2017-03-01

A new scanning system named "Vertex picker" has been developed to rapid collect alpha decay events, which are calibration sources for the range-energy relation in nuclear emulsion. A computer-controlled optical microscope scans emulsion layers exhaustively, and a high-speed and high-resolution camera takes their micrographs. A dedicated image processing picks out vertex-like shapes. Practical operations of alpha decay search were demonstrated by emulsion sheets of the KEK-PS E373 experiment. Alpha decays of nearly 28 events were detected in eye-check work on a PC monitor per hour. This yield is nearly 20 times more effective than that by the conventional eye-scan method. The speed and quality is acceptable for the coming new experiment, J-PARC E07.

Doxorubicin-loaded Zein in situ gel for interstitial chemotherapy.

PubMed

Cao, Xiaoying; Geng, Jianning; Su, Suwen; Zhang, Linan; Xu, Qian; Zhang, Li; Xie, Yinghua; Wu, Shaomei; Sun, Yongjun; Gao, Zibin

2012-01-01

A novel drug delivery system of doxorubicin (DOX)-loaded Zein in situ gel for interstitial chemotherapy was investigated in this study. The possible mechanisms of drug release were described according to morphological analysis by optical microscopy and scanning electronic microscope (SEM). In vitro and in vivo anti-tumor activity studies showed that DOX-loaded Zein in situ gel was superior to DOX solution. Local pharmacokinetics in tumor tissue was studied by quantitative analysis with confocal laser scanning microscopy (CLSM) combined with microdialysis technology. A pharmacokinetics mathematical model of DOX-loaded Zein in situ gel in tumors was then built.

Multiplexed phase-space imaging for 3D fluorescence microscopy.

PubMed

Liu, Hsiou-Yuan; Zhong, Jingshan; Waller, Laura

2017-06-26

Optical phase-space functions describe spatial and angular information simultaneously; examples of optical phase-space functions include light fields in ray optics and Wigner functions in wave optics. Measurement of phase-space enables digital refocusing, aberration removal and 3D reconstruction. High-resolution capture of 4D phase-space datasets is, however, challenging. Previous scanning approaches are slow, light inefficient and do not achieve diffraction-limited resolution. Here, we propose a multiplexed method that solves these problems. We use a spatial light modulator (SLM) in the pupil plane of a microscope in order to sequentially pattern multiplexed coded apertures while capturing images in real space. Then, we reconstruct the 3D fluorescence distribution of our sample by solving an inverse problem via regularized least squares with a proximal accelerated gradient descent solver. We experimentally reconstruct a 101 Megavoxel 3D volume (1010×510×500µm with NA 0.4), demonstrating improved acquisition time, light throughput and resolution compared to scanning aperture methods. Our flexible patterning scheme further allows sparsity in the sample to be exploited for reduced data capture.

A compact Acousto-Optic Lens for 2D and 3D femtosecond based 2-photon microscopy.

PubMed

Kirkby, Paul A; Srinivas Nadella, K M Naga; Silver, R Angus

2010-06-21

We describe a high speed 3D Acousto-Optic Lens Microscope (AOLM) for femtosecond 2-photon imaging. By optimizing the design of the 4 AO Deflectors (AODs) and by deriving new control algorithms, we have developed a compact spherical AOL with a low temporal dispersion that enables 2-photon imaging at 10-fold lower power than previously reported. We show that the AOLM can perform high speed 2D raster-scan imaging (>150 Hz) without scan rate dependent astigmatism. It can deflect and focus a laser beam in a 3D random access sequence at 30 kHz and has an extended focusing range (>137 mum; 40X 0.8NA objective). These features are likely to make the AOLM a useful tool for studying fast physiological processes distributed in 3D space.

Tracking features in retinal images of adaptive optics confocal scanning laser ophthalmoscope using KLT-SIFT algorithm

PubMed Central

Li, Hao; Lu, Jing; Shi, Guohua; Zhang, Yudong

2010-01-01

With the use of adaptive optics (AO), high-resolution microscopic imaging of living human retina in the single cell level has been achieved. In an adaptive optics confocal scanning laser ophthalmoscope (AOSLO) system, with a small field size (about 1 degree, 280 μm), the motion of the eye severely affects the stabilization of the real-time video images and results in significant distortions of the retina images. In this paper, Scale-Invariant Feature Transform (SIFT) is used to abstract stable point features from the retina images. Kanade-Lucas-Tomasi(KLT) algorithm is applied to track the features. With the tracked features, the image distortion in each frame is removed by the second-order polynomial transformation, and 10 successive frames are co-added to enhance the image quality. Features of special interest in an image can also be selected manually and tracked by KLT. A point on a cone is selected manually, and the cone is tracked from frame to frame. PMID:21258443

The Effects of ph on Structural and Optical Characterization of Iron Oxide Thin Films

NASA Astrophysics Data System (ADS)

Tezel, Fatma Meydaneri; Özdemir, Osman; Kariper, I. Afşin

In this study, the iron oxide thin films have been produced by chemical bath deposition (CBD) method as a function of pH onto amorphous glass substrates. The surface images of the films were investigated with scanning electron microscope (SEM). The crystal structures, orientation of crystallization, crystallite sizes, and dislocation density i.e. structural properties of the thin films were analyzed with X-ray diffraction (XRD). The optical band gap (Eg), optical transmission (T%), reflectivity (R%), absorption coefficient (α), refraction index (n), extinction coefficient (k) and dielectric constant (ɛ) of the thin films were investigated depending on pH, deposition time, solution temperature, substrate temperature, thickness of the films by UV-VIS spectrometer.

ScanImage: flexible software for operating laser scanning microscopes.

PubMed

Pologruto, Thomas A; Sabatini, Bernardo L; Svoboda, Karel

2003-05-17

Laser scanning microscopy is a powerful tool for analyzing the structure and function of biological specimens. Although numerous commercial laser scanning microscopes exist, some of the more interesting and challenging applications demand custom design. A major impediment to custom design is the difficulty of building custom data acquisition hardware and writing the complex software required to run the laser scanning microscope. We describe a simple, software-based approach to operating a laser scanning microscope without the need for custom data acquisition hardware. Data acquisition and control of laser scanning are achieved through standard data acquisition boards. The entire burden of signal integration and image processing is placed on the CPU of the computer. We quantitate the effectiveness of our data acquisition and signal conditioning algorithm under a variety of conditions. We implement our approach in an open source software package (ScanImage) and describe its functionality. We present ScanImage, software to run a flexible laser scanning microscope that allows easy custom design.

Confocal Microscopy Imaging with an Optical Transition Edge Sensor

NASA Astrophysics Data System (ADS)

Fukuda, D.; Niwa, K.; Hattori, K.; Inoue, S.; Kobayashi, R.; Numata, T.

2018-05-01

Fluorescence color imaging at an extremely low excitation intensity was performed using an optical transition edge sensor (TES) embedded in a confocal microscope for the first time. Optical TES has the ability to resolve incident single photon energy; therefore, the wavelength of each photon can be measured without spectroscopic elements such as diffraction gratings. As target objects, animal cells labeled with two fluorescent dyes were irradiated with an excitation laser at an intensity below 1 μW. In our confocal system, an optical fiber-coupled TES device is used to detect photons instead of the pinhole and photomultiplier tube used in typical confocal microscopes. Photons emitted from the dyes were collected by the objective lens, and sent to the optical TES via the fiber. The TES measures the wavelength of each photon arriving in an exposure time of 70 ms, and a fluorescent photon spectrum is constructed. This measurement is repeated by scanning the target sample, and finally a two-dimensional RGB-color image is obtained. The obtained image showed that the photons emitted from the dyes of mitochondria and cytoskeletons were clearly resolved at a detection intensity level of tens of photons. TES exhibits ideal performance as a photon detector with a low dark count rate (< 1 Hz) and wavelength resolving power. In the single-mode fiber-coupled system, the confocal microscope can be operated in the super-resolution mode. These features are very promising to realize high-sensitivity and high-resolution photon spectral imaging, and would help avoid cell damage and photobleaching of fluorescence dyes.

Excitation-scanning hyperspectral imaging system for microscopic and endoscopic applications

NASA Astrophysics Data System (ADS)

Mayes, Sam A.; Leavesley, Silas J.; Rich, Thomas C.

2016-04-01

Current microscopic and endoscopic technologies for cancer screening utilize white-light illumination sources. Hyper-spectral imaging has been shown to improve sensitivity while retaining specificity when compared to white-light imaging in both microscopy and in vivo imaging. However, hyperspectral imaging methods have historically suffered from slow acquisition times due to the narrow bandwidth of spectral filters. Often minutes are required to gather a full image stack. We have developed a novel approach called excitation-scanning hyperspectral imaging that provides 2-3 orders of magnitude increased signal strength. This reduces acquisition times significantly, allowing for live video acquisition. Here, we describe a preliminary prototype excitation-scanning hyperspectral imaging system that can be coupled with endoscopes or microscopes for hyperspectral imaging of tissues and cells. Our system is comprised of three subsystems: illumination, transmission, and imaging. The illumination subsystem employs light-emitting diode arrays to illuminate at different wavelengths. The transmission subsystem utilizes a unique geometry of optics and a liquid light guide. Software controls allow us to interface with and control the subsystems and components. Digital and analog signals are used to coordinate wavelength intensity, cycling and camera triggering. Testing of the system shows it can cycle 16 wavelengths at as fast as 1 ms per cycle. Additionally, more than 18% of the light transmits through the system. Our setup should allow for hyperspectral imaging of tissue and cells in real time.

The preparation and characterization of silk fibroin blended with low molecular weight hydroxypropyl methylcellulose (HPMC)

NASA Astrophysics Data System (ADS)

Shetty, G. Rajesha; Rao, B. Lakshmeesha; Gowda, Mahadeva; Shivananda, C. S.; Asha, S.; Sangappa, Y.

2018-04-01

In this work, the structure and optical properties of Silk Fibroin (SF), lower molecular weight Hydroxypropyl Methylcellulose (HPMC(L)) and its blend film of SF-HPMC(L) were studied by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron Microscope (SEM) and UV-Visible spectroscopy (UV-Vis). The results indicates that the homogeneous miscible blend of SF-HPMC(L) has lower crystallite size and lower optical band gap compared to virgin SF and HPMC(L). FTIR study confirms the presence of both SF and HPMC(L) molecules in the prepared blend films.

Multiphoton microscopy for the in-situ investigation of cellular processes and integrity in cryopreservation.

PubMed

Doerr, Daniel; Stark, Martin; Ehrhart, Friederike; Zimmermann, Heiko; Stracke, Frank

2009-08-01

In this study we demonstrate a new noninvasive imaging method to monitor freezing processes in biological samples and to investigate life in the frozen state. It combines a laser scanning microscope with a computer-controlled cryostage. Nearinfrared (NIR) femtosecond laser pulses evoke the fluorescence of endogenous fluorophores and fluorescent labels due to multiphoton absorption.The inherent optical nonlinearity of multiphoton absorption allows 3D fluorescence imaging for optical tomography of frozen biological material in-situ. As an example for functional imaging we use fluorescence lifetime imaging (FLIM) to create images with chemical and physical contrast.

Studies of porphyrin-containing specimens using an optical spectrometer connected to a confocal scanning laser microscope.

PubMed

Trepte, O; Rokahr, I; Andersson-Engels, S; Carlsson, K

1994-12-01

A spectrometer has been developed for use with a confocal scanning laser microscope. With this unit, spectral information from a single point or a user-defined region within the microscope specimen can be recorded. A glass prism is used to disperse the spectral components of the recorded light over a linear CCD photodiode array with 256 elements. A regulated cooling unit keeps the detector at 277 K, thereby allowing integration times of up to 60 s. The spectral resolving power, lambda/delta lambda, ranges from 350 at lambda = 400 nm to 100 at lambda = 700 nm. Since the entrance aperture of the spectrometer has the same size as the detector pinhole used during normal confocal scanning, the three-dimensional spatial resolution is equivalent to that of normal confocal scanning. Light from the specimen is deflected to the spectrometer by a solenoid controlled mirror, allowing fast and easy switching between normal confocal scanning and spectrometer readings. With this equipment, studies of rodent liver specimens containing porphyrins have been made. The subcellular localization is of interest for the mechanisms of photodynamic therapy (PDT) of malignant tumours. Spectroscopic detection is necessary to distinguish the porphyrin signal from other fluorescent components in the specimen. Two different substances were administered to the tissue, Photofrin, a haematoporphyrin derivative (HPD) and delta-amino levulinic acid (ALA), a precursor to protoporphyrin IX and haem in the haem cycle. Both are substances under clinical trials for PDT of malignant tumours. Following administration of these compounds to the tissue, the potent photosensitizer and fluorescent compound Photofrin, or protoporphyrin IX, respectively, is accumulated.(ABSTRACT TRUNCATED AT 250 WORDS)

High temporal and spatial resolution studies of bone cells using real-time confocal reflection microscopy.

PubMed

Boyde, A; Vesely, P; Gray, C; Jones, S J

1994-01-01

Chick and rat bone-derived cells were mounted in sealed coverslip-covered chambers; individual osteoclasts (but also osteoblasts) were selected and studied at 37 degrees C using three different types of high-speed scanning confocal microscopes: (1) A Noran Tandem Scanning Microscope (TSM) was used with a low light level, cooled CCD camera for image transfer to a Noran TN8502 frame store-based image analysing computer to make time lapse movie sequences using 0.1 s exposure periods, thus losing some of the advantage of the high frame rate of the TSM. Rapid focus adjustment using computer controlled piezo drivers permitted two or more focus planes to be imaged sequentially: thus (with additional light-source shuttering) the reflection confocal image could be alternated with the phase contrast image at a different focus. Individual cells were followed for up to 5 days, suggesting no significant irradiation problem. (2) Exceptional temporal and spatial resolution is available in video rate laser confocal scanning microscopes (VRCSLMs). We used the Noran Odyssey unitary beam VRCSLM with an argon ion laser at 488 nm and acousto-optic deflection (AOD) on the line axis: this instrument is truly and adjustably confocal in the reflection mode. (3) We also used the Lasertec 1LM11 line scan instrument, with an He-Ne laser at 633 nm, and AOD for the frame scan. We discuss the technical problems and merits of the different approaches. The VRCSLMs documented rapid, real-time oscillatory motion: all the methods used show rapid net movement of organelles within bone cells. The interference reflection mode gives particularly strong contrasts in confocal instruments. Phase contrast and other interference methods used in the microscopy of living cells can be used simultaneously in the TSM.

Quantitative Near-field Microscopy of Heterogeneous and Correlated Electron Oxides

NASA Astrophysics Data System (ADS)

McLeod, Alexander Swinton

Scanning near-field optical microscopy (SNOM) is a novel scanning probe microscopy technique capable of circumventing the conventional diffraction limit of light, affording unparalleled optical resolution (down to 10 nanometers) even for radiation in the infrared and terahertz energy regimes, with light wavelengths exceeding 10 micrometers. However, although this technique has been developed and employed for more than a decade to a qualitatively impressive effect, researchers have lacked a practically quantitative grasp of its capabilities, and its application scope has so far remained restricted by implementations limited to ambient atmospheric conditions. The two-fold objective of this dissertation work has been to address both these shortcomings. The first half of the dissertation presents a realistic, semi-analytic, and benchmarked theoretical description of probe-sample near-field interactions that form the basis of SNOM. Owing its name to the efficient nano-focusing of light at a sharp metallic apex, the "lightning rod model" of probe-sample near-field interactions is mathematically developed from a flexible and realistic scattering formalism. Powerful and practical applications are demonstrated through the accurate prediction of spectroscopic near-field optical contrasts, as well as the "inversion" of these spectroscopic contrasts into a quantitative description of material optical properties. Thus enabled, this thesis work proceeds to present quantitative applications of infrared near-field spectroscopy to investigate nano-resolved chemical compositions in a diverse host of samples, including technologically relevant lithium ion battery materials, astrophysical planetary materials, and invaluable returned extraterrestrial samples. The second half of the dissertation presents the design, construction, and demonstration of a sophisticated low-temperature scanning near-field infrared microscope. This instrument operates in an ultra-high vacuum environment suitable for the investigation of nano-scale physics in correlated electron matter at cryogenic temperatures, thus vastly expanding the scope of applications for infrared SNOM. Performance of the microscope is demonstrated through quanttiative exploration of the canonical insulator-metal transition occuring in the correlated electron insulator V2O3. The methodology established for this investigation provides a model for ongoing and future nano-optical studies of phase transitions and phase coexistence in correlated electron oxides.

Subwavelenght Light Localization in Nanostructured Surfaces

NASA Astrophysics Data System (ADS)

Coello, V.; Wang, S.; Siqueiros, J.; Bozhevolnyi, S. I.

Using a photon scanning tunneling microscope, we studied near field optical images obtained with a surface plasmon polariton (SPP) being resonantly excited along a surface with a random introduced roughness. The SPP intensity field distributions showed an optical enhancement in the form of round bright spots up to 5 times larger than the background signal. We also show an artificially fabricated SPP curved micromirror along with the corresponding near-field optical image. The recorded optical signal exhibited an enhancement up to 10 times larger than the background, which has been generated for the first time in a controlled form. A numerical simulation of a parabolic micromirror based on isotropic pointlike scatterers is analyzed and compared with experimental results. The potential of creating microstructures able to control SPP optical field enhancement is showed in a novel numerically simulated microcavity for SPP's.

Design and performance of an ultra-flexible two-photon microscope for in vivo research.

PubMed

Mayrhofer, Johannes M; Haiss, Florent; Haenni, Dominik; Weber, Stefan; Zuend, Marc; Barrett, Matthew J P; Ferrari, Kim David; Maechler, Philipp; Saab, Aiman S; Stobart, Jillian L; Wyss, Matthias T; Johannssen, Helge; Osswald, Harald; Palmer, Lucy M; Revol, Vincent; Schuh, Claus-Dieter; Urban, Claus; Hall, Andrew; Larkum, Matthew E; Rutz-Innerhofer, Edith; Zeilhofer, Hanns Ulrich; Ziegler, Urs; Weber, Bruno

2015-11-01

We present a cost-effective in vivo two-photon microscope with a highly flexible frontend for in vivo research. Our design ensures fast and reproducible access to the area of interest, including rotation of imaging plane, and maximizes space for auxiliary experimental equipment in the vicinity of the animal. Mechanical flexibility is achieved with large motorized linear stages that move the objective in the X, Y, and Z directions up to 130 mm. 360° rotation of the frontend (rotational freedom for one axis) is achieved with the combination of a motorized high precision bearing and gearing. Additionally, the modular design of the frontend, based on commercially available optomechanical parts, allows straightforward updates to future scanning technologies. The design exceeds the mobility of previous movable microscope designs while maintaining high optical performance.

Design and performance of an ultra-flexible two-photon microscope for in vivo research

PubMed Central

Mayrhofer, Johannes M.; Haiss, Florent; Haenni, Dominik; Weber, Stefan; Zuend, Marc; Barrett, Matthew J. P.; Ferrari, Kim David; Maechler, Philipp; Saab, Aiman S.; Stobart, Jillian L.; Wyss, Matthias T.; Johannssen, Helge; Osswald, Harald; Palmer, Lucy M.; Revol, Vincent; Schuh, Claus-Dieter; Urban, Claus; Hall, Andrew; Larkum, Matthew E.; Rutz-Innerhofer, Edith; Zeilhofer, Hanns Ulrich; Ziegler, Urs; Weber, Bruno

2015-01-01

We present a cost-effective in vivo two-photon microscope with a highly flexible frontend for in vivo research. Our design ensures fast and reproducible access to the area of interest, including rotation of imaging plane, and maximizes space for auxiliary experimental equipment in the vicinity of the animal. Mechanical flexibility is achieved with large motorized linear stages that move the objective in the X, Y, and Z directions up to 130 mm. 360° rotation of the frontend (rotational freedom for one axis) is achieved with the combination of a motorized high precision bearing and gearing. Additionally, the modular design of the frontend, based on commercially available optomechanical parts, allows straightforward updates to future scanning technologies. The design exceeds the mobility of previous movable microscope designs while maintaining high optical performance. PMID:26600989

A targeted illumination optical fiber probe for high resolution fluorescence imaging and optical switching

NASA Astrophysics Data System (ADS)

Shinde, Anant; Perinchery, Sandeep Menon; Murukeshan, Vadakke Matham

2017-04-01

An optical imaging probe with targeted multispectral and spatiotemporal illumination features has applications in many diagnostic biomedical studies. However, these systems are mostly adapted in conventional microscopes, limiting their use for in vitro applications. We present a variable resolution imaging probe using a digital micromirror device (DMD) with an achievable maximum lateral resolution of 2.7 μm and an axial resolution of 5.5 μm, along with precise shape selective targeted illumination ability. We have demonstrated switching of different wavelengths to image multiple regions in the field of view. Moreover, the targeted illumination feature allows enhanced image contrast by time averaged imaging of selected regions with different optical exposure. The region specific multidirectional scanning feature of this probe has facilitated high speed targeted confocal imaging.

Optimal pupil design for confocal microscopy

NASA Astrophysics Data System (ADS)

Patel, Yogesh G.; Rajadhyaksha, Milind; DiMarzio, Charles A.

2010-02-01

Confocal reflectance microscopy may enable screening and diagnosis of skin cancers noninvasively and in real-time, as an adjunct to biopsy and pathology. Current instruments are large, complex, and expensive. A simpler, confocal line-scanning microscope may accelerate the translation of confocal microscopy in clinical and surgical dermatology. A confocal reflectance microscope may use a beamsplitter, transmitting and detecting through the pupil, or a divided pupil, or theta configuration, with half used for transmission and half for detection. The divided pupil may offer better sectioning and contrast. We present a Fourier optics model and compare the on-axis irradiance of a confocal point-scanning microscope in both pupil configurations, optimizing the profile of a Gaussian beam in a circular or semicircular aperture. We repeat both calculations with a cylindrical lens which focuses the source to a line. The variable parameter is the fillfactor, h, the ratio of the 1/e2 diameter of the Gaussian beam to the diameter of the full aperture. The optimal values of h, for point scanning are 0.90 (full) and 0.66 for the half-aperture. For line-scanning, the fill-factors are 1.02 (full) and 0.52 (half). Additional parameters to consider are the optimal location of the point-source beam in the divided-pupil configuration, the optimal line width for the line-source, and the width of the aperture in the divided-pupil configuration. Additional figures of merit are field-of-view and sectioning. Use of optimal designs is critical in comparing the experimental performance of the different configurations.

Design and performance of a beetle-type double-tip scanning tunneling microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jaschinsky, Philipp; Coenen, Peter; Pirug, Gerhard

2006-09-15

A combination of a double-tip scanning tunneling microscope with a scanning electron microscope in ultrahigh vacuum environment is presented. The compact beetle-type design made it possible to integrate two independently driven scanning tunneling microscopes in a small space. Moreover, an additional level for coarse movement allows the decoupling of the translation and approach of the tunneling tip. The position of the two tips can be controlled from the millimeter scale down to 50 nm with the help of an add-on electron microscope. The instrument is capable of atomic resolution imaging with each tip.

Fast scanning mode and its realization in a scanning acoustic microscope

NASA Astrophysics Data System (ADS)

Ju, Bing-Feng; Bai, Xiaolong; Chen, Jian

2012-03-01

The scanning speed of the two-dimensional stage dominates the efficiency of mechanical scanning measurement systems. This paper focused on a detailed scanning time analysis of conventional raster and spiral scan modes and then proposed two fast alternative scanning modes. Performed on a self-developed scanning acoustic microscope (SAM), the measured images obtained by using the conventional scan mode and fast scan modes are compared. The total scanning time is reduced by 29% of the two proposed fast scan modes. It will offer a better solution for high speed scanning without sacrificing the system stability, and will not introduce additional difficulties to the configuration of scanning measurement systems. They can be easily applied to the mechanical scanning measuring systems with different driving actuators such as piezoelectric, linear motor, dc motor, and so on. The proposed fast raster and square spiral scan modes are realized in SAM, but not specially designed for it. Therefore, they have universal adaptability and can be applied to other scanning measurement systems with two-dimensional mechanical scanning stages, such as atomic force microscope or scanning tunneling microscope.

Interferometer-Controlled Optical Tweezers Constructed for Nanotechnology and Biotechnology

NASA Technical Reports Server (NTRS)

Decker, Arthur J.

2002-01-01

A new method to control microparticles was developed in-house at the NASA Glenn Research Center in support of the nanotechnology project under NASA's Aerospace Propulsion and Power Base Research Program. A prototype interferometer-controlled optical tweezers was constructed to manipulate scanning probe microscope (SPM) tips. A laser beam passed through a Mach-Zehnder interferometer, and a microscope objective then produced an optical trap from the coaxial beams. The trap levitated and generated the coarse motion of a 10-mm polystyrene sphere used to simulate a SPM tip. The interference between the beams provided fine control of the forces and moments on the sphere. The interferometer included a piezoelectric-scanned mirror to modulate the interference pattern. The 10-mm sphere was observed to oscillate about 1 mm as the mirror and fringe pattern oscillated. The prototype tweezers proved the feasibility of constructing a more sophisticated interferometer tweezers to hold and manipulate SPM tips. The SPM tips are intended to interrogate and manipulate nanostructures. A more powerful laser will be used to generate multiple traps to hold nanostructures and SPM tips. The vibrating mirror in the interferometer will be replaced with a spatial light modulator. The modulator will allow the optical phase distribution in one leg of the interferometer to be programmed independently at 640 by 480 points for detailed control of the forces and moments. The interference patterns will be monitored to measure the motion of the SPM tips. Neuralnetwork technology will provide fast analysis of the interference patterns for diagnostic purposes and for local or remote feedback control of the tips. This effort also requires theoretical and modeling support in the form of scattering calculations for twin coherent beams from nonspherical particles.

Integrative advances for OCT-guided ophthalmic surgery and intraoperative OCT: microscope integration, surgical instrumentation, and heads-up display surgeon feedback.

PubMed

Ehlers, Justis P; Srivastava, Sunil K; Feiler, Daniel; Noonan, Amanda I; Rollins, Andrew M; Tao, Yuankai K

2014-01-01

To demonstrate key integrative advances in microscope-integrated intraoperative optical coherence tomography (iOCT) technology that will facilitate adoption and utilization during ophthalmic surgery. We developed a second-generation prototype microscope-integrated iOCT system that interfaces directly with a standard ophthalmic surgical microscope. Novel features for improved design and functionality included improved profile and ergonomics, as well as a tunable lens system for optimized image quality and heads-up display (HUD) system for surgeon feedback. Novel material testing was performed for potential suitability for OCT-compatible instrumentation based on light scattering and transmission characteristics. Prototype surgical instruments were developed based on material testing and tested using the microscope-integrated iOCT system. Several surgical maneuvers were performed and imaged, and surgical motion visualization was evaluated with a unique scanning and image processing protocol. High-resolution images were successfully obtained with the microscope-integrated iOCT system with HUD feedback. Six semi-transparent materials were characterized to determine their attenuation coefficients and scatter density with an 830 nm OCT light source. Based on these optical properties, polycarbonate was selected as a material substrate for prototype instrument construction. A surgical pick, retinal forceps, and corneal needle were constructed with semi-transparent materials. Excellent visualization of both the underlying tissues and surgical instrument were achieved on OCT cross-section. Using model eyes, various surgical maneuvers were visualized, including membrane peeling, vessel manipulation, cannulation of the subretinal space, subretinal intraocular foreign body removal, and corneal penetration. Significant iterative improvements in integrative technology related to iOCT and ophthalmic surgery are demonstrated.

Structural, optical and nonlinear optical studies of AZO thin film prepared by SILAR method for electro-optic applications

NASA Astrophysics Data System (ADS)

Edison, D. Joseph; Nirmala, W.; Kumar, K. Deva Arun; Valanarasu, S.; Ganesh, V.; Shkir, Mohd.; AlFaify, S.

2017-10-01

Aluminium doped (i.e. 3 at%) zinc oxide (AZO) thin films were prepared by simple successive ionic layer adsorption and reaction (SILAR) method with different dipping cycles. The structural and surface morphology of AZO thin films were studied by using X-ray diffraction (XRD) and scanning electron microscopy (SEM). The optical parameters such as, transmittance, band gap, refractive index, extinction coefficient, dielectric constant and nonlinear optical properties of AZO films were investigated. XRD pattern revealed the formation of hexagonal phase ZnO and the intensity of the film was found to increase with increasing dipping cycle. The crystallite size was found to be in the range of 29-37 nm. Scanning Electron Microscope (SEM) images show the presence of small sized grains, revealing that the smoothest surface was obtained at all the films. The EDAX spectrum of AZO conforms the presence of Zn, O and Al. The optical transmittance in the visible region is high 87% and the band gap value is 3.23 eV. The optical transmittance is decreased with respect to dipping cycles. The room temperature PL studies revealed that the AZO films prepared at (30 cycles) has good film quality with lesser defect density. The third order nonlinear optical parameters were also studied using Z-scan technique to know the applications of deposited films in nonlinear devices. The third order nonlinear susceptibility value is found to be 1.69 × 10-7, 3.34 × 10-8, 1.33 × 10-7and 2.52 × 10-7 for AZO films deposited after 15, 20, 25 and 30 dipping cycles.

A next generation positron microscope and a survey of candidate samples for future positron studies

NASA Astrophysics Data System (ADS)

Dull, Terry Lou

A positron microscope has been constructed and is nearing the conclusion of its assembly and testing. The instrument is designed to perform positron and electron microscopy in both scanning and magnifying modes. In scanning mode, a small beam of particles is rastered across the target and the amplitude of a positron or electron related signal is recorded as a function of position. For positrons this signal may come from Doppler Broadening Spectroscopy, Reemitted Positron Spectroscopy or Positron Annihilation Lifetime Spectroscopy. For electrons this signal may come from the number of secondary electrons or Auger Electron Spectroscopy. In magnifying mode an incident beam of particles is directed onto the target and emitted particles, either secondary electrons or reemitted positrons, are magnified to form an image. As a positron microscope the instrument will primarily operate in magnifying mode, as a positron reemission microscope. As an electron microscope the instrument will be able to operate in both magnifying and scanning modes. Depth-profiled Doppler Broadening Spectroscopy studies using a non-microscopic low-energy positron beam have also been performed on a series of samples to ascertain the applicability of positron spectroscopies and/or microscopy to their study. All samples have sub-micron film and/or feature size and thus are only susceptible to positron study with low-energy beams. Several stoichiometries and crystallinities of chalcogenide thin films (which can be optically reversibly switched between crystalline states) were studied and a correlation was found to exist between the amorphous/FCC S-parameter difference and the amorphous/FCC switching time. Amorphous silicon films were studied in an attempt to observe the well-established Staebler-Wronski effect as well as the more controversial photodilatation effect. However, DBS was not able to detect either effect. The passive oxide films on titanium and aluminum were studied in an attempt to verify the Point Defect Model, a detailed, but as yet microscopically unconfirmed, theory of the corrosive breakdown of passive films. DBS results supportive of the PDM were observed. Graphitic carbon fibers were also studied and DBS indicated the presence of a 200 nm thick outer fiber skin possibly characterized by a high degree of graphitic crystallite alignment.

A new Brewster angle microscope

NASA Astrophysics Data System (ADS)

Lheveder, C.; Hénon, S.; Mercier, R.; Tissot, G.; Fournet, P.; Meunier, J.

1998-03-01

We present a new Brewster angle microscope for the study of very thin layers as thin as monolayers, using a custom-made objective. This objective avoids the drawbacks of the models existing at the present time. Its optical axis is perpendicular to the studied layer and consequently gives an image in focus in all the plane contrary to the existing models which give images in focus along a narrow strip. The objective allows one to obtain images with a good resolution (less than 1 μm) without scanning the surface, at the video frequency, allowing for dynamic studies. A large frontal distance associated with a very large aperture is obtained by using a large lens at the entrance of the objective.

Microstructures and properties of rapidly solidified alloys

NASA Technical Reports Server (NTRS)

Shechtman, D.; Horowitz, E.

1984-01-01

The microstructure and properties of rapidly solidified aluminum alloys were researched. The effects of powder and flake chemistry and morphology and alternative consolidation processing parameters are being conducted. Samples of the powders being utilized were obtained for comprehensive metallurgical characterization. Seven aluminum alloys in the form of thin foils were studied by a variety of techniques including optical metallography, scanning electron microscope, and transmission electron microscope. Details of the microstructural characteristics are presented along with a discussion of the solidification process. A better understanding of the microstructure of the rapidly solidified aluminum alloys prepared by a variety of techniques such as roller quenching, the vacuum atomized procedure, ultrasonically atomized in inert atmospheres, and atomized in flue gas was provided.

Plastic deformation at surface during unlubricated sliding

NASA Technical Reports Server (NTRS)

Yamamoto, T.; Buckley, D. H.

1982-01-01

The plastic deformation and wear of 304 stainless-steel surface slid against an aluminum oxide rider were observed by using a scanning electron microscope and an optical microscope. Experiments were conducted in a vacuum of 0.000001 Pa and in an environment of 0.0005 Pa chlorine gas at 25 C. The load was 500 grams and the sliding velocity was 0.5 centimeter per second. The deformed surface layer which accumulates and develops successively is left behind the rider, and step-shaped protuberances are developed even after single pass sliding under both environmental conditions. A fully developed surface layer is gradually torn off leaving a characteristic pattern. These observations result from both adhesion and an adhesive wear mechanism.

A compact acousto-optic lens for 2D and 3D femtosecond based 2-photon microscopy

PubMed Central

Kirkby, Paul A.; Naga Srinivas, N.K.M.; Silver, R. Angus

2010-01-01

We describe a high speed 3D Acousto-Optic Lens Microscope (AOLM) for femtosecond 2-photon imaging. By optimizing the design of the 4 AO Deflectors (AODs) and by deriving new control algorithms, we have developed a compact spherical AOL with a low temporal dispersion that enables 2-photon imaging at 10-fold lower power than previously reported. We show that the AOLM can perform high speed 2D raster-scan imaging (>150 Hz) without scan rate dependent astigmatism. It can deflect and focus a laser beam in a 3D random access sequence at 30 kHz and has an extended focusing range (>137 μm; 40X 0.8NA objective). These features are likely to make the AOLM a useful tool for studying fast physiological processes distributed in 3D space PMID:20588506

Deformation microstructures of Barre granite: An optical, Sem and Tem study

USGS Publications Warehouse

Schedl, A.; Kronenberg, A.K.; Tullis, J.

1986-01-01

New scanning electron microscope techniques have been developed for characterizing ductile deformation microstructures in felsic rocks. In addition, the thermomechanical history of the macroscopically undeformed Barre granite (Vermont, U.S.A.) has been reconstructed based on examination of deformation microstructures using optical microscopy, scanning electron microscopy, and transmission electron microscopy. The microstructures reveal three distinct events: 1. (1) a low-stress, high-temperature event that produced subgrains in feldspars, and subgrains and recrystallized grains in quartz; 2. (2) a high-stress, low-temperature event that produced a high dislocation density in quartz and feldspars; and 3. (3) a lowest-temperature event that produced cracks, oriented primarily along cleavage planes in feldspars, and parallel to the macroscopic rift in quartz. The first two events are believed to reflect various stages in the intrusion and cooling history of the pluton, and the last may be related to the last stages of cooling, or to later tectonism. ?? 1986.

Widely tuneable scattering-type scanning near-field optical microscopy using pulsed quantum cascade lasers

NASA Astrophysics Data System (ADS)

Yoxall, Edward; Navarro-Cía, Miguel; Rahmani, Mohsen; Maier, Stefan A.; Phillips, Chris C.

2013-11-01

We demonstrate the use of a pulsed quantum cascade laser, wavelength tuneable between 6 and 10 μm, with a scattering-type scanning near-field optical microscope (s-SNOM). A simple method for calculating the signal-to-noise ratio (SNR) of the s-SNOM measurement is presented. For pulsed lasers, the SNR is shown to be highly dependent on the degree of synchronization between the laser pulse and the sampling circuitry; in measurements on a gold sample, the SNR is 26 with good synchronization and less than 1 without. Simulations and experimental s-SNOM images, with a resolution of 100 nm, corresponding to λ/80, and an acquisition time of less than 90 s, are presented as proof of concept. They show the change in the field profile of plasmon-resonant broadband antennas when they are excited with wavelengths of 7.9 and 9.5 μm.

Effect of fluorides from various restorative materials on remineralization of adjacent tooth: an in vitro study.

PubMed

Baliga, M S; Bhat, S S

2010-01-01

The aim of the study was to evaluate the extent of surface zone remineralization and the effect of fluoride at the inter-proximal adjacent tooth surface, using restorative materials FusionAlloy, Ketac-Fil and Heliomolar. Ninety extracted molar teeth were used of which 45 were placed in artificial caries for 10 weeks. The remaining 45 teeth were filled with the respective restorative materials, mounted with the artificial carious teeth in proximal contact with plaster and placed in artificial saliva for a period of 28 days. Finally, sectioning of artificially carious teeth was done mesio-distally and observed under the optical microscope and scanning electron microscope. Comparison among the groups was done by one-way analysis of variance [ANOVA] and Fischer's F test. Intercomparison between the groups was done by using Dunnett's t-test. Results obtained from transmitted electron microscopic and scanning electron microscopic observations were almost similar with the Ketac-Fil and Heliomolar showing better results in surface zone remineralization compared to FusionAlloy. Also, Ketac-Fil is a good material in releasing fluoride to remineralize enamel when compared to Heliomolar and FusionAlloy. Thus, it can be used mainly in class II cavity restorations of primary and permanent dentitions due to the potential ability of fluoride containing glass ionomer cements and composite resins to remineralize incipient carious lesions on adjacent teeth.

Soil burial biodegradation studies of palm oil-based UV-curable films

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tajau, Rida, E-mail: rida@nuclearmalaysia.gov.my; Salleh, Mek Zah, E-mail: mekzah@nuclearmalaysia.gov.my; Salleh, Nik Ghazali Nik, E-mail: nik-ghazali@nuclearmalaysia.gov.my

The palm oil-based ultraviolet (uv)-curable films were subjected to an outdoor soil burial test to investigate the biodegradation under natural environment. The films were burial in the soil experiment plot at the Nuclear Malaysia’s Dengkil complex. The uv-curable films were synthesized from the epoxidized palm oil acrylated (EPOLA) resin and the polyurethane palm oil (POBUA) resin, respectively. Biodegradation tests are more specific to burial film in soil experiments for 12 months under natural conditions. The biodegradability of palm oil resin based uv-curable films were investigated and compared with the petrochemical resin based film. The films properties were compared with respectmore » to properties of the thermal characteristic, the crystallinity, the morphology and the weight loss which are analyzed using the thermogravimetric analysis (TGA), the differential scanning calorimetry (DSC), the scanning electron microscope (SEM), an optical microscope and the weight loss of film calculation. These findings suggested that the palm oil-based uv-curable films show quite satisfactory biodegradation levels.« less

Building Practical Apertureless Scanning Near-Field Microscopy

NASA Astrophysics Data System (ADS)

Gungordu, M. Zeki

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

Anisotropic excitation of surface plasmon polaritons on a metal film by a scattering-type scanning near-field microscope with a non-rotationally-symmetric probe tip

NASA Astrophysics Data System (ADS)

Walla, Frederik; Wiecha, Matthias M.; Mecklenbeck, Nicolas; Beldi, Sabri; Keilmann, Fritz; Thomson, Mark D.; Roskos, Hartmut G.

2018-01-01

We investigated the excitation of surface plasmon polaritons on gold films with the metallized probe tip of a scattering-type scanning near-field optical microscope (s-SNOM). The emission of the polaritons from the tip, illuminated by near-infrared laser radiation, was found to be anisotropic and not circularly symmetric as expected on the basis of literature data. We furthermore identified an additional excitation channel via light that was reflected off the tip and excited the plasmon polaritons at the edge of the metal film. Our results, while obtained for a non-rotationally-symmetric type of probe tip and thus specific for this situation, indicate that when an s-SNOM is employed for the investigation of plasmonic structures, the unintentional excitation of surface waves and anisotropic surface wave propagation must be considered in order to correctly interpret the signatures of plasmon polariton generation and propagation.

Soil burial biodegradation studies of palm oil-based UV-curable films

NASA Astrophysics Data System (ADS)

Tajau, Rida; Salleh, Mek Zah; Salleh, Nik Ghazali Nik; Abdurahman, Mohamad Norahiman; Salih, Ashraf Mohammed; Fathy, Siti Farhana; Azman, Anis Asmi; Hamidi, Nur Amira

2016-01-01

The palm oil-based ultraviolet (uv)-curable films were subjected to an outdoor soil burial test to investigate the biodegradation under natural environment. The films were burial in the soil experiment plot at the Nuclear Malaysia's Dengkil complex. The uv-curable films were synthesized from the epoxidized palm oil acrylated (EPOLA) resin and the polyurethane palm oil (POBUA) resin, respectively. Biodegradation tests are more specific to burial film in soil experiments for 12 months under natural conditions. The biodegradability of palm oil resin based uv-curable films were investigated and compared with the petrochemical resin based film. The films properties were compared with respect to properties of the thermal characteristic, the crystallinity, the morphology and the weight loss which are analyzed using the thermogravimetric analysis (TGA), the differential scanning calorimetry (DSC), the scanning electron microscope (SEM), an optical microscope and the weight loss of film calculation. These findings suggested that the palm oil-based uv-curable films show quite satisfactory biodegradation levels.

Optoacoustic Microscopy for Investigation of Material Nanostructures-Embracing the Ultrasmall, Ultrafast, and the Invisible

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nurmikko, Arto; Humphrey, Maris

2014-07-10

The goal of this grant was the development of a new type of scanning acoustic microscope for nanometer resolution ultrasound imaging, based on ultrafast optoacoustics (>GHz). In the microscope, subpicosecond laser pulses was used to generate and detect very high frequency ultrasound with nanometer wavelengths. We report here on the outcome of the 3-year DOE/BES grant which involved the design, multifaceted construction, and proof-of-concept demonstration of an instrument that can be used for quantitative imaging of nanoscale material features – including features that may be buried so as to be inaccessible to conventional lightwave or electron microscopies. The research programmore » has produced a prototype scanning optoacoustic microscope which, in combination with advanced computational modeling, is a system-level new technology (two patents issues) which offer novel means for precision metrology of material nanostructures, particularly those that are of contemporary interest to the frontline micro- and optoelectronics device industry. For accomplishing the ambitious technical goals, the research roadmap was designed and implemented in two phases. In Phase I, we constructed a “non-focusing” optoacoustic microscope instrument (“POAM”), with nanometer vertical (z-) resolution, while limited to approximately 10 micrometer scale lateral recolution. The Phase I version of the instrument which was guided by extensive acoustic and optical numerical modeling of the basic underlying acoustic and optical physics, featured nanometer scale close loop positioning between the optoacoustic transducer element and a nanostructured material sample under investigation. In phase II, we implemented and demonstrated a scanning version of the instrument (“SOAM”) where incident acoustic energy is focused, and scanned on lateral (x-y) spatial scale in the 100 nm range as per the goals of the project. In so doing we developed advanced numerical simulations to provide computational models of the focusing of multi-GHz acoustic waves to the nanometer scale and innovated a series fabrication approaches for a new type of broadband high-frequency acoustic focusing microscope objective by applying methods on nanoimprinting and focused-ion beam techniques. In the following, the Phase I and Phase II instrument development is reported as Section II. The first segment of this section describes the POAM instrument and its development, while including much of the underlying ultrafast acoustic physics which is common to all of our work for this grant. Then, the science and engineering of the SOAM instrument is described, including the methods of fabricating new types of acoustic microlenses. The results section is followed by reports on publications (Section III), Participants (Section IV), and statement of full use of the allocated grant funds (Section V).« less

Hybrid setup for micro- and nano-computed tomography in the hard X-ray range

NASA Astrophysics Data System (ADS)

Fella, Christian; Balles, Andreas; Hanke, Randolf; Last, Arndt; Zabler, Simon

2017-12-01

With increasing miniaturization in industry and medical technology, non-destructive testing techniques are an area of ever-increasing importance. In this framework, X-ray microscopy offers an efficient tool for the analysis, understanding, and quality assurance of microscopic samples, in particular as it allows reconstructing three-dimensional data sets of the whole sample's volume via computed tomography (CT). The following article describes a compact X-ray microscope in the hard X-ray regime around 9 keV, based on a highly brilliant liquid-metal-jet source. In comparison to commercially available instruments, it is a hybrid that works in two different modes. The first one is a micro-CT mode without optics, which uses a high-resolution detector to allow scans of samples in the millimeter range with a resolution of 1 μm. The second mode is a microscope, which contains an X-ray optical element to magnify the sample and allows resolving 150 nm features. Changing between the modes is possible without moving the sample. Thus, the instrument represents an important step towards establishing high-resolution laboratory-based multi-mode X-ray microscopy as a standard investigation method.

High temperature oxidation behavior of austenitic stainless steel AISI 304 in steam of nanofluids contain nanoparticle ZrO2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Prajitno, Djoko Hadi, E-mail: djokohp@batan.go.id; Syarif, Dani Gustaman, E-mail: djokohp@batan.go.id

2014-03-24

The objective of this study is to evaluate high temperature oxidation behavior of austenitic stainless steel SS 304 in steam of nanofluids contain nanoparticle ZrO{sub 2}. The oxidation was performed at high temperatures ranging from 600 to 800°C. The oxidation time was 60 minutes. After oxidation the surface of the samples was analyzed by different methods including, optical microscope, scanning electron microscope (SEM) and X-ray diffraction (XRD). X-ray diffraction examination show that the oxide scale formed during oxidation of stainless steel AISI 304 alloys is dominated by iron oxide, Fe{sub 2}O{sub 3}. Minor element such as Cr{sub 2}O{sub 3} ismore » also appeared in the diffraction pattern. Characterization by optical microscope showed that cross section microstructure of stainless steel changed after oxidized with the oxide scale on the surface stainless steels. SEM and x-ray diffraction examination show that the oxide of ZrO{sub 2} appeared on the surface of stainless steel. Kinetic rate of oxidation of austenite stainless steel AISI 304 showed that increasing oxidation temperature and time will increase oxidation rate.« less

Video-rate confocal microscopy for single-molecule imaging in live cells and superresolution fluorescence imaging.

PubMed

Lee, Jinwoo; Miyanaga, Yukihiro; Ueda, Masahiro; Hohng, Sungchul

2012-10-17

There is no confocal microscope optimized for single-molecule imaging in live cells and superresolution fluorescence imaging. By combining the swiftness of the line-scanning method and the high sensitivity of wide-field detection, we have developed a, to our knowledge, novel confocal fluorescence microscope with a good optical-sectioning capability (1.0 μm), fast frame rates (<33 fps), and superior fluorescence detection efficiency. Full compatibility of the microscope with conventional cell-imaging techniques allowed us to do single-molecule imaging with a great ease at arbitrary depths of live cells. With the new microscope, we monitored diffusion motion of fluorescently labeled cAMP receptors of Dictyostelium discoideum at both the basal and apical surfaces and obtained superresolution fluorescence images of microtubules of COS-7 cells at depths in the range 0-85 μm from the surface of a coverglass. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

An evaluation of a combined scanning probe and optical microscope for lunar regolith studies

NASA Astrophysics Data System (ADS)

Yang, S.; Pike, W. T.; Staufer, U.; Claus, D.; Rodenburg, J. M.

2011-12-01

The microscopic properties of the lunar regolith such as the shape, the surface texture and the size distribution are required for an understanding of both past surface processes and potential hazards for future human exploration [1]. To reveal the particle morphology at the sub micrometer scale, scanning-probe microscopy (SPM), first used on the 2008 Phoenix mission [1], is a proven approach; however, there are two main challenges for the measurement of lunar particles. Firstly, the SPM tip is liable to move particles during scanning, even when using the lower contact forces of the dynamic-mode imaging. Hence the particles need to be stabilised during imaging. Secondly, typically the AFM tip extends about 10 μm from its cantilever, so larger particles protruding more than this height above their substrates cannot be scanned completely. To immobilize particles and eliminate large particles during SPM scanning, micromachined Si substrates, which have been successfully applied in the Phoenix project for Mars investigation in 2008 [2], have been investigated for lunar analogue material. On these substrates micrometer pits are patterned and serve as traps to enhance the stability of the AFM scanning by grasping the particles. In addition, the diameter of pits can determine the size of dusts to be captured and reduce the adhesion for the larger dust and so eliminate the oversized particles. To extend the imaging range and assist in selecting scan areas for the SPM, we use a type of lensless optical imaging (LOM) which uses ptychographic diffractive imaging [3] to eliminate the restrictions and performance limitations of conventional focusing devices. As a reference, scanning electron microscopy (SEM) which minimizes particle-probe interactions and has the advantage of an extended depth of field, is employed to image the same particle fields at resolutions covering both the SPM and LOM. By comparing the differences and the similarities between SEM and LOM images, the ability of LOM for illuminating the details about the lunar particles sample, is demonstrated. The analysis of SEM and SPM images of the same particles of JSC-LunarA analogue soil reveals the potential of the SPM to obtain reliable microscopic images of lunar dusts including detailed morphology with the help of the micromachined Si substrates. [1] J. D. Carpenter, O. Angerer, M. Durante, D. Linnarson, W. T. Pike, "Life Sciences Investigations for ESA's First Lunar Lander," Earth, Moon, and Planets, Vol.107, pp. 11-23, 2010. [2] S. Vijendran, H.Sykulska, and W. T. Pike, "AFM investigation of Martian soil simulant on micromachined Si substrates," Journal of Microscopy, Vol.227, pp.236-245, Sep. 2007. [3] J.M. Rodenburg, "Ptychography and related diffractive imaging techniques," Advances in Imaging and Electron Physics, Vol.150, pp. 87-184, 2008

Scanning evanescent electro-magnetic microscope

DOEpatents

Xiang, Xiao-Dong; Gao, Chen; Schultz, Peter G.; Wei, Tao

2003-01-01

A novel scanning microscope is described that uses near-field evanescent electromagnetic waves to probe sample properties. The novel microscope is capable of high resolution imaging and quantitative measurements of the electrical properties of the sample. The inventive scanning evanescent wave electromagnetic microscope (SEMM) can map dielectric constant, tangent loss, conductivity, complex electrical impedance, and other electrical parameters of materials. The quantitative map corresponds to the imaged detail. The novel microscope can be used to measure electrical properties of both dielectric and electrically conducting materials.

Scanning evanescent electro-magnetic microscope

DOEpatents

Xiang, Xiao-Dong; Gao, Chen

2001-01-01

A novel scanning microscope is described that uses near-field evanescent electromagnetic waves to probe sample properties. The novel microscope is capable of high resolution imaging and quantitative measurements of the electrical properties of the sample. The inventive scanning evanescent wave electromagnetic microscope (SEMM) can map dielectric constant, tangent loss, conductivity, complex electrical impedance, and other electrical parameters of materials. The quantitative map corresponds to the imaged detail. The novel microscope can be used to measure electrical properties of both dielectric and electrically conducting materials.

Acetone sensor based on zinc oxide hexagonal tubes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hastir, Anita, E-mail: anitahastir@gmail.com; Singh, Onkar, E-mail: anitahastir@gmail.com; Anand, Kanika, E-mail: anitahastir@gmail.com

2014-04-24

In this work hexagonal tubes of zinc oxide have been synthesized by co-precipitation method. For structural, morphological, elemental and optical analysis synthesized powders were characterized by using x-ray diffraction, field emission scanning microscope, EDX, UV-visible and FTIR techniques. For acetone sensing thick films of zinc oxide have been deposited on alumina substrate. The fabricated sensors exhibited maximum sensing response towards acetone vapour at an optimum operating temperature of 400°C.

Enhanced weak-signal sensitivity in two-photon microscopy by adaptive illumination.

PubMed

Chu, Kengyeh K; Lim, Daryl; Mertz, Jerome

2007-10-01

We describe a technique to enhance both the weak-signal relative sensitivity and the dynamic range of a laser scanning optical microscope. The technique is based on maintaining a fixed detection power by fast feedback control of the illumination power, thereby transferring high measurement resolution to weak signals while virtually eliminating the possibility of image saturation. We analyze and demonstrate the benefits of adaptive illumination in two-photon fluorescence microscopy.

Panoramic view of human corneal endothelial cell layer observed by a prototype slit-scanning wide-field contact specular microscope.

PubMed

Tanaka, Hiroshi; Okumura, Naoki; Koizumi, Noriko; Sotozono, Chie; Sumii, Yasuhiro; Kinoshita, Shigeru

2017-05-01

To observe the most peripheral region of the corneal endothelial cell (CEC) layer as long as optically recordable by use of a prototype slit-scanning wide-field contact specular microscope and produce a panoramic image to evaluate the variation of CEC density with ageing. Observational case series study. This study involved 15 eyes of 15 normal healthy subjects divided into three groups according to age: A (20-40 years), B (41-60 years) and C (>60 years). The corneal endothelial layer of each eye was recorded in a horizontal direction, from nasal to temporal, with a slit-scanning wide-field contact specular microscope (Konan) and endothelial cell density (ECD) in three specific regions (central, mid-peripheral, and peripheral) was automatically calculated via built-in analysis software. Corneal endothelial images from near the surgical limbus to limbus in all eyes were clearly recorded and panoramic images were made by combining still images. ECD in groups A, B and C were 2809±186, 2717±91 and 2580±129 cells/mm 2 at the centre, 2902±242, 2772±97 and 2604±187 cells/mm 2 at the mid-periphery and 2893±308, 2691±99 and 2533±112 cells/mm 2 at the periphery. Significance differences in ECD was found between groups A and C in all regions and groups between B and C at mid-peripheral region. A prototype slit-scanning wide-field contact specular microscope enabled us to record the endothelial layer from the surgical limbus to limbus of the cornea and compare specific areas among subjects, and showed that ECD in each region of the cornea decreases with ageing. UMIN000021264, Results. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

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

PubMed Central

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. PMID:28819645

Polarization- and wavelength-resolved near-field imaging of complex plasmonic modes in Archimedean nanospirals

DOE PAGES

Hachtel, Jordan A.; Davidson, II, Roderick B.; Kovalik, Elena R.; ...

2018-02-15

Asymmetric nanophotonic structures enable a wide range of opportunities in optical nanotechnology because they support efficient optical nonlinearities mediated by multiple plasmon resonances over a broad spectral range. The Archimedean nanospiral is a canonical example of a chiral plasmonic structure because it supports even-order nonlinearities that are not generally accessible in locally symmetric geometries. However, the complex spiral response makes nanoscale experimental characterization of the plasmonic near-field structure highly desirable. As a result, we employ high-efficiency, high-spatial-resolution cathodoluminescence imaging in a scanning transmission electron microscope to describe the spatial, spectral, and polarization response of plasmon modes in the nanospiral geometry.

Fiber-optic two-photon optogenetic stimulation.

PubMed

Dhakal, K; Gu, L; Black, B; Mohanty, S K

2013-06-01

Optogenetic stimulation of genetically targeted cells is proving to be a powerful tool in the study of cellular systems, both in vitro and in vivo. However, most opsins are activated in the visible spectrum, where significant absorption and scattering of stimulating light occurs, leading to low penetration depth and less precise stimulation. Since we first (to the best of our knowledge) demonstrated two-photon optogenetic stimulation (TPOS), it has gained considerable interest in the probing of cellular circuitry by precise spatial modulation. However, all existing methods use microscope objectives and complex scanning beam geometries. Here, we report a nonscanning method based on multimode fiber to accomplish fiber-optic TPOS of cells.

Fabrication of gradient optical filter containing anisotropic Bragg nanostructure.

PubMed

Cho, Bomin; Um, Sungyong; Woo, Hee-Gweon; Sohn, Honglae

2011-08-01

New gradient optical filters containing asymmetric Bragg structure were prepared from the distributed Bragg reflector (DBR) porous silicon (PSi). Anisotropic DBR PSi displaying a rainbow-colored reflection was generated by using an asymmetric etching configuration. Flexible anisotropic DBR PSi composite films were obtained by casting of polymer solution onto anisotropic DBR PSi thin films. The surface and cross-sectional images images of anisotropic DBR PSi composite films obtained with cold field emission scanning electron microscope indicated that the average pore size and the thickness of porous layer decreased as the lateral distance increased. As lateral distance increased, the reflection resonance shifted to shorter wavelength.

Polarization- and wavelength-resolved near-field imaging of complex plasmonic modes in Archimedean nanospirals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hachtel, Jordan A.; Davidson, II, Roderick B.; Kovalik, Elena R.

Asymmetric nanophotonic structures enable a wide range of opportunities in optical nanotechnology because they support efficient optical nonlinearities mediated by multiple plasmon resonances over a broad spectral range. The Archimedean nanospiral is a canonical example of a chiral plasmonic structure because it supports even-order nonlinearities that are not generally accessible in locally symmetric geometries. However, the complex spiral response makes nanoscale experimental characterization of the plasmonic near-field structure highly desirable. As a result, we employ high-efficiency, high-spatial-resolution cathodoluminescence imaging in a scanning transmission electron microscope to describe the spatial, spectral, and polarization response of plasmon modes in the nanospiral geometry.

Percentage of different aluminum doping influence the morphological and optical properties of ZnO nanostructured growth for sensor application

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mohamed, R., E-mail: ruziana12@gmail.com; NANO-SciTech Centre, Institue of Science, Universiti Teknologi MARA, 40450 Shah Alam, Selangor; Faculty of Applied Sciences, Universiti Teknologi MARA Pahang, 26400 Bandar Tun Razak Jengka, Pahang

In this work, Zinc Oxide (ZnO) with different aluminum (Al) doping percentage was synthesis by sol gel immersion method. Al doped ZnO at various doping percentage from 1, 2, 3, 4 and 5. It was found that with different Al percentage influence the morphological and optical properties of ZnO growth. Field Emission Scanning Electron Microscope (FESEM) image showed the use of different Al doping causes the difference in geometry and size of ZnO nanorods growth. Based on UV-Vis spectroscopy, the transmittance at 1% Al doping has the highest spectrum.

Optimizing pulse compressibility in completely all-fibered Ytterbium chirped pulse amplifiers for in vivo two photon laser scanning microscopy

PubMed Central

Fernández, A.; Grüner-Nielsen, L.; Andreana, M.; Stadler, M.; Kirchberger, S.; Sturtzel, C.; Distel, M.; Zhu, L.; Kautek, W.; Leitgeb, R.; Baltuska, A.; Jespersen, K.; Verhoef, A.

2017-01-01

A simple and completely all-fiber Yb chirped pulse amplifier that uses a dispersion matched fiber stretcher and a spliced-on hollow core photonic bandgap fiber compressor is applied in nonlinear optical microscopy. This stretching-compression approach improves compressibility and helps to maximize the fluorescence signal in two-photon laser scanning microscopy as compared with approaches that use standard single mode fibers as stretcher. We also show that in femtosecond all-fiber systems, compensation of higher order dispersion terms is relevant even for pulses with relatively narrow bandwidths for applications relying on nonlinear optical effects. The completely all-fiber system was applied to image green fluorescent beads, a stained lily-of-the-valley root and rat-tail tendon. We also demonstrated in vivo imaging in zebrafish larvae, where we simultaneously measure second harmonic and fluorescence from two-photon excited red-fluorescent protein. Since the pulses are compressed in a fiber, this source is especially suited for upgrading existing laser scanning (confocal) microscopes with multiphoton imaging capabilities in space restricted settings or for incorporation in endoscope-based microscopy. PMID:28856032

Super-Resolution Scanning Laser Microscopy Based on Virtually Structured Detection

PubMed Central

Zhi, Yanan; Wang, Benquan; Yao, Xincheng

2016-01-01

Light microscopy plays a key role in biological studies and medical diagnosis. The spatial resolution of conventional optical microscopes is limited to approximately half the wavelength of the illumination light as a result of the diffraction limit. Several approaches—including confocal microscopy, stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy, and structured illumination microscopy—have been established to achieve super-resolution imaging. However, none of these methods is suitable for the super-resolution ophthalmoscopy of retinal structures because of laser safety issues and inevitable eye movements. We recently experimentally validated virtually structured detection (VSD) as an alternative strategy to extend the diffraction limit. Without the complexity of structured illumination, VSD provides an easy, low-cost, and phase artifact–free strategy to achieve super-resolution in scanning laser microscopy. In this article we summarize the basic principles of the VSD method, review our demonstrated single-point and line-scan super-resolution systems, and discuss both technical challenges and the potential of VSD-based instrumentation for super-resolution ophthalmoscopy of the retina. PMID:27480461

Optimizing pulse compressibility in completely all-fibered Ytterbium chirped pulse amplifiers for in vivo two photon laser scanning microscopy.

PubMed

Fernández, A; Grüner-Nielsen, L; Andreana, M; Stadler, M; Kirchberger, S; Sturtzel, C; Distel, M; Zhu, L; Kautek, W; Leitgeb, R; Baltuska, A; Jespersen, K; Verhoef, A

2017-08-01

A simple and completely all-fiber Yb chirped pulse amplifier that uses a dispersion matched fiber stretcher and a spliced-on hollow core photonic bandgap fiber compressor is applied in nonlinear optical microscopy. This stretching-compression approach improves compressibility and helps to maximize the fluorescence signal in two-photon laser scanning microscopy as compared with approaches that use standard single mode fibers as stretcher. We also show that in femtosecond all-fiber systems, compensation of higher order dispersion terms is relevant even for pulses with relatively narrow bandwidths for applications relying on nonlinear optical effects. The completely all-fiber system was applied to image green fluorescent beads, a stained lily-of-the-valley root and rat-tail tendon. We also demonstrated in vivo imaging in zebrafish larvae, where we simultaneously measure second harmonic and fluorescence from two-photon excited red-fluorescent protein. Since the pulses are compressed in a fiber, this source is especially suited for upgrading existing laser scanning (confocal) microscopes with multiphoton imaging capabilities in space restricted settings or for incorporation in endoscope-based microscopy.

Comparison of divided and full pupil configurations for line-scanning confocal microscopy in human skin and oral mucosa

NASA Astrophysics Data System (ADS)

Larson, Bjorg; Abeytunge, Sanjeewa; Glazowski, Chris; Rajadhyaksha, Milind

2012-02-01

Confocal point-scanning microscopy has been showing promise in the detection, diagnosing and mapping of skin lesions in clinical settings. The noninvasive technique allows provides optical sectioning and cellular resolution for in vivo diagnosis of melanoma and basal cell carcinoma and pre-operative and intra-operative mapping of margins. The imaging has also enabled more accurate "guided" biopsies while minimizing the otherwise large number of "blind" biopsies. Despite these translational advances, however, point-scanning technology remains relatively complex and expensive. Line-scanning technology may offer an alternative approach to accelerate translation to the clinic. Line-scanning, using fewer optical components, inexpensive linear-array detectors and custom electronics, may enable smaller, simpler and lower-cost confocal microscopes. A line is formed using a cylindrical lens and scanned through the back focal plane of the objective with a galvanometric scanner. A linear CCD is used for detection. Two pupil configurations were compared for performance in imaging human tissue. In the full-pupil configuration, illumination and detection is made through the full objective pupil. In the divided pupil approach, half the pupil is illuminated and the other half is used for detection. The divided pupil configuration loses spatial and axial resolution due to a diminished NA, but the sectioning capability and rejection of background is improved. Imaging in skin and oral mucosa illustrate the performance of the two configurations.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yashchuk, Valeriy V; Conley, Raymond; Anderson, Erik H

Verification of the reliability of metrology data from high quality x-ray optics requires that adequate methods for test and calibration of the instruments be developed. For such verification for optical surface profilometers in the spatial frequency domain, a modulation transfer function (MTF) calibration method based on binary pseudo-random (BPR) gratings and arrays has been suggested [Proc. SPIE 7077-7 (2007), Opt. Eng. 47(7), 073602-1-5 (2008)} and proven to be an effective calibration method for a number of interferometric microscopes, a phase shifting Fizeau interferometer, and a scatterometer [Nucl. Instr. and Meth. A 616, 172-82 (2010)]. Here we describe the details ofmore » development of binary pseudo-random multilayer (BPRML) test samples suitable for characterization of scanning (SEM) and transmission (TEM) electron microscopes. We discuss the results of TEM measurements with the BPRML test samples fabricated from a WiSi2/Si multilayer coating with pseudo randomly distributed layers. In particular, we demonstrate that significant information about the metrological reliability of the TEM measurements can be extracted even when the fundamental frequency of the BPRML sample is smaller than the Nyquist frequency of the measurements. The measurements demonstrate a number of problems related to the interpretation of the SEM and TEM data. Note that similar BPRML test samples can be used to characterize x-ray microscopes. Corresponding work with x-ray microscopes is in progress.« less

Integrated nonlinear optical imaging microscope for on-axis crystal detection and centering at a synchrotron beamline

PubMed Central

Madden, Jeremy T.; Toth, Scott J.; Dettmar, Christopher M.; Newman, Justin A.; Oglesbee, Robert A.; Hedderich, Hartmut G.; Everly, R. Michael; Becker, Michael; Ronau, Judith A.; Buchanan, Susan K.; Cherezov, Vadim; Morrow, Marie E.; Xu, Shenglan; Ferguson, Dale; Makarov, Oleg; Das, Chittaranjan; Fischetti, Robert; Simpson, Garth J.

2013-01-01

Nonlinear optical (NLO) instrumentation has been integrated with synchrotron X-ray diffraction (XRD) for combined single-platform analysis, initially targeting applications for automated crystal centering. Second-harmonic-generation microscopy and two-photon-excited ultraviolet fluorescence microscopy were evaluated for crystal detection and assessed by X-ray raster scanning. Two optical designs were constructed and characterized; one positioned downstream of the sample and one integrated into the upstream optical path of the diffractometer. Both instruments enabled protein crystal identification with integration times between 80 and 150 µs per pixel, representing a ∼103–104-fold reduction in the per-pixel exposure time relative to X-ray raster scanning. Quantitative centering and analysis of phenylalanine hydroxylase from Chromobacterium violaceum cPAH, Trichinella spiralis deubiquitinating enzyme TsUCH37, human κ-opioid receptor complex kOR-T4L produced in lipidic cubic phase (LCP), intimin prepared in LCP, and α-cellulose samples were performed by collecting multiple NLO images. The crystalline samples were characterized by single-crystal diffraction patterns, while α-cellulose was characterized by fiber diffraction. Good agreement was observed between the sample positions identified by NLO and XRD raster measurements for all samples studied. PMID:23765294

Phase Sensitive Demodulation in Multiphoton Microscopy

NASA Astrophysics Data System (ADS)

Fisher, Walt G.; Piston, David W.; Wachter, Eric A.

2002-06-01

Multiphoton laser scanning microscopy offers advantages in depth of penetration into intact samples over other optical sectioning techniques. To achieve these advantages it is necessary to detect the emitted light without spatial filtering. In this nondescanned (nonconfocal) approach, ambient room light can easily contaminate the signal, forcing experiments to be performed in absolute darkness. For multiphoton microscope systems employing mode-locked lasers, signal processing can be used to reduce such problems by taking advantage of the pulsed characteristics of such lasers. Specifically, by recovering fluorescence generated at the mode-locked frequency, interference from stray light and other ambient noise sources can be significantly reduced. This technology can be adapted to existing microscopes by inserting demodulation circuitry between the detector and data collection system. The improvement in signal-to-noise ratio afforded by this approach yields a more robust microscope system and opens the possibility of moving multiphoton microscopy from the research lab to more demanding settings, such as the clinic.

Theory of a Quantum Scanning Microscope for Cold Atoms

NASA Astrophysics Data System (ADS)

Yang, D.; Laflamme, C.; Vasilyev, D. V.; Baranov, M. A.; Zoller, P.

2018-03-01

We propose and analyze a scanning microscope to monitor "live" the quantum dynamics of cold atoms in a cavity QED setup. The microscope measures the atomic density with subwavelength resolution via dispersive couplings to a cavity and homodyne detection within the framework of continuous measurement theory. We analyze two modes of operation. First, for a fixed focal point the microscope records the wave packet dynamics of atoms with time resolution set by the cavity lifetime. Second, a spatial scan of the microscope acts to map out the spatial density of stationary quantum states. Remarkably, in the latter case, for a good cavity limit, the microscope becomes an effective quantum nondemolition device, such that the spatial distribution of motional eigenstates can be measured backaction free in single scans, as an emergent quantum nondemolition measurement.

Theory of a Quantum Scanning Microscope for Cold Atoms.

PubMed

Yang, D; Laflamme, C; Vasilyev, D V; Baranov, M A; Zoller, P

2018-03-30

We propose and analyze a scanning microscope to monitor "live" the quantum dynamics of cold atoms in a cavity QED setup. The microscope measures the atomic density with subwavelength resolution via dispersive couplings to a cavity and homodyne detection within the framework of continuous measurement theory. We analyze two modes of operation. First, for a fixed focal point the microscope records the wave packet dynamics of atoms with time resolution set by the cavity lifetime. Second, a spatial scan of the microscope acts to map out the spatial density of stationary quantum states. Remarkably, in the latter case, for a good cavity limit, the microscope becomes an effective quantum nondemolition device, such that the spatial distribution of motional eigenstates can be measured backaction free in single scans, as an emergent quantum nondemolition measurement.

Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning of thick tissues

NASA Astrophysics Data System (ADS)

Cheng, Li-Chung; Chang, Chia-Yuan; Yen, Wei-Chung; Chen, Shean-Jen

2012-10-01

Conventional multiphoton microscopy employs beam scanning; however, in this study a microscope based on spatiotemporal focusing offering widefield multiphoton excitation has been developed to provide fast optical sectioning images. The microscope integrates a 10 kHz repetition rate ultrafast amplifier featuring strong instantaneous peak power (maximum 400 μJ/pulse at 90 fs pulse width) with a TE-cooled, ultra-sensitive photon detecting, electron multiplying charge-coupled device camera. This configuration can produce multiphoton excited images with an excitation area larger than 200 × 100 μm2 at a frame rate greater than 100 Hz. Brownian motions of fluorescent microbeads as small as 0.5 μm have been instantaneously observed with a lateral spatial resolution of less than 0.5 μm and an axial resolution of approximately 3.5 μm. Moreover, we combine the widefield multiphoton microscopy with structure illuminated technique named HiLo to reject the background scattering noise to get better quality for bioimaging.

Low-power noncontact photoacoustic microscope for bioimaging applications

NASA Astrophysics Data System (ADS)

Sathiyamoorthy, Krishnan; Strohm, Eric M.; Kolios, Michael C.

2017-04-01

An inexpensive noncontact photoacoustic (PA) imaging system using a low-power continuous wave laser and a kilohertz-range microphone has been developed. The system operates in both optical and PA imaging modes and is designed to be compatible with conventional optical microscopes. Aqueous coupling fluids are not required for the detection of the PA signals; air is used as the coupling medium. The main component of the PA system is a custom designed PA imaging sensor that consists of an air-filled sample chamber and a resonator chamber that isolates a standard kilohertz frequency microphone from the input laser. A sample to be examined is placed on the glass substrate inside the chamber. A laser focused to a small spot by a 40× objective onto the substrate enables generation of PA signals from the sample. Raster scanning the laser over the sample with micrometer-sized steps enables high-resolution PA images to be generated. A lateral resolution of 1.37 μm was achieved in this proof of concept study, which can be further improved using a higher numerical aperture objective. The application of the system was investigated on a red blood cell, with a noise-equivalent detection sensitivity of 43,887 hemoglobin molecules (72.88×10-21 mol or 72.88 zeptomol). The minimum pressure detectable limit of the system was 19.1 μPa. This inexpensive, compact noncontact PA sensor is easily integrated with existing commercial optical microscopes, enabling optical and PA imaging of the same sample. Applications include forensic measurements, blood coagulation tests, and monitoring the penetration of drugs into human membrane.

Apertureless near-field optical microscopy

NASA Astrophysics Data System (ADS)

Kazantsev, D. V.; Kuznetsov, E. V.; Timofeev, S. V.; Shelaev, A. V.; Kazantseva, E. A.

2017-05-01

We discuss the operating principles of the apertureless scanning near-field optical microscope (ASNOM), in which the probe acts as a rod antenna and its electromagnetic radiation plays the role of the registered signal. The phase and amplitude of the emitted wave vary depending on the ‘grounding conditions’ of the antenna tip at the sample point under study. Weak radiation from a tiny (2-15 μm long) tip is detected using optical homo- and heterodyning and the nonlinear dependence of the tip polarizability on the tip-surface distance. The lateral resolution of ASNOMs is determined by the tip curvature radius (1- 20 nm), regardless of the wavelength (500 nm-100 μm). ASNOMs are shown to be capable of providing a surface optical map with nanometer resolution and carrying out spectral- and time-resolved measurements at a selected point on the surface.

Near- and far-field spectroscopic imaging investigation of resonant square-loop infrared metasurfaces.

PubMed

D' Archangel, Jeffrey; Tucker, Eric; Kinzel, Ed; Muller, Eric A; Bechtel, Hans A; Martin, Michael C; Raschke, Markus B; Boreman, Glenn

2013-07-15

Optical metamaterials have unique properties which result from geometric confinement of the optical conductivity. We developed a series of infrared metasurfaces based on an array of metallic square loop antennas. The far-field absorption spectrum can be designed with resonances across the infrared by scaling the geometric dimensions. We measure the amplitude and phase of the resonant mode as standing wave patterns within the square loops using scattering-scanning near-field optical microscopy (s-SNOM). Further, using a broad-band synchrotron-based FTIR microscope and s-SNOM at the Advanced Light Source, we are able to correlate far-field spectra to near-field modes of the metasurface as the resonance is tuned between samples. The results highlight the importance of multi-modal imaging for the design and characterization of optical metamaterials.

Development and experimental testing of an optical micro-spectroscopic technique incorporating true line-scan excitation.

PubMed

Biener, Gabriel; Stoneman, Michael R; Acbas, Gheorghe; Holz, Jessica D; Orlova, Marianna; Komarova, Liudmila; Kuchin, Sergei; Raicu, Valerică

2013-12-27

Multiphoton micro-spectroscopy, employing diffraction optics and electron-multiplying CCD (EMCCD) cameras, is a suitable method for determining protein complex stoichiometry, quaternary structure, and spatial distribution in living cells using Förster resonance energy transfer (FRET) imaging. The method provides highly resolved spectra of molecules or molecular complexes at each image pixel, and it does so on a timescale shorter than that of molecular diffusion, which scrambles the spectral information. Acquisition of an entire spectrally resolved image, however, is slower than that of broad-bandwidth microscopes because it takes longer times to collect the same number of photons at each emission wavelength as in a broad bandwidth. Here, we demonstrate an optical micro-spectroscopic scheme that employs a laser beam shaped into a line to excite in parallel multiple sample voxels. The method presents dramatically increased sensitivity and/or acquisition speed and, at the same time, has excellent spatial and spectral resolution, similar to point-scan configurations. When applied to FRET imaging using an oligomeric FRET construct expressed in living cells and consisting of a FRET acceptor linked to three donors, the technique based on line-shaped excitation provides higher accuracy compared to the point-scan approach, and it reduces artifacts caused by photobleaching and other undesired photophysical effects.

Surface plasmon resonance and nonlinear optical behavior of pulsed laser-deposited semitransparent nanostructured copper thin films

NASA Astrophysics Data System (ADS)

Kesarwani, Rahul; Khare, Alika

2018-06-01

In this paper, surface plasmon resonance (SPR) and nonlinear optical properties of semitransparent nanostructured copper thin films fabricated on the glass substrate at 400 °C by pulsed laser deposition technique are reported. The thickness, linear absorption coefficient and linear refractive index of the films were measured by spectroscopic ellipsometer. The average particle size as measured via atomic force microscope was in the range of 12.84-26.02 nm for the deposition time ranging from 5 to 10 min, respectively. X-ray diffraction spectra revealed the formation of Cu (111) and Cu (200) planes. All these thin films exhibited broad SPR peak. The third-order optical nonlinearity of all the samples was investigated via modified z-scan technique using cw laser at a wavelength of 632.8 nm. The open aperture z-scan spectra of Cu thin film deposited for 5 min duration exhibited reverse saturation absorption whereas all the other samples displayed saturation absorption behavior. The nonlinear refractive index coefficient of these films showed a positive sign having the magnitude of the order of 10- 4 cm/W. The real and imaginary parts of susceptibilities were also calculated from the z-scan data and found to be of the order of 10- 6 esu.

Synthesis of poly glycidylmethacrylate grafted azobenzene copolymer: Photosensitivity and nonlinear optical properties

NASA Astrophysics Data System (ADS)

Sousani, Abbas; Moghadam, Peyman Najafi; Hasanzadeh, Reza; Motiei, Hamideh; Bagheri, Massoumeh

2016-01-01

In this work poly glycidylmethacrylate grafted 4-hydroxy-4‧-methoxy-azobenzene (Azo-PGMA) was synthesized. For this propose firstly 4-hydroxy-4‧-methoxy-azobenzene (AZO) was prepared, then poly glycidylmethacrylate was prepared by free radical polymerization of glycidylmethacrylate in the presence of benzoyl peroxide as initiator under inert atmosphere in dry THF. Finally the homopolymer was functionalized by AZO moieties. The characterization of the synthesized copolymer was carried out by 1H NMR, FT-IR, thermal gravimetric analyze (TGA), differential scanning calorimetry (DSC) and optical polarizing microscope (POM) analysis. The UV-vis studies were carried out on Azo-PGMA copolymer and the results showed that the synthesized Azo-PGMA copolymer has ultra-fast response to UV light and has slow relaxation time. Also the third-order nonlinear optical properties of the Azo-PGMA copolymer and AZO were studied by using Z-scan technique. Nonlinear refraction and absorption coefficients of the above mentioned materials were measured by the closed and open aperture Z-scan method using a continuous wave Nd-YAG laser at 532 nm. The positive nonlinear absorption in Azo-PGMA and AZO was investigated at the wavelength of λ = 532 nm, respectively and the measured values of nonlinear refraction in both of the samples were from the order of 10-8 cm2/W.

Laser Scanning In Inspection

NASA Astrophysics Data System (ADS)

West, Patricia; Baker, Lionel R.

1989-03-01

This paper is a review of the applications of laser scanning in inspection. The reasons for the choice of a laser in flying spot scanning and the optical properties of a laser beam which are of value in a scanning instrument will be given. The many methods of scanning laser beams in both one and two dimensions will be described. The use of one dimensional laser scanners for automatic surface inspection for transmitting and reflective products will be covered in detail, with particular emphasis on light collection techniques. On-line inspection applications which will be mentioned include: photographic film web, metal strip products, paper web, glass sheet, car body paint surfaces and internal cylinder bores. Two dimensional laser scanning is employed in applications where increased resolution, increased depth of focus, and better contrast are required compared with conventional vidicon TV or solid state array cameras. Such examples as special microscope laser scanning systems and a TV compatible system for use in restricted areas of a nuclear reactor will be described. The technical and economic benefits and limitations of laser scanning video systems will be compared with conventional TV and CCD array devices.

Eliminating Unwanted Far-Field Excitation in Objective-Type TIRF. Part I. Identifying Sources of Nonevanescent Excitation Light

PubMed Central

Brunstein, Maia; Teremetz, Maxime; Hérault, Karine; Tourain, Christophe; Oheim, Martin

2014-01-01

Total internal reflection fluorescence microscopy (TIRFM) achieves subdiffraction axial sectioning by confining fluorophore excitation to a thin layer close to the cell/substrate boundary. However, it is often unknown how thin this light sheet actually is. Particularly in objective-type TIRFM, large deviations from the exponential intensity decay expected for pure evanescence have been reported. Nonevanescent excitation light diminishes the optical sectioning effect, reduces contrast, and renders TIRFM-image quantification uncertain. To identify the sources of this unwanted fluorescence excitation in deeper sample layers, we here combine azimuthal and polar beam scanning (spinning TIRF), atomic force microscopy, and wavefront analysis of beams passing through the objective periphery. Using a variety of intracellular fluorescent labels as well as negative staining experiments to measure cell-induced scattering, we find that azimuthal beam spinning produces TIRFM images that more accurately portray the real fluorophore distribution, but these images are still hampered by far-field excitation. Furthermore, although clearly measureable, cell-induced scattering is not the dominant source of far-field excitation light in objective-type TIRF, at least for most types of weakly scattering cells. It is the microscope illumination optical path that produces a large cell- and beam-angle invariant stray excitation that is insensitive to beam scanning. This instrument-induced glare is produced far from the sample plane, inside the microscope illumination optical path. We identify stray reflections and high-numerical aperture aberrations of the TIRF objective as one important source. This work is accompanied by a companion paper (Pt.2/2). PMID:24606927

Video-rate volumetric functional imaging of the brain at synaptic resolution.

PubMed

Lu, Rongwen; Sun, Wenzhi; Liang, Yajie; Kerlin, Aaron; Bierfeld, Jens; Seelig, Johannes D; Wilson, Daniel E; Scholl, Benjamin; Mohar, Boaz; Tanimoto, Masashi; Koyama, Minoru; Fitzpatrick, David; Orger, Michael B; Ji, Na

2017-04-01

Neurons and neural networks often extend hundreds of micrometers in three dimensions. Capturing the calcium transients associated with their activity requires volume imaging methods with subsecond temporal resolution. Such speed is a challenge for conventional two-photon laser-scanning microscopy, because it depends on serial focal scanning in 3D and indicators with limited brightness. Here we present an optical module that is easily integrated into standard two-photon laser-scanning microscopes to generate an axially elongated Bessel focus, which when scanned in 2D turns frame rate into volume rate. We demonstrated the power of this approach in enabling discoveries for neurobiology by imaging the calcium dynamics of volumes of neurons and synapses in fruit flies, zebrafish larvae, mice and ferrets in vivo. Calcium signals in objects as small as dendritic spines could be resolved at video rates, provided that the samples were sparsely labeled to limit overlap in their axially projected images.

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

NASA Astrophysics Data System (ADS)

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.

The Optical Properties of Ion Implanted Silica

NASA Technical Reports Server (NTRS)

Smith, Cydale C.; Ila, D.; Sarkisov, S.; Williams, E. K.; Poker, D. B.; Hensley, D. K.

1997-01-01

We will present our investigation on the change in the optical properties of silica, 'suprasil', after keV through MeV implantation of copper, tin, silver and gold and after annealing. Suprasil-1, name brand of silica glass produced by Hereaus Amerisil, which is chemically pure with well known optical properties. Both linear nonlinear optical properties of the implanted silica were investigated before and after thermal annealing. All implants, except for Sn, showed strong optical absorption bands in agreement with Mie's theory. We have also used Z-scan to measure the strength of the third order nonlinear optical properties of the produced thin films, which is composed of the host material and the metallic nanoclusters. For implants with a measurable optical absorption band we used Doyle's theory and the full width half maximum of the absorption band to calculate the predicted size of the formed nanoclusters at various heat treatment temperatures. These results are compared with those obtained from direct observation using transmission electron microscopic techniques.

Purchase of a Laser Scanning Confocal Microscope at Xavier University of Louisiana

DTIC Science & Technology

2016-05-04

SECURITY CLASSIFICATION OF: The purpose of this grant was to purchase a laser scanning confocal microscope to be used by multiple laboratories at...was being developed for undergraduate education. Over the course of the funding period, the microscope was purchased and installed, multiple training...Distribution Unlimited UU UU UU UU 04-05-2016 1-Feb-2015 31-Jan-2016 Final Report: Purchase of a Laser Scanning Confocal Microscope at Xavier

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sato, Chikara, E-mail: ti-sato@aist.go.jp; Manaka, Sachie; Nakane, Daisuke

Highlights: Black-Right-Pointing-Pointer Mycoplasma mobile was observed in buffer with the Atmospheric Scanning Electron Microscope. Black-Right-Pointing-Pointer Characteristic protein localizations were visualized using immuno-labeling. Black-Right-Pointing-Pointer M. mobile attached to sialic acid on the SiN film surface within minutes. Black-Right-Pointing-Pointer Cells were observed at low concentrations. Black-Right-Pointing-Pointer ASEM should promote study and early-stage diagnosis of mycoplasma. -- Abstract: Mycoplasma is a genus of bacterial pathogen that causes disease in vertebrates. In humans, the species Mycoplasma pneumoniae causes 15% or more of community-acquired pneumonia. Because this bacterium is tiny, corresponding in size to a large virus, diagnosis using optical microscopy is not easy. Inmore » current methods, chest X-rays are usually the first action, followed by serology, PCR amplification, and/or culture, but all of these are particularly difficult at an early stage of the disease. Using Mycoplasma mobile as a model species, we directly observed mycoplasma in buffer with the newly developed Atmospheric Scanning Electron Microscope (ASEM). This microscope features an open sample dish with a pressure-resistant thin film window in its base, through which the SEM beam scans samples in solution, from below. Because of its 2-3 {mu}m-deep scanning capability, it can observe the whole internal structure of mycoplasma cells stained with metal solutions. Characteristic protein localizations were visualized using immuno-labeling. Cells were observed at low concentrations, because suspended cells concentrate in the observable zone by attaching to sialic acid on the silicon nitride (SiN) film surface within minutes. These results suggest the applicability of the ASEM for the study of mycoplasmas as well as for early-stage mycoplasma infection diagnosis.« less

An interchangeable scanning Hall probe/scanning SQUID microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tang, Chiu-Chun; Lin, Hui-Ting; Wu, Sing-Lin

2014-08-15

We have constructed a scanning probe microscope for magnetic imaging, which can function as a scanning Hall probe microscope (SHPM) and as a scanning SQUID microscope (SSM). The scanning scheme, applicable to SHPM and SSM, consists of a mechanical positioning (sub) micron-XY stage and a flexible direct contact to the sample without a feedback control system for the Z-axis. With the interchangeable capability of operating two distinct scanning modes, our microscope can incorporate the advantageous functionalities of the SHPM and SSM with large scan range up to millimeter, high spatial resolution (⩽4 μm), and high field sensitivity in a widemore » range of temperature (4.2 K-300 K) and magnetic field (10{sup −7} T-1 T). To demonstrate the capabilities of the system, we present magnetic images scanned with SHPM and SSM, including a RbFeB magnet and a nickel grid pattern at room temperature, surface magnetic domain structures of a La{sub 2/3}Ca{sub 1/3}MnO{sub 3} thin film at 77 K, and superconducting vortices in a striped niobium film at 4.2 K.« less

Negatively-chirped laser enables nonlinear excitation and nanoprocessing with sub-20-fs pulses

NASA Astrophysics Data System (ADS)

Uchugonova, A.; Müller, J.; Bückle, R.; Tempea, G.; Isemann, A.; Stingl, A.; König, K.

2008-02-01

It has long been considered that the advantages emerging from employing chirp pre-compensation in nonlinear microscopy were overweighed by the complexity of prism- or grating-based compressors. These concerns were refuted with the advent of dispersive-mirrors-based compressors that are compact, user-friendly and sufficiently accurate to support sub-20-fs pulse delivery. Recent advances in the design of dispersive multilayer mirrors resulted in improved bandwidth (covering now as much as half of the gain bandwidth of Ti:Sapphire) and increased dispersion per bounce (one reflection off a state-of-the-art dispersive mirror pre-compensates the dispersion corresponding to >10mm of glass). The compressor built with these mirrors is sufficiently compact to be integrated in the housing of a sub-12-fs Ti:Sapphire oscillator. A complete scanning nonlinear microscope (FemtOgene, JenLab GmbH) equipped with highly-dispersive, large-NA objectives (Zeiss EC Plan-Neofluoar 40x/1.3, Plan-Neofluar 63x/1,25 Oil) was directly seeded with this negatively chirped laser. The pulse duration was measured at the focus of the objectives by inserting a scanning autocorrelator in the beam path between the laser and the microscope and recording the second order interferometric autocorrelation traces with the detector integrated in the microscope. Pulse durations <20fs were measured with both objectives. The system has been applied for two-photon imaging, transfection and optical manipulation of stem cells. Here we report on the successful transfection of human stem cells by transient optoporation of the cell membrane with a low mean power of < 7 mW and a short μs beam dwell time. Optically transfected cells were able to reproduce. The daughter cell expressed also green fluorescent proteins (GFP) indicating the successful modification of the cellular DNA.

Image-guided feedback for ophthalmic microsurgery using multimodal intraoperative swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography

NASA Astrophysics Data System (ADS)

Li, Jianwei D.; Malone, Joseph D.; El-Haddad, Mohamed T.; Arquitola, Amber M.; Joos, Karen M.; Patel, Shriji N.; Tao, Yuankai K.

2017-02-01

Surgical interventions for ocular diseases involve manipulations of semi-transparent structures in the eye, but limited visualization of these tissue layers remains a critical barrier to developing novel surgical techniques and improving clinical outcomes. We addressed limitations in image-guided ophthalmic microsurgery by using microscope-integrated multimodal intraoperative swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography (iSS-SESLO-OCT). We previously demonstrated in vivo human ophthalmic imaging using SS-SESLO-OCT, which enabled simultaneous acquisition of en face SESLO images with every OCT cross-section. Here, we integrated our new 400 kHz iSS-SESLO-OCT, which used a buffered Axsun 1060 nm swept-source, with a surgical microscope and TrueVision stereoscopic viewing system to provide image-based feedback. In vivo human imaging performance was demonstrated on a healthy volunteer, and simulated surgical maneuvers were performed in ex vivo porcine eyes. Denselysampled static volumes and volumes subsampled at 10 volumes-per-second were used to visualize tissue deformations and surgical dynamics during corneal sweeps, compressions, and dissections, and retinal sweeps, compressions, and elevations. En face SESLO images enabled orientation and co-registration with the widefield surgical microscope view while OCT imaging enabled depth-resolved visualization of surgical instrument positions relative to anatomic structures-of-interest. TrueVision heads-up display allowed for side-by-side viewing of the surgical field with SESLO and OCT previews for real-time feedback, and we demonstrated novel integrated segmentation overlays for augmented-reality surgical guidance. Integration of these complementary imaging modalities may benefit surgical outcomes by enabling real-time intraoperative visualization of surgical plans, instrument positions, tissue deformations, and image-based surrogate biomarkers correlated with completion of surgical goals.

Electron Beam/Laser Glazing of Iron-Base Materials.

DTIC Science & Technology

1981-07-01

alloy (-l. 5wt %Cr) steels after laser and electron beam glazing. In this work it is shown that the dramatic difference in microstructure and hardness...highly alloyed tool steels can be critical in determining the complexity of the solidification route. The analyses of M2, M42 and M7 are given in...the type described in Fe- Ni alloys (1). This con- clusion is based on optical and scanning electron microscope observation unambig- uously showing

Microanalysis of tool steel and glass with laser-induced breakdown spectroscopy

NASA Astrophysics Data System (ADS)

Loebe, Klaus; Uhl, Arnold; Lucht, Hartmut

2003-10-01

A laser microscope system for the microanalytical characterization of complex materials is described. The universal measuring principle of laser-induced breakdown spectroscopy (LIBS) in combination with echelle optics permits a fast simultaneous multielement analysis with a possible spatial resolution below 10 pm. The developed system features completely UV-transparent optics for the laser-microscope coupling and the emission beam path and enables parallel signal detection within the wavelength range of 200-800 nm with a spectral resolution of a few picometers. Investigations of glass defects and tool steels were performed. The characterization of a glass defect in a tumbler by a micro-LIBS line scan, with use of a 266-nm diode-pumped Nd:YAG laser for excitation, is possible by simple comparison of plasma spectra of the defect and the surrounding area. Variations in the main elemental composition as well as impurities by trace elements are detected at the same time. Through measurement of the calibration samples with the known concentration of the corresponding element, a correlation between the intensity of spectral lines and the element concentration was also achieved. The change of elemental composition at the transient stellite solder of tool steels has been determined by an area scan. The two-dimensional pictures show abrupt changes of the element distribution along the solder edge and allow fundamental researches of dynamic modifications (e.g., diffusion) in steel.

Seeing with the nano-eye: accessing structure, function, and dynamics of matter on its natural length and time scales

NASA Astrophysics Data System (ADS)

Raschke, Markus

2015-03-01

To understand and ultimately control the properties of most functional materials, from molecular soft-matter to quantum materials, requires access to the structure, coupling, and dynamics on the elementary time and length scales that define the microscopic interactions in these materials. To gain the desired nanometer spatial resolution with simultaneous spectroscopic specificity we combine scanning probe microscopy with different optical, including coherent, nonlinear, and ultrafast spectroscopies. The underlying near-field interaction mediated by the atomic-force or scanning tunneling microscope tip provides the desired deep-sub wavelength nano-focusing enabling few-nm spatial resolution. I will introduce our generalization of the approach in terms of the near-field impedance matching to a quantum system based on special optical antenna-tip designs. The resulting enhanced and qualitatively new forms of light-matter interaction enable measurements of quantum dynamics in an interacting environment or to image the electromagnetic local density of states of thermal radiation. Other applications include the inter-molecular coupling and dynamics in soft-matter hetero-structures, surface plasmon interferometry as a probe of electronic structure and dynamics in graphene, and quantum phase transitions in correlated electron materials. These examples highlight the general applicability of the new near-field microscopy approach, complementing emergent X-ray and electron imaging tools, aiming towards the ultimate goal of probing matter on its most elementary spatio-temporal level.

Integrative Advances for OCT-Guided Ophthalmic Surgery and Intraoperative OCT: Microscope Integration, Surgical Instrumentation, and Heads-Up Display Surgeon Feedback

PubMed Central

Ehlers, Justis P.; Srivastava, Sunil K.; Feiler, Daniel; Noonan, Amanda I.; Rollins, Andrew M.; Tao, Yuankai K.

2014-01-01

Purpose To demonstrate key integrative advances in microscope-integrated intraoperative optical coherence tomography (iOCT) technology that will facilitate adoption and utilization during ophthalmic surgery. Methods We developed a second-generation prototype microscope-integrated iOCT system that interfaces directly with a standard ophthalmic surgical microscope. Novel features for improved design and functionality included improved profile and ergonomics, as well as a tunable lens system for optimized image quality and heads-up display (HUD) system for surgeon feedback. Novel material testing was performed for potential suitability for OCT-compatible instrumentation based on light scattering and transmission characteristics. Prototype surgical instruments were developed based on material testing and tested using the microscope-integrated iOCT system. Several surgical maneuvers were performed and imaged, and surgical motion visualization was evaluated with a unique scanning and image processing protocol. Results High-resolution images were successfully obtained with the microscope-integrated iOCT system with HUD feedback. Six semi-transparent materials were characterized to determine their attenuation coefficients and scatter density with an 830 nm OCT light source. Based on these optical properties, polycarbonate was selected as a material substrate for prototype instrument construction. A surgical pick, retinal forceps, and corneal needle were constructed with semi-transparent materials. Excellent visualization of both the underlying tissues and surgical instrument were achieved on OCT cross-section. Using model eyes, various surgical maneuvers were visualized, including membrane peeling, vessel manipulation, cannulation of the subretinal space, subretinal intraocular foreign body removal, and corneal penetration. Conclusions Significant iterative improvements in integrative technology related to iOCT and ophthalmic surgery are demonstrated. PMID:25141340

Laser Cladding of Ti-6Al-4V Alloy with Ti-Al2O3 Coating for Biomedical Applications

NASA Astrophysics Data System (ADS)

Mthisi, A.; Popoola, A. P. I.; Adebiyi, D. I.; Popoola, O. M.

2018-05-01

The indispensable properties of Ti-6Al-4V alloy coupled with poor tribological properties and delayed bioactivity make it a subject of interest to explore in biomedical application. A quite number of numerous coatings have been employed on titanium alloys, with aim to overcome the poor properties exhibited by this alloy. In this work, the possibility of laser cladding different ad-mixed powders (Ti - 5 wt.% Al2O3 and Ti - 8wt.% Al2O3) on Ti-6Al-4V at various laser scan speed (0.6 and 0.8 m/min) were investigated. The microstructure, phase constituents and corrosion of the resultant coatings were characterized by scanning electron microscope (SEM), Optical microscope, X-Ray diffractometer (XRD) and potentiostat respectively. The electrochemical behaviour of the produced coatings was studied in a simulated body fluid (Hanks solution). The microstructural results show that a defect free coating is achieved at low scan speed and ad-mixed of Ti-5 wt. % Al2O3. Cladding of Ti - Al2O3 improved the corrosion resistance of Ti-6Al-4V alloy regardless of varying neither scan speed nor ad-mixed percentage. However, Ti-5 wt.% Al2O3 coating produced at low scan speed revealed the highest corrosion resistance among the coatings due to better quality coating layer. Henceforth, this coating may be suitable for biomedical applications.

Dark-field microscopic image stitching method for surface defects evaluation of large fine optics.

PubMed

Liu, Dong; Wang, Shitong; Cao, Pin; Li, Lu; Cheng, Zhongtao; Gao, Xin; Yang, Yongying

2013-03-11

One of the challenges in surface defects evaluation of large fine optics is to detect defects of microns on surfaces of tens or hundreds of millimeters. Sub-aperture scanning and stitching is considered to be a practical and efficient method. But since there are usually few defects on the large aperture fine optics, resulting in no defects or only one run-through line feature in many sub-aperture images, traditional stitching methods encounter with mismatch problem. In this paper, a feature-based multi-cycle image stitching algorithm is proposed to solve the problem. The overlapping areas of sub-apertures are categorized based on the features they contain. Different types of overlapping areas are then stitched in different cycles with different methods. The stitching trace is changed to follow the one that determined by the features. The whole stitching procedure is a region-growing like process. Sub-aperture blocks grow bigger after each cycle and finally the full aperture image is obtained. Comparison experiment shows that the proposed method is very suitable to stitch sub-apertures that very few feature information exists in the overlapping areas and can stitch the dark-field microscopic sub-aperture images very well.

Mirrorlike pulsed laser deposited tungsten thin film.

PubMed

Mostako, A T T; Rao, C V S; Khare, Alika

2011-01-01

Mirrorlike tungsten thin films on stainless steel substrate deposited via pulsed laser deposition technique in vacuum (10(-5) Torr) is reported, which may find direct application as first mirror in fusion devices. The crystal structure of tungsten film is analyzed using x-ray diffraction pattern, surface morphology of the tungsten films is studied with scanning electron microscope and atomic force microscope. The film composition is identified using energy dispersive x-ray. The specular and diffuse reflectivities with respect to stainless steel substrate of the tungsten films are recorded with FTIR spectra. The thickness and the optical quality of pulsed laser deposition deposited films are tested via interferometric technique. The reflectivity is approaching about that of the bulk for the tungsten film of thickness ∼782 nm.

Effects of Laser Shock Processing on Morphologies and Mechanical Properties of ANSI 304 Stainless Steel Weldments Subjected to Cavitation Erosion

PubMed Central

Zhang, Lei; Lu, Jin-Zhong; Zhang, Yong-Kang; Ma, Hai-Le; Luo, Kai-Yu; Dai, Feng-Ze

2017-01-01

Effects of laser shock processing (LSP) on the cavitation erosion resistance of laser weldments were investigated by optical microscope (OM), scanning electron microscope (SEM) observations, roughness tester, micro hardness tester, and X-ray diffraction (XRD) technology. The morphological microstructures were characterized. Cumulative mass loss, incubation period, erosion rate, and damaged surface areas were monitored during cavitation erosion. Surface roughness, micro-hardness, and residual stress were measured in different zones. Results showed that LSP could improve the damage of morphological microstructures and mechanical properties after cavitation erosion. The compressive residual stresses were generated during the process of LSP, which was an effective guarantee for the improvement of the above mentioned properties. PMID:28772652

Microstructure and wear property of Fe-Cr13-C hardfacing alloy reinforced by WC particles

NASA Astrophysics Data System (ADS)

Yang, Ke; Li, Jiaqi; Bao, Yefeng; Jiang, Yongfeng

2017-07-01

Tungsten as the most effective carbide-forming element was added in the Fe-Cr13-C hardfacing alloy to precipitate WC particles. Optical microscope (OM), scanning electron microscope (SEM) and energy-dispersive spectrometer (EDS) were used to investigate the microstructures of the hardfacing alloy. The wear resistance was tested through a slurry rubber wheel abrasion test machine, and the wear behavior was also studied. The results indicate that the microstructures of the hardfacing alloy consist of lath martensite, residual austenite and WC particles. The wear resistance can be significantly improved through the addition of tungsten element being provided by the precipitation of WC particles. And the predominant wear mechanism was microcutting with shallow grooves and spalling.

A cell-free assay to determine the stoichiometry of plasma membrane proteins.

PubMed

Trigo, Cesar; Vivar, Juan P; Gonzalez, Carlos B; Brauchi, Sebastian

2013-04-01

Plasma membrane receptors, transporters, and ion channel molecules are often found as oligomeric structures that participate in signaling cascades essential for cell survival. Different states of protein oligomerization may play a role in functional control and allosteric regulation. Stochastic GFP-photobleaching (SGP) has emerged as an affordable and simple method to determine the stoichiometry of proteins at the plasma membrane. This non-invasive optical approach can be useful for total internal reflection of fluorescence microscopy (TIRFM), where signal-to-noise ratio is very high at the plasma membrane. Here, we report an alternative methodology implemented on a standard laser scanning confocal microscope (LSCM). The simplicity of our method will allow for its implementation in any epifluorescence microscope of choice.

Motion compensation for in vivo subcellular optical microscopy.

PubMed

Lucotte, B; Balaban, R S

2014-04-01

In this review, we focus on the impact of tissue motion on attempting to conduct subcellular resolution optical microscopy, in vivo. Our position is that tissue motion is one of the major barriers in conducting these studies along with light induced damage, optical probe loading as well as absorbing and scattering effects on the excitation point spread function and collection of emitted light. Recent developments in the speed of image acquisition have reached the limit, in most cases, where the signal from a subcellular voxel limits the speed and not the scanning rate of the microscope. Different schemes for compensating for tissue displacements due to rigid body and deformation are presented from tissue restriction, gating, adaptive gating and active tissue tracking. We argue that methods that minimally impact the natural physiological motion of the tissue are desirable because the major reason to perform in vivo studies is to evaluate normal physiological functions. Towards this goal, active tracking using the optical imaging data itself to monitor tissue displacement and either prospectively or retrospectively correct for the motion without affecting physiological processes is desirable. Critical for this development was the implementation of near real time image processing in conjunction with the control of the microscope imaging parameters. Clearly, the continuing development of methods of motion compensation as well as significant technological solutions to the other barriers to tissue subcellular optical imaging in vivo, including optical aberrations and overall signal-to-noise ratio, will make major contributions to the understanding of cell biology within the body.

Compact, single-tube scanning tunneling microscope with thermoelectric cooling.

PubMed

Jobbins, Matthew M; Agostino, Christopher J; Michel, Jolai D; Gans, Ashley R; Kandel, S Alex

2013-10-01

We have designed and built a scanning tunneling microscope with a compact inertial-approach mechanism that fits inside the piezoelectric scanner tube. Rigid construction allows the microscope to be operated without the use of external vibration isolators or acoustic enclosures. Thermoelectric cooling and a water-ice bath are used to increase temperature stability when scanning under ambient conditions.

Surface topography characterization using 3D stereoscopic reconstruction of SEM images

NASA Astrophysics Data System (ADS)

Vedantha Krishna, Amogh; Flys, Olena; Reddy, Vijeth V.; Rosén, B. G.

2018-06-01

A major drawback of the optical microscope is its limitation to resolve finer details. Many microscopes have been developed to overcome the limitations set by the diffraction of visible light. The scanning electron microscope (SEM) is one such alternative: it uses electrons for imaging, which have much smaller wavelength than photons. As a result high magnification with superior image resolution can be achieved. However, SEM generates 2D images which provide limited data for surface measurements and analysis. Often many research areas require the knowledge of 3D structures as they contribute to a comprehensive understanding of microstructure by allowing effective measurements and qualitative visualization of the samples under study. For this reason, stereo photogrammetry technique is employed to convert SEM images into 3D measurable data. This paper aims to utilize a stereoscopic reconstruction technique as a reliable method for characterization of surface topography. Reconstructed results from SEM images are compared with coherence scanning interferometer (CSI) results obtained by measuring a roughness reference standard sample. This paper presents a method to select the most robust/consistent surface texture parameters that are insensitive to the uncertainties involved in the reconstruction technique itself. Results from the two-stereoscopic reconstruction algorithms are also documented in this paper.

High-contrast 3D image acquisition using HiLo microscopy with an electrically tunable lens

NASA Astrophysics Data System (ADS)

Philipp, Katrin; Smolarski, André; Fischer, Andreas; Koukourakis, Nektarios; Stürmer, Moritz; Wallrabe, Ulricke; Czarske, Jürgen

2016-04-01

We present a HiLo microscope with an electrically tunable lens for high-contrast three-dimensional image acquisition. HiLo microscopy combines wide field and speckled illumination images to create optically sectioned images. Additionally, the depth-of-field is not fixed, but can be adjusted between wide field and confocal-like axial resolution. We incorporate an electrically tunable lens in the HiLo microscope for axial scanning, to obtain three-dimensional data without the need of moving neither the sample nor the objective. The used adaptive lens consists of a transparent polydimethylsiloxane (PDMS) membrane into which an annular piezo bending actuator is embedded. A transparent fluid is filled between the membrane and the glass substrate. When actuated, the piezo generates a pressure in the lens which deflects the membrane and thus changes the refractive power. This technique enables a large tuning range of the refractive power between 1/f = (-24 . . . 25) 1/m. As the NA of the adaptive lens is only about 0.05, a fixed high-NA lens is included in the setup to provide high resolution. In this contribution, the scan properties and capabilities of the tunable lens in the HiLo microscope are analyzed. Eventually, exemplary measurements are presented and discussed.

A STED-FLIM microscope applied to imaging the natural killer cell immune synapse

NASA Astrophysics Data System (ADS)

Lenz, M. O.; Brown, A. C. N.; Auksorius, E.; Davis, D. M.; Dunsby, C.; Neil, M. A. A.; French, P. M. W.

2011-03-01

We present a stimulated emission depletion (STED) fluorescence lifetime imaging (FLIM) microscope, excited by a microstructured optical fibre supercontinuum source that is pumped by a femtosecond Ti:Sapphire-laser, which is also used for depletion. Implemented using a piezo-scanning stage on a laser scanning confocal fluorescence microscope system with FLIM realised using time correlated single photon counting (TCSPC), this provides convenient switching between confocal and STED-FLIM with spatial resolution down to below 60 nm. We will present our design considerations to make a robust instrument for biological applications including a comparison between fixed phase plate and spatial light modulator (SLM) approaches to shape the STED beam and the correlation of STED and confocal FLIM microscopy. Following our previous application of FLIM-FRET to study intercellular signalling at the immunological synapse (IS), we are employing STED microscopy to characterize the spatial distribution of cellular molecules with subdiffraction resolution at the IS. In particular, we are imaging cytoskeletal structure at the Natural Killer cell activated immune synapse. We will also present our progress towards multilabel STED microscopy to determine how relative spatial molecular organization, previously undetectable by conventional microscopy techniques, is important for NK cell cytotoxic function. Keywords: STED, Stimulated Emission Depletion Microscopy, Natural Killer (NK) cell, Fluorescence lifetime imaging, FLIM, Super resolution microscopy.

Imaging mouse cerebellum with serial optical coherence scanner (Conference Presentation)

NASA Astrophysics Data System (ADS)

Liu, Chao J.; Williams, Kristen; Orr, Harry; Taner, Akkin

2017-02-01

We present the serial optical coherence scanner (SOCS), which consists of a polarization sensitive optical coherence tomography and a vibratome with associated controls for serial imaging, to visualize the cerebellum and adjacent brainstem of mouse. The cerebellar cortical layers and white matter are distinguished by using intrinsic optical contrasts. Images from serial scans reveal the large-scale anatomy in detail and map the nerve fiber pathways in the cerebellum and adjacent brainstem. The optical system, which has 5.5 μm axial resolution, utilizes a scan lens or a water-immersion microscope objective resulting in 10 μm or 4 μm lateral resolution, respectively. The large-scale brain imaging at high resolution requires an efficient way to collect large datasets. It is important to improve the SOCS system to deal with large-scale and large number of samples in a reasonable time. The imaging and slicing procedure for a section took about 4 minutes due to a low speed of the vibratome blade to maintain slicing quality. SOCS has potential to investigate pathological changes and monitor the effects of therapeutic drugs in cerebellar diseases such as spinocerebellar ataxia 1 (SCA1). The SCA1 is a neurodegenerative disease characterized by atrophy and eventual loss of Purkinje cells from the cerebellar cortex, and the optical contrasts provided by SOCS is being evaluated for biomarkers of the disease.

Thickness dependent structural, optical and electrical properties of Se85In12Bi3 nanochalcogenide thin films

NASA Astrophysics Data System (ADS)

Tripathi, Ravi P.; Zulfequar, M.; Khan, Shamshad A.

2018-04-01

Our aim is to study the thickness dependent effects on structure, electrical and optical properties of Se85In12Bi3 nanochalcogenide thin films. Bulk alloy of Se85In12Bi3 was synthesized by melt-quenching technique. The amorphous as well as glassy nature of Se85In12Bi3 chalcogenide was confirmed by non-isothermal Differential Scanning Calorimetry (DSC) measurements. The nanochalcogenide thin films of thickness 30, 60 and 90 nm were prepared on glass/Si wafer substrate using Physical Vapour Condensation Technique (PVCT). From XRD studies it was found that thin films have amorphous texture. The surface morphology and particle size of films were studied by Field Emission Scanning Electron Microscope (FESEM). From optical studies, different optical parameters were estimated for Se85In12Bi3 thin films at different thickness. It was found that the absorption coefficient (α) and extinction coefficient (k) increases with photon energy and decreases with film thickness. The optical absorption process followed the rule of indirect transitions and optical band gap were found to be increase with film thickness. The value of Urbach energy (Et) and steepness parameter (σ) were also calculated for different film thickness. For electrical studies, dc-conductivity measurement was done at different temperature and activation energy (ΔEc) were determined and found to be increase with film thickness.

Development of large field-of-view two photon microscopy for imaging mouse cortex (Conference Presentation)

NASA Astrophysics Data System (ADS)

Bumstead, Jonathan; Côté, Daniel C.; Culver, Joseph P.

2017-02-01

Spontaneous neuronal activity has been measured at cellular resolution in mice, zebrafish, and C. elegans using optical sectioning microscopy techniques, such as light sheet microscopy (LSM) and two photon microscopy (TPM). Recent improvements in these modalities and genetically encoded calcium indicators (GECI's) have enabled whole brain imaging of calcium dynamics in zebrafish and C. elegans. However, these whole brain microscopy studies have not been extended to mice due to the limited field of view (FOV) of TPM and the cumbersome geometry of LSM. Conventional TPM is restricted to diffraction limited imaging over this small FOV (around 500 x 500 microns) due to the use of high magnification objectives (e.g. 1.0 NA; 20X) and the aberrations introduced by relay optics used in scanning the beam across the sample. To overcome these limitations, we have redesigned the entire optical path of the two photon microscope (scanning optics and objective lens) to support a field of view of Ø7 mm with relatively high spatial resolution (<10 microns). Using optical engineering software Zemax, we designed our system with commercially available optics that minimize astigmatism, field curvature, chromatic focal shift, and vignetting. Performance of the system was also tested experimentally with fluorescent beads in agarose, fixed samples, and in vivo structural imaging. Our large-FOV TPM provides a modality capable of studying distributed brain networks in mice at cellular resolution.

AOSLO: from benchtop to clinic

NASA Astrophysics Data System (ADS)

Zhang, Yuhua; Poonja, Siddharth; Roorda, Austin

2006-08-01

We present a clinically deployable adaptive optics scanning laser ophthalmoscope (AOSLO) that features micro-electro-mechanical (MEMS) deformable mirror (DM) based adaptive optics (AO) and low coherent light sources. With the miniaturized optical aperture of a μDMS-Multi TM MEMS DM (Boston Micromachines Corporation, Watertown, MA), we were able to develop a compact and robust AOSLO optical system that occupies a 50 cm X 50 cm area on a mobile optical table. We introduced low coherent light sources, which are superluminescent laser diodes (SLD) at 680 nm with 9 nm bandwidth and 840 nm with 50 nm bandwidth, in confocal scanning ophthalmoscopy to eliminate interference artifacts in the images. We selected a photo multiplier tube (PMT) for photon signal detection and designed low noise video signal conditioning circuits. We employed an acoustic-optical (AOM) spatial light modulator to modulate the light beam so that we could avoid unnecessary exposure to the retina or project a specific stimulus pattern onto the retina. The MEMS DM based AO system demonstrated robust performance. The use of low coherent light sources effectively mitigated the interference artifacts in the images and yielded high-fidelity retinal images of contiguous cone mosaic. We imaged patients with inherited retinal degenerations including cone-rod dystrophy (CRD) and retinitis pigmentosa (RP). We have produced high-fidelity, real-time, microscopic views of the living human retina for healthy and diseased eyes.

SIMS analysis of extended impact features on LDEF experiment

NASA Technical Reports Server (NTRS)

Amari, S.; Foote, J.; Jessberger, E. K.; Simon, C.; Stadermann, F. J.; Swan, P.; Walker, R.; Zinner, E.

1991-01-01

Discussed here are the first Secondary Ion Mass Spectroscopy (SIMS) analysis of projectile material deposited in extended impact features on Ge wafers from the trailing edge. Although most capture cells lost their plastic film covers, they contain extended impact features that apparently were produced by high velocity impacts when the plastic foils were still intact. Detailed optical scanning of all bare capture cells from the trailing edge revealed more than 100 impacts. Fifty-eight were selected by scanning electron microscope (SEM) inspection as prime candidates for SIMS analysis. Preliminary SIMS measurements were made on 15 impacts. More than half showed substantial enhancements of Mg, Al, Si, Ca, and Fe in the impact region, indicating micrometeorites as the projectiles.

From Single Atoms to Nanoparticles — Spectroscopy on the Atomic Level

NASA Astrophysics Data System (ADS)

Nilius, Niklas

2003-12-01

The scanning tunneling microscope is not only a well-established tool for a topographic characterization of the sample surface on the atomic scale. It also provides a variety of spectroscopic techniques to examine electronic, magnetic, vibrational and optical properties of a localized system. The following presentation gives an overview, how scanning tunneling spectroscopy, inelastic electron tunneling spectroscopy and photon emission spectroscopy with the STM can be employed to investigate spatially confined metal systems and their interaction with molecular gases. The experiments were performed on single Pd and Au atoms, mono-atomic chains and individual Ag clusters on a NiAl support and a Al2O3 thin film.

Optical coherence tomography - principles and applications

NASA Astrophysics Data System (ADS)

Fercher, A. F.; Drexler, W.; Hitzenberger, C. K.; Lasser, T.

2003-02-01

There have been three basic approaches to optical tomography since the early 1980s: diffraction tomography, diffuse optical tomography and optical coherence tomography (OCT). Optical techniques are of particular importance in the medical field, because these techniques promise to be safe and cheap and, in addition, offer a therapeutic potential. Advances in OCT technology have made it possible to apply OCT in a wide variety of applications but medical applications are still dominating. Specific advantages of OCT are its high depth and transversal resolution, the fact, that its depth resolution is decoupled from transverse resolution, high probing depth in scattering media, contact-free and non-invasive operation, and the possibility to create various function dependent image contrasting methods. This report presents the principles of OCT and the state of important OCT applications. OCT synthesises cross-sectional images from a series of laterally adjacent depth-scans. At present OCT is used in three different fields of optical imaging, in macroscopic imaging of structures which can be seen by the naked eye or using weak magnifications, in microscopic imaging using magnifications up to the classical limit of microscopic resolution and in endoscopic imaging, using low and medium magnification. First, OCT techniques, like the reflectometry technique and the dual beam technique were based on time-domain low coherence interferometry depth-scans. Later, Fourier-domain techniques have been developed and led to new imaging schemes. Recently developed parallel OCT schemes eliminate the need for lateral scanning and, therefore, dramatically increase the imaging rate. These schemes use CCD cameras and CMOS detector arrays as photodetectors. Video-rate three-dimensional OCT pictures have been obtained. Modifying interference microscopy techniques has led to high-resolution optical coherence microscopy that achieved sub-micrometre resolution. This report is concluded with a short presentation of important OCT applications. Ophthalmology is, due to the transparent ocular structures, still the main field of OCT application. The first commercial instrument too has been introduced for ophthalmic diagnostics (Carl Zeiss Meditec AG). Advances in using near-infrared light, however, opened the path for OCT imaging in strongly scattering tissues. Today, optical in vivo biopsy is one of the most challenging fields of OCT application. High resolution, high penetration depth, and its potential for functional imaging attribute to OCT an optical biopsy quality, which can be used to assess tissue and cell function and morphology in situ. OCT can already clarify the relevant architectural tissue morphology. For many diseases, however, including cancer in its early stages, higher resolution is necessary. New broad-bandwidth light sources, like photonic crystal fibres and superfluorescent fibre sources, and new contrasting techniques, give access to new sample properties and unmatched sensitivity and resolution.

Chemical mapping and quantification at the atomic scale by scanning transmission electron microscopy.

PubMed

Chu, Ming-Wen; Chen, Cheng Hsuan

2013-06-25

With innovative modern material-growth methods, a broad spectrum of fascinating materials with reduced dimensions-ranging from single-atom catalysts, nanoplasmonic and nanophotonic materials to two-dimensional heterostructural interfaces-is continually emerging and extending the new frontiers of materials research. A persistent central challenge in this grand scientific context has been the detailed characterization of the individual objects in these materials with the highest spatial resolution, a problem prompting the need for experimental techniques that integrate both microscopic and spectroscopic capabilities. To date, several representative microscopy-spectroscopy combinations have become available, such as scanning tunneling microscopy, tip-enhanced scanning optical microscopy, atom probe tomography, scanning transmission X-ray microscopy, and scanning transmission electron microscopy (STEM). Among these tools, STEM boasts unique chemical and electronic sensitivity at unparalleled resolution. In this Perspective, we elucidate the advances in STEM and chemical mapping applications at the atomic scale by energy-dispersive X-ray spectroscopy and electron energy loss spectroscopy with a focus on the ultimate challenge of chemical quantification with atomic accuracy.

Effect of etching time on morphological, optical, and electronic properties of silicon nanowires.

PubMed

Nafie, Nesma; Lachiheb, Manel Abouda; Bouaicha, Mongi

2012-07-16

Owing to their interesting electronic, mechanical, optical, and transport properties, silicon nanowires (SiNWs) have attracted much attention, giving opportunities to several potential applications in nanoscale electronic, optoelectronic devices, and silicon solar cells. For photovoltaic application, a superficial film of SiNWs could be used as an efficient antireflection coating. In this work we investigate the morphological, optical, and electronic properties of SiNWs fabricated at different etching times. Characterizations of the formed SiNWs films were performed using a scanning electron microscope, ultraviolet-visible-near-infrared spectroscopy, and light-beam-induced-current technique. The latter technique was used to determine the effective diffusion length in SiNWs films. From these investigations, we deduce that the homogeneity of the SiNWs film plays a key role on the electronic properties.

Multimodal backside imaging of a microcontroller using confocal laser scanning and optical-beam-induced current imaging

NASA Astrophysics Data System (ADS)

Finkeldey, Markus; Göring, Lena; Schellenberg, Falk; Brenner, Carsten; Gerhardt, Nils C.; Hofmann, Martin

2017-02-01

Microscopy imaging with a single technology is usually restricted to a single contrast mechanism. Multimodal imaging is a promising technique to improve the structural information that could be obtained about a device under test (DUT). Due to the different contrast mechanisms of laser scanning microscopy (LSM), confocal laser scanning microscopy (CLSM) and optical beam induced current microscopy (OBICM), a combination could improve the detection of structures in integrated circuits (ICs) and helps to reveal their layout. While OBIC imaging is sensitive to the changes between differently doped areas and to semiconductor-metal transitions, CLSM imaging is mostly sensitive to changes in absorption and reflection. In this work we present the implementation of OBIC imaging into a CLSM. We show first results using industry standard Atmel microcontrollers (MCUs) with a feature size of about 250nm as DUTs. Analyzing these types of microcontrollers helps to improve in the field of side-channel attacks to find hardware Trojans, possible spots for laser fault attacks and for reverse engineering. For the experimental results the DUT is placed on a custom circuit board that allows us to measure the current while imaging it in our in-house built stage scanning microscope using a near infrared (NIR) laser diode as light source. The DUT is thinned and polished, allowing backside imaging through the Si-substrate. We demonstrate the possibilities using this optical setup by evaluating OBIC, LSM and CLSM images above and below the threshold of the laser source.

Effect of Nickel Contents on the Microstructure and Mechanical Properties for Low-Carbon Bainitic Weld Metals

NASA Astrophysics Data System (ADS)

Mao, Gaojun; Cao, Rui; Yang, Jun; Jiang, Yong; Wang, Shuai; Guo, Xili; Yuan, Junjun; Zhang, Xiaobo; Chen, Jianhong

2017-05-01

Multi-pass weld metals were deposited on Q345 base steel using metal powder-flux-cored wire with various Ni contents to investigate the effects of the Ni content on the weld microstructure and property. The types of the microstructures were identified by optical microscope, scanning electron microscope, transmission electron microscope, and micro-hardness tests. As a focusing point, the lath bainite and lath martensite were distinguished by their compositions, morphologies, and hardness. In particular, a number of black plane facets appearing between lath bainite or lath martensite packets were characterized by laser scanning confocal microscope. The results indicated that with the increase in Ni contents in the range of 0, 2, 4, and 6%, the microstructures in the weld-deposited metal were changed from the domination of the granular bainite to the majority of the lath bainite and/or the lath martensite and the micro-hardness of the weld-deposited metal increased. Meanwhile, the average width of columnar grain displays a decreasing trend and prior austenite grain size decreases while increases with higher Ni content above 4%. Yield strength and ultimate tensile strength decrease, while the reduction in fracture area increases with the decreasing Ni mass fraction and the increasing test temperature, respectively. And poor yield strength in Ni6 specimen can be attributed to elements segregation caused by weld defect. Finally, micro-hardness distribution in correspondence with specimens presents as a style of cloud-map.

A compilation of cold cases using scanning electron microscopy at the University of Rhode Island

NASA Astrophysics Data System (ADS)

Platek, Michael J.; Gregory, Otto J.

2015-10-01

Scanning electron microscopy combined with microchemical analysis has evolved into one of the most widely used instruments in forensic science today. In particular, the environmental scanning electron microscope (SEM) in conjunction with energy dispersive spectroscopy (EDS), has created unique opportunities in forensic science in regard to the examination of trace evidence; i.e. the examination of evidence without altering the evidence with conductive coatings, thereby enabling criminalists to solve cases that were previously considered unsolvable. Two cold cases were solved at URI using a JEOL 5900 LV SEM in conjunction with EDS. A cold case murder and a cold missing person case will be presented from the viewpoint of the microscopist and will include sample preparation, as well as image and chemical analysis of the trace evidence using electron microscopy and optical microscopy.

Photodynamic therapy and knocking out of single tumor cells by multiphoton excitation processes

NASA Astrophysics Data System (ADS)

Riemann, Iris; Fischer, Peter; Koenig, Karsten

2004-09-01

Near infrared (NIR) ultrashort laser pulses of 780 nm have been used to induce intracellular photodynamic reactions by nonlinear excitation of porphyrin photosensitizers. Intracellular accumulation and photobleaching of the fluorescent photosensitizers protoporphyrin IX and Photofrin (PF) have been studied by non-resonant two-photon fluorescence excitation of PF and aminolevulinic acid (ALA)-labeled Chinese hamster ovary (CHO) cells. To testify the efficacy of both substrates to induce irreversible destructive effects, the cloning efficiency (CE) of cells exposed to femtosecond pulses of a multiphoton laser scanning microscope (40x/1.3) was determined. In the case of Photofrin accumulation, CEs of 50% and 0% were obtained after 17 laserscans (2 mW?, 16 s/ frame) and 50 scans, respectively. All cells exposed to 50 scans died within 48h after laser exposure. 100 scans were required to induce lethal effects in ALA labeled cells. Sensitizer-free control cells could be scanned 250 times (1.1 h) and more without impact on the reproduction behavior, morphology, and vitality. In addition to the slow phototoxic effect by photooxidation processes, another destructive but immediate effect based on optical breakdown was induced when employing high intense NIR femtosecond laser beams. This was used to optically knock out single tumor cells in living mice (solid Ehrlich-Carcinoma) in a depth of 10 to 100 μm.

Spectroscopic and fiber optic ethanol sensing properties Gd doped ZnO nanoparticles.

PubMed

Noel, J L; Udayabhaskar, R; Renganathan, B; Muthu Mariappan, S; Sastikumar, D; Karthikeyan, B

2014-11-11

We report the structural, optical and gas sensing properties of prepared pure and Gd doped ZnO nanoparticles through solgel method at moderate temperature. Structural studies are carried out by X-ray diffraction method confirms hexagonal wurtzite structure and doping induced changes in lattice parameters is observed. Optical absorption spectral studies shows red shift in the absorption peak corresponds to band-gap from 3.42 eV to 3.05 eV and broad absorption in the visible range after Gd doping is observed. Scanning electron microscopic studies shows increase in particle size where the particle diameters increase from few nm to micrometers after Gd doping. The clad modified ethanol fiber-optic sensor studies for ethanol sensing exhibits best sensitivity for the 3% Gd doped ZnO nanoparticles and the sensitivity get lowered incase of higher percentage of Gd doped ZnO sample. Copyright © 2014 Elsevier B.V. All rights reserved.

Low cost paths to binary optics

NASA Technical Reports Server (NTRS)

Nelson, Arthur; Domash, Lawrence

1993-01-01

Application of binary optics has been limited to a few major laboratories because of the limited availability of fabrication facilities such as e-beam machines and the lack of standardized design software. Foster-Miller has attempted to identify low cost approaches to medium-resolution binary optics using readily available computer and fabrication tools, primarily for the use of students and experimenters in optical computing. An early version of our system, MacBEEP, made use of an optimized laser film recorder from the commercial typesetting industry with 10 micron resolution. This report is an update on our current efforts to design and build a second generation MacBEEP, which aims at 1 micron resolution and multiple phase levels. Trails included a low cost scanning electron microscope in microlithography mode, and alternative laser inscribers or photomask generators. Our current software approach is based on Mathematica and PostScript compatibility.

Evanescent wave assisted nanomaterial coating.

PubMed

Mondal, Samir K; Pal, Sudipta Sarkar; Kumbhakar, Dharmadas; Tiwari, Umesh; Bhatnagar, Randhir

2013-08-01

In this work we present a novel nanomaterial coating technique using evanescent wave (EW). The gradient force in the EW is used as an optical tweezer for tweezing and self-assembling nanoparticles on the source of EW. As a proof of the concept, we have used a laser coupled etched multimode optical fiber, which generates EW for the EW assisted coating. The section-wise etched multimode optical fiber is horizontally and superficially dipped into a silver/gold nanoparticles solution while the laser is switched on. The fiber is left until the solution recedes due to evaporation leaving the fiber in air. The coating time usually takes 40-50 min at room temperature. The scanning electron microscope image shows uniform and thin coating of self-assembled nanoparticles due to EW around the etched section. A coating thickness <200 nm is achieved. The technique could be useful for making surface-plasmon-resonance-based optical fiber probes and other plasmonic circuits.

Fabrication of near-field optical apertures in aluminium by a highly selective corrosion process in the evanescent field.

PubMed

Haefliger, D; Stemmer, A

2003-03-01

A simple, one-step process to fabricate high-quality apertures for scanning near-field optical microscope probes based on aluminium-coated silicon nitride cantilevers is presented. A thin evanescent optical field at a glass-water interface was used to heat the aluminium at the tip apex due to light absorption. The heat induced a breakdown of the passivating oxide layer and local corrosion of the metal, which selectively exposed the front-most part of the probe tip from the aluminium. Apertures with a protruding silicon nitride tip up to 72 nm in height were fabricated. The height of the protrusion was controlled by the extent of the evanescent field, whereas the diameter depended on the geometry of the probe substrate. The corrosion process proved to be self-terminating, yielding highly reproducible tip heights. Near-field optical resolution in a transmission mode of 85 nm was demonstrated.

Giant Kerr response of ultrathin gold films from quantum size effect.

PubMed

Qian, Haoliang; Xiao, Yuzhe; Liu, Zhaowei

2016-10-10

With the size of plasmonic devices entering into the nanoscale region, the impact of quantum physics needs to be considered. In the past, the quantum size effect on linear material properties has been studied extensively. However, the nonlinear aspects have not been explored much so far. On the other hand, much effort has been put into the field of integrated nonlinear optics and a medium with large nonlinearity is desirable. Here we study the optical nonlinear properties of a nanometre scale gold quantum well by using the z-scan method and nonlinear spectrum broadening technique. The quantum size effect results in a giant optical Kerr susceptibility, which is four orders of magnitude higher than the intrinsic value of bulk gold and several orders larger than traditional nonlinear media. Such high nonlinearity enables efficient nonlinear interaction within a microscopic footprint, making quantum metallic films a promising candidate for integrated nonlinear optical applications.

Physics and engineering aspects of cell and tissue imaging systems: microscopic devices and computer assisted diagnosis.

PubMed

Chen, Xiaodong; Ren, Liqiang; Zheng, Bin; Liu, Hong

2013-01-01

The conventional optical microscopes have been used widely in scientific research and in clinical practice. The modern digital microscopic devices combine the power of optical imaging and computerized analysis, archiving and communication techniques. It has a great potential in pathological examinations for improving the efficiency and accuracy of clinical diagnosis. This chapter reviews the basic optical principles of conventional microscopes, fluorescence microscopes and electron microscopes. The recent developments and future clinical applications of advanced digital microscopic imaging methods and computer assisted diagnosis schemes are also discussed.

Classification of Human Retinal Microaneurysms Using Adaptive Optics Scanning Light Ophthalmoscope Fluorescein Angiography

PubMed Central

Dubow, Michael; Pinhas, Alexander; Shah, Nishit; Cooper, Robert F.; Gan, Alexander; Gentile, Ronald C.; Hendrix, Vernon; Sulai, Yusufu N.; Carroll, Joseph; Chui, Toco Y. P.; Walsh, Joseph B.; Weitz, Rishard; Dubra, Alfredo; Rosen, Richard B.

2014-01-01

Purpose. Microaneurysms (MAs) are considered a hallmark of retinal vascular disease, yet what little is known about them is mostly based upon histology, not clinical observation. Here, we use the recently developed adaptive optics scanning light ophthalmoscope (AOSLO) fluorescein angiography (FA) to image human MAs in vivo and to expand on previously described MA morphologic classification schemes. Methods. Patients with vascular retinopathies (diabetic, hypertensive, and branch and central retinal vein occlusion) were imaged with reflectance AOSLO and AOSLO FA. Ninety-three MAs, from 14 eyes, were imaged and classified according to appearance into six morphologic groups: focal bulge, saccular, fusiform, mixed, pedunculated, and irregular. The MA perimeter, area, and feret maximum and minimum were correlated to morphology and retinal pathology. Select MAs were imaged longitudinally in two eyes. Results. Adaptive optics scanning light ophthalmoscope fluorescein angiography imaging revealed microscopic features of MAs not appreciated on conventional images. Saccular MAs were most prevalent (47%). No association was found between the type of retinal pathology and MA morphology (P = 0.44). Pedunculated and irregular MAs were among the largest MAs with average areas of 4188 and 4116 μm2, respectively. Focal hypofluorescent regions were noted in 30% of MAs and were more likely to be associated with larger MAs (3086 vs. 1448 μm2, P = 0.0001). Conclusions. Retinal MAs can be classified in vivo into six different morphologic types, according to the geometry of their two-dimensional (2D) en face view. Adaptive optics scanning light ophthalmoscope fluorescein angiography imaging of MAs offers the possibility of studying microvascular change on a histologic scale, which may help our understanding of disease progression and treatment response. PMID:24425852

Coherent anti-Stokes Raman scattering microscope with a high-signal-to-noise ratio, high stability, and high-speed imaging for live cell observation

NASA Astrophysics Data System (ADS)

Hayashi, Shinichi; Takimoto, Shinichi; Hashimoto, Takeshi

2007-02-01

Coherent anti-Stokes Raman scattering (CARS) microscopy, which can produce images of specific molecules without staining, has attracted the attention of researchers, as it matches the need for molecular imaging and pathway analysis of live cells. In particular, there have been an increasing number of CARS experimental results regarding lipids in live cells, which cannot be fluorescently tagged while keeping the cells alive. One of the important applications of lipid research is for the metabolic syndrome. Since the metabolic syndrome is said to be related to the lipids in lipocytes, blood, arterial vessels, and so on, the CARS technique is expected to find application in this field. However, CARS microscopy requires a pair of picosecond laser pulses, which overlap both temporally and spatially. This makes the optical adjustments of a CARS microscope challenging. The authors developed a CARS unit that includes optics for easy and stable adjustment of the overlap of these laser pulses. Adding the CARS unit to a laser scanning microscope provides CARS images of a high signal-to-noise ratio, with an acquisition rate as high as 2 microseconds per pixel. Thus, images of fast-moving lipid droplets in Hela cells were obtained.

A Multiscale Material Testing System for In Situ Optical and Electron Microscopes and Its Application

PubMed Central

Ye, Xuan; Cui, Zhiguo; Fang, Huajun; Li, Xide

2017-01-01

We report a novel material testing system (MTS) that uses hierarchical designs for in-situ mechanical characterization of multiscale materials. This MTS is adaptable for use in optical microscopes (OMs) and scanning electron microscopes (SEMs). The system consists of a microscale material testing module (m-MTM) and a nanoscale material testing module (n-MTM). The MTS can measure mechanical properties of materials with characteristic lengths ranging from millimeters to tens of nanometers, while load capacity can vary from several hundred micronewtons to several nanonewtons. The m-MTM is integrated using piezoelectric motors and piezoelectric stacks/tubes to form coarse and fine testing modules, with specimen length from millimeters to several micrometers, and displacement distances of 12 mm with 0.2 µm resolution for coarse level and 8 µm with 1 nm resolution for fine level. The n-MTM is fabricated using microelectromechanical system technology to form active and passive components and realizes material testing for specimen lengths ranging from several hundred micrometers to tens of nanometers. The system’s capabilities are demonstrated by in-situ OM and SEM testing of the system’s performance and mechanical properties measurements of carbon fibers and metallic microwires. In-situ multiscale deformation tests of Bacillus subtilis filaments are also presented. PMID:28777341

Nanoscale characterization of local structures and defects in photonic crystals using synchrotron-based transmission soft X-ray microscopy

PubMed Central

Nho, Hyun Woo; Kalegowda, Yogesh; Shin, Hyun-Joon; Yoon, Tae Hyun

2016-01-01

For the structural characterization of the polystyrene (PS)-based photonic crystals (PCs), fast and direct imaging capabilities of full field transmission X-ray microscopy (TXM) were demonstrated at soft X-ray energy. PS-based PCs were prepared on an O2-plasma treated Si3N4 window and their local structures and defects were investigated using this label-free TXM technique with an image acquisition speed of ~10 sec/frame and marginal radiation damage. Micro-domains of face-centered cubic (FCC (111)) and hexagonal close-packed (HCP (0001)) structures were dominantly found in PS-based PCs, while point and line defects, FCC (100), and 12-fold symmetry structures were also identified as minor components. Additionally, in situ observation capability for hydrated samples and 3D tomographic reconstruction of TXM images were also demonstrated. This soft X-ray full field TXM technique with faster image acquisition speed, in situ observation, and 3D tomography capability can be complementally used with the other X-ray microscopic techniques (i.e., scanning transmission X-ray microscopy, STXM) as well as conventional characterization methods (e.g., electron microscopic and optical/fluorescence microscopic techniques) for clearer structure identification of self-assembled PCs and better understanding of the relationship between their structures and resultant optical properties. PMID:27087141

Analysis of Phase Separation in Czochralski Grown Single Crystal Ilmenite

NASA Technical Reports Server (NTRS)

Wilkins, R.; Powell, Kirk St. A.; Loregnard, Kieron R.; Lin, Sy-Chyi; Muthusami, Jayakumar; Zhou, Feng; Pandey, R. K.; Brown, Geoff; Hawley, M. E.

1998-01-01

Ilmenite (FeTiOs) is a wide bandgap semiconductor with an energy gap of 2.58 eV. Ilmenite has properties suited for radiation tolerant applications, as well as a variety of other electronic applications. Single crystal ilmenite has been grown from the melt using the Czochralski method. Growth conditions have a profound effect on the microstructure of the samples. Here we present data from a variety of analytical techniques which indicate that some grown crystals exhibit distinct phase separation during growth. This phase separation is apparent for both post-growth annealed and unannealed samples. Under optical microscopy, there appear two distinct areas forming a matrix with an array of dots on order of 5 pm diameter. While appearing bright in the optical micrograph, atomic force microscope (AFM) shows the dots to be shallow pits on the surface. Magnetic force microscope (MFM) shows the dots to be magnetic. Phase identification via electron microprobe analysis (EMPA) indicates two major phases in the unannealed samples and four in the annealed samples, where the dots appear to be almost pure iron. This is consistent with micrographs taken with a scanning probe microscope used in the magnetic force mode. Samples that do not exhibit the phase separation have little or no discernible magnetic structure detectable by the MFM.

ZnO nanoparticles based fiber optic gas sensor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Narasimman, S.; Sivacoumar, R.; Alex, Z. C.

In this work, ZnO nanoparticles were synthesized by simple aqueous chemical route method. The synthesized ZnO nanoparticles were characterized by X-ray diffraction and scanning electron microscope. The sensitivity of the nanoparticles was studied for different gases like acetone, ammonia and ethanol in terms of variation in spectral light intensity. The XRD and SEM analysis confirms the formation of hexagonal wurtzite structure with the grain size of 11.2 nm. The small cladding region of the optical fiber was replaced with the synthesized nanoparticles. The light spectrum was recorded for different gas concentrations. The synthesized nanoparticles showed high sensitivity towards ammonia in lowmore » ppm level and acetone in high ppm level.« less