Enhancing the performance of the light field microscope using wavefront coding

PubMed Central

Cohen, Noy; Yang, Samuel; Andalman, Aaron; Broxton, Michael; Grosenick, Logan; Deisseroth, Karl; Horowitz, Mark; Levoy, Marc

2014-01-01

Light field microscopy has been proposed as a new high-speed volumetric computational imaging method that enables reconstruction of 3-D volumes from captured projections of the 4-D light field. Recently, a detailed physical optics model of the light field microscope has been derived, which led to the development of a deconvolution algorithm that reconstructs 3-D volumes with high spatial resolution. However, the spatial resolution of the reconstructions has been shown to be non-uniform across depth, with some z planes showing high resolution and others, particularly at the center of the imaged volume, showing very low resolution. In this paper, we enhance the performance of the light field microscope using wavefront coding techniques. By including phase masks in the optical path of the microscope we are able to address this non-uniform resolution limitation. We have also found that superior control over the performance of the light field microscope can be achieved by using two phase masks rather than one, placed at the objective’s back focal plane and at the microscope’s native image plane. We present an extended optical model for our wavefront coded light field microscope and develop a performance metric based on Fisher information, which we use to choose adequate phase masks parameters. We validate our approach using both simulated data and experimental resolution measurements of a USAF 1951 resolution target; and demonstrate the utility for biological applications with in vivo volumetric calcium imaging of larval zebrafish brain. PMID:25322056

Optimising electron microscopy experiment through electron optics simulation.

PubMed

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

2017-04-01

We developed a new type of electron trajectories simulation inside a complete model of a modern transmission electron microscope (TEM). Our model incorporates the precise and real design of each element constituting a TEM, i.e. the field emission (FE) cathode, the extraction optic and acceleration stages of a 300kV cold field emission gun, the illumination lenses, the objective lens, the intermediate and projection lenses. Full trajectories can be computed using magnetically saturated or non-saturated round lenses, magnetic deflectors and even non-cylindrical symmetry elements like electrostatic biprism. This multi-scale model gathers nanometer size components (FE tip) with parts of meter length (illumination and projection systems). We demonstrate that non-trivial TEM experiments requiring specific and complex optical configurations can be simulated and optimized prior to any experiment using such model. We show that all the currents set in all optical elements of the simulated column can be implemented in the real column (I2TEM in CEMES) and used as starting alignment for the requested experiment. We argue that the combination of such complete electron trajectory simulations in the whole TEM column with automatic optimization of the microscope parameters for optimal experimental data (images, diffraction, spectra) allows drastically simplifying the implementation of complex experiments in TEM and will facilitate the development of advanced use of the electron microscope in the near future. Copyright © 2017 Elsevier B.V. All rights reserved.

Mathematical model of a DIC position sensing system within an optical trap

NASA Astrophysics Data System (ADS)

Wulff, Kurt D.; Cole, Daniel G.; Clark, Robert L.

2005-08-01

The quantitative study of displacements and forces of motor proteins and processes that occur at the microscopic level and below require a high level of sensitivity. For optical traps, two techniques for position sensing have been accepted and used quite extensively: quadrant photodiodes and an interferometric position sensing technique based on DIC imaging. While quadrant photodiodes have been studied in depth and mathematically characterized, a mathematical characterization of the interferometric position sensor has not been presented to the authors' knowledge. The interferometric position sensing method works off of the DIC imaging capabilities of a microscope. Circularly polarized light is sent into the microscope and the Wollaston prism used for DIC imaging splits the beam into its orthogonal components, displacing them by a set distance determined by the user. The distance between the axes of the beams is set so the beams overlap at the specimen plane and effectively share the trapped microsphere. A second prism then recombines the light beams and the exiting laser light's polarization is measured and related to position. In this paper we outline the mathematical characterization of a microsphere suspended in an optical trap using a DIC position sensing method. The sensitivity of this mathematical model is then compared to the QPD model. The mathematical model of a microsphere in an optical trap can serve as a calibration curve for an experimental setup.

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

Augmented microscopy with near-infrared fluorescence detection

NASA Astrophysics Data System (ADS)

Watson, Jeffrey R.; Martirosyan, Nikolay; Skoch, Jesse; Lemole, G. Michael; Anton, Rein; Romanowski, Marek

2015-03-01

Near-infrared (NIR) fluorescence has become a frequently used intraoperative technique for image-guided surgical interventions. In procedures such as cerebral angiography, surgeons use the optical surgical microscope for the color view of the surgical field, and then switch to an electronic display for the NIR fluorescence images. However, the lack of stereoscopic, real-time, and on-site coregistration adds time and uncertainty to image-guided surgical procedures. To address these limitations, we developed the augmented microscope, whereby the electronically processed NIR fluorescence image is overlaid with the anatomical optical image in real-time within the optical path of the microscope. In vitro, the augmented microscope can detect and display indocyanine green (ICG) concentrations down to 94.5 nM, overlaid with the anatomical color image. We prepared polyacrylamide tissue phantoms with embedded polystyrene beads, yielding scattering properties similar to brain matter. In this model, 194 μM solution of ICG was detectable up to depths of 5 mm. ICG angiography was then performed in anesthetized rats. A dynamic process of ICG distribution in the vascular system overlaid with anatomical color images was observed and recorded. In summary, the augmented microscope demonstrates NIR fluorescence detection with superior real-time coregistration displayed within the ocular of the stereomicroscope. In comparison to other techniques, the augmented microscope retains full stereoscopic vision and optical controls including magnification and focus, camera capture, and multiuser access. Augmented microscopy may find application in surgeries where the use of traditional microscopes can be enhanced by contrast agents and image guided delivery of therapeutics, including oncology, neurosurgery, and ophthalmology.

Measuring optical properties of a blood vessel model using optical coherence tomography

NASA Astrophysics Data System (ADS)

Levitz, David; Hinds, Monica T.; Tran, Noi; Vartanian, Keri; Hanson, Stephen R.; Jacques, Steven L.

2006-02-01

In this paper we develop the concept of a tissue-engineered optical phantom that uses engineered tissue as a phantom for calibration and optimization of biomedical optics instrumentation. With this method, the effects of biological processes on measured signals can be studied in a well controlled manner. To demonstrate this concept, we attempted to investigate how the cellular remodeling of a collagen matrix affected the optical properties extracted from optical coherence tomography (OCT) images of the samples. Tissue-engineered optical phantoms of the vascular system were created by seeding smooth muscle cells in a collagen matrix. Four different optical properties were evaluated by fitting the OCT signal to 2 different models: the sample reflectivity ρ and attenuation parameter μ were extracted from the single scattering model, and the scattering coefficient μ s and root-mean-square scattering angle θ rms were extracted from the extended Huygens-Fresnel model. We found that while contraction of the smooth muscle cells was clearly evident macroscopically, on the microscopic scale very few cells were actually embedded in the collagen. Consequently, no significant difference between the cellular and acellular samples in either set of measured optical properties was observed. We believe that further optimization of our tissue-engineering methods is needed in order to make the histology and biochemistry of the cellular samples sufficiently different from the acellular samples on the microscopic level. Once these methods are optimized, we can better verify whether the optical properties of the cellular and acellular collagen samples differ.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2010-06-29

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P.; Chernobrod, Boris M.

2009-11-10

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of impaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P.; Chernobrod, Boris M.

2007-12-11

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2010-07-13

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2009-10-27

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Sensing of Streptococcus mutans by microscopic imaging ellipsometry

NASA Astrophysics Data System (ADS)

Khaleel, Mai Ibrahim; Chen, Yu-Da; Chien, Ching-Hang; Chang, Yia-Chung

2017-05-01

Microscopic imaging ellipsometry is an optical technique that uses an objective and sensing procedure to measure the ellipsometric parameters Ψ and Δ in the form of microscopic maps. This technique is well known for being noninvasive and label-free. Therefore, it can be used to detect and characterize biological species without any impact. Microscopic imaging ellipsometry was used to measure the optical response of dried Streptococcus mutans cells on a glass substrate. The ellipsometric Ψ and Δ maps were obtained with the Optrel Multiskop system for specular reflection in the visible range (λ=450 to 750 nm). The Ψ and Δ images at 500, 600, and 700 nm were analyzed using three different theoretical models with single-bounce, two-bounce, and multibounce light paths to obtain the optical constants and height distribution. The obtained images of the optical constants show different aspects when comparing the single-bounce analysis with the two-bounce or multibounce analysis in detecting S. mutans samples. Furthermore, the height distributions estimated by two-bounce and multibounce analyses of S. mutans samples were in agreement with the thickness values measured by AFM, which implies that the two-bounce and multibounce analyses can provide information complementary to that obtained by a single-bounce light path.

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.

Sub-nanosecond time-resolved near-field scanning magneto-optical microscope.

PubMed

Rudge, J; Xu, H; Kolthammer, J; Hong, Y K; Choi, B C

2015-02-01

We report on the development of a new magnetic microscope, time-resolved near-field scanning magneto-optical microscope, which combines a near-field scanning optical microscope and magneto-optical contrast. By taking advantage of the high temporal resolution of time-resolved Kerr microscope and the sub-wavelength spatial resolution of a near-field microscope, we achieved a temporal resolution of ∼50 ps and a spatial resolution of <100 nm. In order to demonstrate the spatiotemporal magnetic imaging capability of this microscope, the magnetic field pulse induced gyrotropic vortex dynamics occurring in 1 μm diameter, 20 nm thick CoFeB circular disks has been investigated. The microscope provides sub-wavelength resolution magnetic images of the gyrotropic motion of the vortex core at a resonance frequency of ∼240 MHz.

The Scanning Optical Microscope.

ERIC Educational Resources Information Center

Sheppard, C. J. R.

1978-01-01

Describes the principle of the scanning optical microscope and explains its advantages over the conventional microscope in the improvement of resolution and contrast, as well as the possibility of producing a picture from optical harmonies generated within the specimen.

Sub-micron materials characterization using near-field optics

NASA Astrophysics Data System (ADS)

Blodgett, David Wesley

1998-12-01

High-resolution sub-surface materials characterization and inspection are critical in the microelectronics and thin films industries. To this end, a technique is described that couples the bulk property measurement capabilities of high-frequency ultrasound with the high-resolution surface imaging capabilities of the near-field optical microscope. Sensing bulk microstructure variations in the material, such as grain boundaries, requires a detection footprint smaller than the variation itself. The near-field optical microscope, with the ability to exceed the diffraction limit in optical resolution, meets this requirement. Two apertureless near-field optical microscopes, on-axis and off-axis illumination, have been designed and built. Near-field and far-field approach curves for both microscopes are presented. The sensitivity of the near-field approach curve was 8.3 muV/nm. Resolution studies for the near-field microscope indicate optical resolutions on the order of 50 nm, which exceeds the diffraction limit. The near-field microscope has been adapted to detect both contact-transducer-generated and laser-generated ultrasound. The successful detection of high-frequency ultrasound with the near-field optical microscope demonstrates the potential of this technique.

Microscope collision protection apparatus

DOEpatents

DeNure, Charles R.

2001-10-23

A microscope collision protection apparatus for a remote control microscope which protects the optical and associated components from damage in the event of an uncontrolled collision with a specimen, regardless of the specimen size or shape. In a preferred embodiment, the apparatus includes a counterbalanced slide for mounting the microscope's optical components. This slide replaces the rigid mounts on conventional upright microscopes with a precision ball bearing slide. As the specimen contacts an optical component, the contacting force will move the slide and the optical components mounted thereon. This movement will protect the optical and associated components from damage as the movement causes a limit switch to be actuated, thereby stopping all motors responsible for the collision.

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

Optical second harmonic images of 90 deg domain structure in BaTiO3 and periodically inverted antiparallel domains in LiTaO3

NASA Astrophysics Data System (ADS)

Uesu, Y.; Kurimura, S.; Yamamoto, Y.

1995-04-01

Applied is a microscope to observations of 90 deg ferroelectric domain structure in BaTiO3 and inverted periodically are ferroelectric domains in LiTaO3. It is founded that the second harmonic generation microscope gives information which cannot be obtained by ordinary optical microscopes. The developed nonlinear optical microscope builds two dimensional second harmonic image of a specimen with inhomogenous distribution of d(sub ijk) and applied the microscope to observations of inhomogeneity in some nonlinear-optical organic microcrystals.

Transmissive Nanohole Arrays for Massively-Parallel Optical Biosensing

PubMed Central

2015-01-01

A high-throughput optical biosensing technique is proposed and demonstrated. This hybrid technique combines optical transmission of nanoholes with colorimetric silver staining. The size and spacing of the nanoholes are chosen so that individual nanoholes can be independently resolved in massive parallel using an ordinary transmission optical microscope, and, in place of determining a spectral shift, the brightness of each nanohole is recorded to greatly simplify the readout. Each nanohole then acts as an independent sensor, and the blocking of nanohole optical transmission by enzymatic silver staining defines the specific detection of a biological agent. Nearly 10000 nanoholes can be simultaneously monitored under the field of view of a typical microscope. As an initial proof of concept, biotinylated lysozyme (biotin-HEL) was used as a model analyte, giving a detection limit as low as 0.1 ng/mL. PMID:25530982

Enhancing microscopic cascading contributions to higher-order nonlinear-optical responses through forced geometric constraints

NASA Astrophysics Data System (ADS)

Dawson, Nathan J.; Andrews, James H.; Crescimanno, Michael

2012-10-01

We review a model that was developed to take into account all possible microscopic cascading schemes in a single species system out to the fifth order using a self-consistent field approach. This model was designed to study the effects of boundaries in mesoscopic systems with constrained boundaries. These geometric constraints on the macroscopic structure show how the higher-ordered susceptibilities are manipulated by increasing the surface to volume ratio, while the microscopic structure influences the local field from all other molecules in the system. In addition to the review, we discuss methods of modeling real systems of molecules, where efforts are currently underway.

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.

Model wavefront sensor for adaptive confocal microscopy

NASA Astrophysics Data System (ADS)

Booth, Martin J.; Neil, Mark A. A.; Wilson, Tony

2000-05-01

A confocal microscope permits 3D imaging of volume objects by the inclusion of a pinhole in the detector path which eliminates out of focus light. This configuration is however very sensitive to aberrations induced by the specimen or the optical system and would therefore benefit from an adaptive optics approach. We present a wavefront sensor capable of measuring directly the Zernike components of an aberrated wavefront and show that it is particularly applicable to the confocal microscope since only those wavefronts originating in the focal region contribute to the measured aberration.

Adaptive optics plug-and-play setup for high-resolution microscopes with multi-actuator adaptive lens

NASA Astrophysics Data System (ADS)

Quintavalla, M.; Pozzi, P.; Verhaegen, Michelle; Bijlsma, Hielke; Verstraete, Hans; Bonora, S.

2018-02-01

Adaptive Optics (AO) has revealed as a very promising technique for high-resolution microscopy, where the presence of optical aberrations can easily compromise the image quality. Typical AO systems however, are almost impossible to implement on commercial microscopes. We propose a simple approach by using a Multi-actuator Adaptive Lens (MAL) that can be inserted right after the objective and works in conjunction with an image optimization software allowing for a wavefront sensorless correction. We presented the results obtained on several commercial microscopes among which a confocal microscope, a fluorescence microscope, a light sheet microscope and a multiphoton microscope.

Microscopic Optical Projection Tomography In Vivo

PubMed Central

Meyer, Heiko; Ripoll, Jorge; Tavernarakis, Nektarios

2011-01-01

We describe a versatile optical projection tomography system for rapid three-dimensional imaging of microscopic specimens in vivo. Our tomographic setup eliminates the in xy and z strongly asymmetric resolution, resulting from optical sectioning in conventional confocal microscopy. It allows for robust, high resolution fluorescence as well as absorption imaging of live transparent invertebrate animals such as C. elegans. This system offers considerable advantages over currently available methods when imaging dynamic developmental processes and animal ageing; it permits monitoring of spatio-temporal gene expression and anatomical alterations with single-cell resolution, it utilizes both fluorescence and absorption as a source of contrast, and is easily adaptable for a range of small model organisms. PMID:21559481

Feedback effects in optical communication systems: characteristic curve for single-mode InGaAsP lasers.

PubMed

Brivio, F; Reverdito, C; Sacchi, G; Chiaretti, G; Milani, M

1992-08-20

An experimental analysis of InGaAsP injection lasers shows an unexpected decrease of the differential quantum efficiency as a function of injected current when optical power is fed back into the active cavity of a diode inserted into a long transmission line. To investigate the response of laser diodes to optical feedback, we base our analysis on a microscopic model, resulting in a set of coupled equations that include the microscopic parameters that characterize the material and the device. This description takes into account the nonlinear dependence of the interband carrier lifetime on the level of optical feedback. Good agreement between the analytical description and experimental data is obtained for threshold current and differential quantum efficiency as functions of the feedback ratio.

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.

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.

Noise induced chaos in optically driven colloidal rings.

NASA Astrophysics Data System (ADS)

Roichman, Yael; Zaslavsky, George; Grier, David G.

2007-03-01

Given a constant flux of energy, many driven dissipative systems rapidly organize themselves into configurations that support steady state motion. Examples include swarming of bacterial colonies, convection in shaken sandpiles, and synchronization in flowing traffic. How simple objects interacting in simple ways self-organize generally is not understood, mainly because so few of the available experimental systems afford the necessary access to their microscopic degrees of freedom. This talk introduces a new class of model driven dissipative systems typified by three colloidal spheres circulating around a ring-like optical trap known as an optical vortex. By controlling the interplay between hydrodynamic interactions and fixed disorder we are able to drive a transition from a previously predicted periodic steady state to fully developed chaos. In addition, by tracking both microscopic trajectories and macroscopic collective fluctuations the relation between the onset of microscopic weak chaos and the evolution of space-time self-similarity in macroscopic transport properties is revealed. In a broader scope, several optical vortices can be coupled to create a large dissipative system where each building block has internal degrees of freedom. In such systems the little understood dynamics of processes like frustration and jamming, fluctuation-dissipation relations and the propagation of collective motion can be tracked microscopically.

Development and application of a ray-based model of light propagation through a spherical acousto-optic lens

PubMed Central

Evans, Geoffrey J.; Kirkby, Paul A.; Nadella, K. M. Naga Srinivas; Marin, Bóris; Silver, R. Angus

2016-01-01

A spherical acousto-optic lens (AOL) consists of four acousto-optic deflectors (AODs) that can rapidly and precisely control the focal position of an optical beam in 3D space. Development and application of AOLs has increased the speed at which 3D random access point measurements can be performed with a two-photon microscope. This has been particularly useful for measuring brain activity with fluorescent reporter dyes because neuronal signalling is rapid and sparsely distributed in 3D space. However, a theoretical description of light propagation through AOLs has lagged behind their development, resulting in only a handful of simplified principles to guide AOL design and optimization. To address this we have developed a ray-based computer model of an AOL incorporating acousto-optic diffraction and refraction by anisotropic media. We extended an existing model of a single AOD with constant drive frequency to model a spherical AOL: four AODs in series driven with linear chirps. AOL model predictions of the relationship between optical transmission efficiency and acoustic drive frequency including second order diffraction effects closely matched experimental measurements from a 3D two-photon AOL microscope. Moreover, exploration of different AOL drive configurations identified a new simple rule for maximizing the field of view of our compact AOL design. By providing a theoretical basis for understanding optical transmission through spherical AOLs, our open source model is likely to be useful for comparing and improving different AOL designs, as well as identifying the acoustic drive configurations that provide the best transmission performance over the 3D focal region. PMID:26368449

Development and application of a ray-based model of light propagation through a spherical acousto-optic lens.

PubMed

Evans, Geoffrey J; Kirkby, Paul A; Naga Srinivas Nadella, K M; Marin, Bóris; Angus Silver, R

2015-09-07

A spherical acousto-optic lens (AOL) consists of four acousto-optic deflectors (AODs) that can rapidly and precisely control the focal position of an optical beam in 3D space. Development and application of AOLs has increased the speed at which 3D random access point measurements can be performed with a two-photon microscope. This has been particularly useful for measuring brain activity with fluorescent reporter dyes because neuronal signalling is rapid and sparsely distributed in 3D space. However, a theoretical description of light propagation through AOLs has lagged behind their development, resulting in only a handful of simplified principles to guide AOL design and optimization. To address this we have developed a ray-based computer model of an AOL incorporating acousto-optic diffraction and refraction by anisotropic media. We extended an existing model of a single AOD with constant drive frequency to model a spherical AOL: four AODs in series driven with linear chirps. AOL model predictions of the relationship between optical transmission efficiency and acoustic drive frequency including second order diffraction effects closely matched experimental measurements from a 3D two-photon AOL microscope. Moreover, exploration of different AOL drive configurations identified a new simple rule for maximizing the field of view of our compact AOL design. By providing a theoretical basis for understanding optical transmission through spherical AOLs, our open source model is likely to be useful for comparing and improving different AOL designs, as well as identifying the acoustic drive configurations that provide the best transmission performance over the 3D focal region.

Limits of agreement between the optical pachymeter and a noncontact specular microscope.

PubMed

Ogbuehi, Kelechi C; Almubrad, Turki M

2005-07-01

To determine the limits of agreement between central corneal thickness (CCT) measurements made with the slit lamp-attached optical pachymeter and the SP2000P noncontact specular microscope. Triplicate readings for CCT were obtained for each of 130 (right) eyes of 130 patients, using the slit lamp-attached optical pachymeter and then the SP2000P noncontact specular microscope. The average CCT measured by each method was compared. Subsequently, the mean difference between both sets of measurements was assessed, and the 95% confidence interval (limits of agreement) between both techniques was determined. The mean +/- SD CCT measured by the optical pachymeter was 543 +/- 34 microm and 532 +/- 34 microm for the specular microscope. We found a statistically significant (P < 0.001) mean bias of 10 mum between CCT values measured with both types of equipment, with the optical pachymeter returning the higher values. The coefficient of variation was 6.3% for the optical pachymeter and 6.4% for the specular microscope. The right eye CCT measurements made by the optical pachymeter are, on average, 10 mum thicker than those made with the SP2000P specular microscope, which suggests that both pieces of equipment cannot be used interchangeably to monitor CCT changes in patients. Excluding left eye measurements, the reliability of the optical pachymeter is identical to that of the noncontact specular microscope.

Imaging Schwarzschild multilayer X-ray microscope

NASA Technical Reports Server (NTRS)

Hoover, Richard B.; Baker, Phillip C.; Shealy, David L.; Core, David B.; Walker, Arthur B. C., Jr.; Barbee, Troy W., Jr.; Kerstetter, Ted

1993-01-01

We have designed, analyzed, fabricated, and tested Schwarzschild multilayer X-ray microscopes. These instruments use flow-polished Zerodur mirror substrates which have been coated with multilayers optimized for maximum reflectivity at normal incidence at 135 A. They are being developed as prototypes for the Water Window Imaging X-Ray Microscope. Ultrasmooth mirror sets of hemlite grade sapphire have been fabricated and they are now being coated with multilayers to reflect soft X-rays at 38 A, within the biologically important 'water window'. In this paper, we discuss the fabrication of the microscope optics and structural components as well as the mounting of the optics and assembly of the microscopes. We also describe the optical alignment, interferometric and visible light testing of the microscopes, present interferometrically measured performance data, and provide the first results of optical imaging tests.

Science 101: How Does an Electron Microscope Work?

ERIC Educational Resources Information Center

Robertson, Bill

2013-01-01

Contrary to popular opinion, electron microscopes are not used to look at electrons. They are used to look for structure in things that are too small to observe with an optical microscope, or to obtain images that are magnified much more than is obtainable with an optical microscope. To understand how electron microscopes work, it will help to go…

Theoretical and Experimental Studies of the Electro-Optic Effect: Toward a Microscopic Understanding.

DTIC Science & Technology

1981-08-01

electro - optic effect is investigated both theoretically and experimentally. The theoretical approach is based upon W.A. Harrison’s ’Bond-Orbital Model’. The separate electronic and lattice contributions to the second-order, electro - optic susceptibility are examined within the context of this model and formulae which can accommodate any crystal structure are presented. In addition, a method for estimating the lattice response to a low frequency (dc) electric field is outlined. Finally, experimental measurements of the electro -

Microscopic few-body and Gaussian-shaped density distributions for the analysis of the 6He exotic nucleus with different target nuclei

NASA Astrophysics Data System (ADS)

Aygun, M.; Kucuk, Y.; Boztosun, I.; Ibraheem, Awad A.

2010-12-01

The elastic scattering angular distributions of 6He projectile on different medium and heavy mass target nuclei including 12C, 27Al, 58Ni, 64Zn, 65Cu, 197Au, 208Pb and 209Bi have been examined by using the few-body and Gaussian-shaped density distributions at various energies. The microscopic real parts of the complex nuclear optical potential have been obtained by using the double-folding model for each of the density distributions and the phenomenological imaginary potentials have been taken as the Woods-Saxon type. Comparative results of the few-body and Gaussian-shaped density distributions together with the experimental data are presented within the framework of the optical model.

Optimal model-based sensorless adaptive optics for epifluorescence microscopy.

PubMed

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

2018-01-01

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

The petrographic microscope: Evolution of a mineralogical research instrument

USGS Publications Warehouse

Kile, D.E.

2003-01-01

The petrographic microscope, designed to observe and measure the optical properties of minerals as a means of identifying them, has provided a foundation for mineralogical and petrological research for more than 120 years. Much of what is known today in these fields is attributable to this instrument, the development of which paralleled an evolution of fundamental optical theory and its correlation with mineral structure and composition. This instrument and its related accessories have evolved through a range of models and designs, which are in themselves distinctive for their scientific function and elegant construction, and are today prized by collectors of scientific instruments.

Volumetric Light-field Encryption at the Microscopic Scale

PubMed Central

Li, Haoyu; Guo, Changliang; Muniraj, Inbarasan; Schroeder, Bryce C.; Sheridan, John T.; Jia, Shu

2017-01-01

We report a light-field based method that allows the optical encryption of three-dimensional (3D) volumetric information at the microscopic scale in a single 2D light-field image. The system consists of a microlens array and an array of random phase/amplitude masks. The method utilizes a wave optics model to account for the dominant diffraction effect at this new scale, and the system point-spread function (PSF) serves as the key for encryption and decryption. We successfully developed and demonstrated a deconvolution algorithm to retrieve both spatially multiplexed discrete data and continuous volumetric data from 2D light-field images. Showing that the method is practical for data transmission and storage, we obtained a faithful reconstruction of the 3D volumetric information from a digital copy of the encrypted light-field image. The method represents a new level of optical encryption, paving the way for broad industrial and biomedical applications in processing and securing 3D data at the microscopic scale. PMID:28059149

Volumetric Light-field Encryption at the Microscopic Scale

NASA Astrophysics Data System (ADS)

Li, Haoyu; Guo, Changliang; Muniraj, Inbarasan; Schroeder, Bryce C.; Sheridan, John T.; Jia, Shu

2017-01-01

We report a light-field based method that allows the optical encryption of three-dimensional (3D) volumetric information at the microscopic scale in a single 2D light-field image. The system consists of a microlens array and an array of random phase/amplitude masks. The method utilizes a wave optics model to account for the dominant diffraction effect at this new scale, and the system point-spread function (PSF) serves as the key for encryption and decryption. We successfully developed and demonstrated a deconvolution algorithm to retrieve both spatially multiplexed discrete data and continuous volumetric data from 2D light-field images. Showing that the method is practical for data transmission and storage, we obtained a faithful reconstruction of the 3D volumetric information from a digital copy of the encrypted light-field image. The method represents a new level of optical encryption, paving the way for broad industrial and biomedical applications in processing and securing 3D data at the microscopic scale.

Microscopic optical model potential based on a Dirac Brueckner Hartree Fock approach and the relevant uncertainty analysis

NASA Astrophysics Data System (ADS)

Xu, Ruirui; Ma, Zhongyu; Muether, Herbert; van Dalen, E. N. E.; Liu, Tinjin; Zhang, Yue; Zhang, Zhi; Tian, Yuan

2017-09-01

A relativistic microscopic optical model potential, named CTOM, for nucleon-nucleus scattering is investigated in the framework of Dirac-Brueckner-Hartree-Fock approach. The microscopic feature of CTOM is guaranteed through rigorously adopting the isospin dependent DBHF calculation within the subtracted T matrix scheme. In order to verify its prediction power, a global study n, p+ A scattering are carried out. The predicted scattering observables coincide with experimental data within a good accuracy over a broad range of targets and a large region of energies only with two free items, namely the free-range factor t in the applied improved local density approximation and minor adjustments of the scalar and vector potentials in the low-density region. In addition, to estimate the uncertainty of the theoretical results, the deterministic simple least square approach is preliminarily employed to derive the covariance of predicted angular distributions, which is also briefly contained in this paper.

New design of a cryostat-mounted scanning near-field optical microscope for single molecule spectroscopy

NASA Astrophysics Data System (ADS)

Durand, Yannig; Woehl, Jörg C.; Viellerobe, Bertrand; Göhde, Wolfgang; Orrit, Michel

1999-02-01

Due to the weakness of the fluorescence signal from a single fluorophore, a scanning near-field optical microscope for single molecule spectroscopy requires a very efficient setup for the collection and detection of emitted photons. We have developed a home-built microscope for operation in a l-He cryostat which uses a solid parabolic mirror in order to optimize the fluorescence collection efficiency. This microscope works with Al-coated, tapered optical fibers in illumination mode. The tip-sample separation is probed by an optical shear-force detection. First results demonstrate the capability of the microscope to image single molecules and achieve a topographical resolution of a few nanometers vertically and better than 50 nm laterally.

A stereo-compound hybrid microscope for combined intracellular and optical recording of invertebrate neural network activity.

PubMed

Frost, William N; Wang, Jean; Brandon, Christopher J

2007-05-15

Optical recording studies of invertebrate neural networks with voltage-sensitive dyes seldom employ conventional intracellular electrodes. This may in part be due to the traditional reliance on compound microscopes for such work. While such microscopes have high light-gathering power, they do not provide depth of field, making working with sharp electrodes difficult. Here we describe a hybrid microscope design, with switchable compound and stereo objectives, that eases the use of conventional intracellular electrodes in optical recording experiments. We use it, in combination with a voltage-sensitive dye and photodiode array, to identify neurons participating in the swim motor program of the marine mollusk Tritonia. This microscope design should be applicable to optical recording studies in many preparations.

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.

X-ray laser microscope apparatus

DOEpatents

Suckewer, Szymon; DiCicco, Darrell S.; Hirschberg, Joseph G.; Meixler, Lewis D.; Sathre, Robert; Skinner, Charles H.

1990-01-01

A microscope consisting of an x-ray contact microscope and an optical microscope. The optical, phase contrast, microscope is used to align a target with respect to a source of soft x-rays. The source of soft x-rays preferably comprises an x-ray laser but could comprise a synchrotron or other pulse source of x-rays. Transparent resist material is used to support the target. The optical microscope is located on the opposite side of the transparent resist material from the target and is employed to align the target with respect to the anticipated soft x-ray laser beam. After alignment with the use of the optical microscope, the target is exposed to the soft x-ray laser beam. The x-ray sensitive transparent resist material whose chemical bonds are altered by the x-ray beam passing through the target mater GOVERNMENT LICENSE RIGHTS This invention was made with government support under Contract No. De-FG02-86ER13609 awarded by the Department of Energy. The Government has certain rights in this invention.

A stereo-compound hybrid microscope for combined intracellular and optical recording of invertebrate neural network activity

PubMed Central

Frost, William N.; Wang, Jean; Brandon, Christopher J.

2007-01-01

Optical recording studies of invertebrate neural networks with voltage-sensitive dyes seldom employ conventional intracellular electrodes. This may in part be due to the traditional reliance on compound microscopes for such work. While such microscopes have high light-gathering power, they do not provide depth of field, making working with sharp electrodes difficult. Here we describe a hybrid microscope design, with switchable compound and stereo objectives, that eases the use of conventional intracellular electrodes in optical recording experiments. We use it, in combination with a voltage-sensitive dye and photodiode array, to identify neurons participating in the swim motor program of the marine mollusk Tritonia. This microscope design should be applicable to optical recording studies in many preparations. PMID:17306887

Fractal propagation method enables realistic optical microscopy simulations in biological tissues

PubMed Central

Glaser, Adam K.; Chen, Ye; Liu, Jonathan T.C.

2017-01-01

Current simulation methods for light transport in biological media have limited efficiency and realism when applied to three-dimensional microscopic light transport in biological tissues with refractive heterogeneities. We describe here a technique which combines a beam propagation method valid for modeling light transport in media with weak variations in refractive index, with a fractal model of refractive index turbulence. In contrast to standard simulation methods, this fractal propagation method (FPM) is able to accurately and efficiently simulate the diffraction effects of focused beams, as well as the microscopic heterogeneities present in tissue that result in scattering, refractive beam steering, and the aberration of beam foci. We validate the technique and the relationship between the FPM model parameters and conventional optical parameters used to describe tissues, and also demonstrate the method’s flexibility and robustness by examining the steering and distortion of Gaussian and Bessel beams in tissue with comparison to experimental data. We show that the FPM has utility for the accurate investigation and optimization of optical microscopy methods such as light-sheet, confocal, and nonlinear microscopy. PMID:28983499

Combined magnetic resonance, fluorescence, and histology imaging strategy in a human breast tumor xenograft model

PubMed Central

Jiang, Lu; Greenwood, Tiffany R.; Amstalden van Hove, Erika R.; Chughtai, Kamila; Raman, Venu; Winnard, Paul T.; Heeren, Ron; Artemov, Dmitri; Glunde, Kristine

2014-01-01

Applications of molecular imaging in cancer and other diseases frequently require combining in vivo imaging modalities, such as magnetic resonance and optical imaging, with ex vivo optical, fluorescence, histology, and immunohistochemical (IHC) imaging, to investigate and relate molecular and biological processes to imaging parameters within the same region of interest. We have developed a multimodal image reconstruction and fusion framework that accurately combines in vivo magnetic resonance imaging (MRI) and magnetic resonance spectroscopic imaging (MRSI), ex vivo brightfield and fluorescence microscopic imaging, and ex vivo histology imaging. Ex vivo brightfield microscopic imaging was used as an intermediate modality to facilitate the ultimate link between ex vivo histology and in vivo MRI/MRSI. Tissue sectioning necessary for optical and histology imaging required generation of a three-dimensional (3D) reconstruction module for 2D ex vivo optical and histology imaging data. We developed an external fiducial marker based 3D reconstruction method, which was able to fuse optical brightfield and fluorescence with histology imaging data. Registration of 3D tumor shape was pursued to combine in vivo MRI/MRSI and ex vivo optical brightfield and fluorescence imaging data. This registration strategy was applied to in vivo MRI/MRSI, ex vivo optical brightfield/fluorescence, as well as histology imaging data sets obtained from human breast tumor models. 3D human breast tumor data sets were successfully reconstructed and fused with this platform. PMID:22945331

Stereovision-based integrated system for point cloud reconstruction and simulated brain shift validation.

PubMed

Yang, Xiaochen; Clements, Logan W; Luo, Ma; Narasimhan, Saramati; Thompson, Reid C; Dawant, Benoit M; Miga, Michael I

2017-07-01

Intraoperative soft tissue deformation, referred to as brain shift, compromises the application of current image-guided surgery navigation systems in neurosurgery. A computational model driven by sparse data has been proposed as a cost-effective method to compensate for cortical surface and volumetric displacements. We present a mock environment developed to acquire stereoimages from a tracked operating microscope and to reconstruct three-dimensional point clouds from these images. A reconstruction error of 1 mm is estimated by using a phantom with a known geometry and independently measured deformation extent. The microscope is tracked via an attached tracking rigid body that facilitates the recording of the position of the microscope via a commercial optical tracking system as it moves during the procedure. Point clouds, reconstructed under different microscope positions, are registered into the same space to compute the feature displacements. Using our mock craniotomy device, realistic cortical deformations are generated. When comparing our tracked microscope stereo-pair measure of mock vessel displacements to that of the measurement determined by the independent optically tracked stylus marking, the displacement error was [Formula: see text] on average. These results demonstrate the practicality of using tracked stereoscopic microscope as an alternative to laser range scanners to collect sufficient intraoperative information for brain shift correction.

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.

Failure analysis on optical fiber on swarm flight payload

NASA Astrophysics Data System (ADS)

Bourcier, Frédéric; Fratter, Isabelle; Teyssandier, Florent; Barenes, Magali; Dhenin, Jérémie; Peyriguer, Marie; Petre-Bordenave, Romain

2017-11-01

Failure analysis on optical components is usually carried-out, on standard testing devices such as optical/electronic microscopes and spectrometers, on isolated but representative samples. Such analyses are not contactless and not totally non-invasive, so they cannot be used easily on flight models. Furthermore, for late payload or satellite integration/validation phases with tight schedule issues, it could be necessary to carry out a failure analysis directly on the flight hardware, in cleanroom.

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

PubMed

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

2015-01-01

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

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

PubMed

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

2015-09-01

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

Hyperlens-array-implemented optical microscopy

NASA Astrophysics Data System (ADS)

Iwanaga, Masanobu

2014-08-01

Limit of resolution of conventional optical microscopes has never reached below 100 nm under visible light illumination. We show that numerically designed high-transmittance hyperlens array (HLA) is implemented in an optical microscope and works in practice for achieving one-shot-recording optical images of in-situ placed objects with sub 50 nm resolution in lateral direction. Direct resolution test employing well-defined nanopatterns proves that the HLA-implemented imaging is super-resolution optical microscopy, which works even under nW/mm2 visible illumination for objects. The HLA implementation makes the resolution of conventional microscopes one-scale higher, leading to the 1/10 illumination wavelength range, that is, mesoscopic range.

Optical design and system characterization of an imaging microscope at 121.6 nm

NASA Astrophysics Data System (ADS)

Gao, Weichuan; Finan, Emily; Kim, Geon-Hee; Kim, Youngsik; Milster, Thomas D.

2018-03-01

We present the optical design and system characterization of an imaging microscope prototype at 121.6 nm. System engineering processes are demonstrated through the construction of a Schwarzschild microscope objective, including tolerance analysis, fabrication, alignment, and testing. Further improvements on the as-built system with a correction phase plate are proposed and analyzed. Finally, the microscope assembly and the imaging properties of the prototype are demonstrated.

Large area fabrication of plasmonic nanoparticle grating structure by conventional scanning electron microscope

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

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

Plasmonic nanoparticle grating (PNG) structure of different periods has been fabricated by electron beam lithography using silver halide based transmission electron microscope film as a substrate. Conventional scanning electron microscope is used as a fabrication tool for electron beam lithography. Optical microscope and energy dispersive spectroscopy (EDS) have been used for its morphological and elemental characterization. Optical characterization is performed by UV-Vis absorption spectroscopic technique.

Application of classical models of chirality to optical rectification

NASA Astrophysics Data System (ADS)

Wang, Xiao-Ou; Gong, Li-Jing; Li, Chun-Fei

2008-08-01

Classical models of chirality are used to investigate the optical rectification effect in chiral molecular media. Calculation of the zero frequency first hyperpolarizabilities of chiral molecules with different structures is performed and applied to the derivation of a dc electric-dipole polarization. The expression of second-order nonlinear static-electric-dipole susceptibilities is obtained by theoretical derivation in the isotropic chiral thin films. The microscopic mechanism producing optical rectification is analyzed in view of this calculation. We find that optical rectification is derived from interaction between the electric field gradient (spatial dispersion) and chiral molecules in optically active liquids and solution by our calculation, which is consistent with the result given by Woźniak and Wagnière [Opt. Commun. 114, 131 (1995)]: The optical rectification depends on the fourth-order electric-dipole susceptibilities.

Optical path difference microscopy with a Shack-Hartmann wavefront sensor.

PubMed

Gong, Hai; Agbana, Temitope E; Pozzi, Paolo; Soloviev, Oleg; Verhaegen, Michel; Vdovin, Gleb

2017-06-01

In this Letter, we show that a Shack-Hartmann wavefront sensor can be used for the quantitative measurement of the specimen optical path difference (OPD) in an ordinary incoherent optical microscope, if the spatial coherence of the illumination light in the plane of the specimen is larger than the microscope resolution. To satisfy this condition, the illumination numerical aperture should be smaller than the numerical aperture of the imaging lens. This principle has been successfully applied to build a high-resolution reference-free instrument for the characterization of the OPD of micro-optical components and microscopic biological samples.

Design of small confocal endo-microscopic probe working under multiwavelength environment

NASA Astrophysics Data System (ADS)

Kim, Young-Duk; Ahn, MyoungKi; Gweon, Dae-Gab

2010-02-01

Recently, optical imaging system is widely used in medical purpose. By using optical imaging system specific diseases can be easily diagnosed at early stage because optical imaging system has high resolution performance and various imaging method. These methods are used to get high resolution image of human body and can be used to verify whether the cell is infected by virus. Confocal microscope is one of the famous imaging systems which is used for in-vivo imaging. Because most of diseases are accompanied with cellular level changes, doctors can diagnosis at early stage by observing the cellular image of human organ. Current research is focused in the development of endo-microscope that has great advantage in accessibility to human body. In this research, I designed small probe that is connected to confocal microscope through optical fiber bundle and work as endo-microscope. And this small probe is mainly designed to correct chromatic aberration to use various laser sources for both fluorescence type and reflection type confocal images. By using two kinds of laser sources at the same time we demonstrated multi-modality confocal endo-microscope.

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2015-11-24

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2016-10-25

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2016-11-22

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2017-04-25

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Simultaneous imaging of cellular morphology and multiple biomarkers using an acousto-optic tunable filter-based bright field microscope.

PubMed

Wachman, Elliot S; Geyer, Stanley J; Recht, Joel M; Ward, Jon; Zhang, Bill; Reed, Murray; Pannell, Chris

2014-05-01

An acousto-optic tunable filter (AOTF)-based multispectral imaging microscope system allows the combination of cellular morphology and multiple biomarker stainings on a single microscope slide. We describe advances in AOTF technology that have greatly improved spectral purity, field uniformity, and image quality. A multispectral imaging bright field microscope using these advances demonstrates pathology results that have great potential for clinical use.

Apertureless near-field/far-field CW two-photon microscope for biological and material imaging and spectroscopic applications.

PubMed

Nowak, Derek B; Lawrence, A J; Sánchez, Erik J

2010-12-10

We present the development of a versatile spectroscopic imaging tool to allow for imaging with single-molecule sensitivity and high spatial resolution. The microscope allows for near-field and subdiffraction-limited far-field imaging by integrating a shear-force microscope on top of a custom inverted microscope design. The instrument has the ability to image in ambient conditions with optical resolutions on the order of tens of nanometers in the near field. A single low-cost computer controls the microscope with a field programmable gate array data acquisition card. High spatial resolution imaging is achieved with an inexpensive CW multiphoton excitation source, using an apertureless probe and simplified optical pathways. The high-resolution, combined with high collection efficiency and single-molecule sensitive optical capabilities of the microscope, are demonstrated with a low-cost CW laser source as well as a mode-locked laser source.

Images from Phoenix's MECA Instruments

NASA Technical Reports Server (NTRS)

2008-01-01

The image on the upper left is from NASA's Phoenix Mars Lander's Optical Microscope after a sample informally called 'Sorceress' was delivered to its silicon substrate on the 38th Martian day, or sol, of the mission (July 2, 2008).

A 3D representation of the same sample is on the right, as seen by Phoenix's Atomic Force Microscope. This is 100 times greater magnification than the view from the Optical Microscope, and the most highly magnified image ever seen from another world.

The Optical Microscope and the Atomic Force Microscope are part of Phoenix's Microscopy, Electrochemistry and Conductivity Analyzer instrument.

The Atomic Force Microscope was developed by a Swiss-led consortium in collaboration with Imperial College London.

The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

On the influence of lipid-induced optical anisotropy for the bioimaging of exo- or endocytosis with interference microscopic imaging.

PubMed

Marques, D; Miranda, A; Silva, A G; Munro, P R T; DE Beule, P A A

2018-05-01

Some implementations of interference microscopy imaging use digital holographic measurements of complex scattered fields to reconstruct three-dimensional refractive index maps of weakly scattering, semi-transparent objects, frequently encountered in biological investigations. Reconstruction occurs through application of the object scattering potential which assumes an isotropic refractive index throughout the object. Here, we demonstrate that this assumption can in some circumstances be invalid for biological imaging due to the presence of lipid-induced optical anisotropy. We show that the nanoscale organization of lipids in the observation of cellular endocytosis with polarized light induces a significant change in far-field scattering. We obtain this result by presenting a general solution to Maxwell's equations describing light scattering of core-shell particles near an isotropic substrate covered with an anisotropic thin film. This solution is based on an extension of the Bobbert-Vlieger solution for particle scattering near a substrate delivering an exact solution to the scattering problem in the near field as well as far field. By applying this solution to study light scattering by a lipid vesicle near a lipid bilayer, whereby the lipids are represented through a biaxial optical model, we conclude through ellipsometry concepts that effective amounts of lipid-induced optical anisotropy significantly alter far-field optical scattering in respect to an equivalent optical model that neglects the presence of optical anisotropy. © 2018 The Authors Journal of Microscopy © 2018 Royal Microscopical Society.

Advantages of microscope-integrated intraoperative online optical coherence tomography: usage in Boston keratoprosthesis type I surgery

NASA Astrophysics Data System (ADS)

Siebelmann, Sebastian; Steven, Philipp; Hos, Deniz; Hüttmann, Gereon; Lankenau, Eva; Bachmann, Björn; Cursiefen, Claus

2016-01-01

Boston keratoprosthesis (KPro) type I is a technique to treat patients with corneal diseases that are not amenable to conventional keratoplasty. Correct assembly and central implantation of the prosthesis are crucial for postoperative visual recovery. This study investigates the potential benefit of intraoperative optical coherence tomography (OCT) to monitor KPro surgery. Retrospective case series are presented for two patients who underwent Boston KPro type I implantation. The surgery in both patients was monitored intraoperatively using a commercially available intraoperative OCT (iOCT) device mounted on a surgical microscope. Microscope-integrated intraoperative OCT was able to evaluate the correct assembly and implantation of the KPro. All parts of the prosthesis were visible, and interfaces between the corneal graft and titanium backplate or anterior optics were clearly depictable. Moreover, iOCT visualized a gap between the backplate and graft in one case, and in the other case, a gap between the anterior optic and graft. Neither gap was visible with a conventional surgical microscope. The gap between the anterior optic and the graft could easily be corrected. Microscope-integrated iOCT delivers enhanced information, adding to the normal surgical microscope view during KPro surgery. Correct assembly can be controlled as well as the correct placement of the Boston KPro into the anterior chamber.

Design of a normal incidence multilayer imaging X-ray microscope

NASA Astrophysics Data System (ADS)

Shealy, David L.; Gabardi, David R.; Hoover, Richard B.; Walker, Arthur B. C., Jr.; Lindblom, Joakim F.

Normal incidence multilayer Cassegrain X-ray telescopes were flown on the Stanford/MSFC Rocket X-ray Spectroheliograph. These instruments produced high spatial resolution images of the sun and conclusively demonstrated that doubly reflecting multilayer X-ray optical systems are feasible. The images indicated that aplanatic imaging soft X-ray/EUV microscopes should be achievable using multilayer optics technology. A doubly reflecting normal incidence multilayer imaging X-ray microscope based on the Schwarzschild configuration has been designed. The design of the microscope and the results of the optical system ray trace analysis are discussed. High resolution aplanatic imaging X-ray microscopes using normal incidence multilayer X-ray mirrors should have many important applications in advanced X-ray astronomical instrumentation, X-ray lithography, biological, biomedical, metallurgical, and laser fusion research.

Chromatic confocal microscope using hybrid aspheric diffractive lenses

NASA Astrophysics Data System (ADS)

Rayer, Mathieu; Mansfield, Daniel

2014-05-01

A chromatic confocal microscope is a single point non-contact distance measurement sensor. For three decades the vast majority of the chromatic confocal microscope use refractive-based lenses to code the measurement axis chromatically. However, such an approach is limiting the range of applications. In this paper the performance of refractive, diffractive and Hybrid aspheric diffractive are compared. Hybrid aspheric diffractive lenses combine the low geometric aberration of a diffractive lens with the high optical power of an aspheric lens. Hybrid aspheric diffractive lenses can reduce the number of elements in an imaging system significantly or create large hyper- chromatic lenses for sensing applications. In addition, diffractive lenses can improve the resolution and the dynamic range of a chromatic confocal microscope. However, to be suitable for commercial applications, the diffractive optical power must be significant. Therefore, manufacturing such lenses is a challenge. We show in this paper how a theoretical manufacturing model can demonstrate that the hybrid aspheric diffractive configuration with the best performances is achieved by step diffractive surface. The high optical quality of step diffractive surface is then demonstrated experimentally. Publisher's Note: This paper, originally published on 5/10/14, was replaced with a corrected/revised version on 5/19/14. If you downloaded the original PDF but are unable to access the revision, please contact SPIE Digital Library Customer Service for assistance.

Integrated system for point cloud reconstruction and simulated brain shift validation using tracked surgical microscope

NASA Astrophysics Data System (ADS)

Yang, Xiaochen; Clements, Logan W.; Luo, Ma; Narasimhan, Saramati; Thompson, Reid C.; Dawant, Benoit M.; Miga, Michael I.

2017-03-01

Intra-operative soft tissue deformation, referred to as brain shift, compromises the application of current imageguided surgery (IGS) navigation systems in neurosurgery. A computational model driven by sparse data has been used as a cost effective method to compensate for cortical surface and volumetric displacements. Stereoscopic microscopes and laser range scanners (LRS) are the two most investigated sparse intra-operative imaging modalities for driving these systems. However, integrating these devices in the clinical workflow to facilitate development and evaluation requires developing systems that easily permit data acquisition and processing. In this work we present a mock environment developed to acquire stereo images from a tracked operating microscope and to reconstruct 3D point clouds from these images. A reconstruction error of 1 mm is estimated by using a phantom with a known geometry and independently measured deformation extent. The microscope is tracked via an attached tracking rigid body that facilitates the recording of the position of the microscope via a commercial optical tracking system as it moves during the procedure. Point clouds, reconstructed under different microscope positions, are registered into the same space in order to compute the feature displacements. Using our mock craniotomy device, realistic cortical deformations are generated. Our experimental results report approximately 2mm average displacement error compared with the optical tracking system. These results demonstrate the practicality of using tracked stereoscopic microscope as an alternative to LRS to collect sufficient intraoperative information for brain shift correction.

Observation of three-dimensional internal structure of steel materials by means of serial sectioning with ultrasonic elliptical vibration cutting.

PubMed

Fujisaki, K; Yokota, H; Nakatsuchi, H; Yamagata, Y; Nishikawa, T; Udagawa, T; Makinouchi, A

2010-01-01

A three-dimensional (3D) internal structure observation system based on serial sectioning was developed from an ultrasonic elliptical vibration cutting device and an optical microscope combined with a high-precision positioning device. For bearing steel samples, the cutting device created mirrored surfaces suitable for optical metallography, even for long-cutting distances during serial sectioning of these ferrous materials. Serial sectioning progressed automatically by means of numerical control. The system was used to observe inclusions in steel materials on a scale of several tens of micrometers. Three specimens containing inclusions were prepared from bearing steels. These inclusions could be detected as two-dimensional (2D) sectional images with resolution better than 1 mum. A three-dimensional (3D) model of each inclusion was reconstructed from the 2D serial images. The microscopic 3D models had sharp edges and complicated surfaces.

Distributed microscopic actuation analysis of deformable plate membrane mirrors

NASA Astrophysics Data System (ADS)

Lu, Yifan; Yue, Honghao; Deng, Zongquan; Tzou, Hornsen

2018-02-01

To further reduce the areal density of optical mirrors used in space telescopes and other space-borne optical structures, the concept of flexible membrane deformable mirror has been proposed. Because of their high flexibility, poor stiffness and low damping properties, environmental excitations such as orbital maneuver, path changing, and non-uniform heating may induce unexpected vibrations and thus reduce working performance. Therefore, active vibration control is essential for these membrane mirrors. In this paper, two different mirror models, i.e., the plate membrane model and pure membrane model, are studied respectively. In order to investigate the modal vibration characteristics of the mirror, a piezoelectric layer is fully laminated on its non-reflective side to serve as actuators. Dynamic equations of the mirror laminated with piezoelectric actuators are presented first. Then, the actuator induced modal control force is defined. When the actuator area shrinks to infinitesimal, the expressions of microscopic local modal control force and its two components are obtained to predict the spatial microscopic actuation behavior of the mirror. Different membrane pretension forces are also applied to reveal the tension effects on the actuation of the mirror. Analyses indicate that the spatial distribution of modal micro-control forces is exactly the same with the sensing signals distribution of the mirror, which provides crucial guidelines for optimal actuator placement of membrane deformable mirrors.

Tunable thin-film optical filters for hyperspectral microscopy

NASA Astrophysics Data System (ADS)

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

2013-02-01

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

In Vivo Near Infrared Virtual Intraoperative Surgical Photoacoustic Optical Coherence Tomography

PubMed Central

Lee, Donghyun; Lee, Changho; Kim, Sehui; Zhou, Qifa; Kim, Jeehyun; Kim, Chulhong

2016-01-01

Since its first implementation in otolaryngological surgery nearly a century ago, the surgical microscope has improved the accuracy and the safety of microsurgeries. However, the microscope shows only a magnified surface view of the surgical region. To overcome this limitation, either optical coherence tomography (OCT) or photoacoustic microscopy (PAM) has been independently combined with conventional surgical microscope. Herein, we present a near-infrared virtual intraoperative photoacoustic optical coherence tomography (NIR-VISPAOCT) system that combines both PAM and OCT with a conventional surgical microscope. Using optical scattering and absorption, the NIR-VISPAOCT system simultaneously provides surgeons with real-time comprehensive biological information such as tumor margins, tissue structure, and a magnified view of the region of interest. Moreover, by utilizing a miniaturized beam projector, it can back-project 2D cross-sectional PAM and OCT images onto the microscopic view plane. In this way, both microscopic and cross-sectional PAM and OCT images are concurrently displayed on the ocular lens of the microscope. To verify the usability of the NIR-VISPAOCT system, we demonstrate simulated surgeries, including in vivo image-guided melanoma resection surgery and in vivo needle injection of carbon particles into a mouse thigh. The proposed NIR-VISPAOCT system has potential applications in neurosurgery, ophthalmological surgery, and other microsurgeries. PMID:27731390

In vivo fiber-optic confocal reflectance microscope with an injection-molded plastic miniature objective lens.

PubMed

Carlson, Kristen; Chidley, Matthew; Sung, Kung-Bin; Descour, Michael; Gillenwater, Ann; Follen, Michele; Richards-Kortum, Rebecca

2005-04-01

For in vivo optical diagnostic technologies to be distributed to the developed and developing worlds, optical imaging systems must be constructed of inexpensive components. We present a fiber-optic confocal reflectance microscope with a cost-effective injection-molded plastic miniature objective lens for in vivo imaging of human tissues in near real time. The measured lateral resolution is less than 2.2 microm, and the measured axial resolution is 10 microm. Confocal images of ex vivo cervical tissue biopsies and in vivo human lip taken at 15 frames/s demonstrate the microscope's capability of imaging cell morphology and tissue architecture.

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.

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.

Fibre-optic nonlinear optical microscopy and endoscopy.

PubMed

Fu, L; Gu, M

2007-06-01

Nonlinear optical microscopy has been an indispensable laboratory tool of high-resolution imaging in thick tissue and live animals. Rapid developments of fibre-optic components in terms of growing functionality and decreasing size provide enormous opportunities for innovations in nonlinear optical microscopy. Fibre-based nonlinear optical endoscopy is the sole instrumentation to permit the cellular imaging within hollow tissue tracts or solid organs that are inaccessible to a conventional optical microscope. This article reviews the current development of fibre-optic nonlinear optical microscopy and endoscopy, which includes crucial technologies for miniaturized nonlinear optical microscopy and their embodiments of endoscopic systems. A particular attention is given to several classes of photonic crystal fibres that have been applied to nonlinear optical microscopy due to their unique properties for ultrashort pulse delivery and signal collection. Furthermore, fibre-optic nonlinear optical imaging systems can be classified into portable microscopes suitable for imaging behaving animals, rigid endoscopes that allow for deep tissue imaging with minimally invasive manners, and flexible endoscopes enabling imaging of internal organs. Fibre-optic nonlinear optical endoscopy is coming of age and a paradigm shift leading to optical microscope tools for early cancer detection and minimally invasive surgery.

Optical Tweezer Assembly and Calibration

NASA Technical Reports Server (NTRS)

Collins, Timothy M.

2004-01-01

An Optical Tweezer, as the name implies, is a useful tool for precision manipulation of micro and nano scale objects. Using the principle of electromagnetic radiation pressure, an optical tweezer employs a tightly focused laser beam to trap and position objects of various shapes and sizes. These devices can trap micrometer and nanometer sized objects. An exciting possibility for optical tweezers is its future potential to manipulate and assemble micro and nano sized sensors. A typical optical tweezer makes use of the following components: laser, mirrors, lenses, a high quality microscope, stage, Charge Coupled Device (CCD) camera, TV monitor and Position Sensitive Detectors (PSDs). The laser wavelength employed is typically in the visible or infrared spectrum. The laser beam is directed via mirrors and lenses into the microscope. It is then tightly focused by a high magnification, high numerical aperture microscope objective into the sample slide, which is mounted on a translating stage. The sample slide contains a sealed, small volume of fluid that the objects are suspended in. The most common objects trapped by optical tweezers are dielectric spheres. When trapped, a sphere will literally snap into and center itself in the laser beam. The PSD s are mounted in such a way to receive the backscatter after the beam has passed through the trap. PSD s used with the Differential Interference Contrast (DIC) technique provide highly precise data. Most optical tweezers employ lasers with power levels ranging from 10 to 100 miliwatts. Typical forces exerted on trapped objects are in the pico-newton range. When PSDs are employed, object movement can be resolved on a nanometer scale in a time range of milliseconds. Such accuracy, however, can only by utilized by calibrating the optical tweezer. Fortunately, an optical tweezer can be modeled accurately as a simple spring. This allows Hook s Law to be used. My goal this summer at NASA Glenn Research Center is the assembly and calibration of an optical tweezer setup in the Instrumentation and Controls Division (5520). I am utilizing a custom LabVIEW Virtual Instrument program for data collection and microscope stage control. Helping me in my assignment are the following people: Mentor Susan Wrbanek (5520), Dr. Baha Jassemnejad (UCO) and Technicians Ken Weiland (7650) and James Williams (7650). Without their help, my task would not be possible.

Multi-scale image segmentation and numerical modeling in carbonate rocks

NASA Astrophysics Data System (ADS)

Alves, G. C.; Vanorio, T.

2016-12-01

Numerical methods based on computational simulations can be an important tool in estimating physical properties of rocks. These can complement experimental results, especially when time constraints and sample availability are a problem. However, computational models created at different scales can yield conflicting results with respect to the physical laboratory. This problem is exacerbated in carbonate rocks due to their heterogeneity at all scales. We developed a multi-scale approach performing segmentation of the rock images and numerical modeling across several scales, accounting for those heterogeneities. As a first step, we measured the porosity and the elastic properties of a group of carbonate samples with varying micrite content. Then, samples were imaged by Scanning Electron Microscope (SEM) as well as optical microscope at different magnifications. We applied three different image segmentation techniques to create numerical models from the SEM images and performed numerical simulations of the elastic wave-equation. Our results show that a multi-scale approach can efficiently account for micro-porosities in tight micrite-supported samples, yielding acoustic velocities comparable to those obtained experimentally. Nevertheless, in high-porosity samples characterized by larger grain/micrite ratio, results show that SEM scale images tend to overestimate velocities, mostly due to their inability to capture macro- and/or intragranular- porosity. This suggests that, for high-porosity carbonate samples, optical microscope images would be more suited for numerical simulations.

Integrating the Advanced Human Eye Model (AHEM) and optical instrument models to model complete visual optical systems inclusive of the typical or atypical eye

NASA Astrophysics Data System (ADS)

Donnelly, William J., III

2012-06-01

PURPOSE: To present a commercially available optical modeling software tool to assist the development of optical instrumentation and systems that utilize and/or integrate with the human eye. METHODS: A commercially available flexible eye modeling system is presented, the Advanced Human Eye Model (AHEM). AHEM is a module that the engineer can use to perform rapid development and test scenarios on systems that integrate with the eye. Methods include merging modeled systems initially developed outside of AHEM and performing a series of wizard-type operations that relieve the user from requiring an optometric or ophthalmic background to produce a complete eye inclusive system. Scenarios consist of retinal imaging of targets and sources through integrated systems. Uses include, but are not limited to, optimization, telescopes, microscopes, spectacles, contact and intraocular lenses, ocular aberrations, cataract simulation and scattering, and twin eye model (binocular) systems. RESULTS: Metrics, graphical data, and exportable CAD geometry are generated from the various modeling scenarios.

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.

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.

Foveal light exposure is increased at the time of removal of silicone oil with the potential for phototoxicity.

PubMed

Dogramaci, Mahmut; Williams, Katie; Lee, Ed; Williamson, Tom H

2013-01-01

There is sudden and dramatic visual function deterioration in 1-10 % of eyes filled with silicone oil at the time of removal of silicon oil. Transmission of high-energy blue light is increased in eyes filled with silicone oil. We sought to identify if increased foveal light exposure is a potential factor in the pathophysiology of the visual loss at the time of removal of silicone oil. A graphic ray tracing computer program and laboratory models were used to determine the effect of the intraocular silicone oil bubble size on the foveal illuminance at the time of removal of silicone oil under direct microscope light. The graphic ray tracing computer program revealed a range of optical vignetting effects created by different sizes of silicone oil bubble within the vitreous cavity giving rise to an uneven macular illumination. The laboratory model was used to quantify the variation of illuminance at the foveal region with different sizes of silicone oil bubble with in the vitreous cavity at the time of removal of silicon oil under direct microscope light. To substantiate the hypothesis of the light toxicity during removal of silicone oil, The outcome of oil removal procedures performed under direct microscope illumination in compared to those performed under blocked illumination. The computer program showed that the optical vignetting effect at the macula was dependent on the size of the intraocular silicone oil bubble. The laboratory eye model showed that the foveal illuminance followed a bell-shaped curve with 70 % greater illuminance demonstrated at with 50-60 % silicone oil fill. The clinical data identified five eyes with unexplained vision loss out of 114 eyes that had the procedure performed under direct microscope illumination compared to none out of 78 eyes that had the procedure under blocked illumination. Foveal light exposure, and therefore the potential for phototoxicity, is transiently increased at the time of removal of silicone oil. This is due to uneven macular illumination resulting from the optical vignetting effect of different silicone oil bubble sizes. The increase in foveal light exposure may be significant when the procedure is performed under bright operating microscope light on already stressed photoreceptors of an eye filled with silicon oil. We advocate the use of precautions, such as central shadow filter on the operating microscope light source to reduce foveal light exposure and the risk of phototoxicity at the time of removal of silicone oil. The graphic ray tracing computer program used in this study shows promise in eye modeling for future studies.

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.

Simulation and experimental results of optical and thermal modeling of gold nanoshells.

PubMed

Ghazanfari, Lida; Khosroshahi, Mohammad E

2014-09-01

This paper proposes a generalized method for optical and thermal modeling of synthesized magneto-optical nanoshells (MNSs) for biomedical applications. Superparamagnetic magnetite nanoparticles with diameter of 9.5 ± 1.4 nm are fabricated using co-precipitation method and subsequently covered by a thin layer of gold to obtain 15.8 ± 3.5 nm MNSs. In this paper, simulations and detailed analysis are carried out for different nanoshell geometry to achieve a maximum heat power. Structural, magnetic and optical properties of MNSs are assessed using vibrating sample magnetometer (VSM), X-ray diffraction (XRD), UV-VIS spectrophotometer, dynamic light scattering (DLS), and transmission electron microscope (TEM). Magnetic saturation of synthesized magnetite nanoparticles are reduced from 46.94 to 11.98 emu/g after coating with gold. The performance of the proposed optical-thermal modeling technique is verified by simulation and experimental results. Copyright © 2014 Elsevier B.V. All rights reserved.

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

Naulleau, Patrick; Mochi, Iacopo; Goldberg, Kenneth A.

Defect free masks remain one of the most significant challenges facing the commercialization of extreme ultraviolet (EUV) lithography. Progress on this front requires high-performance wavelength-specific metrology of EUV masks, including high-resolution and aerial-image microscopy performed near the 13.5 nm wavelength. Arguably the most cost-effective and rapid path to proliferating this capability is through the development of Fresnel zoneplate-based microscopes. Given the relative obscurity of such systems, however, modeling tools are not necessarily optimized to deal with them and their imaging properties are poorly understood. Here we present a modeling methodology to analyze zoneplate microscopes based on commercially available optical modelingmore » software and use the technique to investigate the imaging performance of an off-axis EUV microscope design. The modeling predicts that superior performance can be achieved by tilting the zoneplate, making it perpendicular to the chief ray at the center of the field, while designing the zoneplate to explicitly work in that tilted plane. Although the examples presented here are in the realm of EUV mask inspection, the methods described and analysis results are broadly applicable to zoneplate microscopes in general, including full-field soft-x-ray microscopes rou tinely used in the synchrotron community.« less

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

Naulleau, Patrick P.; Mochi, Iacopo; Goldberg, Kenneth A.

Defect free masks remain one of the most significant challenges facing the commercialization of extreme ultraviolet (EUV) lithography. Progress on this front requires high-performance wavelength-specific metrology of EUV masks, including high-resolution and aerial-image microscopy performed near the 13.5 nm wavelength. Arguably the most cost-effective and rapid path to proliferating this capability is through the development of Fresnel zoneplate-based microscopes. Given the relative obscurity of such systems, however, modeling tools are not necessarily optimized to deal with them and their imaging properties are poorly understood. Here we present a modeling methodology to analyze zoneplate microscopes based on commercially available optical modelingmore » software and use the technique to investigate the imaging performance of an off-axis EUV microscope design. The modeling predicts that superior performance can be achieved by tilting the zoneplate, making it perpendicular to the chief ray at the center of the field, while designing the zoneplate to explicitly work in that tilted plane. Although the examples presented here are in the realm of EUV mask inspection, the methods described and analysis results are broadly applicable to zoneplate microscopes in general, including full-field soft-x-ray microscopes routinely used in the synchrotron community.« less

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.

Surface defects evaluation system based on electromagnetic model simulation and inverse-recognition calibration method

NASA Astrophysics Data System (ADS)

Yang, Yongying; Chai, Huiting; Li, Chen; Zhang, Yihui; Wu, Fan; Bai, Jian; Shen, Yibing

2017-05-01

Digitized evaluation of micro sparse defects on large fine optical surfaces is one of the challenges in the field of optical manufacturing and inspection. The surface defects evaluation system (SDES) for large fine optical surfaces is developed based on our previously reported work. In this paper, the electromagnetic simulation model based on Finite-Difference Time-Domain (FDTD) for vector diffraction theory is firstly established to study the law of microscopic scattering dark-field imaging. Given the aberration in actual optical systems, point spread function (PSF) approximated by a Gaussian function is introduced in the extrapolation from the near field to the far field and the scatter intensity distribution in the image plane is deduced. Analysis shows that both diffraction-broadening imaging and geometrical imaging should be considered in precise size evaluation of defects. Thus, a novel inverse-recognition calibration method is put forward to avoid confusion caused by diffraction-broadening effect. The evaluation method is applied to quantitative evaluation of defects information. The evaluation results of samples of many materials by SDES are compared with those by OLYMPUS microscope to verify the micron-scale resolution and precision. The established system has been applied to inspect defects on large fine optical surfaces and can achieve defects inspection of surfaces as large as 850 mm×500 mm with the resolution of 0.5 μm.

Optical properties of graphene superlattices.

PubMed

Le, H Anh; Ho, S Ta; Nguyen, D Chien; Do, V Nam

2014-10-08

In this work, the optical responses of graphene superlattices, i.e. graphene subjected to a periodic scalar potential, are theoretically reported. The optical properties were studied by investigating the optical conductivity, which was calculated using the Kubo formalism. It was found that the optical conductivity becomes dependent on the photon polarization and is suppressed in the photon energy range of (0, Ub), where Ub is the potential barrier height. In the higher photon energy range, i.e. Ω > Ub, the optical conductivity is, however, almost identical to that of pristine graphene. Such behaviors of the optical conductivity are explained microscopically through the analysis of the elements of optical matrices and effectively through a simple model, which is based on the Pauli blocking mechanism.

Mie scattering off coated microbubbles

NASA Astrophysics Data System (ADS)

Nelissen, Radboud; Koene, Elmer; Hilgenfeldt, Sascha; Versluis, Michel

2002-11-01

The acoustic behavior of coated microbubbles depends on parameters of the shell coating, which are in turn dependent on bubble size. More intimate knowledge of this size dependence is required for an improved modeling of a distribution of coated microbubbles such as found in ultrasound contrast agents (UCA). Here a setup is designed to simultaneously measure the optical and acoustic response of an ultrasound-driven single bubble contained in a capillary or levitated by the pressure field of a focused transducer. Optical detection is done by Mie scattering through an inverted microscope. Acoustical detection of the single bubble by a receiving transducer is made possible because of the large working distance of the microscope. For Mie scattering investigation of excited bubbles, two regimes can be distinguished, which require different detection techniques: Conventional wide-angle detection through the microscope objective is sufficient for bubbles of radius exceeding 10 mum. For smaller bubbles, two narrow-aperture detectors are used to reconstruct the bubble dynamics from the complex angle-dependence of the scattered light.

On the importance of image formation optics in the design of infrared spectroscopic imaging systems

PubMed Central

Mayerich, David; van Dijk, Thomas; Walsh, Michael; Schulmerich, Matthew; Carney, P. Scott

2014-01-01

Infrared spectroscopic imaging provides micron-scale spatial resolution with molecular contrast. While recent work demonstrates that sample morphology affects the recorded spectrum, considerably less attention has been focused on the effects of the optics, including the condenser and objective. This analysis is extremely important, since it will be possible to understand effects on recorded data and provides insight for reducing optical effects through rigorous microscope design. Here, we present a theoretical description and experimental results that demonstrate the effects of commonly-employed cassegranian optics on recorded spectra. We first combine an explicit model of image formation and a method for quantifying and visualizing the deviations in recorded spectra as a function of microscope optics. We then verify these simulations with measurements obtained from spatially heterogeneous samples. The deviation of the computed spectrum from the ideal case is quantified via a map which we call a deviation map. The deviation map is obtained as a function of optical elements by systematic simulations. Examination of deviation maps demonstrates that the optimal optical configuration for minimal deviation is contrary to prevailing practice in which throughput is maximized for an instrument without a sample. This report should be helpful for understanding recorded spectra as a function of the optics, the analytical limits of recorded data determined by the optical design, and potential routes for optimization of imaging systems. PMID:24936526

On the importance of image formation optics in the design of infrared spectroscopic imaging systems.

PubMed

Mayerich, David; van Dijk, Thomas; Walsh, Michael J; Schulmerich, Matthew V; Carney, P Scott; Bhargava, Rohit

2014-08-21

Infrared spectroscopic imaging provides micron-scale spatial resolution with molecular contrast. While recent work demonstrates that sample morphology affects the recorded spectrum, considerably less attention has been focused on the effects of the optics, including the condenser and objective. This analysis is extremely important, since it will be possible to understand effects on recorded data and provides insight for reducing optical effects through rigorous microscope design. Here, we present a theoretical description and experimental results that demonstrate the effects of commonly-employed cassegranian optics on recorded spectra. We first combine an explicit model of image formation and a method for quantifying and visualizing the deviations in recorded spectra as a function of microscope optics. We then verify these simulations with measurements obtained from spatially heterogeneous samples. The deviation of the computed spectrum from the ideal case is quantified via a map which we call a deviation map. The deviation map is obtained as a function of optical elements by systematic simulations. Examination of deviation maps demonstrates that the optimal optical configuration for minimal deviation is contrary to prevailing practice in which throughput is maximized for an instrument without a sample. This report should be helpful for understanding recorded spectra as a function of the optics, the analytical limits of recorded data determined by the optical design, and potential routes for optimization of imaging systems.

Measuring Roughnesses Of Optical Surfaces

NASA Technical Reports Server (NTRS)

Coulter, Daniel R.; Al-Jumaily, Gahnim A.; Raouf, Nasrat A.; Anderson, Mark S.

1994-01-01

Report discusses use of scanning tunneling microscopy and atomic force microscopy to measure roughnesses of optical surfaces. These techniques offer greater spatial resolution than other techniques. Report notes scanning tunneling microscopes and atomic force microscopes resolve down to 1 nm.

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.

The optics of microscope image formation.

PubMed

Wolf, David E

2013-01-01

Although geometric optics gives a good understanding of how the microscope works, it fails in one critical area, which is explaining the origin of microscope resolution. To accomplish this, one must consider the microscope from the viewpoint of physical optics. This chapter describes the theory of the microscope-relating resolution to the highest spatial frequency that a microscope can collect. The chapter illustrates how Huygens' principle or construction can be used to explain the propagation of a plane wave. It is shown that this limit increases with increasing numerical aperture (NA). As a corollary to this, resolution increases with decreasing wavelength because of how NA depends on wavelength. The resolution is higher for blue light than red light. Resolution is dependent on contrast, and the higher the contrast, the higher the resolution. This last point relates to issues of signal-to-noise and dynamic range. The use of video and new digital cameras has necessitated redefining classical limits such as those of Rayleigh's criterion. Copyright © 2007 Elsevier Inc. All rights reserved.

Comparisons between conventional optical imaging and parametric indirect microscopic imaging on human skin detection

NASA Astrophysics Data System (ADS)

Liu, Guoyan; Gao, Kun; Liu, Xuefeng; Ni, Guoqiang

2016-10-01

We report a new method, polarization parameters indirect microscopic imaging with a high transmission infrared light source, to detect the morphology and component of human skin. A conventional reflection microscopic system is used as the basic optical system, into which a polarization-modulation mechanics is inserted and a high transmission infrared light source is utilized. The near-field structural characteristics of human skin can be delivered by infrared waves and material coupling. According to coupling and conduction physics, changes of the optical wave parameters can be calculated and curves of the intensity of the image can be obtained. By analyzing the near-field polarization parameters in nanoscale, we can finally get the inversion images of human skin. Compared with the conventional direct optical microscope, this method can break diffraction limit and achieve a super resolution of sub-100nm. Besides, the method is more sensitive to the edges, wrinkles, boundaries and impurity particles.

Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.

PubMed

Hayashi, Shinichi; Okada, Yasushi

2015-05-01

Most current superresolution (SR) microscope techniques surpass the diffraction limit at the expense of temporal resolution, compromising their applications to live-cell imaging. Here we describe a new SR fluorescence microscope based on confocal microscope optics, which we name the spinning disk superresolution microscope (SDSRM). Theoretically, the SDSRM is equivalent to a structured illumination microscope (SIM) and achieves a spatial resolution of 120 nm, double that of the diffraction limit of wide-field fluorescence microscopy. However, the SDSRM is 10 times faster than a conventional SIM because SR signals are recovered by optical demodulation through the stripe pattern of the disk. Therefore a single SR image requires only a single averaged image through the rotating disk. On the basis of this theory, we modified a commercial spinning disk confocal microscope. The improved resolution around 120 nm was confirmed with biological samples. The rapid dynamics of micro-tubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30-100 frames/s. Because our method requires only small optical modifications, it will enable an easy upgrade from an existing spinning disk confocal to a SR microscope for live-cell imaging. © 2015 Hayashi and Okada. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

HIGH TEMPERATURE MICROSCOPE AND FURNACE

DOEpatents

Olson, D.M.

1961-01-31

A high-temperature microscope is offered. It has a reflecting optic situated above a molten specimen in a furnace and reflecting the image of the same downward through an inert optic member in the floor of the furnace, a plurality of spaced reflecting plane mirrors defining a reflecting path around the furnace, a standard microscope supported in the path of and forming the end terminus of the light path.

Sedimentological Investigations of the Martian Surface using the Mars 2001 Robotic Arm Camera and MECA Optical Microscope

NASA Technical Reports Server (NTRS)

Rice, J. W., Jr.; Smith, P. H.; Marshall, J. R.

1999-01-01

The first microscopic sedimentological studies of the Martian surface will commence with the landing of the Mars Polar Lander (MPL) December 3, 1999. The Robotic Arm Camera (RAC) has a resolution of 25 um/p which will permit detailed micromorphological analysis of surface and subsurface materials. The Robotic Ann will be able to dig up to 50 cm below the surface. The walls of the trench will also be inspected by RAC to look for evidence of stratigraphic and / or sedimentological relationships. The 2001 Mars Lander will build upon and expand the sedimentological research begun by the RAC on MPL. This will be accomplished by: (1) Macroscopic (dm to cm): Descent Imager, Pancam, RAC; (2) Microscopic (mm to um RAC, MECA Optical Microscope (Figure 2), AFM This paper will focus on investigations that can be conducted by the RAC and MECA Optical Microscope.

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.

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.

Optical anisotropy of the human cornea determined with a polarizing microscope.

PubMed

Bone, Richard A; Draper, Grenville

2007-12-01

We have investigated the optical anisotropy of the human cornea using a polarizing microscope normally used for optical mineralogy studies. The central part of the cornea was removed from 14 eyes (seven donors). With the sample placed on the microscope stage, we consistently observed hyperbolic isogyres characteristic of a negative biaxial material. The angle between the optic axes, generally similar in both eyes, ranged from 12 degrees to 40 degrees (mean+/-SD=31 degrees +/-8 degrees ). The optic axial plane always inclined downward in the nasal direction at 1 degrees -45 degrees below the horizontal (mean+/-SD=22+/-13 degrees ). The retardance produced by the corneas was estimated to be less than 200 nm. In conclusion, the human cornea possesses the anisotropy of a negative biaxial material. Both the angle between the optic axes and the retardance were fairly constant among the majority of samples, suggestive of uniformity in corneal structure.

The long road to the use of microscope in clinical medicine in vivo: from early pioneering proposals to the modern perspectives of optical biopsy.

PubMed

Ponti, Giovanni; Muscatello, Umberto; Sgantzos, Markos

2015-01-01

For a long period the scientists did not recognized the potentialities of the compound microscope in medicine. Only few scientists recognized the potentialities of the microscope for the medicine; among them G. Campani who proposed the utilization of his microscope to investigate the skin lesions directly on the patient. The proposal was illustrated in a letter Acta Eruditorum of 1686. The recent development of optical techniques, capable of providing in-focus images of an object from different planes with high spatial resolution, significantly increased the diagnostic potential of the microscope directly on the patient.

Design of an imaging microscope for soft X-ray applications

NASA Astrophysics Data System (ADS)

Hoover, Richard B.; Shealy, David L.; Gabardi, David R.; Walker, Arthur B. C., Jr.; Lindblom, Joakim F.

1988-01-01

An imaging soft X-ray microscope with a spatial resolution of 0.1 micron and normal incidence multilayer optics is discussed. The microscope has a Schwarzschild configuration, which consists of two concentric spherical mirrors with radii of curvature which minimize third-order spherical aberration, coma, and astigmatism. The performance of the Stanford/MSFC Cassegrain X-ray telescope and its relevance to the present microscope are addressed. A ray tracing analysis of the optical system indicates that diffraction-limited performance can be expected for an object height of 0.2 mm.

Sub-25-nm laboratory x-ray microscopy using a compound Fresnel zone plate.

PubMed

von Hofsten, Olov; Bertilson, Michael; Reinspach, Julia; Holmberg, Anders; Hertz, Hans M; Vogt, Ulrich

2009-09-01

Improving the resolution in x-ray microscopes is of high priority to enable future applications in nanoscience. However, high-resolution zone-plate optics often have low efficiency, which makes implementation in laboratory microscopes difficult. We present a laboratory x-ray microscope based on a compound zone plate. The compound zone plate utilizes multiple diffraction orders to achieve high resolution while maintaining reasonable efficiency. We analyze the illumination conditions necessary for this type of optics in order to suppress stray light and demonstrate microscopic imaging resolving 25 nm features.

Measurement of Trap Length for an Optical Trap

NASA Technical Reports Server (NTRS)

Wrbanek, Susan Y.

2009-01-01

The trap length along the beam axis for an optical trap formed with an upright, oil-immersion microscope was measured. The goals for this effort were twofold. It was deemed useful to understand the depth to which an optical trap can reach for purposes of developing a tool to assist in the fabrication of miniature devices. Additionally, it was desired to know whether the measured trap length favored one or the other of two competing theories to model an optical trap. The approach was to trap a microsphere of known size and mass and raise it from its initial trap position. The microsphere was then dropped by blocking the laser beam for a pre-determined amount of time. Dropping the microsphere in a free-fall mode from various heights relative to the coverslip provides an estimate of how the trapping length changes with depth in water in a sample chamber on a microscope slide. While it was not possible to measure the trap length with sufficient precision to support any particular theory of optical trap formation, it was possible to find regions where the presence of physical boundaries influenced optical traps, and determine that the trap length, for the apparatus studied, is between 6 and 7 m. These results allow more precise control using optical micromanipulation to assemble miniature devices by providing information about the distance over which an optical trap is effective.

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

PubMed

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

2013-10-21

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

Surface properties for α-cluster nuclear matter

NASA Astrophysics Data System (ADS)

Castro, J. J.; Soto, J. R.; Yépez, E.

2013-03-01

We introduce a new microscopic model for α-cluster matter, which simulates the properties of ordinary nuclear matter and α-clustering in a curved surface of a large but finite nucleus. The model is based on a nested icosahedral fullerene-like multiple-shell structure, where each vertex is occupied by a microscopic α-particle. The novel aspect of this model is that it allows a consistent description of nuclear surface properties from microscopic parameters to be made without using the leptodermous expansion. In particular, we show that the calculated surface energy is in excellent agreement with the corresponding coefficient of the Bethe-Weizäcker semi-empirical mass formula. We discuss the properties of the surface α-cluster state, which resembles an ultra cold bosonic quantum gas trapped in an optical lattice. By comparing the surface and interior states we are able to estimate the α preformation probability. Possible extensions of this model to study nuclear dynamics through surface vibrations and departures from approximate sphericity are mentioned.

Classification of Salmonella serotypes with hyperspectral microscope imagery

USDA-ARS?s Scientific Manuscript database

Previous research has demonstrated an optical method with acousto-optic tunable filter (AOTF) based hyperspectral microscope imaging (HMI) had potential for classifying gram-negative from gram-positive foodborne pathogenic bacteria rapidly and nondestructively with a minimum sample preparation. In t...

Internal scanning method as unique imaging method of optical vortex scanning microscope

NASA Astrophysics Data System (ADS)

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

2018-06-01

The internal scanning method is specific for the optical vortex microscope. It allows to move the vortex point inside the focused vortex beam with nanometer resolution while the whole beam stays in place. Thus the sample illuminated by the focused vortex beam can be scanned just by the vortex point. We show that this method enables high resolution imaging. The paper presents the preliminary experimental results obtained with the first basic image recovery procedure. A prospect of developing more powerful tools for topography recovery with the optical vortex scanning microscope is discussed shortly.

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.

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

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

Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.

2016-10-01

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

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

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

Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.

2016-03-30

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

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.

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.

Design and Implementation of Harmful Algal Bloom Diagnosis System Based on J2EE Platform

NASA Astrophysics Data System (ADS)

Guo, Chunfeng; Zheng, Haiyong; Ji, Guangrong; Lv, Liang

According to the shortcomings which are time consuming and laborious of the traditional HAB (Harmful Algal Bloom) diagnosis by the experienced experts using microscope, all kinds of methods and technologies to identify HAB emerged such as microscopic images, molecular biology, characteristics of pigments analysis, fluorescence spectra, inherent optical properties, etc. This paper proposes the design and implementation of a web-based diagnosis system integrating the popular methods for HAB identification. This system is designed with J2EE platform based on MVC (Model-View-Controller) model as well as technologies such as JSP, Servlets, EJB and JDBC.

Three Dimensional Imaging of Cold Atoms in a Magneto Optical Trap with a Light Field Microscope

DTIC Science & Technology

2017-09-14

dimensional (3D) volume of the atoms is reconstructed using a modeled point spread function (PSF), taking into consideration the low magnification (1.25...axis fluorescence image. Optical axis separation between two atom clouds is measured to a 100µm accuracy in a 3mm deep volume , with a 16µm in-focus...79 vi Page 4.5 Phase Term Effects on the 3D Volume

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

Microscopic theory of linear light scattering from mesoscopic media and in near-field optics.

PubMed

Keller, Ole

2005-08-01

On the basis of quantum mechanical response theory a microscopic propagator theory of linear light scattering from mesoscopic systems is presented. The central integral equation problem is transferred to a matrix equation problem by discretization in transitions between pairs of (many-body) energy eigenstates. The local-field calculation which appears from this approach is valid down to the microscopic region. Previous theories based on the (macroscopic) dielectric constant concept make use of spatial (geometrical) discretization and cannot in general be trusted on the mesoscopic length scale. The present theory can be applied to light scattering studies in near-field optics. After a brief discussion of the macroscopic integral equation problem a microscopic potential description of the scattering process is established. In combination with the use of microscopic electromagnetic propagators the formalism allows one to make contact to the macroscopic theory of light scattering and to the spatial photon localization problem. The quantum structure of the microscopic conductivity response tensor enables one to establish a clear physical picture of the origin of local-field phenomena in mesoscopic and near-field optics. The Huygens scalar propagator formalism is revisited and its generality in microscopic physics pointed out.

Water window imaging x ray microscope

NASA Technical Reports Server (NTRS)

Hoover, Richard B. (Inventor)

1992-01-01

A high resolution x ray microscope for imaging microscopic structures within biological specimens has an optical system including a highly polished primary and secondary mirror coated with identical multilayer coatings, the mirrors acting at normal incidence. The coatings have a high reflectivity in the narrow wave bandpass between 23.3 and 43.7 angstroms and have low reflectivity outside of this range. The primary mirror has a spherical concave surface and the secondary mirror has a spherical convex surface. The radii of the mirrors are concentric about a common center of curvature on the optical axis of the microscope extending from the object focal plane to the image focal plane. The primary mirror has an annular configuration with a central aperture and the secondary mirror is positioned between the primary mirror and the center of curvature for reflecting radiation through the aperture to a detector. An x ray filter is mounted at the stage end of the microscope, and film sensitive to x rays in the desired band width is mounted in a camera at the image plane of the optical system. The microscope is mounted within a vacuum chamber for minimizing the absorption of x rays in air from a source through the microscope.

In vivo cellular imaging with microscopes enabled by MEMS scanners

NASA Astrophysics Data System (ADS)

Ra, Hyejun

High-resolution optical imaging plays an important role in medical diagnosis and biomedical research. Confocal microscopy is a widely used imaging method for obtaining cellular and sub-cellular images of biological tissue in reflectance and fluorescence modes. Its characteristic optical sectioning capability also enables three-dimensional (3-D) image reconstruction. However, its use has mostly been limited to excised tissues due to the requirement of high numerical aperture (NA) lenses for cellular resolution. Microscope miniaturization can enable in vivo imaging to make possible early cancer diagnosis and biological studies in the innate environment. In this dissertation, microscope miniaturization for in vivo cellular imaging is presented. The dual-axes confocal (DAC) architecture overcomes limitations of the conventional single-axis confocal (SAC) architecture to allow for miniaturization with high resolution. A microelectromechanical systems (MEMS) scanner is the central imaging component that is key in miniaturization of the DAC architecture. The design, fabrication, and characterization of the two-dimensional (2-D) MEMS scanner are presented. The gimbaled MEMS scanner is fabricated on a double silicon-on-insulator (SOI) wafer and is actuated by self-aligned vertical electrostatic combdrives. The imaging performance of the MEMS scanner in a DAC configuration is shown in a breadboard microscope setup, where reflectance and fluorescence imaging is demonstrated. Then, the MEMS scanner is integrated into a miniature DAC microscope. The whole imaging system is integrated into a portable unit for research in small animal models of human biology and disease. In vivo 3-D imaging is demonstrated on mouse skin models showing gene transfer and siRNA silencing. The siRNA silencing process is sequentially imaged in one mouse over time.

In vivo imaging of middle-ear and inner-ear microstructures of a mouse guided by SD-OCT combined with a surgical microscope

PubMed Central

Cho, Nam Hyun; Jang, Jeong Hun; Jung, Woonggyu; Kim, Jeehyun

2014-01-01

We developed an augmented-reality system that combines optical coherence tomography (OCT) with a surgical microscope. By sharing the common optical path in the microscope and OCT, we could simultaneously acquire OCT and microscope views. The system was tested to identify the middle-ear and inner-ear microstructures of a mouse. Considering the probability of clinical application including otorhinolaryngology, diseases such as middle-ear effusion were visualized using in vivo mouse and OCT images simultaneously acquired through the eyepiece of the surgical microscope during surgical manipulation using the proposed system. This system is expected to realize a new practical area of OCT application. PMID:24787787

(Gene sequencing by scanning molecular exciton microscopy)

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

Not Available

1991-01-01

This report details progress made in setting up a laboratory for optical microscopy of genes. The apparatus including a fluorescence microscope, a scanning optical microscope, various spectrometers, and supporting computers is described. Results in developing photon and exciton tips, and in preparing samples are presented. (GHH)

Design, assembly, and optical bench testing of a high-numerical-aperture miniature injection-molded objective for fiber-optic confocal reflectance microscopy.

PubMed

Chidley, Matthew D; Carlson, Kristen D; Richards-Kortum, Rebecca R; Descour, Michael R

2006-04-10

The design, analysis, assembly methods, and optical-bench test results for a miniature injection-molded plastic objective lens used in a fiber-optic confocal reflectance microscope are presented. The five-lens plastic objective was tested as a stand-alone optical system before its integration into a confocal microscope for in vivo imaging of cells and tissue. Changing the spacing and rotation of the individual optical elements can compensate for fabrication inaccuracies and improve performance. The system performance of the miniature objective lens is measured by use of an industry-accepted slanted-edge modulation transfer function (MTF) metric. An estimated Strehl ratio of 0.61 and a MTF value of 0.66 at the fiber-optic bundle Nyquist frequency have been obtained. The optical bench testing system is configured to permit interactive optical alignment during testing to optimize performance. These results are part of an effort to demonstrate the manufacturability of low-cost, high-performance biomedical optics for high-resolution in vivo imaging. Disposable endoscopic microscope objectives could help in vivo confocal microscopy technology mature to permit wide-scale clinical screening and detection of early cancers and precancerous lesions.

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.

Modeling of coherent ultrafast magneto-optical experiments: Light-induced molecular mean-field model

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

Hinschberger, Y.; Hervieux, P.-A.

2015-12-28

We present calculations which aim to describe coherent ultrafast magneto-optical effects observed in time-resolved pump-probe experiments. Our approach is based on a nonlinear semi-classical Drude-Voigt model and is used to interpret experiments performed on nickel ferromagnetic thin film. Within this framework, a phenomenological light-induced coherent molecular mean-field depending on the polarizations of the pump and probe pulses is proposed whose microscopic origin is related to a spin-orbit coupling involving the electron spins of the material sample and the electric field of the laser pulses. Theoretical predictions are compared to available experimental data. The model successfully reproduces the observed experimental trendsmore » and gives meaningful insight into the understanding of magneto-optical rotation behavior in the ultrafast regime. Theoretical predictions for further experimental studies are also proposed.« less

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.

Wide spectral range confocal microscope based on endlessly single-mode fiber.

PubMed

Hubbard, R; Ovchinnikov, Yu B; Hayes, J; Richardson, D J; Fu, Y J; Lin, S D; See, P; Sinclair, A G

2010-08-30

We report an endlessly single mode, fiber-optic confocal microscope, based on a large mode area photonic crystal fiber. The microscope confines a very broad spectral range of excitation and emission wavelengths to a single spatial mode in the fiber. Single-mode operation over an optical octave is feasible. At a magnification of 10 and λ = 900 nm, its resolution was measured to be 1.0 μm (lateral) and 2.5 μm (axial). The microscope's use is demonstrated by imaging single photons emitted by individual InAs quantum dots in a pillar microcavity.

Generation-3 programmable array microscope (PAM) with digital micro-mirror device (DMD)

NASA Astrophysics Data System (ADS)

De Beule, Pieter A. A.; de Vries, Anthony H. B.; Arndt-Jovin, Donna J.; Jovin, Thomas M.

2011-03-01

We report progress on the construction of an optical sectioning programmable array microscope (PAM) implemented with a digital micro-mirror device (DMD) spatial light modulator (SLM) utilized for both fluorescence illumination and detection. The introduction of binary intensity modulation at the focal plane of a microscope objective in a computer controlled pixilated mode allows the recovery of an optically sectioned image. Illumination patterns can be changed very quickly, in contrast to static Nipkow disk or aperture correlation implementations, thereby creating an optical system that can be optimized to the optical specimen in a convenient manner, e.g. for patterned photobleaching, photobleaching reduction, or spatial superresolution. We present a third generation (Gen-3) dual path PAM module incorporating the 25 kHz binary frame rate TI 1080p DMD and a newly developed optical system that offers diffraction limited imaging with compensation of tilt angle distortion.

Image formation of thick three-dimensional objects in differential-interference-contrast microscopy.

PubMed

Trattner, Sigal; Kashdan, Eugene; Feigin, Micha; Sochen, Nir

2014-05-01

The differential-interference-contrast (DIC) microscope is of widespread use in life sciences as it enables noninvasive visualization of transparent objects. The goal of this work is to model the image formation process of thick three-dimensional objects in DIC microscopy. The model is based on the principles of electromagnetic wave propagation and scattering. It simulates light propagation through the components of the DIC microscope to the image plane using a combined geometrical and physical optics approach and replicates the DIC image of the illuminated object. The model is evaluated by comparing simulated images of three-dimensional spherical objects with the recorded images of polystyrene microspheres. Our computer simulations confirm that the model captures the major DIC image characteristics of the simulated object, and it is sensitive to the defocusing effects.

Microscopic modeling of direct pre-equilibrium emission from neutron induced reactions on even and odd actinides

NASA Astrophysics Data System (ADS)

Dupuis, M.; Hilaire, S.; Péru, S.; Bauge, E.; Kerveno, M.; Dessagne, P.; Henning, G.

2017-09-01

Direct inelastic scattering to discrete excitations and pre-equilibrium emission are described within a microscopic model. Nuclear structure information are obtained in the (Quasi) Random Phase Approximation ((Q)RPA) framework implemented with the Gogny force. The relevant optical and transition potentials are build considering the JLM folding model. Various successful applications are shown for (n,n), (n,n'), (n,xn) and (n,xnγ) reactions for spherical and axially deformed even-even or odd targets. The rearrangement corrections to transition potentials and the contribution of unnatural parity excitations to pre-equilibrium emission are discussed. Our model predictions for (n,n'γ) reactions, for intra- and inter-band transitions in 238U, and for the 239Pu(n,2n) cross section are analyzed.

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

Bourdon, Christopher Jay; Olsen, Michael G.; Gorby, Allen D.

The analytical model for the depth of correlation (measurement depth) of a microscopic particle image velocimetry (micro-PIV) experiment derived by Olsen and Adrian (Exp. Fluids, 29, pp. S166-S174, 2000) has been modified to be applicable to experiments using high numerical aperture optics. A series of measurements are presented that experimentally quantify the depth of correlation of micro-PIV velocity measurements which employ high numerical aperture and magnification optics. These measurements demonstrate that the modified analytical model is quite accurate in estimating the depth of correlation in micro-PIV measurements using this class of optics. Additionally, it was found that the Gaussian particlemore » approximation made in this model does not significantly affect the model's performance. It is also demonstrated that this modified analytical model easily predicts the depth of correlation when viewing into a medium of a different index of refraction than the immersion medium.« less

Assessment of a liquid lens enabled in vivo optical coherence microscope.

PubMed

Murali, Supraja; Meemon, Panomsak; Lee, Kye-Sung; Kuhn, William P; Thompson, Kevin P; Rolland, Jannick P

2010-06-01

The optical aberrations induced by imaging through skin can be predicted using formulas for Seidel aberrations of a plane-parallel plate. Knowledge of these aberrations helps to guide the choice of numerical aperture (NA) of the optics we can use in an implementation of Gabor domain optical coherence microscopy (GD-OCM), where the focus is the only aberration adjustment made through depth. On this basis, a custom-designed, liquid-lens enabled dynamic focusing optical coherence microscope operating at 0.2 NA is analyzed and validated experimentally. As part of the analysis, we show that the full width at half-maximum metric, as a characteristic descriptor for the point spread function, while commonly used, is not a useful metric for quantifying resolution in non-diffraction-limited systems. Modulation transfer function (MTF) measurements quantify that the liquid lens performance is as predicted by design, even when accounting for the effect of gravity. MTF measurements in a skinlike scattering medium also quantify the performance of the microscope in its potential applications. To guide the fusion of images across the various focus positions of the microscope, as required in GD-OCM, we present depth of focus measurements that can be used to determine the effective number of focusing zones required for a given goal resolution. Subcellular resolution in an onion sample, and high-definition in vivo imaging in human skin are demonstrated with the custom-designed and built microscope.

Nanoscale Optical Imaging and Spectroscopy from Visible to Mid-Infrared

DTIC Science & Technology

2015-11-13

field characterization of nanoscale materials, it also complements the near- field scanning optical microscope currently available in the PI’s lab...field scanning optical microscope currently available in the PI’s lab. This equipment will begin making major impacts on at least three current DoD...SECURITY CLASSIFICATION OF: 1. REPORT DATE (DD-MM-YYYY) 4. TITLE AND SUBTITLE 13. SUPPLEMENTARY NOTES 12. DISTRIBUTION AVAILIBILITY STATEMENT 6

Microscopic Engine Powered by Critical Demixing

NASA Astrophysics Data System (ADS)

Schmidt, Falko; Magazzù, Alessandro; Callegari, Agnese; Biancofiore, Luca; Cichos, Frank; Volpe, Giovanni

2018-02-01

We experimentally demonstrate a microscopic engine powered by the local reversible demixing of a critical mixture. We show that, when an absorbing microsphere is optically trapped by a focused laser beam in a subcritical mixture, it is set into rotation around the optical axis of the beam because of the emergence of diffusiophoretic propulsion. This behavior can be controlled by adjusting the optical power, the temperature, and the criticality of the mixture.

Tapered fiber optical tweezers for microscopic particle trapping: fabrication and application

NASA Astrophysics Data System (ADS)

Liu, Zhihai; Guo, Chengkai; Yang, Jun; Yuan, Libo

2006-12-01

A novel single tapered fiber optical tweezers is proposed and fabricated by heating and drawing technology. The microscopic particle tapping performance of this special designed tapered fiber probe is demonstrated and investigated. The distribution of the optical field emerging from the tapered fiber tip is numerically calculated based on the beam propagation method. The trapping force FDTD analysis results, both axial and transverse, are also given.

White matter segmentation by estimating tissue optical attenuation from volumetric OCT massive histology of whole rodent brains

NASA Astrophysics Data System (ADS)

Lefebvre, Joël.; Castonguay, Alexandre; Lesage, Frédéric

2017-02-01

A whole rodent brain was imaged using an automated massive histology setup and an Optical Coherence Tomography (OCT) microscope. Thousands of OCT volumetric tiles were acquired, each covering a size of about 2.5x2.5x0.8 mm3 with a sampling resolution of 4.9x4.9x6.5 microns. This paper shows the techniques for reconstruction, attenuation compensation and segmentation of the sliced brains. The tile positions within the mosaic were evaluated using a displacement model of the motorized stage and pairwise coregistration. Volume blending was then performed by solving the 3D Laplace equation, and consecutive slices were assembled using the cross-correlation of their 2D image gradient. This reconstruction algorithm resulted in a 3D map of optical reflectivity for the whole brain at micrometric resolution. OCT tissue slices were then used to estimate the local attenuation coefficient based on a single scattering photon model. The attenuation map obtained exhibits a high contrast for all white matter fibres, regardless of their orientation. The tissue optical attenuation from the intrinsic OCT reflectivity contributes to better white matter tissue segmentation. The combined 3D maps of reflectivity and attenuation is a step toward the study of white matter at a microscopic scale for the whole brain in small animals.

Integration of a Spectral Domain Optical Coherence Tomography System into a Surgical Microscope for Intraoperative Imaging

PubMed Central

Ehlers, Justis P.; Tao, Yuankai K.; Farsiu, Sina; Maldonado, Ramiro; Izatt, Joseph A.

2011-01-01

Purpose. To demonstrate an operating microscope-mounted spectral domain optical coherence tomography (MMOCT) system for human retinal and model surgery imaging. Methods. A prototype MMOCT system was developed to interface directly with an ophthalmic surgical microscope, to allow SDOCT imaging during surgical viewing. Nonoperative MMOCT imaging was performed in an Institutional Review Board–approved protocol in four healthy volunteers. The effect of surgical instrument materials on MMOCT imaging was evaluated while performing retinal surface, intraretinal, and subretinal maneuvers in cadaveric porcine eyes. The instruments included forceps, metallic and polyamide subretinal needles, and soft silicone-tipped instruments, with and without diamond dusting. Results. High-resolution images of the human retina were successfully obtained with the MMOCT system. The optical properties of surgical instruments affected the visualization of the instrument and the underlying retina. Metallic instruments (e.g., forceps and needles) showed high reflectivity with total shadowing below the instrument. Polyamide material had a moderate reflectivity with subtotal shadowing. Silicone instrumentation showed moderate reflectivity with minimal shadowing. Summed voxel projection MMOCT images provided clear visualization of the instruments, whereas the B-scans from the volume revealed details of the interactions between the tissues and the instrumentation (e.g., subretinal space cannulation, retinal elevation, or retinal holes). Conclusions. High-quality retinal imaging is feasible with an MMOCT system. Intraoperative imaging with model eyes provides high-resolution depth information including visualization of the instrument and intraoperative tissue manipulation. This study demonstrates a key component of an interactive platform that could provide enhanced information for the vitreoretinal surgeon. PMID:21282565

Integration of a spectral domain optical coherence tomography system into a surgical microscope for intraoperative imaging.

PubMed

Ehlers, Justis P; Tao, Yuankai K; Farsiu, Sina; Maldonado, Ramiro; Izatt, Joseph A; Toth, Cynthia A

2011-05-16

To demonstrate an operating microscope-mounted spectral domain optical coherence tomography (MMOCT) system for human retinal and model surgery imaging. A prototype MMOCT system was developed to interface directly with an ophthalmic surgical microscope, to allow SDOCT imaging during surgical viewing. Nonoperative MMOCT imaging was performed in an Institutional Review Board-approved protocol in four healthy volunteers. The effect of surgical instrument materials on MMOCT imaging was evaluated while performing retinal surface, intraretinal, and subretinal maneuvers in cadaveric porcine eyes. The instruments included forceps, metallic and polyamide subretinal needles, and soft silicone-tipped instruments, with and without diamond dusting. High-resolution images of the human retina were successfully obtained with the MMOCT system. The optical properties of surgical instruments affected the visualization of the instrument and the underlying retina. Metallic instruments (e.g., forceps and needles) showed high reflectivity with total shadowing below the instrument. Polyamide material had a moderate reflectivity with subtotal shadowing. Silicone instrumentation showed moderate reflectivity with minimal shadowing. Summed voxel projection MMOCT images provided clear visualization of the instruments, whereas the B-scans from the volume revealed details of the interactions between the tissues and the instrumentation (e.g., subretinal space cannulation, retinal elevation, or retinal holes). High-quality retinal imaging is feasible with an MMOCT system. Intraoperative imaging with model eyes provides high-resolution depth information including visualization of the instrument and intraoperative tissue manipulation. This study demonstrates a key component of an interactive platform that could provide enhanced information for the vitreoretinal surgeon.

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.

The potential for early and rapid pathogen detection within poultry processing through hyperspectral microscopy

USDA-ARS?s Scientific Manuscript database

The acquisition of hyperspectral microscopic images containing both spatial and spectral data has shown potential for the early and rapid optical classification of foodborne pathogens. A hyperspectral microscope with a metal halide light source and acousto-optical tunable filter (AOTF) collects 89 ...

Acousto-Optic Tunable Filter Hyperspectral Microscope Imaging Method for Characterizing Spectra from Foodborne Pathogens.

USDA-ARS?s Scientific Manuscript database

Hyperspectral microscope imaging (HMI) method, which provides both spatial and spectral characteristics of samples, can be effective for foodborne pathogen detection. The acousto-optic tunable filter (AOTF)-based HMI method can be used to characterize spectral properties of biofilms formed by Salmon...

In vivo time-serial multi-modality optical imaging in a mouse model of ovarian tumorigenesis

PubMed Central

Watson, Jennifer M; Marion, Samuel L; Rice, Photini F; Bentley, David L; Besselsen, David G; Utzinger, Urs; Hoyer, Patricia B; Barton, Jennifer K

2014-01-01

Identification of the early microscopic changes associated with ovarian cancer may lead to development of a diagnostic test for high-risk women. In this study we use optical coherence tomography (OCT) and multiphoton microscopy (MPM) (collecting both two photon excited fluorescence [TPEF] and second harmonic generation [SHG]) to image mouse ovaries in vivo at multiple time points. We demonstrate the feasibility of imaging mouse ovaries in vivo during a long-term survival study and identify microscopic changes associated with early tumor development. These changes include alterations in tissue microstructure, as seen by OCT, alterations in cellular fluorescence and morphology, as seen by TPEF, and remodeling of collagen structure, as seen by SHG. These results suggest that a combined OCT-MPM system may be useful for early detection of ovarian cancer. PMID:24145178

Neural imaging in songbirds using fiber optic fluorescence microscopy

NASA Astrophysics Data System (ADS)

Nooshabadi, Fatemeh; Hearn, Gentry; Lints, Thierry; Maitland, Kristen C.

2012-02-01

The song control system of juvenile songbirds is an important model for studying the developmental acquisition and generation of complex learned vocal motor sequences, two processes that are fundamental to human speech and language. To understand the neural mechanisms underlying song production, it is critical to characterize the activity of identified neurons in the song control system when the bird is singing. Neural imaging in unrestrained singing birds, although technically challenging, will advance our understanding of neural ensemble coding mechanisms in this system. We are exploring the use of a fiber optic microscope for functional imaging in the brain of behaving and singing birds in order to better understand the contribution of a key brain nucleus (high vocal center nucleus; HVC) to temporal aspects of song motor control. We have constructed a fluorescence microscope with LED illumination, a fiber bundle for transmission of fluorescence excitation and emission light, a ~2x GRIN lens, and a CCD for image acquisition. The system has 2 μm resolution, 375 μm field of view, 200 μm working distance, and 1 mm outer diameter. As an initial characterization of this setup, neurons in HVC were imaged using the fiber optic microscope after injection of quantum dots or fluorescent retrograde tracers into different song nuclei. A Lucid Vivascope confocal microscope was used to confirm the imaging results. Long-term imaging of the activity of these neurons in juvenile birds during singing may lead us to a better understanding of the central motor codes for song and the central mechanism by which auditory experience modifies song motor commands to enable vocal learning and imitation.

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 (

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

PubMed

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

2016-03-15

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

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.

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

Design of a normal incidence multilayer imaging x-ray microscope.

PubMed

Shealy, D L; Gabardi, D R; Hoover, R B; Walker, A B; Lindblom, J F; Barbee, T W

1989-01-01

Normal incidence multilayer Cassegrain x-ray telescopes were flown on the Stanford/MSFC Rocket X-Ray Spectroheliograph. These instruments produced high spatial resolution images of the Sun and conclusively demonstrated that doubly reflecting multilayer x-ray optical systems are feasible. The images indicated that aplanatic imaging soft x-ray /EUV microscopes should be achievable using multilayer optics technology. We have designed a doubly reflecting normal incidence multilayer imaging x-ray microscope based on the Schwarzschild configuration. The Schwarzschild microscope utilizes two spherical mirrors with concentric radii of curvature which are chosen such that the third-order spherical aberration and coma are minimized. We discuss the design of the microscope and the results of the optical system ray trace analysis which indicates that diffraction-limited performance with 600 Å spatial resolution should be obtainable over a 1 mm field of view at a wavelength of 100 Å. Fabrication of several imaging soft x-ray microscopes based upon these designs, for use in conjunction with x-ray telescopes and laser fusion research, is now in progress. High resolution aplanatic imaging x-ray microscopes using normal incidence multilayer x-ray mirrors should have many important applications in advanced x-ray astronomical instrumentation, x-ray lithography, biological, biomedical, metallurgical, and laser fusion research.

Extending Single-Molecule Microscopy Using Optical Fourier Processing

PubMed Central

2015-01-01

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

Extending single-molecule microscopy using optical Fourier processing.

PubMed

Backer, Adam S; Moerner, W E

2014-07-17

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

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.

Microscope basics.

PubMed

Sluder, Greenfield; Nordberg, Joshua J

2013-01-01

This chapter provides information on how microscopes work and discusses some of the microscope issues to be considered in using a video camera on the microscope. There are two types of microscopes in use today for research in cell biology-the older finite tube-length (typically 160mm mechanical tube length) microscopes and the infinity optics microscopes that are now produced. The objective lens forms a magnified, real image of the specimen at a specific distance from the objective known as the intermediate image plane. All objectives are designed to be used with the specimen at a defined distance from the front lens element of the objective (the working distance) so that the image formed is located at a specific location in the microscope. Infinity optics microscopes differ from the finite tube-length microscopes in that the objectives are designed to project the image of the specimen to infinity and do not, on their own, form a real image of the specimen. Three types of objectives are in common use today-plan achromats, plan apochromats, and plan fluorite lenses. The concept of mounting video cameras on the microscope is also presented in the chapter. Copyright © 2003 Elsevier Inc. All rights reserved.

Design and construction of a modular low-cost epifluorescence upright microscope for neuron visualized recording and fluorescence detection.

PubMed

Beltran-Parrazal, Luis; Morgado-Valle, Consuelo; Serrano, Raul E; Manzo, Jorge; Vergara, Julio L

2014-03-30

One of the limitations when establishing an electrophysiology setup, particularly in low resource settings, is the high cost of microscopes. The average cost for a microscope equipped with the optics for infrared (IR) contrast or microfluorometry is $40,000. We hypothesized that optical elements and features included in commercial microscopes are not necessary to IR video-visualize neurons or for microfluorometry. We present instructions for building a low-cost epifluorescence upright microscope suitable for visualized patch-clamp recording and fluorescence detection using mostly catalog-available parts. This microscope supports applications such as visualized whole-cell recording using IR oblique illumination (IR-OI), or more complex applications such as microfluorometry using a photodiode. In both IR-OI and fluorescence, actual resolution measured with 2-μm latex beads is close to theoretical resolution. The lack of movable parts to switch configurations ensures stability when doing intracellular recording. The low cost is a significant advantage of this microscope compared to existent custom-built microscopes. The cost of the simplest configuration with IR-OI is ∼$2000, whereas the cost of the configuration with epifluorescence is ∼$5000. Since this design does not use pieces discarded from commercial microscopes, it is completely reproducible. We suggest that this microscope is a viable alternative for doing in vitro electrophysiology and microfluorometry in low-resource settings. Characteristics such as an open box design, easy assembly, and low-cost make this microscope a useful instrument for science education and teaching for topics such as optics, biology, neuroscience, and for scientific "hands-on" workshops. Copyright © 2014 Elsevier B.V. All rights reserved.

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.

Modified Linnik microscopic interferometry for quantitative depth evaluation of diffraction-limited microgroove

NASA Astrophysics Data System (ADS)

Ye, Shiwei; Takahashi, Satoru; Michihata, Masaki; Takamasu, Kiyoshi

2018-05-01

The quality control of microgrooves is extremely crucial to ensure the performance and stability of microstructures and improve their fabrication efficiency. This paper introduces a novel optical inspection method and a modified Linnik microscopic interferometer measurement system to detect the depth of microgrooves with a width less than the diffraction limit. Using this optical method, the depth of diffraction-limited microgrooves can be related to the near-field optical phase difference, which cannot be practically observed but can be computed from practical far-field observations. Thus, a modified Linnik microscopic interferometer system based on three identical objective lenses and an optical path reversibility principle were developed. In addition, experiments for standard grating microgrooves on the silicon surface were carried out to demonstrate the feasibility and repeatability of the proposed method and developed measurement system.

Understanding the optics to aid microscopy image segmentation.

PubMed

Yin, Zhaozheng; Li, Kang; Kanade, Takeo; Chen, Mei

2010-01-01

Image segmentation is essential for many automated microscopy image analysis systems. Rather than treating microscopy images as general natural images and rushing into the image processing warehouse for solutions, we propose to study a microscope's optical properties to model its image formation process first using phase contrast microscopy as an exemplar. It turns out that the phase contrast imaging system can be relatively well explained by a linear imaging model. Using this model, we formulate a quadratic optimization function with sparseness and smoothness regularizations to restore the "authentic" phase contrast images that directly correspond to specimen's optical path length without phase contrast artifacts such as halo and shade-off. With artifacts removed, high quality segmentation can be achieved by simply thresholding the restored images. The imaging model and restoration method are quantitatively evaluated on two sequences with thousands of cells captured over several days.

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.

[Gene sequencing by scanning molecular exciton microscopy]. Progress report, October 1, 1990--September 30, 1991

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

Not Available

1991-12-31

This report details progress made in setting up a laboratory for optical microscopy of genes. The apparatus including a fluorescence microscope, a scanning optical microscope, various spectrometers, and supporting computers is described. Results in developing photon and exciton tips, and in preparing samples are presented. (GHH)

Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry.

PubMed

Schwertner, M; Booth, M J; Neil, M A A; Wilson, T

2004-01-01

Confocal or multiphoton microscopes, which deliver optical sections and three-dimensional (3D) images of thick specimens, are widely used in biology. These techniques, however, are sensitive to aberrations that may originate from the refractive index structure of the specimen itself. The aberrations cause reduced signal intensity and the 3D resolution of the instrument is compromised. It has been suggested to correct for aberrations in confocal microscopes using adaptive optics. In order to define the design specifications for such adaptive optics systems, one has to know the amount of aberrations present for typical applications such as with biological samples. We have built a phase stepping interferometer microscope that directly measures the aberration of the wavefront. The modal content of the wavefront is extracted by employing Zernike mode decomposition. Results for typical biological specimens are presented. It was found for all samples investigated that higher order Zernike modes give only a small contribution to the overall aberration. Therefore, these higher order modes can be neglected in future adaptive optics sensing and correction schemes implemented into confocal or multiphoton microscopes, leading to more efficient designs.

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.

Novel microscope-integrated stereoscopic heads-up display for intrasurgical optical coherence tomography

PubMed Central

Shen, Liangbo; Carrasco-Zevallos, Oscar; Keller, Brenton; Viehland, Christian; Waterman, Gar; Hahn, Paul S.; Kuo, Anthony N.; Toth, Cynthia A.; Izatt, Joseph A.

2016-01-01

Intra-operative optical coherence tomography (OCT) requires a display technology which allows surgeons to visualize OCT data without disrupting surgery. Previous research and commercial intrasurgical OCT systems have integrated heads-up display (HUD) systems into surgical microscopes to provide monoscopic viewing of OCT data through one microscope ocular. To take full advantage of our previously reported real-time volumetric microscope-integrated OCT (4D MIOCT) system, we describe a stereoscopic HUD which projects a stereo pair of OCT volume renderings into both oculars simultaneously. The stereoscopic HUD uses a novel optical design employing spatial multiplexing to project dual OCT volume renderings utilizing a single micro-display. The optical performance of the surgical microscope with the HUD was quantitatively characterized and the addition of the HUD was found not to substantially effect the resolution, field of view, or pincushion distortion of the operating microscope. In a pilot depth perception subject study, five ophthalmic surgeons completed a pre-set dexterity task with 50.0% (SD = 37.3%) higher success rate and in 35.0% (SD = 24.8%) less time on average with stereoscopic OCT vision compared to monoscopic OCT vision. Preliminary experience using the HUD in 40 vitreo-retinal human surgeries by five ophthalmic surgeons is reported, in which all surgeons reported that the HUD did not alter their normal view of surgery and that live surgical maneuvers were readily visible in displayed stereoscopic OCT volumes. PMID:27231616

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.

Proper alignment of the microscope.

PubMed

Rottenfusser, Rudi

2013-01-01

The light microscope is merely the first element of an imaging system in a research facility. Such a system may include high-speed and/or high-resolution image acquisition capabilities, confocal technologies, and super-resolution methods of various types. Yet more than ever, the proverb "garbage in-garbage out" remains a fact. Image manipulations may be used to conceal a suboptimal microscope setup, but an artifact-free image can only be obtained when the microscope is optimally aligned, both mechanically and optically. Something else is often overlooked in the quest to get the best image out of the microscope: Proper sample preparation! The microscope optics can only do its job when its design criteria are matched to the specimen or vice versa. The specimen itself, the mounting medium, the cover slip, and the type of immersion medium (if applicable) are all part of the total optical makeup. To get the best results out of a microscope, understanding the functions of all of its variable components is important. Only then one knows how to optimize these components for the intended application. Different approaches might be chosen to discuss all of the microscope's components. We decided to follow the light path which starts with the light source and ends at the camera or the eyepieces. To add more transparency to this sequence, the section up to the microscope stage was called the "Illuminating Section", to be followed by the "Imaging Section" which starts with the microscope objective. After understanding the various components, we can start "working with the microscope." To get the best resolution and contrast from the microscope, the practice of "Koehler Illumination" should be understood and followed by every serious microscopist. Step-by-step instructions as well as illustrations of the beam path in an upright and inverted microscope are included in this chapter. A few practical considerations are listed in Section 3. Copyright © 2013 Elsevier Inc. All rights reserved.

Optical and Probe Diagnostics Applied to Reacting Flows

NASA Technical Reports Server (NTRS)

Ticich, Thomas M.

2003-01-01

The general theme of the research my NASA colleague and I have planned is "Optical and probe diagnostics applied to reacting flows". We plan to explore three major threads during the fellowship period. The first interrogates the flame synthesis of carbon nanotubes using aerosol catalysts. Having demonstrated the viability of the technique for nanotube synthesis, we seek to understand the details of this reacting system which are important to its practical application. Laser light scattering will reveal changes in particle size at various heights above the burner. Analysis of the flame gas by mass spectroscopy will reveal the chemical composition of the mixture. Finally, absorption measurements will map the nanotube concentration within the flow. The second thread explores soot oxidation kinetics. Despite the impact of soot on engine performance, fire safety and pollution, models for its oxidation are inhibited by uncertainty in the values of the oxidation rate. We plan to employ both optical and microscopic measurements to refine this rate. Cavity ring-down absorption measurements of the carbonaceous aerosol can provide a measure of the mass concentration with time and, hence, an oxidation rate. Spectroscopic and direct probe measurements will provide the temperature of the system needed for subsequent modeling. These data will be benchmarked against changes in soot nanostructures as revealed by transmission electron microscopic images from directly sampled material.

Study of 11Li+p elastic scattering using BHF formalism with three body force

NASA Astrophysics Data System (ADS)

Sharma, Manjari; Haider, W.

2018-04-01

In the present work we have analyzed the elastic scattering data of 11Li + p at 62, 68.4 and 75 MeV/nucleon, using the microscopic optical potential calculated within the framework of Brueckner-Hartree-Fock formalism (BHF). The calculation uses Argonne v18 and Urbana v14 inter-nucleon potentials and the Urbana IX (UVIX) model of three body force. The required nucleon-density distributions for 11Li are obtained using the semi-phenomenological model for nuclear density distributions. The optical potential has been obtained by folding the g-matrices as calculated in BHF (with and without three body forces) over the nucleon density distributions. We have used the exact method for calculating both the direct and the exchange parts of the spin-orbit potential. Our results reveal that the spin-orbit potential significantly contributes to 11Li+p elastic scattering at all three incident energies. Further, the calculated spin-orbit potential in BHF is much smaller and more diffused as compared with the phenomenological spin-orbit potential. The analysis reveals that the calculated microscopic optical potentials, with and without three body force using BHF approach with phenomenological form of density distribution, provides satisfactory agreement with the elastic scattering data for 11Li+p.

The measurement of ultrasound scattering from individual micron-sized objects and its application in single cell scattering.

PubMed

Falou, Omar; Rui, Min; El Kaffas, Ahmed; Kumaradas, J Carl; Kolios, Michael C

2010-08-01

The measurement of the ultrasound backscatter from individual micron-sized objects such as cells is required for various applications such as tissue characterization. However, performing such a measurement remains a challenge. For example, the presence of air bubbles in a suspension of cells during the measurements may lead to the incorrect interpretation of the acoustic signals. This work introduces a technique for measuring the ultrasound backscatter from individual micron-sized objects by combining a microinjection system with a co-registered optical microscope and an ultrasound imaging device. This allowed the measurement of the ultrasound backscatter response from a single object under optical microscope guidance. The optical and ultrasonic data were used to determine the size of the object and to deduce its backscatter responses, respectively. In order to calibrate the system, the backscatter frequency responses from polystyrene microspheres were measured and compared to theoretical predictions. A very good agreement was found between the measured backscatter responses of individual microspheres and theoretical predictions of an elastic sphere. The backscatter responses from single OCI-AML-5 cells were also investigated. It was found that the backscatter responses from AML cells are best modeled using the fluid sphere model. The advantages, limitations, and future applications of the developed technique are discussed.

Three-dimensional automated nanoparticle tracking using Mie scattering in an optical microscope.

PubMed

Gineste, J-M; Macko, P; Patterson, E A; Whelan, M P

2011-08-01

The forward scattering of light in a conventional inverted optical microscope by nanoparticles ranging in diameter from 10 to 50nm has been used to automatically and quantitatively identify and track their location in three-dimensions with a temporal resolution of 200ms. The standard deviation of the location of nominally stationary 50-nm-diameter nanoparticles was found to be about 50nm along the light path and about 5nm in the plane perpendicular to the light path. The method is based on oscillating the microscope objective along the light path using a piezo actuator and acquiring images with the condenser aperture closed to a minimum to enhance the effects of diffraction. Data processing in the time and spatial domains allowed the location of particles to be obtained automatically so that the technique has potential applications both in the processing of nanoparticles and in their use in a variety of fields including nanobiotechnology, pharmaceuticals and food processing where a simple optical microscope maybe preferred for a variety of reasons. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

Programmable Colored Illumination Microscopy (PCIM): A practical and flexible optical staining approach for microscopic contrast enhancement

NASA Astrophysics Data System (ADS)

Zuo, Chao; Sun, Jiasong; Feng, Shijie; Hu, Yan; Chen, Qian

2016-03-01

Programmable colored illumination microscopy (PCIM) has been proposed as a flexible optical staining technique for microscopic contrast enhancement. In this method, we replace the condenser diaphragm of a conventional microscope with a programmable thin film transistor-liquid crystal display (TFT-LCD). By displaying different patterns on the LCD, numerous established imaging modalities can be realized, such as bright field, dark field, phase contrast, oblique illumination, and Rheinberg illuminations, which conventionally rely on intricate alterations in the respective microscope setups. Furthermore, the ease of modulating both the color and the intensity distribution at the aperture of the condenser opens the possibility to combine multiple microscopic techniques, or even realize completely new methods for optical color contrast staining, such as iridescent dark-field and iridescent phase-contrast imaging. The versatility and effectiveness of PCIM is demonstrated by imaging of several transparent colorless specimens, such as unstained lung cancer cells, diatom, textile fibers, and a cryosection of mouse kidney. Finally, the potentialities of PCIM for RGB-splitting imaging with stained samples are also explored by imaging stained red blood cells and a histological section.

A dark-field microscope for background-free detection of resonance fluorescence from single semiconductor quantum dots operating in a set-and-forget mode

NASA Astrophysics Data System (ADS)

Kuhlmann, Andreas V.; Houel, Julien; Brunner, Daniel; Ludwig, Arne; Reuter, Dirk; Wieck, Andreas D.; Warburton, Richard J.

2013-07-01

Optically active quantum dots, for instance self-assembled InGaAs quantum dots, are potentially excellent single photon sources. The fidelity of the single photons is much improved using resonant rather than non-resonant excitation. With resonant excitation, the challenge is to distinguish between resonance fluorescence and scattered laser light. We have met this challenge by creating a polarization-based dark-field microscope to measure the resonance fluorescence from a single quantum dot at low temperature. We achieve a suppression of the scattered laser exceeding a factor of 107 and background-free detection of resonance fluorescence. The same optical setup operates over the entire quantum dot emission range (920-980 nm) and also in high magnetic fields. The major development is the outstanding long-term stability: once the dark-field point has been established, the microscope operates for days without alignment. The mechanical and optical designs of the microscope are presented, as well as exemplary resonance fluorescence spectroscopy results on individual quantum dots to underline the microscope's excellent performance.

Sensing cantilever beam bending by the optical lever technique and its application to surface stress.

PubMed

Evans, Drew R; Craig, Vincent S J

2006-03-23

Cantilever beams, both microscopic and macroscopic, are used as sensors in a great variety of applications. An optical lever system is commonly employed to determine the deflection and thereby the profile of the cantilever under load. The sensitivity of the optical lever must be calibrated, and this is usually achieved by application of a known load or deflection to the free end of the cantilever. When the sensing operation involves a different type of load or a combination of types of loadings, the calibration and the deflection values derived from it become invalid. Here we develop a master equation that permits the true deflection of the cantilever to be obtained simply from the measurement of the apparent deflection for uniformly distributed loadings and end-moment loadings. These loadings are relevant to the uniform adsorption or application of material to the cantilever or the application of a surface stress to the cantilever and should assist experimentalists using the optical lever, such as in the atomic force microscope, to measure cantilever deflections in a great variety of sensing applications. We then apply this treatment to the experimental evaluation of surface stress. Three forms of Stoney's equation that relate the apparent deflection to the surface stress, which is valid for both macroscopic and microscopic experiments, are derived. Analysis of the errors arising from incorrect modeling of the loading conditions of the cantilever currently applied in experiments is also presented. It is shown that the reported literature values for surface stress in microscopic experiments are typically 9% smaller than their true value. For macroscopic experiments, we demonstrate that the added mass of the film or coating generally dominates the measured deflection and must be accounted for accurately if surface stress measurements are to be made. Further, the reported measurements generally use a form of Stoney's equation that is in error, resulting in an overestimation of surface stress by a factor >5.

Variable Magnification With Kirkpatrick-Baez Optics for Synchrotron X-Ray Microscopy

PubMed Central

Jach, Terrence; Bakulin, Alex S.; Durbin, Stephen M.; Pedulla, Joseph; Macrander, Albert

2006-01-01

We describe the distinction between the operation of a short focal length x-ray microscope forming a real image with a laboratory source (convergent illumination) and with a highly collimated intense beam from a synchrotron light source (Köhler illumination). We demonstrate the distinction with a Kirkpatrick-Baez microscope consisting of short focal length multilayer mirrors operating at an energy of 8 keV. In addition to realizing improvements in the resolution of the optics, the synchrotron radiation microscope is not limited to the usual single magnification at a fixed image plane. Higher magnification images are produced by projection in the limit of geometrical optics with a collimated beam. However, in distinction to the common method of placing the sample behind the optical source of a diverging beam, we describe the situation in which the sample is located in the collimated beam before the optical element. The ultimate limits of this magnification result from diffraction by the specimen and are determined by the sample position relative to the focal point of the optic. We present criteria by which the diffraction is minimized. PMID:27274930

Enhancing performance of LCoS-SLM as adaptive optics by using computer-generated holograms modulation software

NASA Astrophysics Data System (ADS)

Tsai, Chun-Wei; Lyu, Bo-Han; Wang, Chen; Hung, Cheng-Chieh

2017-05-01

We have already developed multi-function and easy-to-use modulation software that was based on LabVIEW system. There are mainly four functions in this modulation software, such as computer generated holograms (CGH) generation, CGH reconstruction, image trimming, and special phase distribution. Based on the above development of CGH modulation software, we could enhance the performance of liquid crystal on silicon - spatial light modulator (LCoSSLM) as similar as the diffractive optical element (DOE) and use it on various adaptive optics (AO) applications. Through the development of special phase distribution, we are going to use the LCoS-SLM with CGH modulation software into AO technology, such as optical microscope system. When the LCOS-SLM panel is integrated in an optical microscope system, it could be placed on the illumination path or on the image forming path. However, LCOS-SLM provides a program-controllable liquid crystal array for optical microscope. It dynamically changes the amplitude or phase of light and gives the obvious advantage, "Flexibility", to the system

Microscopic Studies of Quantum Phase Transitions in Optical Lattices

NASA Astrophysics Data System (ADS)

Bakr, Waseem S.

2011-12-01

In this thesis, I report on experiments that microscopically probe quantum phase transitions of ultracold atoms in optical lattices. We have developed a "quantum gas microscope" that allowed, for the first time, optical imaging and manipulation of single atoms in a quantum-degenerate gas on individual sites of an optical lattice. This system acts as a quantum simulator of strongly correlated materials, which are currently the subject of intense research because of the technological potential of high--T c superconductors and spintronic materials. We have used our microscope to study the superfluid to Mott insulator transition in bosons and a magnetic quantum phase transition in a spin system. In our microscopic study of the superfluid-insulator transition, we have characterized the on-site number statistics in a space- and time-resolved manner. We observed Mott insulators with fidelities as high as 99%, corresponding to entropies of 0.06kB per particle. We also measured local quantum dynamics and directly imaged the shell structure of the Mott insulator. I report on the first quantum magnetism experiments in optical lattices. We have realized a quantum Ising chain in a magnetic field, and observed a quantum phase transition between a paramagnet and antiferromagnet. We achieved strong spin interactions by encoding spins in excitations of a Mott insulator in a tilted lattice. We detected the transition by measuring the total magnetization of the system across the transition using in-situ measurements as well as the Neel ordering in the antiferromagnetic state using noise-correlation techniques. We characterized the dynamics of domain formation in the system. The spin mapping introduced opens up a new path to realizing more exotic states in optical lattices including spin liquids and quantum valence bond solids. As our system sizes become larger, simulating their physics on classical computers will require exponentially larger resources because of entanglement build-up near a quantum phase transition. We have demonstrated a quantum simulator in which all degrees of freedom can be read out microscopically, allowing the simulation of quantum many-body systems with manageable resources. More generally, the ability to image and manipulate individual atoms in optical lattices opens an avenue towards scalable quantum computation.

Development and Optical Testing of the Camera, Hand Lens, and Microscope Probe with Scannable Laser Spectroscopy (CHAMP-SLS)

NASA Technical Reports Server (NTRS)

Mungas, Greg S.; Gursel, Yekta; Sepulveda, Cesar A.; Anderson, Mark; La Baw, Clayton; Johnson, Kenneth R.; Deans, Matthew; Beegle, Luther; Boynton, John

2008-01-01

Conducting high resolution field microscopy with coupled laser spectroscopy that can be used to selectively analyze the surface chemistry of individual pixels in a scene is an enabling capability for next generation robotic and manned spaceflight missions, civil, and military applications. In the laboratory, we use a range of imaging and surface preparation tools that provide us with in-focus images, context imaging for identifying features that we want to investigate at high magnification, and surface-optical coupling that allows us to apply optical spectroscopic analysis techniques for analyzing surface chemistry particularly at high magnifications. The camera, hand lens, and microscope probe with scannable laser spectroscopy (CHAMP-SLS) is an imaging/spectroscopy instrument capable of imaging continuously from infinity down to high resolution microscopy (resolution of approx. 1 micron/pixel in a final camera format), the closer CHAMP-SLS is placed to a feature, the higher the resultant magnification. At hand lens to microscopic magnifications, the imaged scene can be selectively interrogated with point spectroscopic techniques such as Raman spectroscopy, microscopic Laser Induced Breakdown Spectroscopy (micro-LIBS), laser ablation mass-spectrometry, Fluorescence spectroscopy, and/or Reflectance spectroscopy. This paper summarizes the optical design, development, and testing of the CHAMP-SLS optics.

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.

Optical Coherence Tomography–Enhanced Microlaryngoscopy: Preliminary Report of a Noncontact Optical Coherence Tomography System Integrated With a Surgical Microscope

PubMed Central

Vokes, David E.; Jackson, Ryan; Guo, Shuguang; Perez, Jorge A.; Su, Jianping; Ridgway, James M.; Armstrong, William B.; Chen, Zhongping; Wong, Brian J. F.

2014-01-01

Objectives Optical coherence tomography (OCT) is a new imaging modality that uses near-infrared light to produce cross-sectional images of tissue with a resolution approaching that of light microscopy. We have previously reported use of OCT imaging of the vocal folds (VFs) during direct laryngoscopy with a probe held in contact or near-contact with the VFs. This aim of this study was to develop and evaluate a novel OCT system integrated with a surgical microscope to allow hands-free OCT imaging of the VFs, which could be performed simultaneously with microscopic visualization. Methods We performed a prospective evaluation of a new method of acquiring OCT images of the VFs. Results An OCT system was successfully integrated with a surgical microscope to permit noncontact OCT imaging of the VFs of 10 patients. With this novel device we were able to identify VF epithelium and lamina propria; however, the resolution was reduced compared to that achieved with the standard contact or near-contact OCT. Conclusions Optical coherence tomography is able to produce high-resolution images of vocal fold mucosa to a maximum depth of 1.6 mm. It may be used in the diagnosis of VF lesions, particularly early squamous cell carcinoma, in which OCT can show disruption of the basement membrane. Mounting the OCT device directly onto the operating microscope allows hands-free noncontact OCT imaging and simultaneous conventional microscopic visualization of the VFs. However, the lateral resolution of the OCT microscope system is 50 µm, in contrast to the conventional handheld probe system (10 µm). Although such images at this resolution are still useful clinically, improved resolution would enhance the system’s performance, potentially enabling real-time OCT-guided microsurgery of the larynx. PMID:18700431

Structural Investigation of Biological and Semiconductor Nanostructures with Nonlinear Multicontrast Microscopy

NASA Astrophysics Data System (ADS)

Cisek, Richard

Physical and functional properties of advanced nano-composite materials and biological structures are determined by self-organized atoms and molecules into nanostructures and in turn by microscopic organization of the nanostructures into assemblies of higher structural complexity. Therefore, microscopes are indispensable tools for structural investigations at various levels of organization. In this work, novel nonlinear optical microscopy methods were developed to non-invasively study structural organization at the nanoscopic and microscopic levels. Atomic organization of semiconductor nanowires, molecular organization of amylose biocrystallites in starch granules, and microscopic organization of several photosynthetic organisms was elucidated. The structure of ZnSe nanowires, key components in many modern nanodevices, was investigated using polarization harmonic generation microscopy. Based on nonlinear optical properties of the different crystal lattices, zinc blende and wurtzite nanowires were differentiated, and the three-dimensional orientation of the zinc blende nanowires could be found. The structure of starch granules, a model biocrystal, important in food as well as health sciences, was also investigated using polarization harmonic microscopy. The study was combined with ab initio calculations using the crystal structures of amylose A and B, revealing that second harmonic signals originate from the hydroxide and hydrogen bonds in the starch granules. Visualization of several photosynthetic organisms including the green algae, Chlamydomonas reinhardtii, two species of cyanobacteria, Leptolyngbya sp. and Anabaena sp., aggregates of light-harvesting pigment-protein complexes as well as chloroplasts from green plants were also explored, revealing that future nonlinear microscopy applications could include structural studies of cell walls, the Chlamydomonas eyespot, and photosynthetic membranes. In this study, several nonlinear optical microscopy modalities were developed for quantitative structural investigations of nano and micro-sized architectures. Non-invasive extraction of crystallographic information in microscopic samples will have a number of potential benefits, for example, in clinical applications, allowing observations of disease states inside tissues without the need for biopsy. Industrial nanotechnology will benefit from fast determination of nanostructures with nonlinear microscopy that will improve quality of nanodevices.

Portable and cost-effective pixel super-resolution on-chip microscope for telemedicine applications.

PubMed

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

2011-01-01

We report a field-portable lensless on-chip microscope with a lateral resolution of <1 μm and a large field-of-view of ~24 mm(2). This microscope is based on digital in-line holography and a pixel super-resolution algorithm to process multiple lensfree holograms and obtain a single high-resolution hologram. In its compact and cost-effective design, we utilize 23 light emitting diodes butt-coupled to 23 multi-mode optical fibers, and a simple optical filter, with no moving parts. Weighing only ~95 grams, we demonstrate the performance of this field-portable microscope by imaging various objects including human malaria parasites in thin blood smears.

Optics of high-performance electron microscopes*

PubMed Central

Rose, H H

2008-01-01

During recent years, the theory of charged particle optics together with advances in fabrication tolerances and experimental techniques has lead to very significant advances in high-performance electron microscopes. Here, we will describe which theoretical tools, inventions and designs have driven this development. We cover the basic theory of higher-order electron optics and of image formation in electron microscopes. This leads to a description of different methods to correct aberrations by multipole fields and to a discussion of the most advanced design that take advantage of these techniques. The theory of electron mirrors is developed and it is shown how this can be used to correct aberrations and to design energy filters. Finally, different types of energy filters are described. PMID:27877933

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.

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

Reflection of a polarized light cone

NASA Astrophysics Data System (ADS)

Brody, Jed; Weiss, Daniel; Berland, Keith

2013-01-01

We introduce a visually appealing experimental demonstration of Fresnel reflection. In this simple optical experiment, a polarized light beam travels through a high numerical-aperture microscope objective, reflects off a glass slide, and travels back through the same objective lens. The return beam is sampled with a polarizing beam splitter and produces a surprising geometric pattern on an observation screen. Understanding the origin of this pattern requires careful attention to geometry and an understanding of the Fresnel coefficients for S and P polarized light. We demonstrate that in addition to a relatively simple experimental implementation, the shape of the observed pattern can be computed both analytically and by using optical modeling software. The experience of working through complex mathematical computations and demonstrating their agreement with a surprising experimental observation makes this a highly educational experiment for undergraduate optics or advanced-lab courses. It also provides a straightforward yet non-trivial system for teaching students how to use optical modeling software.

Semimicroscopic, Lane-consistent nucleon-nucleus optical model potential up to 200 MeV

NASA Astrophysics Data System (ADS)

Bauge, Eric; Delaroche, Jean-Paul; Girod, Michel

2000-10-01

Our semimicroscopic optical model potential (E. Bauge et al., Phys. Rev. C 58), 1118 (1998). is re-evaluated in order to obtain a Lane-consistent description of (p,p), (n,n) and (p,n IAS) elastic scattering and reaction observables. The re-assessed nuclear matter interaction (which includes sizable renormalizations of the isovector potentials) is folded with microscopic HFB nuclear densities, producing OMPs that are free of adjustable parameters for nuclei with A >= 40. With Lane-consistency of the interaction, and the predictive nature of our HFB calculations, this scheme can be used to calculate observables for nuclei far from the stability line with good predictivity.

Design, Fabrication and Testing of Multilayer Coated X-Ray Optics for the Water Window Imaging X-Ray Microscope

NASA Technical Reports Server (NTRS)

Spencer, Dwight C.

1996-01-01

Hoover et. al. built and tested two imaging Schwarzschild multilayer microscopes. These instruments were constructed as prototypes for the "Water Window Imaging X-Ray Microscope," which is a doubly reflecting, multilayer x-ray microscope configured to operate within the "water window." The "water window" is the narrow region of the x-ray spectrum between the K absorption edges of oxygen (lamda = 23.3 Angstroms) and of carbon (lamda = 43.62 Angstroms), where water is relatively highly transmissive and carbon is highly absorptive. This property of these materials, thus permits the use of high resolution multilayer x-ray microscopes for producing high contrast images of carbon-based structures within the aqueous physiological environments of living cells. We report the design, fabrication and testing of multilayer optics that operate in this regime.

Modeling a Miniaturized Scanning Electron Microscope Focusing Column - Lessons Learned in Electron Optics Simulation

NASA Technical Reports Server (NTRS)

Loyd, Jody; Gregory, Don; Gaskin, Jessica

2016-01-01

This presentation discusses work done to assess the design of a focusing column in a miniaturized Scanning Electron Microscope (SEM) developed at the NASA Marshall Space Flight Center (MSFC) for use in-situ on the Moon-in particular for mineralogical analysis. The MSFC beam column design uses purely electrostatic fields for focusing, because of the severe constraints on mass and electrical power consumption imposed by the goals of lunar exploration and of spaceflight in general. The resolution of an SEM ultimately depends on the size of the focused spot of the scanning beam probe, for which the stated goal here is a diameter of 10 nanometers. Optical aberrations are the main challenge to this performance goal, because they blur the ideal geometrical optical image of the electron source, effectively widening the ideal spot size of the beam probe. In the present work the optical aberrations of the mini SEM focusing column were assessed using direct tracing of non-paraxial rays, as opposed to mathematical estimates of aberrations based on paraxial ray-traces. The geometrical ray-tracing employed here is completely analogous to ray-tracing as conventionally understood in the realm of photon optics, with the major difference being that in electron optics the lens is simply a smoothly varying electric field in vacuum, formed by precisely machined electrodes. Ray-tracing in this context, therefore, relies upon a model of the electrostatic field inside the focusing column to provide the mathematical description of the "lens" being traced. This work relied fundamentally on the boundary element method (BEM) for this electric field model. In carrying out this research the authors discovered that higher accuracy in the field model was essential if aberrations were to be reliably assessed using direct ray-tracing. This led to some work in testing alternative techniques for modeling the electrostatic field. Ultimately, the necessary accuracy was attained using a BEM/Fourier series hybrid approach. The presentation will give background remarks about the MSFC mini Lunar SEM concept and electron optics modeling, followed by a description of the alternate field modeling techniques that were tried, along with their incorporation into a ray-trace simulation. Next, the validation of this simulation against commercially available software will be discussed using an example lens as a test case. Then, the efficacy of aberration assessment using direct ray-tracing will be demonstrated, using this same validation case. The discussion will include practical error checks of the field solution. Finally, the ray-trace assessment of the MSFC mini Lunar SEM concept will be shown and discussed. The authors believe this presentation will be of general interest to practitioners of modeling and simulation, as well as those with a general optics background. Because electron optics and photon optics share many basic concepts (e.g., lenses, images, aberrations, etc.), the appeal of this presentation need not be restricted to just those interested in charged particle optics.

Control algorithms and applications of the wavefront sensorless adaptive optics

NASA Astrophysics Data System (ADS)

Ma, Liang; Wang, Bin; Zhou, Yuanshen; Yang, Huizhen

2017-10-01

Compared with the conventional adaptive optics (AO) system, the wavefront sensorless (WFSless) AO system need not to measure the wavefront and reconstruct it. It is simpler than the conventional AO in system architecture and can be applied to the complex conditions. Based on the analysis of principle and system model of the WFSless AO system, wavefront correction methods of the WFSless AO system were divided into two categories: model-free-based and model-based control algorithms. The WFSless AO system based on model-free-based control algorithms commonly considers the performance metric as a function of the control parameters and then uses certain control algorithm to improve the performance metric. The model-based control algorithms include modal control algorithms, nonlinear control algorithms and control algorithms based on geometrical optics. Based on the brief description of above typical control algorithms, hybrid methods combining the model-free-based control algorithm with the model-based control algorithm were generalized. Additionally, characteristics of various control algorithms were compared and analyzed. We also discussed the extensive applications of WFSless AO system in free space optical communication (FSO), retinal imaging in the human eye, confocal microscope, coherent beam combination (CBC) techniques and extended objects.

Hyperspectral microscope for in vivo imaging of microstructures and cells in tissues

DOEpatents

Demos,; Stavros, G [Livermore, CA

2011-05-17

An optical hyperspectral/multimodal imaging method and apparatus is utilized to provide high signal sensitivity for implementation of various optical imaging approaches. Such a system utilizes long working distance microscope objectives so as to enable off-axis illumination of predetermined tissue thereby allowing for excitation at any optical wavelength, simplifies design, reduces required optical elements, significantly reduces spectral noise from the optical elements and allows for fast image acquisition enabling high quality imaging in-vivo. Such a technology provides a means of detecting disease at the single cell level such as cancer, precancer, ischemic, traumatic or other type of injury, infection, or other diseases or conditions causing alterations in cells and tissue micro structures.

An improved segmentation method for defects inspection on steel roller surface

NASA Astrophysics Data System (ADS)

Xu, Jirui; Li, Xuekun; Cao, Yuzhong; Shi, Depeng; Yang, Jun; Jiang, Sheng; Rong, Yiming

2018-05-01

In the field of metal rolling, the quality of the steel roller's surface is significant for the final rolling products, e.g. metal sheets or foils. Besides the dimensional accuracy and surface roughness, the optical uniformity of the roller surface is also required for high quality rolling application. The typical optical defects of rollers after finish grinding include speckles, chatter marks, feed traces, and combination of all above. Unlike surface roughness, the optical defects can hardly be characterized by the topography or scanning electrical microscope measurement. Only the inspection by bared eyes of experienced engineers appears to be the effective manner for surface optical defects examination for large steel rollers. In this paper, an on-site machine vision system is designed to add on to the roller grinding machine to capture the surface image, and then an improved optical defects segmentation algorithm is developed based on the active contour model. Finally, experiments are carried out to verify the efficacy of the improved model.

Skin suturing and cortical surface viral infusion improves imaging of neuronal ensemble activity with head-mounted miniature microscopes.

PubMed

Li, Xinjian; Cao, Vania Y; Zhang, Wenyu; Mastwal, Surjeet S; Liu, Qing; Otte, Stephani; Wang, Kuan Hong

2017-11-01

In vivo optical imaging of neural activity provides important insights into brain functions at the single-cell level. Cranial windows and virally delivered calcium indicators are commonly used for imaging cortical activity through two-photon microscopes in head-fixed animals. Recently, head-mounted one-photon microscopes have been developed for freely behaving animals. However, minimizing tissue damage from the virus injection procedure and maintaining window clarity for imaging can be technically challenging. We used a wide-diameter glass pipette at the cortical surface for infusing the viral calcium reporter AAV-GCaMP6 into the cortex. After infusion, the scalp skin over the implanted optical window was sutured to facilitate postoperative recovery. The sutured scalp was removed approximately two weeks later and a miniature microscope was attached above the window to image neuronal activity in freely moving mice. We found that cortical surface virus infusion efficiently labeled neurons in superficial layers, and scalp skin suturing helped to maintain the long-term clarity of optical windows. As a result, several hundred neurons could be recorded in freely moving animals. Compared to intracortical virus injection and open-scalp postoperative recovery, our methods minimized tissue damage and dura overgrowth underneath the optical window, and significantly increased the experimental success rate and the yield of identified neurons. Our improved cranial surgery technique allows for high-yield calcium imaging of cortical neurons with head-mounted microscopes in freely behaving animals. This technique may be beneficial for other optical applications such as two-photon microscopy, multi-site imaging, and optogenetic modulation. Published by Elsevier B.V.

A Jones matrix formalism for simulating three-dimensional polarized light imaging of brain tissue.

PubMed

Menzel, M; Michielsen, K; De Raedt, H; Reckfort, J; Amunts, K; Axer, M

2015-10-06

The neuroimaging technique three-dimensional polarized light imaging (3D-PLI) provides a high-resolution reconstruction of nerve fibres in human post-mortem brains. The orientations of the fibres are derived from birefringence measurements of histological brain sections assuming that the nerve fibres—consisting of an axon and a surrounding myelin sheath—are uniaxial birefringent and that the measured optic axis is oriented in the direction of the nerve fibres (macroscopic model). Although experimental studies support this assumption, the molecular structure of the myelin sheath suggests that the birefringence of a nerve fibre can be described more precisely by multiple optic axes oriented radially around the fibre axis (microscopic model). In this paper, we compare the use of the macroscopic and the microscopic model for simulating 3D-PLI by means of the Jones matrix formalism. The simulations show that the macroscopic model ensures a reliable estimation of the fibre orientations as long as the polarimeter does not resolve structures smaller than the diameter of single fibres. In the case of fibre bundles, polarimeters with even higher resolutions can be used without losing reliability. When taking the myelin density into account, the derived fibre orientations are considerably improved. © 2015 The Author(s).

HOMER: the Holographic Optical Microscope for Education and Research

NASA Astrophysics Data System (ADS)

Luviano, Anali

Holography was invented in 1948 by Dennis Gabor and has undergone major advancements since the 2000s leading to the development of commercial digital holographic microscopes (DHM). This noninvasive form of microscopy produces a three-dimensional (3-D) digital model of a sample without altering or destroying the sample, thus allowing the same sample to be studied multiple times. HOMER-the Holographic Optical Microscope for Education and Research-produces a 3-D image from a two-dimensional (2-D) interference pattern captured by a camera that is then put through reconstruction software. This 2-D pattern is created when a reference wave interacts with the sample to produce a secondary wave that interferes with the unaltered part of the reference wave. I constructed HOMER to be an efficient, portable in-line DHM using inexpensive material and free reconstruction software. HOMER uses three different-colored LEDs as light sources. I am testing the performance of HOMER with the goal of producing tri-color images of samples. I'm using small basic biological samples to test the effectiveness of HOMER and plan to transition to complex cellular and biological specimens as I pursue my interest in biophysics. Norwich University.

Model of cohesive properties and structural phase transitions in non-metallic solids

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

Majewski, J.A.; Vogl, P.

1986-01-01

We have developed a simple, yet microscopic and universal model for cohesive properties of solids. This model explains the physical mechanisms determining the chemical and predicts semiquantitatively static and dynamic cohesive properties. It predicts a substantial softening of the long-wavelength transverse optical phonons across the pressure induced phase transition from the zincblenda to rocksalt structure in II-VI compounds. The origin of this softening is shown to be closely related to ferroelectricity.

In vivo imaging of oral neoplasia using a miniaturized fiber optic confocal reflectance microscope.

PubMed

Maitland, Kristen C; Gillenwater, Ann M; Williams, Michelle D; El-Naggar, Adel K; Descour, Michael R; Richards-Kortum, Rebecca R

2008-11-01

The purpose of this study was to determine whether in vivo images of oral mucosa obtained with a fiber optic confocal reflectance microscope could be used to differentiate normal and neoplastic tissues. We imaged 20 oral sites in eight patients undergoing surgery for squamous cell carcinoma. Normal and abnormal areas within the oral cavity were identified clinically, and real-time videos of each site were obtained in vivo using a fiber optic confocal reflectance microscope. Following imaging, each site was biopsied and submitted for histopathologic examination. We identified distinct features, such as nuclear irregularity and spacing, which can be used to qualitatively differentiate between normal and abnormal tissue. Representative confocal images of normal, pre-neoplastic, and neoplastic oral tissue are presented. Previous work using much larger microscopes has demonstrated the ability of confocal reflectance microscopy to image cellular and tissue architecture in situ. New advances in technology have enabled miniaturization of imaging systems for in vivo use.

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

Kuhlmann, Andreas V.; Houel, Julien; Warburton, Richard J.

Optically active quantum dots, for instance self-assembled InGaAs quantum dots, are potentially excellent single photon sources. The fidelity of the single photons is much improved using resonant rather than non-resonant excitation. With resonant excitation, the challenge is to distinguish between resonance fluorescence and scattered laser light. We have met this challenge by creating a polarization-based dark-field microscope to measure the resonance fluorescence from a single quantum dot at low temperature. We achieve a suppression of the scattered laser exceeding a factor of 10{sup 7} and background-free detection of resonance fluorescence. The same optical setup operates over the entire quantum dotmore » emission range (920–980 nm) and also in high magnetic fields. The major development is the outstanding long-term stability: once the dark-field point has been established, the microscope operates for days without alignment. The mechanical and optical designs of the microscope are presented, as well as exemplary resonance fluorescence spectroscopy results on individual quantum dots to underline the microscope's excellent performance.« less

Directional sensitivity of the retina: A layered scattering model of outer-segment photoreceptor pigments

PubMed Central

Vohnsen, Brian

2014-01-01

Photoreceptor outer segments have been modeled as stacked arrays of discs or membrane infoldings containing visual pigments with light-induced dipole moments. Waveguiding has been excluded so fields diffract beyond the physical boundaries of each photoreceptor cell. Optical reciprocity is used to argue for identical radiative and light gathering properties of pigments to model vision. Two models have been introduced: one a macroscopic model that assumes a uniform pigment density across each layer and another microscopic model that includes the spatial location of each pigment molecule within each layer. Both models result in highly similar directionality at the pupil plane which proves to be insensitive to the exact details of the outer-segment packing being predominantly determined by the first and last contributing layers as set by the fraction of bleaching. The versatility of the microscopic model is demonstrated with an array of examples that includes the Stiles-Crawford effect, visibility of a focused beam of light and the role of defocus. PMID:24877016

A Review of Adaptive Optics Optical Coherence Tomography: Technical Advances, Scientific Applications, and the Future.

PubMed

Jonnal, Ravi S; Kocaoglu, Omer P; Zawadzki, Robert J; Liu, Zhuolin; Miller, Donald T; Werner, John S

2016-07-01

Optical coherence tomography (OCT) has enabled "virtual biopsy" of the living human retina, revolutionizing both basic retina research and clinical practice over the past 25 years. For most of those years, in parallel, adaptive optics (AO) has been used to improve the transverse resolution of ophthalmoscopes to foster in vivo study of the retina at the microscopic level. Here, we review work done over the last 15 years to combine the microscopic transverse resolution of AO with the microscopic axial resolution of OCT, building AO-OCT systems with the highest three-dimensional resolution of any existing retinal imaging modality. We surveyed the literature to identify the most influential antecedent work, important milestones in the development of AO-OCT technology, its applications that have yielded new knowledge, research areas into which it may productively expand, and nascent applications that have the potential to grow. Initial efforts focused on demonstrating three-dimensional resolution. Since then, many improvements have been made in resolution and speed, as well as other enhancements of acquisition and postprocessing techniques. Progress on these fronts has produced numerous discoveries about the anatomy, function, and optical properties of the retina. Adaptive optics OCT continues to evolve technically and to contribute to our basic and clinical knowledge of the retina. Due to its capacity to reveal cellular and microscopic detail invisible to clinical OCT systems, it is an ideal companion to those instruments and has the demonstrable potential to produce images that can guide the interpretation of clinical findings.

Optical analysis of a compound quasi-microscope for planetary landers

NASA Technical Reports Server (NTRS)

Wall, S. D.; Burcher, E. E.; Huck, F. O.

1974-01-01

A quasi-microscope concept, consisting of facsimile camera augmented with an auxiliary lens as a magnifier, was introduced and analyzed. The performance achievable with this concept was primarily limited by a trade-off between resolution and object field; this approach leads to a limiting resolution of 20 microns when used with the Viking lander camera (which has an angular resolution of 0.04 deg). An optical system is analyzed which includes a field lens between camera and auxiliary lens to overcome this limitation. It is found that this system, referred to as a compound quasi-microscope, can provide improved resolution (to about 2 microns ) and a larger object field. However, this improvement is at the expense of increased complexity, special camera design requirements, and tighter tolerances on the distances between optical components.

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

NASA Astrophysics Data System (ADS)

Kettel, Johannes; Reinecke, Holger; Müller, Claas

2016-04-01

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

RIPL - Reference Input Parameter Library for Calculation of Nuclear Reactions and Nuclear Data Evaluations

NASA Astrophysics Data System (ADS)

Capote, R.; Herman, M.; Obložinský, P.; Young, P. G.; Goriely, S.; Belgya, T.; Ignatyuk, A. V.; Koning, A. J.; Hilaire, S.; Plujko, V. A.; Avrigeanu, M.; Bersillon, O.; Chadwick, M. B.; Fukahori, T.; Ge, Zhigang; Han, Yinlu; Kailas, S.; Kopecky, J.; Maslov, V. M.; Reffo, G.; Sin, M.; Soukhovitskii, E. Sh.; Talou, P.

2009-12-01

We describe the physics and data included in the Reference Input Parameter Library, which is devoted to input parameters needed in calculations of nuclear reactions and nuclear data evaluations. Advanced modelling codes require substantial numerical input, therefore the International Atomic Energy Agency (IAEA) has worked extensively since 1993 on a library of validated nuclear-model input parameters, referred to as the Reference Input Parameter Library (RIPL). A final RIPL coordinated research project (RIPL-3) was brought to a successful conclusion in December 2008, after 15 years of challenging work carried out through three consecutive IAEA projects. The RIPL-3 library was released in January 2009, and is available on the Web through http://www-nds.iaea.org/RIPL-3/. This work and the resulting database are extremely important to theoreticians involved in the development and use of nuclear reaction modelling (ALICE, EMPIRE, GNASH, UNF, TALYS) both for theoretical research and nuclear data evaluations. The numerical data and computer codes included in RIPL-3 are arranged in seven segments: MASSES contains ground-state properties of nuclei for about 9000 nuclei, including three theoretical predictions of masses and the evaluated experimental masses of Audi et al. (2003). DISCRETE LEVELS contains 117 datasets (one for each element) with all known level schemes, electromagnetic and γ-ray decay probabilities available from ENSDF in October 2007. NEUTRON RESONANCES contains average resonance parameters prepared on the basis of the evaluations performed by Ignatyuk and Mughabghab. OPTICAL MODEL contains 495 sets of phenomenological optical model parameters defined in a wide energy range. When there are insufficient experimental data, the evaluator has to resort to either global parameterizations or microscopic approaches. Radial density distributions to be used as input for microscopic calculations are stored in the MASSES segment. LEVEL DENSITIES contains phenomenological parameterizations based on the modified Fermi gas and superfluid models and microscopic calculations which are based on a realistic microscopic single-particle level scheme. Partial level densities formulae are also recommended. All tabulated total level densities are consistent with both the recommended average neutron resonance parameters and discrete levels. GAMMA contains parameters that quantify giant resonances, experimental gamma-ray strength functions and methods for calculating gamma emission in statistical model codes. The experimental GDR parameters are represented by Lorentzian fits to the photo-absorption cross sections for 102 nuclides ranging from 51V to 239Pu. FISSION includes global prescriptions for fission barriers and nuclear level densities at fission saddle points based on microscopic HFB calculations constrained by experimental fission cross sections.

RIPL - Reference Input Parameter Library for Calculation of Nuclear Reactions and Nuclear Data Evaluations

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

Capote, R.; Herman, M.; Oblozinsky, P.

We describe the physics and data included in the Reference Input Parameter Library, which is devoted to input parameters needed in calculations of nuclear reactions and nuclear data evaluations. Advanced modelling codes require substantial numerical input, therefore the International Atomic Energy Agency (IAEA) has worked extensively since 1993 on a library of validated nuclear-model input parameters, referred to as the Reference Input Parameter Library (RIPL). A final RIPL coordinated research project (RIPL-3) was brought to a successful conclusion in December 2008, after 15 years of challenging work carried out through three consecutive IAEA projects. The RIPL-3 library was released inmore » January 2009, and is available on the Web through (http://www-nds.iaea.org/RIPL-3/). This work and the resulting database are extremely important to theoreticians involved in the development and use of nuclear reaction modelling (ALICE, EMPIRE, GNASH, UNF, TALYS) both for theoretical research and nuclear data evaluations. The numerical data and computer codes included in RIPL-3 are arranged in seven segments: MASSES contains ground-state properties of nuclei for about 9000 nuclei, including three theoretical predictions of masses and the evaluated experimental masses of Audi et al. (2003). DISCRETE LEVELS contains 117 datasets (one for each element) with all known level schemes, electromagnetic and {gamma}-ray decay probabilities available from ENSDF in October 2007. NEUTRON RESONANCES contains average resonance parameters prepared on the basis of the evaluations performed by Ignatyuk and Mughabghab. OPTICAL MODEL contains 495 sets of phenomenological optical model parameters defined in a wide energy range. When there are insufficient experimental data, the evaluator has to resort to either global parameterizations or microscopic approaches. Radial density distributions to be used as input for microscopic calculations are stored in the MASSES segment. LEVEL DENSITIES contains phenomenological parameterizations based on the modified Fermi gas and superfluid models and microscopic calculations which are based on a realistic microscopic single-particle level scheme. Partial level densities formulae are also recommended. All tabulated total level densities are consistent with both the recommended average neutron resonance parameters and discrete levels. GAMMA contains parameters that quantify giant resonances, experimental gamma-ray strength functions and methods for calculating gamma emission in statistical model codes. The experimental GDR parameters are represented by Lorentzian fits to the photo-absorption cross sections for 102 nuclides ranging from {sup 51}V to {sup 239}Pu. FISSION includes global prescriptions for fission barriers and nuclear level densities at fission saddle points based on microscopic HFB calculations constrained by experimental fission cross sections.« less

RIPL-Reference Input Parameter Library for Calculation of Nuclear Reactions and Nuclear Data Evaluations

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

Capote, R.; Herman, M.; Capote,R.

We describe the physics and data included in the Reference Input Parameter Library, which is devoted to input parameters needed in calculations of nuclear reactions and nuclear data evaluations. Advanced modelling codes require substantial numerical input, therefore the International Atomic Energy Agency (IAEA) has worked extensively since 1993 on a library of validated nuclear-model input parameters, referred to as the Reference Input Parameter Library (RIPL). A final RIPL coordinated research project (RIPL-3) was brought to a successful conclusion in December 2008, after 15 years of challenging work carried out through three consecutive IAEA projects. The RIPL-3 library was released inmore » January 2009, and is available on the Web through http://www-nds.iaea.org/RIPL-3/. This work and the resulting database are extremely important to theoreticians involved in the development and use of nuclear reaction modelling (ALICE, EMPIRE, GNASH, UNF, TALYS) both for theoretical research and nuclear data evaluations. The numerical data and computer codes included in RIPL-3 are arranged in seven segments: MASSES contains ground-state properties of nuclei for about 9000 nuclei, including three theoretical predictions of masses and the evaluated experimental masses of Audi et al. (2003). DISCRETE LEVELS contains 117 datasets (one for each element) with all known level schemes, electromagnetic and {gamma}-ray decay probabilities available from ENSDF in October 2007. NEUTRON RESONANCES contains average resonance parameters prepared on the basis of the evaluations performed by Ignatyuk and Mughabghab. OPTICAL MODEL contains 495 sets of phenomenological optical model parameters defined in a wide energy range. When there are insufficient experimental data, the evaluator has to resort to either global parameterizations or microscopic approaches. Radial density distributions to be used as input for microscopic calculations are stored in the MASSES segment. LEVEL DENSITIES contains phenomenological parameterizations based on the modified Fermi gas and superfluid models and microscopic calculations which are based on a realistic microscopic single-particle level scheme. Partial level densities formulae are also recommended. All tabulated total level densities are consistent with both the recommended average neutron resonance parameters and discrete levels. GAMMA contains parameters that quantify giant resonances, experimental gamma-ray strength functions and methods for calculating gamma emission in statistical model codes. The experimental GDR parameters are represented by Lorentzian fits to the photo-absorption cross sections for 102 nuclides ranging from {sup 51}V to {sup 239}Pu. FISSION includes global prescriptions for fission barriers and nuclear level densities at fission saddle points based on microscopic HFB calculations constrained by experimental fission cross sections.« less

Effects of source shape on the numerical aperture factor with a geometrical-optics model.

PubMed

Wan, Der-Shen; Schmit, Joanna; Novak, Erik

2004-04-01

We study the effects of an extended light source on the calibration of an interference microscope, also referred to as an optical profiler. Theoretical and experimental numerical aperture (NA) factors for circular and linear light sources along with collimated laser illumination demonstrate that the shape of the light source or effective aperture cone is critical for a correct NA factor calculation. In practice, more-accurate results for the NA factor are obtained when a linear approximation to the filament light source shape is used in a geometric model. We show that previously measured and derived NA factors show some discrepancies because a circular rather than linear approximation to the filament source was used in the modeling.

Optical constants of electroplated gold from spectroscopic ellipsometry

NASA Astrophysics Data System (ADS)

Synowicki, R. A.; Herzinger, Craig M.; Hall, James T.; Malingowski, Andrew

2017-11-01

The optical constants of an opaque electroplated gold film (Laser Gold from Epner Technology Inc.), were determined by spectroscopic ellipsometry at room temperature over the spectral range from 0.142 μm in the vacuum ultraviolet to 36 μm in the infrared (photon energy range 0.034-8.75 eV). Data from two separate ellipsometer instruments covering different spectral ranges were analyzed simultaneously. The optical constants n&k or ε1&ε2 were determined by fitting an oscillator dispersion model combining Drude, Gaussian, and Sellmeier dispersion functions to the experimental Ψ and Δ data. The data were analyzed using both an ideal bulk substrate model and a simple overlayer model to account for surface roughness. Including the optical surface roughness layer improved ellipsometric data fits in the UV, and using a separate Drude function for the surface layer improved fits in the infrared. The surface roughness was also characterized using an Atomic Force Microscope. Using an oscillator dispersion model for the optical constants determined in this work allows for more realistic extrapolation to longer infrared wavelengths. Extending optical constants out to 50 μm and beyond is important for calibrating far-infrared reflectance measurements. Applications include understanding the thermal performance of cryogenic space-based instruments, such as the James Webb Space Telescope (JWST).

A new computerized moving stage for optical microscopes

NASA Astrophysics Data System (ADS)

Hatiboglu, Can Ulas; Akin, Serhat

2004-06-01

Measurements of microscope stage movements in the x and y directions are of importance for some stereological methods. Traditionally, the length of stage movements is measured with differing precision and accuracy using a suitable motorized stage, a microscope and software. Such equipment is generally expensive and not readily available in many laboratories. One other challenging problem is the adaptability to available microscope systems which weakens the possibility of the equipment to be used with any kind of light microscope. This paper describes a simple and cheap programmable moving stage that can be used with the available microscopes in the market. The movements of the stage are controlled by two servo-motors and a controller chip via a Java-based image processing software. With the developed motorized stage and a microscope equipped with a CCD camera, the software allows complete coverage of the specimens with minimum overlap, eliminating the optical strain associated with counting hundreds of images through an eyepiece, in a quick and precise fashion. The uses and the accuracy of the developed stage are demonstrated using thin sections obtained from a limestone core plug.

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

NASA Astrophysics Data System (ADS)

Lorenzen, Manfred; Campbell, Duncan R.; Johnson, Craig W.

1991-03-01

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 array 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.

Scanning optical microscope with long working distance objective

DOEpatents

Cloutier, Sylvain G.

2010-10-19

A scanning optical microscope, including: a light source to generate a beam of probe light; collimation optics to substantially collimate the probe beam; a probe-result beamsplitter; a long working-distance, infinity-corrected objective; scanning means to scan a beam spot of the focused probe beam on or within a sample; relay optics; and a detector. The collimation optics are disposed in the probe beam. The probe-result beamsplitter is arranged in the optical paths of the probe beam and the resultant light from the sample. The beamsplitter reflects the probe beam into the objective and transmits resultant light. The long working-distance, infinity-corrected objective is also arranged in the optical paths of the probe beam and the resultant light. It focuses the reflected probe beam onto the sample, and collects and substantially collimates the resultant light. The relay optics are arranged to relay the transmitted resultant light from the beamsplitter to the detector.

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

Microscopic optical potentials derived from ab initio translationally invariant nonlocal one-body densities

NASA Astrophysics Data System (ADS)

Gennari, Michael; Vorabbi, Matteo; Calci, Angelo; Navrátil, Petr

2018-03-01

Background: The nuclear optical potential is a successful tool for the study of nucleon-nucleus elastic scattering and its use has been further extended to inelastic scattering and other nuclear reactions. The nuclear density of the target nucleus is a fundamental ingredient in the construction of the optical potential and thus plays an important role in the description of the scattering process. Purpose: In this paper we derive a microscopic optical potential for intermediate energies using ab initio translationally invariant nonlocal one-body nuclear densities computed within the no-core shell model (NCSM) approach utilizing two- and three-nucleon chiral interactions as the only input. Methods: The optical potential is derived at first order within the spectator expansion of the nonrelativistic multiple scattering theory by adopting the impulse approximation. Nonlocal nuclear densities are derived from the NCSM one-body densities calculated in the second quantization. The translational invariance is generated by exactly removing the spurious center-of-mass (COM) component from the NCSM eigenstates. Results: The ground-state local and nonlocal densities of He 4 ,6 ,8 , 12C, and 16O are calculated and applied to optical potential construction. The differential cross sections and the analyzing powers for the elastic proton scattering off these nuclei are then calculated for different values of the incident proton energy. The impact of nonlocality and the COM removal is discussed. Conclusions: The use of nonlocal densities has a substantial impact on the differential cross sections and improves agreement with experiment in comparison to results generated with the local densities especially for light nuclei. For the halo nuclei 6He and 8He, the results for the differential cross section are in a reasonable agreement with the data although a more sophisticated model for the optical potential is required to properly describe the analyzing powers.

Aqueous carrier waveguide in a flow cytometer

DOEpatents

Mariella, Jr., Raymond P.; van den Engh, Gerrit; Northrup, M. Allen

1995-01-01

The liquid of a flow cytometer itself acts as an optical waveguide, thus transmitting the light to an optical filter/detector combination. This alternative apparatus and method for detecting scattered light in a flow cytometer is provided by a device which views and detects the light trapped within the optical waveguide formed by the flow stream. A fiber optic or other light collecting device is positioned within the flow stream. This provides enormous advantages over the standard light collection technique which uses a microscope objective. The signal-to-noise ratio is greatly increased over that for right-angle-scattered light collected by a microscope objective, and the alignment requirements are simplified.

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.

Optical microscope and tapered fiber coupling apparatus for a dilution refrigerator.

PubMed

MacDonald, A J R; Popowich, G G; Hauer, B D; Kim, P H; Fredrick, A; Rojas, X; Doolin, P; Davis, J P

2015-01-01

We have developed a system for tapered fiber measurements of optomechanical resonators inside a dilution refrigerator, which is compatible with both on- and off-chip devices. Our apparatus features full three-dimensional control of the taper-resonator coupling conditions enabling critical coupling, with an overall fiber transmission efficiency of up to 70%. Notably, our design incorporates an optical microscope system consisting of a coherent bundle of 37,000 optical fibers for real-time imaging of the experiment at a resolution of ∼1 μm. We present cryogenic optical and optomechanical measurements of resonators coupled to tapered fibers at temperatures as low as 9 mK.

Differential polarization nonlinear optical microscopy with adaptive optics controlled multiplexed beams.

PubMed

Samim, Masood; Sandkuijl, Daaf; Tretyakov, Ian; Cisek, Richard; Barzda, Virginijus

2013-09-09

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

Design and analysis of a fast, two-mirror soft-x-ray microscope

NASA Technical Reports Server (NTRS)

Shealy, D. L.; Wang, C.; Jiang, W.; Jin, L.; Hoover, R. B.

1992-01-01

During the past several years, a number of investigators have addressed the design, analysis, fabrication, and testing of spherical Schwarzschild microscopes for soft-x-ray applications using multilayer coatings. Some of these systems have demonstrated diffraction limited resolution for small numerical apertures. Rigorously aplanatic, two-aspherical mirror Head microscopes can provide near diffraction limited resolution for very large numerical apertures. The relationships between the numerical aperture, mirror radii and diameters, magnifications, and total system length for Schwarzschild microscope configurations are summarized. Also, an analysis of the characteristics of the Head-Schwarzschild surfaces will be reported. The numerical surface data predicted by the Head equations were fit by a variety of functions and analyzed by conventional optical design codes. Efforts have been made to determine whether current optical substrate and multilayer coating technologies will permit construction of a very fast Head microscope which can provide resolution approaching that of the wavelength of the incident radiation.

Microscopic theory of multiple-phonon-mediated dephasing and relaxation of quantum dots near a photonic band gap

NASA Astrophysics Data System (ADS)

Roy, Chiranjeeb; John, Sajeev

2010-02-01

We derive a quantum theory of the role of acoustic and optical phonons in modifying the optical absorption line shape, polarization dynamics, and population dynamics of a two-level atom (quantum dot) in the “colored” electromagnetic vacuum of a photonic band-gap (PBG) material. This is based on a microscopic Hamiltonian describing both radiative and vibrational processes quantum mechanically. We elucidate the extent to which phonon-assisted decay limits the lifetime of a single photon-atom bound state and derive the modified spontaneous emission dynamics due to coupling to various phonon baths. We demonstrate that coherent interaction with undamped phonons can lead to an enhanced lifetime of a photon-atom bound state in a PBG. This results in reduction of the steady-state atomic polarization but an increase in the fractionalized upper state population in the photon-atom bound state. We demonstrate, on the other hand, that the lifetime of the photon-atom bound state in a PBG is limited by the lifetime of phonons due to lattice anharmonicities (breakup of phonons into lower energy phonons) and purely nonradiative decay. We also derive the modified polarization decay and dephasing rates in the presence of such damping. This leads to a microscopic, quantum theory of the optical absorption line shapes. Our model and formalism provide a starting point for describing dephasing and relaxation in the presence of external coherent fields and multiple quantum dot interactions in electromagnetic reservoirs with radiative memory effects.

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.

4Pi Microscopy.

PubMed

Schmidt, Roman; Engelhardt, Johann; Lang, Marion

2013-01-01

Optical microscopy has become a key technology in the life sciences today. Its noninvasive nature provides access to the interior of intact and even living cells, where specific molecules can be precisely localized by fluorescent tagging. However, the attainable 3D resolution of an optical microscope has long been hampered by a comparatively poor resolution along the optic axis. By coherent focusing through two objective lenses, 4Pi microscopy improves the axial resolution by three- to fivefold. This primer is intended as a starting point for the design and operation of a 4Pi microscope of type A.

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.

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, 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.

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.

Shape oscillations of microparticles on an optical microscope stage.

PubMed

Zhu, Z M; Apfel, R E

1985-11-01

A modulated acoustic radiation pressure technique to produce quadrupole shape oscillations of drops ranging in diameter from 50-220 micron has been used by us. These drops have been suspended by acoustic levitation in a small chamber mounted on a stage of an optical microscope, which allowed easy viewing. The fission of drops and the deformation of sea urchin eggs were also observed.

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

Hyperspectral microscope imaging methods to classify gram-positive and gram-negative foodborne pathogenic bacteria

USDA-ARS?s Scientific Manuscript database

An acousto-optic tunable filter-based hyperspectral microscope imaging method has potential for identification of foodborne pathogenic bacteria from microcolony rapidly with a single cell level. We have successfully developed the method to acquire quality hyperspectral microscopic images from variou...

Laser-induced surface deformation microscope for the study of the dynamic viscoelasticity of plasma membrane in a living cell.

PubMed

Morisaku, Toshinori; Yui, Hiroharu

2018-05-15

A laser-induced surface deformation (LISD) microscope is developed and applied to measurement of the dynamic relaxation responses of the plasma membrane in a living cell. A laser beam is tightly focused on an optional area of cell surface and the focused light induces microscopic deformation on the surface via radiation pressure. The LISD microscope not only allows non-contact and destruction-free measurement but provides power spectra of the surface responses depending on the frequency of the intensity of the laser beam. An optical system for the LISD is equipped via a microscope, allowing us to measure the relaxation responses in sub-cellular-sized regions of the plasma membrane. In addition, the forced oscillation caused by the radiation pressure for surface deformation extends the upper limit of the frequency range in the obtained power spectra to 106 Hz, which enables us to measure relaxation responses in local regions within the plasma membrane. From differences in power-law exponents at higher frequencies, it is realized that a cancerous cell obeys a weaker single power-law than a normal fibroblast cell. Furthermore, the power spectrum of a keratinocyte cell obeys a power-law with two exponents, indicating that alternative mechanical models to a conventional soft glassy rheology model (where single power-laws explain cells' responses below about 103 Hz) are needed for the understanding over a wider frequency range. The LISD microscope would contribute to investigation of microscopic cell rheology, which is important for clarifying the mechanisms of cell migration and tissue construction.

Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope

PubMed Central

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-01-01

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications. PMID:26525841

Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope.

PubMed

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-11-03

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.

Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope

NASA Astrophysics Data System (ADS)

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-11-01

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.

Label-free hyperspectral dark-field microscopy for quantitative scatter imaging

NASA Astrophysics Data System (ADS)

Cheney, Philip; McClatchy, David; Kanick, Stephen; Lemaillet, Paul; Allen, David; Samarov, Daniel; Pogue, Brian; Hwang, Jeeseong

2017-03-01

A hyperspectral dark-field microscope has been developed for imaging spatially distributed diffuse reflectance spectra from light-scattering samples. In this report, quantitative scatter spectroscopy is demonstrated with a uniform scattering phantom, namely a solution of polystyrene microspheres. A Monte Carlo-based inverse model was used to calculate the reduced scattering coefficients of samples of different microsphere concentrations from wavelength-dependent backscattered signal measured by the dark-field microscope. The results are compared to the measurement results from a NIST double-integrating sphere system for validation. Ongoing efforts involve quantitative mapping of scattering and absorption coefficients in samples with spatially heterogeneous optical properties.

Measurement of the Resolution of the Optical Microscope.

ERIC Educational Resources Information Center

Bowlt, C.

1983-01-01

Outlines procedures demonstrating that the aperture of a microscope objective limits resolving power and then, by using ancillary measurements made with a calibrated graticule in the microscope eyepiece, that the experimentally determined value for the maximum resolving power of a given objective is close to the value predicted by theory. (JN)

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.

Polarization microscopy by use of digital holography: application to optical-fiber birefringence measurements.

PubMed

Colomb, Tristan; Dürr, Florian; Cuche, Etienne; Marquet, Pierre; Limberger, Hans G; Salathé, René-Paul; Depeursinge, Christian

2005-07-20

We present a digital holographic microscope that permits one to image polarization state. This technique results from the coupling of digital holographic microscopy and polarization digital holography. The interference between two orthogonally polarized reference waves and the wave transmitted by a microscopic sample, magnified by a microscope objective, is recorded on a CCD camera. The off-axis geometry permits one to reconstruct separately from this single hologram two wavefronts that are used to image the object-wave Jones vector. We applied this technique to image the birefringence of a bent fiber. To evaluate the precision of the phase-difference measurement, the birefringence induced by internal stress in an optical fiber is measured and compared to the birefringence profile captured by a standard method, which had been developed to obtain high-resolution birefringence profiles of optical fibers.

Modeling and validation of spectral BRDF on material surface of space target

NASA Astrophysics Data System (ADS)

Hou, Qingyu; Zhi, Xiyang; Zhang, Huili; Zhang, Wei

2014-11-01

The modeling and the validation methods of the spectral BRDF on the material surface of space target were presented. First, the microscopic characteristics of the space targets' material surface were analyzed based on fiber-optic spectrometer using to measure the direction reflectivity of the typical materials surface. To determine the material surface of space target is isotropic, atomic force microscopy was used to measure the material surface structure of space target and obtain Gaussian distribution model of microscopic surface element height. Then, the spectral BRDF model based on that the characteristics of the material surface were isotropic and the surface micro-facet with the Gaussian distribution which we obtained was constructed. The model characterizes smooth and rough surface well for describing the material surface of the space target appropriately. Finally, a spectral BRDF measurement platform in a laboratory was set up, which contains tungsten halogen lamp lighting system, fiber optic spectrometer detection system and measuring mechanical systems with controlling the entire experimental measurement and collecting measurement data by computers automatically. Yellow thermal control material and solar cell were measured with the spectral BRDF, which showed the relationship between the reflection angle and BRDF values at three wavelengths in 380nm, 550nm, 780nm, and the difference between theoretical model values and the measured data was evaluated by relative RMS error. Data analysis shows that the relative RMS error is less than 6%, which verified the correctness of the spectral BRDF model.

Hartmann characterization of the PEEM-3 aberration-corrected X-ray photoemission electron microscope.

PubMed

Scholl, A; Marcus, M A; Doran, A; Nasiatka, J R; Young, A T; MacDowell, A A; Streubel, R; Kent, N; Feng, J; Wan, W; Padmore, H A

2018-05-01

Aberration correction by an electron mirror dramatically improves the spatial resolution and transmission of photoemission electron microscopes. We will review the performance of the recently installed aberration corrector of the X-ray Photoemission Electron Microscope PEEM-3 and show a large improvement in the efficiency of the electron optics. Hartmann testing is introduced as a quantitative method to measure the geometrical aberrations of a cathode lens electron microscope. We find that aberration correction leads to an order of magnitude reduction of the spherical aberrations, suggesting that a spatial resolution of below 100 nm is possible at 100% transmission of the optics when using x-rays. We demonstrate this improved performance by imaging test patterns employing element and magnetic contrast. Published by Elsevier B.V.

A colinear backscattering Mueller matrix microscope for reflection Muller matrix imaging

NASA Astrophysics Data System (ADS)

Chen, Zhenhua; Yao, Yue; Zhu, Yuanhuan; Ma, Hui

2018-02-01

In a recent attempt, we developed a colinear backscattering Mueller matrix microscope by adding polarization state generator (PSG) and polarization state analyzer (PSA) into the illumination and detection optical paths of a commercial metallurgical microscope. It is found that specific efforts have to be made to reduce the artifacts due to the intrinsic residual polarizations of the optical system, particularly the dichroism due to the 45 degrees beam splitter. In this paper, we present a new calibration method based on numerical reconstruction of the instrument matrix to remove the artifacts introduced by beam splitter. Preliminary tests using a mirror as a standard sample show that the maximum Muller matrix element error of the colinear backscattering Muller matrix microscope can be reduced to a few percent.

The construction and characterization of optical traps for manipulating microscopic particles

NASA Astrophysics Data System (ADS)

Thompson, Tiffany; Behringer, Ernest

2011-04-01

Optical traps use tightly focused laser light to manipulate microscopic particles and have applications in nanofabrication, characterizing DNA, and in vitro fertilization [1]. We will describe the design, construction, and characterization of an optical trap that is capable of trapping and imaging 3 μm polystyrene spheres using a 12 mW HeNe laser. The design was based on previous work [2,3] describing how to build affordable optical traps. We will discuss trapping forces and their calibration. [4pt] [1] D.G. Grier, "A Revolution in Optical Manipulation," Nature 424, 810-816 (2003). [0pt] [2] S.P. Smith et al., "Inexpensive optical tweezers for undergraduate laboratories," Am. J. Phys. 67 (1), 26-35 (1999).[0pt] [3] J. Bechhoefer et al., "Faster, cheaper, safer optical tweezers for the undergraduate laboratory," Am. J. Phys. 70 (4), 393-400 (2001).

A Review of Adaptive Optics Optical Coherence Tomography: Technical Advances, Scientific Applications, and the Future

PubMed Central

Jonnal, Ravi S.; Kocaoglu, Omer P.; Zawadzki, Robert J.; Liu, Zhuolin; Miller, Donald T.; Werner, John S.

2016-01-01

Purpose Optical coherence tomography (OCT) has enabled “virtual biopsy” of the living human retina, revolutionizing both basic retina research and clinical practice over the past 25 years. For most of those years, in parallel, adaptive optics (AO) has been used to improve the transverse resolution of ophthalmoscopes to foster in vivo study of the retina at the microscopic level. Here, we review work done over the last 15 years to combine the microscopic transverse resolution of AO with the microscopic axial resolution of OCT, building AO-OCT systems with the highest three-dimensional resolution of any existing retinal imaging modality. Methods We surveyed the literature to identify the most influential antecedent work, important milestones in the development of AO-OCT technology, its applications that have yielded new knowledge, research areas into which it may productively expand, and nascent applications that have the potential to grow. Results Initial efforts focused on demonstrating three-dimensional resolution. Since then, many improvements have been made in resolution and speed, as well as other enhancements of acquisition and postprocessing techniques. Progress on these fronts has produced numerous discoveries about the anatomy, function, and optical properties of the retina. Conclusions Adaptive optics OCT continues to evolve technically and to contribute to our basic and clinical knowledge of the retina. Due to its capacity to reveal cellular and microscopic detail invisible to clinical OCT systems, it is an ideal companion to those instruments and has the demonstrable potential to produce images that can guide the interpretation of clinical findings. PMID:27409507

Integrated Micro-Optics for Microfluidic Detection.

PubMed

Kazama, Yuto; Hibara, Akihide

2016-01-01

A method of embedding micro-optics into a microfluidic device was proposed and demonstrated. First, the usefulness of embedded right-angle prisms was demonstrated in microscope observation. Lateral-view microscopic observation of an aqueous dye flow in a 100-μm-sized microchannel was demonstrated. Then, the embedded right-angle prisms were utilized for multi-beam laser spectroscopy. Here, crossed-beam thermal lens detection of a liquid sample was applied to glucose detection.

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.

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.

High-Definition Infrared Spectroscopic Imaging

PubMed Central

Reddy, Rohith K.; Walsh, Michael J.; Schulmerich, Matthew V.; Carney, P. Scott; Bhargava, Rohit

2013-01-01

The quality of images from an infrared (IR) microscope has traditionally been limited by considerations of throughput and signal-to-noise ratio (SNR). An understanding of the achievable quality as a function of instrument parameters, from first principals is needed for improved instrument design. Here, we first present a model for light propagation through an IR spectroscopic imaging system based on scalar wave theory. The model analytically describes the propagation of light along the entire beam path from the source to the detector. The effect of various optical elements and the sample in the microscope is understood in terms of the accessible spatial frequencies by using a Fourier optics approach and simulations are conducted to gain insights into spectroscopic image formation. The optimal pixel size at the sample plane is calculated and shown much smaller than that in current mid-IR microscopy systems. A commercial imaging system is modified, and experimental data are presented to demonstrate the validity of the developed model. Building on this validated theoretical foundation, an optimal sampling configuration is set up. Acquired data were of high spatial quality but, as expected, of poorer SNR. Signal processing approaches were implemented to improve the spectral SNR. The resulting data demonstrated the ability to perform high-definition IR imaging in the laboratory by using minimally-modified commercial instruments. PMID:23317676

High-definition infrared spectroscopic imaging.

PubMed

Reddy, Rohith K; Walsh, Michael J; Schulmerich, Matthew V; Carney, P Scott; Bhargava, Rohit

2013-01-01

The quality of images from an infrared (IR) microscope has traditionally been limited by considerations of throughput and signal-to-noise ratio (SNR). An understanding of the achievable quality as a function of instrument parameters, from first principals is needed for improved instrument design. Here, we first present a model for light propagation through an IR spectroscopic imaging system based on scalar wave theory. The model analytically describes the propagation of light along the entire beam path from the source to the detector. The effect of various optical elements and the sample in the microscope is understood in terms of the accessible spatial frequencies by using a Fourier optics approach and simulations are conducted to gain insights into spectroscopic image formation. The optimal pixel size at the sample plane is calculated and shown much smaller than that in current mid-IR microscopy systems. A commercial imaging system is modified, and experimental data are presented to demonstrate the validity of the developed model. Building on this validated theoretical foundation, an optimal sampling configuration is set up. Acquired data were of high spatial quality but, as expected, of poorer SNR. Signal processing approaches were implemented to improve the spectral SNR. The resulting data demonstrated the ability to perform high-definition IR imaging in the laboratory by using minimally-modified commercial instruments.

Aqueous carrier waveguide in a flow cytometer

DOEpatents

Mariella, R.P. Jr.; Engh, G. van den; Northrup, M.A.

1995-12-12

The liquid of a flow cytometer itself acts as an optical waveguide, thus transmitting the light to an optical filter/detector combination. This alternative apparatus and method for detecting scattered light in a flow cytometer is provided by a device which views and detects the light trapped within the optical waveguide formed by the flow stream. A fiber optic or other light collecting device is positioned within the flow stream. This provides enormous advantages over the standard light collection technique which uses a microscope objective. The signal-to-noise ratio is greatly increased over that for right-angle-scattered light collected by a microscope objective, and the alignment requirements are simplified. 6 figs.

Images of intravitreal objects projected onto posterior surface of model eye.

PubMed

Kawamura, Ryosuke; Shinoda, Kei; Inoue, Makoto; Noda, Toru; Ohnuma, Kazuhiko; Hirakata, Akito

2013-11-01

To try to recreate the images reported by patients during vitreous surgery in a model eye. A fluid-filled model eye with a posterior frosted translucent surface which corresponded to the retina was used. Three holes were made in the model eye through which an endoillumination pipe and intraocular forceps could be inserted. A thin plastic sheet simulating an epiretinal membrane and an intraocular lens (IOL) simulating a dislocated IOL were placed on the retina. The images falling on the posterior surface were photographed from the rear. The images seen through the surgical microscope were also recorded. The images from the rear were mirror images of those seen through the surgical microscope. Intraocular instruments were seen as black shafts from the rear. When the plastic sheet was picked up, the tip of the forceps was seen more sharply on the posterior surface. The images of the dislocated IOL from the posterior were similar to that seen through the surgical microscope, including the yellow optics and blue haptics. Intravitreal objects can form images on the surface of a model eye. Objects located closer to the surface are seen more sharply, and the colour of the objects can be identified. © 2013 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Reliable measurement of E. coli single cell fluorescence distribution using a standard microscope set-up.

PubMed

Cortesi, Marilisa; Bandiera, Lucia; Pasini, Alice; Bevilacqua, Alessandro; Gherardi, Alessandro; Furini, Simone; Giordano, Emanuele

2017-01-01

Quantifying gene expression at single cell level is fundamental for the complete characterization of synthetic gene circuits, due to the significant impact of noise and inter-cellular variability on the system's functionality. Commercial set-ups that allow the acquisition of fluorescent signal at single cell level (flow cytometers or quantitative microscopes) are expensive apparatuses that are hardly affordable by small laboratories. A protocol that makes a standard optical microscope able to acquire quantitative, single cell, fluorescent data from a bacterial population transformed with synthetic gene circuitry is presented. Single cell fluorescence values, acquired with a microscope set-up and processed with custom-made software, are compared with results that were obtained with a flow cytometer in a bacterial population transformed with the same gene circuitry. The high correlation between data from the two experimental set-ups, with a correlation coefficient computed over the tested dynamic range > 0.99, proves that a standard optical microscope- when coupled with appropriate software for image processing- might be used for quantitative single-cell fluorescence measurements. The calibration of the set-up, together with its validation, is described. The experimental protocol described in this paper makes quantitative measurement of single cell fluorescence accessible to laboratories equipped with standard optical microscope set-ups. Our method allows for an affordable measurement/quantification of intercellular variability, whose better understanding of this phenomenon will improve our comprehension of cellular behaviors and the design of synthetic gene circuits. All the required software is freely available to the synthetic biology community (MUSIQ Microscope flUorescence SIngle cell Quantification).

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.

Coherent scattering of near-resonant light by a dense, microscopic cloud of cold two-level atoms: Experiment versus theory

NASA Astrophysics Data System (ADS)

Jennewein, Stephan; Brossard, Ludovic; Sortais, Yvan R. P.; Browaeys, Antoine; Cheinet, Patrick; Robert, Jacques; Pillet, Pierre

2018-05-01

We measure the coherent scattering of low-intensity, near-resonant light by a cloud of laser-cooled two-level rubidium atoms with a size comparable to the wavelength of light. We isolate a two-level atomic structure by applying a 300-G magnetic field. We measure both the temporal and the steady-state coherent optical response of the cloud for various detunings of the laser and for atom numbers ranging from 5 to 100. We compare our results to a microscopic coupled-dipole model and to a multimode, paraxial Maxwell-Bloch model. In the low-intensity regime, both models are in excellent agreement, thus validating the Maxwell-Bloch model. Comparing to the data, the models are found in very good agreement for relatively low densities (n /k3≲0.1 ), while significant deviations start to occur at higher density. This disagreement indicates that light scattering in dense, cold atomic ensembles is still not quantitatively understood, even in pristine experimental conditions.

LIPS database with LIPService: a microscopic image database of intracellular structures in Arabidopsis guard cells.

PubMed

Higaki, Takumi; Kutsuna, Natsumaro; Hasezawa, Seiichiro

2013-05-16

Intracellular configuration is an important feature of cell status. Recent advances in microscopic imaging techniques allow us to easily obtain a large number of microscopic images of intracellular structures. In this circumstance, automated microscopic image recognition techniques are of extreme importance to future phenomics/visible screening approaches. However, there was no benchmark microscopic image dataset for intracellular organelles in a specified plant cell type. We previously established the Live Images of Plant Stomata (LIPS) database, a publicly available collection of optical-section images of various intracellular structures of plant guard cells, as a model system of environmental signal perception and transduction. Here we report recent updates to the LIPS database and the establishment of a database table, LIPService. We updated the LIPS dataset and established a new interface named LIPService to promote efficient inspection of intracellular structure configurations. Cell nuclei, microtubules, actin microfilaments, mitochondria, chloroplasts, endoplasmic reticulum, peroxisomes, endosomes, Golgi bodies, and vacuoles can be filtered using probe names or morphometric parameters such as stomatal aperture. In addition to the serial optical sectional images of the original LIPS database, new volume-rendering data for easy web browsing of three-dimensional intracellular structures have been released to allow easy inspection of their configurations or relationships with cell status/morphology. We also demonstrated the utility of the new LIPS image database for automated organelle recognition of images from another plant cell image database with image clustering analyses. The updated LIPS database provides a benchmark image dataset for representative intracellular structures in Arabidopsis guard cells. The newly released LIPService allows users to inspect the relationship between organellar three-dimensional configurations and morphometrical parameters.

Real-Time Nanoscopy by Using Blinking Enhanced Quantum Dots

PubMed Central

Watanabe, Tomonobu M.; Fukui, Shingo; Jin, Takashi; Fujii, Fumihiko; Yanagida, Toshio

2010-01-01

Superresolution optical microscopy (nanoscopy) is of current interest in many biological fields. Superresolution optical fluctuation imaging, which utilizes higher-order cumulant of fluorescence temporal fluctuations, is an excellent method for nanoscopy, as it requires neither complicated optics nor illuminations. However, it does need an impractical number of images for real-time observation. Here, we achieved real-time nanoscopy by modifying superresolution optical fluctuation imaging and enhancing the fluctuation of quantum dots. Our developed quantum dots have higher blinking than commercially available ones. The fluctuation of the blinking improved the resolution when using a variance calculation for each pixel instead of a cumulant calculation. This enabled us to obtain microscopic images with 90-nm and 80-ms spatial-temporal resolution by using a conventional fluorescence microscope without any optics or devices. PMID:20923631

Measurement of anchoring coefficient of homeotropically aligned nematic liquid crystal using a polarizing optical microscope in reflective mode

NASA Astrophysics Data System (ADS)

Baek, Sang-In; Kim, Sung-Jo; Kim, Jong-Hyun

2015-09-01

Although the homeotropic alignment of liquid crystals is widely used in LCD TVs, no easy method exists to measure its anchoring coefficient. In this study, we propose an easy and convenient measurement technique in which a polarizing optical microscope is used in the reflective mode with an objective lens having a low depth of focus. All measurements focus on the reflection of light near the interface between the liquid crystal and alignment layer. The change in the reflected light is measured by applying an electric field. We model the response of the director of the liquid crystal to the electric field and, thus, the change in reflectance. By adjusting the extrapolation length in the calculation, we match the experimental and calculated results and obtain the anchoring coefficient. In our experiment, the extrapolation lengths were 0.31 ± 0.04 μm, 0.32 ± 0.08 μm, and 0.23 ± 0.05 μm for lecithin, AL-64168, and SE-5662, respectively.

Kerr microscopy studies of the effects of bending stress on galfenola)

NASA Astrophysics Data System (ADS)

Raghunath, Ganesh; Marana, Michael; Na, Suok-Min; Flatau, Alison

2014-05-01

This work deals with using a magneto-optic Kerr effect (MOKE) microscope to optically analyze the evolution of magnetic domains in a rolled and Goss textured galfenol (Fe81Ga19 + 1.0% NbC) sample when subjected to a bending stress. The initial magnetization state of the cantilevered sample was fixed along its length by a 0.3 T permanent magnet. The magnetic state was monitored with the MOKE microscope as a tip load was applied to bend the sample. The magnetic state of galfenol depends on its magneto-elastic properties. A finite element model that incorporates an energy based formulation of magnetostriction [W. D. Armstrong, J. Magn. Magn. Mater. 263(1-2), 208-218 (2003)] was used to investigate the stresses in the sample and the corresponding change in the magnetic induction as bending occurred. A qualitative comparison with the domain pictures is presented, and the experimental micromagnetic behavior results are shown to correlate well to the macro scale bending stress and magnetization results obtained in the FEM simulations.

Engineering of frustration in colloidal artificial ice (Conference Presentation)

NASA Astrophysics Data System (ADS)

Ortiz-Ambriz, Antonio; Tierno, Pietro

2016-09-01

Artificial spin-ice systems have been used to date as microscopic models of frustration induced by lattice topology, as they allow for the direct visualization of spin arrangements and textures. However, the engineering of frustrated ice states in which individual spins can be manipulated in situ and the real-time observation of their collective dynamics remain both challenging tasks. Recently, an analogue system has been proposed theoretically, where an optical landscape confined colloidal particles that interacted electrostatically. Here we realize experimentally another version of a colloidal artificial ice system using interacting magnetically polarizable particles confined to lattices of bistable gravitational traps. We show quantitatively that ice-selection rules emerge in this frustrated soft matter system by tuning the strength of the pair-interactions between the microscopic units. By using optical tweezers, we can control particle positioning and dipolar coupling, we introduce monopole-like defects and strings and use loops with defined chirality as an elementary unit to store binary information.

Microscopic origin of the optical processes in blue sapphire.

PubMed

Bristow, Jessica K; Parker, Stephen C; Catlow, C Richard A; Woodley, Scott M; Walsh, Aron

2013-06-11

Al2O3 changes from transparent to a range of intense colours depending on the chemical impurities present. In blue sapphire, Fe and Ti are incorporated; however, the chemical process that gives rise to the colour has long been debated. Atomistic modelling identifies charge transfer from Ti(III) to Fe(III) as being responsible for the characteristic blue appearance.

Two-probe atomic-force microscope manipulator and its applications.

PubMed

Zhukov, A A; Stolyarov, V S; Kononenko, O V

2017-06-01

We report on a manipulator based on a two-probe atomic force microscope (AFM) with an individual feedback system for each probe. This manipulator works under an upright optical microscope with 3 mm focal distance. The design of the microscope helps us tomanipulate nanowires using the microscope probes as a two-prong fork. The AFM feedback is realized based on the dynamic full-time contact mode. The applications of the manipulator and advantages of its two-probe design are presented.

SIL-STED microscopy technique enhancing super-resolution of fluorescence microscopy

NASA Astrophysics Data System (ADS)

Park, No-Cheol; Lim, Geon; Lee, Won-sup; Moon, Hyungbae; Choi, Guk-Jong; Park, Young-Pil

2017-08-01

We have characterized a new type STED microscope which combines a high numerical aperture (NA) optical head with a solid immersion lens (SIL), and we call it as SIL-STED microscope. The advantage of a SIL-STED microscope is that its high NA of the SIL makes it superior to a general STED microscope in lateral resolution, thus overcoming the optical diffraction limit at the macromolecular level and enabling advanced super-resolution imaging of cell surface or cell membrane structure and function Do. This study presents the first implementation of higher NA illumination in a STED microscope limiting the fluorescence lateral resolution to about 40 nm. The refractive index of the SIL which is made of material KTaO3 is about 2.23 and 2.20 at a wavelength of 633 nm and 780 nm which are used for excitation and depletion in STED imaging, respectively. Based on the vector diffraction theory, the electric field focused by the SILSTED microscope is numerically calculated so that the numerical results of the point dispersion function of the microscope and the expected resolution could be analyzed. For further investigation, fluorescence imaging of nano size fluorescent beads is fulfilled to show improved performance of the technique.

Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains.

PubMed

Müllenbroich, M Caroline; Silvestri, Ludovico; Onofri, Leonardo; Costantini, Irene; Hoff, Marcel Van't; Sacconi, Leonardo; Iannello, Giulio; Pavone, Francesco S

2015-10-01

Comprehensive mapping and quantification of neuronal projections in the central nervous system requires high-throughput imaging of large volumes with microscopic resolution. To this end, we have developed a confocal light-sheet microscope that has been optimized for three-dimensional (3-D) imaging of structurally intact clarified whole-mount mouse brains. We describe the optical and electromechanical arrangement of the microscope and give details on the organization of the microscope management software. The software orchestrates all components of the microscope, coordinates critical timing and synchronization, and has been written in a versatile and modular structure using the LabVIEW language. It can easily be adapted and integrated to other microscope systems and has been made freely available to the light-sheet community. The tremendous amount of data routinely generated by light-sheet microscopy further requires novel strategies for data handling and storage. To complete the full imaging pipeline of our high-throughput microscope, we further elaborate on big data management from streaming of raw images up to stitching of 3-D datasets. The mesoscale neuroanatomy imaged at micron-scale resolution in those datasets allows characterization and quantification of neuronal projections in unsectioned mouse brains.

The contributions of Otto Scherzer (1909-1982) to the development of the electron microscope.

PubMed

Marko, Michael; Rose, Harald

2010-08-01

Otto Scherzer was one of the pioneers of theoretical electron optics. He was coauthor of the first comprehensive book on electron optics and was the first to understand that round electron lenses could not be combined to correct aberrations, as is the case in light optics. He subsequently was the first to describe several alternative means to correct spherical and chromatic aberration of electron lenses. These ideas were put into practice by his laboratory and students at Darmstadt and their successors, leading to the fully corrected electron microscopes now in operation.

Remote Histology Learning from Static versus Dynamic Microscopic Images

ERIC Educational Resources Information Center

Mione, Sylvia; Valcke, Martin; Cornelissen, Maria

2016-01-01

Histology is the study of microscopic structures in normal tissue sections. Curriculum redesign in medicine has led to a decrease in the use of optical microscopes during practical classes. Other imaging solutions have been implemented to facilitate remote learning. With advancements in imaging technologies, learning material can now be digitized.…

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

A Novel, Real-Time, In Vivo Mouse Retinal Imaging System.

PubMed

Butler, Mark C; Sullivan, Jack M

2015-11-01

To develop an efficient, low-cost instrument for robust real-time imaging of the mouse retina in vivo, and assess system capabilities by evaluating various animal models. Following multiple disappointing attempts to visualize the mouse retina during a subretinal injection using commercially available systems, we identified the key limitation to be inadequate illumination due to off axis illumination and poor optical train optimization. Therefore, we designed a paraxial illumination system for Greenough-type stereo dissecting microscope incorporating an optimized optical launch and an efficiently coupled fiber optic delivery system. Excitation and emission filters control spectral bandwidth. A color coupled-charged device (CCD) camera is coupled to the microscope for image capture. Although, field of view (FOV) is constrained by the small pupil aperture, the high optical power of the mouse eye, and the long working distance (needed for surgical manipulations), these limitations can be compensated by eye positioning in order to observe the entire retina. The retinal imaging system delivers an adjustable narrow beam to the dilated pupil with minimal vignetting. The optic nerve, vasculature, and posterior pole are crisply visualized and the entire retina can be observed through eye positioning. Normal and degenerative retinal phenotypes can be followed over time. Subretinal or intraocular injection procedures are followed in real time. Real-time, intravenous fluorescein angiography for the live mouse has been achieved. A novel device is established for real-time viewing and image capture of the small animal retina during subretinal injections for preclinical gene therapy studies.

Quantitative Analysis of Intracellular Motility Based on Optical Flow Model

PubMed Central

Li, Heng

2017-01-01

Analysis of cell mobility is a key issue for abnormality identification and classification in cell biology research. However, since cell deformation induced by various biological processes is random and cell protrusion is irregular, it is difficult to measure cell morphology and motility in microscopic images. To address this dilemma, we propose an improved variation optical flow model for quantitative analysis of intracellular motility, which not only extracts intracellular motion fields effectively but also deals with optical flow computation problem at the border by taking advantages of the formulation based on L1 and L2 norm, respectively. In the energy functional of our proposed optical flow model, the data term is in the form of L2 norm; the smoothness of the data changes with regional features through an adaptive parameter, using L1 norm near the edge of the cell and L2 norm away from the edge. We further extract histograms of oriented optical flow (HOOF) after optical flow field of intracellular motion is computed. Then distances of different HOOFs are calculated as the intracellular motion features to grade the intracellular motion. Experimental results show that the features extracted from HOOFs provide new insights into the relationship between the cell motility and the special pathological conditions. PMID:29065574

Chiral Nucleon-Nucleus Potentials at N3LO

NASA Astrophysics Data System (ADS)

Finelli, Paolo; Vorabbi, Matteo; Giusti, Carlotta

2018-03-01

Elastic scattering is probably one of the most relevant tools to study nuclear interactions. In this contribution we study the domain of applicability of microscopic two-body chiral potentials in the construction of an optical potential. A microscopic complex optical potential is derived and tested performing calculations on 16O at different energies. Good agreement with empirical data is obtained if a Lippmann-Schwinger cutoff at relatively high energies (above 500 MeV) is employed.

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

Analysis system of submicron particle tracks in the fine-grained nuclear emulsion by a combination of hard x-ray and optical microscopy

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

Naka, T., E-mail: naka@flab.phys.nagoya-u.ac.jp; Institute for Advanced Research, Nagoya University, Aichi 464-8602; Asada, T.

Analyses of nuclear emulsion detectors that can detect and identify charged particles or radiation as tracks have typically utilized optical microscope systems because the targets have lengths from several μm to more than 1000 μm. For recent new nuclear emulsion detectors that can detect tracks of submicron length or less, the current readout systems are insufficient due to their poor resolution. In this study, we developed a new system and method using an optical microscope system for rough candidate selection and the hard X-ray microscope system at SPring-8 for high-precision analysis with a resolution of better than 70 nm resolution.more » Furthermore, we demonstrated the analysis of submicron-length tracks with a matching efficiency of more than 99% and position accuracy of better than 5 μm. This system is now running semi-automatically.« less

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.

High resolution tip-tilt positioning system for a next generation MLL-based x-ray microscope

DOE PAGES

Xu, Weihe; Schlossberger, Noah; Xu, Wei; ...

2017-11-15

Multilayer Laue lenses (MLLs) are x-ray focusing optics with the potential to focus hard x-rays down to a single nanometer level. In order to achieve point focus, an MLL microscope needs to have the capability to perform tip-tilt motion of MLL optics and to hold the angular position for an extended period of time. Here, we present a 2D tip-tilt system that can achieve an angular resolution of over 100 microdegree with a working range of 4°, by utilizing a combination of laser interferometer and mini retroreflector. The linear dimensions of the developed system are about 30 mm in allmore » directions, and the thermal dissipation of the system during operation is negligible. Compact design and high angular resolution make the developed system suitable for MLL optics alignment in the next generation of MLL-based x-ray microscopes.« less

High resolution tip-tilt positioning system for a next generation MLL-based x-ray microscope

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

Xu, Weihe; Schlossberger, Noah; Xu, Wei

Multilayer Laue lenses (MLLs) are x-ray focusing optics with the potential to focus hard x-rays down to a single nanometer level. In order to achieve point focus, an MLL microscope needs to have the capability to perform tip-tilt motion of MLL optics and to hold the angular position for an extended period of time. Here, we present a 2D tip-tilt system that can achieve an angular resolution of over 100 microdegree with a working range of 4°, by utilizing a combination of laser interferometer and mini retroreflector. The linear dimensions of the developed system are about 30 mm in allmore » directions, and the thermal dissipation of the system during operation is negligible. Compact design and high angular resolution make the developed system suitable for MLL optics alignment in the next generation of MLL-based x-ray microscopes.« less

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.

Theoretical study of ferroelectric nanoparticles using phase reconstructed electron microscopy

NASA Astrophysics Data System (ADS)

Phatak, C.; Petford-Long, A. K.; Beleggia, M.; De Graef, M.

2014-06-01

Ferroelectric nanostructures are important for a variety of applications in electronic and electro-optical devices, including nonvolatile memories and thin-film capacitors. These applications involve stability and switching of polarization using external stimuli, such as electric fields. We present a theoretical model describing how the shape of a nanoparticle affects its polarization in the absence of screening charges, and quantify the electron-optical phase shift for detecting ferroelectric signals with phase-sensitive techniques in a transmission electron microscope. We provide an example phase shift computation for a uniformly polarized prolate ellipsoid with varying aspect ratio in the absence of screening charges.

Second-order nonlinear optical microscopy of spider silk

NASA Astrophysics Data System (ADS)

Zhao, Yue; Hien, Khuat Thi Thu; Mizutani, Goro; Rutt, Harvey N.

2017-06-01

Asymmetric β-sheet protein structures in spider silk should induce nonlinear optical interaction such as second harmonic generation (SHG) which is experimentally observed for a radial line and dragline spider silk using an imaging femtosecond laser SHG microscope. By comparing different spider silks, we found that the SHG signal correlates with the existence of the protein β-sheets. Measurements of the polarization dependence of SHG from the dragline indicated that the β-sheet has a nonlinear response depending on the direction of the incident electric field. We propose a model of what orientation the β-sheet takes in spider silk.

Multi-modality optical imaging of ovarian cancer in a post-menopausal mouse model

NASA Astrophysics Data System (ADS)

Watson, Jennifer M.; Rice, Photini Faith; Marion, Samuel L.; Bentley, David L.; Brewer, Molly A.; Utzinger, Urs; Hoyer, Patricia B.; Barton, Jennifer K.

2011-03-01

Our goal is to use optical imaging to detect cancer development on the sub cellular scale. By determining the microscopic changes that precede ovarian cancer we hope to develop a minimally invasive screening test for high risk patients. A mouse ovarian cancer model has been developed by treating mice with 4-Vinylcyclohexene Diepoxide to induce ovarian failure and 7, 12-Dimethylbenz[a]anthracene (DMBA) to induce ovarian cancer. Using optical coherence tomography (OCT) and multiphoton microscopy (MPM) we have obtained co-registered en face images of sixty-seven mouse ovaries ex vivo and forty-two ovaries in vivo. Preliminary analysis indicates that OCT and MPM can visualize ovarian microstructure. During the next year we will be completing a long term survival study using post-menopausal mice that have been treated with DMBA to induce cancer and imaged in vivo at time points before and after treatment.

The effect of mechanical drawing on optical and structural properties of nylon 6 fibres

NASA Astrophysics Data System (ADS)

El-Bakary, M. A.

2007-09-01

The Pluta polarizing double-refracting interference microscope was attached to a mechanical drawing device to study the effect of cold drawing on the optical and structural properties of nylon 6 fibres. The microscope was used in its two positions for determining the refractive indices and birefringence of fibres. Different applied stresses and strain rates were obtained using the mechanical-drawing device. The effect of the applied stresses on the optical and physical parameters was investigated. The resulting optical parameters were utilized to investigate the polarizability per unit volume, the optical orientation factor, the orientation angle and the average work per chain. The refractive index and birefringence profiles were measured. Relationships between the average work per chain and optical parameters at different strains rates were determined. An empirical formula was deduced for these fibres. Micro-interferograms are given for illustration.

Spatiotemporal polarization modulation microscopy with a microretarder array

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

Differentiating characteristic microstructural features of cancerous tissues using Mueller matrix microscope.

PubMed

Wang, Ye; He, Honghui; Chang, Jintao; Zeng, Nan; Liu, Shaoxiong; Li, Migao; Ma, Hui

2015-12-01

Polarized light imaging can provide rich microstructural information of samples, and has been applied to the detections of various abnormal tissues. In this paper, we report a polarized light microscope based on Mueller matrix imaging by adding the polarization state generator and analyzer (PSG and PSA) to a commercial transmission optical microscope. The maximum errors for the absolute values of Mueller matrix elements are reduced to 0.01 after calibration. This Mueller matrix microscope has been used to examine human cervical and liver cancerous tissues with fibrosis. Images of the transformed Mueller matrix parameters provide quantitative assessment on the characteristic features of the pathological tissues. Contrast mechanism of the experimental results are backed up by Monte Carlo simulations based on the sphere-cylinder birefringence model, which reveal the relationship between the pathological features in the cancerous tissues at the cellular level and the polarization parameters. Both the experimental and simulated data indicate that the microscopic transformed Mueller matrix parameters can distinguish the breaking down of birefringent normal tissues for cervical cancer, or the formation of birefringent surrounding structures accompanying the inflammatory reaction for liver cancer. With its simple structure, fast measurement and high precision, polarized light microscope based on Mueller matrix shows a good diagnosis application prospect. Copyright © 2015 Elsevier Ltd. All rights reserved.

Operating microscopes: past, present, and future.

PubMed

Uluç, Kutluay; Kujoth, Gregory C; Başkaya, Mustafa K

2009-09-01

The operating microscope is a fixture of modern surgical facilities, and it is a critically important factor in the success of many of the most complex and difficult surgical interventions used in medicine today. The rise of this key surgical tool reflects advances in understanding the principles of optics and vision that have occurred over centuries. The development of reading spectacles in the late 13th century led to the construction of early compound microscopes in the 16th and 17th centuries by Lippershey, Janssen, Galileo, Hooke, and others. Perhaps surprisingly, Leeuwenhoek's simple microscopes of this era offered improved performance over his contemporaries' designs. The intervening years saw improvements that reduced the spherical and chromatic aberrations present in compound microscopes. By the late 19th century, Carl Zeiss and Ernst Abbe ushered the compound microscope into the beginnings of the modern era of commercial design and production. The introduction of the microscope into the operating room by Nylén in 1921 initiated a revolution in surgical practice that gained momentum throughout the 1950s with multiple refinements, the introduction of the Zeiss OPMI series, and Kurze's application of the microscope to neurosurgery in 1957. Many of the refinements of the last 50 years have greatly improved the handling and practical operation of the surgical microscope, considerations which are equally important to its optical performance. Today's sophisticated operating microscopes allow for advanced real-time angiographic and tumor imaging. In this paper the authors discuss what might be found in the operating rooms of tomorrow.

A fiber-optic fluorescence microscope using a consumer-grade digital camera for in vivo cellular imaging.

PubMed

Shin, Dongsuk; Pierce, Mark C; Gillenwater, Ann M; Williams, Michelle D; Richards-Kortum, Rebecca R

2010-06-23

Early detection is an essential component of cancer management. Unfortunately, visual examination can often be unreliable, and many settings lack the financial capital and infrastructure to operate PET, CT, and MRI systems. Moreover, the infrastructure and expense associated with surgical biopsy and microscopy are a challenge to establishing cancer screening/early detection programs in low-resource settings. Improvements in performance and declining costs have led to the availability of optoelectronic components, which can be used to develop low-cost diagnostic imaging devices for use at the point-of-care. Here, we demonstrate a fiber-optic fluorescence microscope using a consumer-grade camera for in vivo cellular imaging. The fiber-optic fluorescence microscope includes an LED light, an objective lens, a fiber-optic bundle, and a consumer-grade digital camera. The system was used to image an oral cancer cell line labeled with 0.01% proflavine. A human tissue specimen was imaged following surgical resection, enabling dysplastic and cancerous regions to be evaluated. The oral mucosa of a healthy human subject was imaged in vivo, following topical application of 0.01% proflavine. The fiber-optic microscope resolved individual nuclei in all specimens and tissues imaged. This capability allowed qualitative and quantitative differences between normal and precancerous or cancerous tissues to be identified. The optical efficiency of the system permitted imaging of the human oral mucosa in real time. Our results indicate this device as a useful tool to assist in the identification of early neoplastic changes in epithelial tissues. This portable, inexpensive unit may be particularly appropriate for use at the point-of-care in low-resource settings.

Fully microscopic analysis of laser-driven finite plasmas using the example of clusters

NASA Astrophysics Data System (ADS)

Peltz, Christian; Varin, Charles; Brabec, Thomas; Fennel, Thomas

2012-06-01

We discuss a microscopic particle-in-cell (MicPIC) approach that allows bridging of the microscopic and macroscopic realms of laser-driven plasma physics. The simultaneous resolution of collisions and electromagnetic field propagation in MicPIC enables the investigation of processes that have been inaccessible to rigorous numerical scrutiny so far. This is illustrated by the two main findings of our analysis of pre-ionized, resonantly laser-driven clusters, which can be realized experimentally in pump-probe experiments. In the linear response regime, MicPIC data are used to extract the individual microscopic contributions to the dielectric cluster response function, such as surface and bulk collision frequencies. We demonstrate that the competition between surface collisions and radiation damping is responsible for the maximum in the size-dependent lifetime of the Mie surface plasmon. The capacity to determine the microscopic underpinning of optical material parameters opens new avenues for modeling nano-plasmonics and nano-photonics systems. In the non-perturbative regime, we analyze the formation and evolution of recollision-induced plasma waves in laser-driven clusters. The resulting dynamics of the electron density and local field hot spots opens a new research direction for the field of attosecond science.

Transmission electron microscope CCD camera

DOEpatents

Downing, Kenneth H.

1999-01-01

In order to improve the performance of a CCD camera on a high voltage electron microscope, an electron decelerator is inserted between the microscope column and the CCD. This arrangement optimizes the interaction of the electron beam with the scintillator of the CCD camera while retaining optimization of the microscope optics and of the interaction of the beam with the specimen. Changing the electron beam energy between the specimen and camera allows both to be optimized.

Assessment of Petrological Microscopes.

ERIC Educational Resources Information Center

Mathison, Charter Innes

1990-01-01

Presented is a set of procedures designed to check the design, ergonomics, illumination, function, optics, accessory equipment, and image quality of a microscope being considered for purchase. Functions for use in a petrology or mineralogy laboratory are stressed. (CW)

Electrons and Phonons in Semiconductor Multilayers

NASA Astrophysics Data System (ADS)

Ridley, B. K.

1996-11-01

This book provides a detailed description of the quantum confinement of electrons and phonons in semiconductor wells, superlattices and quantum wires, and shows how this affects their mutual interactions. It discusses the transition from microscopic to continuum models, emphasizing the use of quasi-continuum theory to describe the confinement of optical phonons and electrons. The hybridization of optical phonons and their interactions with electrons are treated, as are other electron scattering mechanisms. The book concludes with an account of the electron distribution function in three-, two- and one-dimensional systems, in the presence of electrical or optical excitation. This text will be of great use to graduate students and researchers investigating low-dimensional semiconductor structures, as well as to those developing new devices based on these systems.

Assessment of nerve ultrastructure by fibre-optic confocal microscopy.

PubMed

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

1996-01-01

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

Micro axial tomography: A miniaturized, versatile stage device to overcome resolution anisotropy in fluorescence light microscopy

NASA Astrophysics Data System (ADS)

Staier, Florian; Eipel, Heinz; Matula, Petr; Evsikov, Alexei V.; Kozubek, Michal; Cremer, Christoph; Hausmann, Michael

2011-09-01

With the development of novel fluorescence techniques, high resolution light microscopy has become a challenging technique for investigations of the three-dimensional (3D) micro-cosmos in cells and sub-cellular components. So far, all fluorescence microscopes applied for 3D imaging in biosciences show a spatially anisotropic point spread function resulting in an anisotropic optical resolution or point localization precision. To overcome this shortcoming, micro axial tomography was suggested which allows object tilting on the microscopic stage and leads to an improvement in localization precision and spatial resolution. Here, we present a miniaturized device which can be implemented in a motor driven microscope stage. The footprint of this device corresponds to a standard microscope slide. A special glass fiber can manually be adjusted in the object space of the microscope lens. A stepwise fiber rotation can be controlled by a miniaturized stepping motor incorporated into the device. By means of a special mounting device, test particles were fixed onto glass fibers, optically localized with high precision, and automatically rotated to obtain views from different perspective angles under which distances of corresponding pairs of objects were determined. From these angle dependent distance values, the real 3D distance was calculated with a precision in the ten nanometer range (corresponding here to an optical resolution of 10-30 nm) using standard microscopic equipment. As a proof of concept, the spindle apparatus of a mature mouse oocyte was imaged during metaphase II meiotic arrest under different perspectives. Only very few images registered under different rotation angles are sufficient for full 3D reconstruction. The results indicate the principal advantage of the micro axial tomography approach for many microscopic setups therein and also those of improved resolutions as obtained by high precision localization determination.

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.

Four-dimensional Microscope-Integrated Optical Coherence Tomography to Visualize Suture Depth in Strabismus Surgery.

PubMed

Pasricha, Neel D; Bhullar, Paramjit K; Shieh, Christine; Carrasco-Zevallos, Oscar M; Keller, Brenton; Izatt, Joseph A; Toth, Cynthia A; Freedman, Sharon F; Kuo, Anthony N

2017-02-14

The authors report the use of swept-source microscope-integrated optical coherence tomography (SS-MIOCT), capable of live four-dimensional (three-dimensional across time) intraoperative imaging, to directly visualize suture depth during lateral rectus resection. Key surgical steps visualized in this report included needle depth during partial and full-thickness muscle passes along with scleral passes. [J Pediatr Ophthalmol Strabismus. 2017;54:e1-e5.]. Copyright 2017, SLACK Incorporated.

Microscope Integrated Intraoperative Spectral Domain Optical Coherence Tomography for Cataract Surgery: Uses and Applications.

PubMed

Das, Sudeep; Kummelil, Mathew Kurian; Kharbanda, Varun; Arora, Vishal; Nagappa, Somshekar; Shetty, Rohit; Shetty, Bhujang K

2016-05-01

To demonstrate the uses and applications of a microscope integrated intraoperative Optical Coherence Tomography in Micro Incision Cataract Surgery (MICS) and Femtosecond Laser Assisted Cataract Surgery (FLACS). Intraoperative real time imaging using the RESCAN™ 700 (Carl Zeiss Meditec, Oberkochen, Germany) was done for patients undergoing MICS as well as FLACS. The OCT videos were reviewed at each step of the procedure and the findings were noted and analyzed. Microscope Integrated Intraoperative Optical Coherence Tomography was found to be beneficial during all the critical steps of cataract surgery. We were able to qualitatively assess wound morphology in clear corneal incisions, in terms of subclinical Descemet's detachments, tears in the inner or outer wound lips, wound gaping at the end of surgery and in identifying the adequacy of stromal hydration, for both FLACS as well as MICS. It also enabled us to segregate true posterior polar cataracts from suspected cases intraoperatively. Deciding the adequate depth of trenching was made simpler with direct visualization. The final position of the intraocular lens in the capsular bag and the lack of bioadhesivity of hydrophobic acrylic lenses were also observed. Even though Microscope Integrated Intraoperative Optical Coherence Tomography is in its early stages for its application in cataract surgery, this initial assessment does show a very promising role for this technology in the future for cataract surgery both in intraoperative decision making as well as for training purposes.

VizieR Online Data Catalog: Brussels nuclear reaction rate library (Aikawa+, 2005)

NASA Astrophysics Data System (ADS)

Aikawa, M.; Arnould, M.; Goriely, S.; Jorissen, A.; Takahashi, K.

2005-07-01

The present data is part of the Brussels nuclear reaction rate library (BRUSLIB) for astrophysics applications and concerns nuclear reaction rate predictions calculated within the statistical Hauser-Feshbach approximation and making use of global and coherent microscopic nuclear models for the quantities (nuclear masses, nuclear structure properties, nuclear level densities, gamma-ray strength functions, optical potentials) entering the rate calculations. (4 data files).

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

Study of a quasi-microscope design for planetary landers

NASA Technical Reports Server (NTRS)

Giat, O.; Brown, E. B.

1973-01-01

The Viking Lander fascimile camera, in its present form, provides for a minimum object distance of 1.9 meters, at which distance its resolution of 0.0007 radian provides an object resolution of 1.33 millimeters. It was deemed desirable, especially for follow-on Viking missions, to provide means for examing Martian terrain at resolutions considerably higher than that now provided. This led to the concept of quasi-microscope, an attachment to be used in conjunction with the fascimile camera to convert it to a low power microscope. The results are reported of an investigation to consider alternate optical configurations for the quasi-microscope and to develop optical designs for the selected system or systems. Initial requirements included consideration of object resolutions in the range of 2 to 50 micrometers, an available field of view of the order of 500 pixels, and no significant modifications to the fascimile camera.

Mosaic of Commemorative Microscope Substrate

NASA Technical Reports Server (NTRS)

2008-01-01

Written by electron beam lithography in the Microdevices Laboratory of NASA's Jet Propulsion Laboratory, this Optical Microscope substrate helps the Phoenix Mars Mission science team learn how to assemble individual microscope images into a mosaic by aligning rows of text.

Each line is about 0.1 millimeter tall, the average thickness of a human hair. Except for the Mogensen twins, the names are of babies born and team members lost during the original development of MECA (the Microscopy, Electrochemistry and Conductivity Analyzer) for the canceled 2001 Mars lander mission. The plaque also acknowledges the MECA 2001 principal investigator, now retired.

This image was taken by the MECA Optical Microscope on Sol 111, or the 111th day of the Phoenix mission (Sept. 16, 2008).

The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by JPL, Pasadena, Calif. Spacecraft development was by Lockheed Martin Space Systems, Denver.

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.

Studies on transmitted beam modulation effect from laser induced damage on fused silica optics.

PubMed

Zheng, Yi; Ma, Ping; Li, Haibo; Liu, Zhichao; Chen, Songlin

2013-07-15

UV laser induced damage (LID) on exit surface of fused silica could cause modulation effect to transmitted beam and further influence downstream propagation properties. This paper presents our experimental and analytical studies on this topic. In experiment, a series of measurement instruments are applied, including beam profiler, interferometer, microscope, and optical coherent tomography (OCT). Creating and characterizing of LID on fused silica sample have been implemented. Morphological features are studied based on their particular modulation effects on transmitted beam. In theoretical investigation, analytical modeling and numerical simulation are performed. Modulation effects from amplitude, phase, and size factors are analyzed respectively. Furthermore, we have novelly designed a simplified polygon model to simulate actual damage site with multiform modulation features, and the simulation results demonstrate that the modeling is usable and representative.

Optical droplet vaporization of nanoparticle-loaded stimuli-responsive microbubbles

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

Si, Ting; Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio 43210; Li, Guangbin

2016-03-14

A capillary co-flow focusing process is developed to generate stimuli-responsive microbubbles (SRMs) that comprise perfluorocarbon (PFC) suspension of silver nanoparticles (SNPs) in a lipid shell. Upon continuous laser irradiation at around their surface plasmon resonance band, the SNPs effectively absorb electromagnetic energy, induce heat accumulation in SRMs, trigger PFC vaporization, and eventually lead to thermal expansion and fragmentation of the SRMs. This optical droplet vaporization (ODV) process is further simulated by a theoretical model that combines heat generation of SNPs, phase change of PFC, and thermal expansion of SRMs. The model is validated by benchtop experiments, where the ODV processmore » is monitored by microscopic imaging. The effects of primary process parameters on behaviors of ODV are predicted by the theoretical model, indicating the technical feasibility for process control and optimization in future drug delivery applications.« less

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.

Microscope-on-Chip Using Micro-Channel and Solid State Image Sensors

NASA Technical Reports Server (NTRS)

Wang, Yu

2000-01-01

Recently, Jet Propulsion Laboratory has invented and developed a miniature optical microscope, microscope-on-chip using micro-channel and solid state image sensors. It is lightweight, low-power, fast speed instrument, it has no image lens, does not need focus adjustment, and the total mass is less than 100g. A prototype has been built and demonstrated at JPL.

The PC9A Filter Screening Tool

DTIC Science & Technology

2016-02-01

conjunction with an optical microscope for identification of other important debris such as glass beads. The FST has now been installed at RAAF East...conservative screening limits need to be sent for detailed laboratory analysis. Laboratory analysis has traditionally involved a manual microscopic ...Electron Microscope with Energy Dispersive Spectroscopy (SEM EDS) to determine the composition and likely source. The Engine Maintenance Manual

A high-resolution multimode digital microscope system.

PubMed

Salmon, Edward D; Shaw, Sidney L; Waters, Jennifer C; Waterman-Storer, Clare M; Maddox, Paul S; Yeh, Elaine; Bloom, Kerry

2013-01-01

This chapter describes the development of a high-resolution, multimode digital imaging system based on a wide-field epifluorescent and transmitted light microscope, and a cooled charge-coupled device (CCD) camera. The three main parts of this imaging system are Nikon FXA microscope, Hamamatsu C4880 cooled CCD camera, and MetaMorph digital imaging system. This chapter presents various design criteria for the instrument and describes the major features of the microscope components-the cooled CCD camera and the MetaMorph digital imaging system. The Nikon FXA upright microscope can produce high resolution images for both epifluorescent and transmitted light illumination without switching the objective or moving the specimen. The functional aspects of the microscope set-up can be considered in terms of the imaging optics, the epi-illumination optics, the transillumination optics, the focus control, and the vibration isolation table. This instrument is somewhat specialized for microtubule and mitosis studies, and it is also applicable to a variety of problems in cellular imaging, including tracking proteins fused to the green fluorescent protein in live cells. The instrument is also valuable for correlating the assembly dynamics of individual cytoplasmic microtubules (labeled by conjugating X-rhodamine to tubulin) with the dynamics of membranes of the endoplasmic reticulum (labeled with DiOC6) and the dynamics of the cell cortex (by differential interference contrast) in migrating vertebrate epithelial cells. This imaging system also plays an important role in the analysis of mitotic mutants in the powerful yeast genetic system Saccharomyces cerevisiae. Copyright © 1998 Elsevier Inc. All rights reserved.

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.

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

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.

Auricular burns associated with operating microscope use during otologic surgery.

PubMed

Latuska, Richard F; Carlson, Matthew L; Neff, Brian A; Driscoll, Colin L; Wanna, George B; Haynes, David S

2014-02-01

To raise awareness of the potential hazard of auricular burns associated with operating microscope use during otologic surgery. Retrospective case series and summary of the Food and Drug Administration's (FDA) Manufacturer and User Facility Device Experience (MAUDE) database of voluntary adverse event reports pertaining to microscope related auricular thermal injuries. All patients who sustained auricular burns while using the operating microscope during otologic surgery at 2 tertiary academic referral centers. Surgical procedure, microscope model, intensity of illumination, length of procedure, focal length, location and severity of burn, and patient outcome. A total of 4 microscope-related auricular thermal injuries were identified from the authors' institutions. Additionally, 82 unique cases of soft tissue burns associated with the use of an operative microscope have been voluntarily reported to the FDA since 2004. A disproportionately large percent (∼ 30%) of these occurred within the field of otology, the majority of which were during tympanoplasty or tympanomastoidectomy procedures at focal length distances of 300 mm or less with xenon light source microscopes. Simultaneous advancements in light delivery technologies and lens optics have continued to improve the efficiency of the operating microscope; however, these improvements also increase the potential for thermal injuries. Although rare, a review of the FDA MAUDE database suggests that microscope-related soft tissue burns occur more frequently in otology than any other surgical specialty. A variety of factors may help explain this finding, including the unique anatomy of the external ear with thin skin and limited underlying adipose tissue. Preventative measures should be taken to decrease the risk of thermal injuries including use of the lowest comfortable light intensity, adjusting the aperture width to match the operative field, frequent wound irrigation, and covering exposed portions of the pinna with a moist surgical sponge.

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.

Photonic-crystal membranes for optical detection of single nano-particles, designed for biosensor application.

PubMed

Grepstad, Jon Olav; Kaspar, Peter; Solgaard, Olav; Johansen, Ib-Rune; Sudbø, Aasmund S

2012-03-26

A sensor designed to detect bio-molecules is presented. The sensor exploits a planar 2D photonic crystal (PC) membrane with sub-micron thickness and through holes, to induce high optical fields that allow detection of nano-particles smaller than the diffraction limit of an optical microscope. We report on our design and fabrication of a PC membrane with a nano-particle trapped inside. We have also designed and built an imaging system where an optical microscope and a CCD camera are used to take images of the PC membrane. Results show how the trapped nano-particle appears as a bright spot in the image. In a first experimental realization of the imaging system, single particles with a radius of 75 nm can be detected.

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.

Simulation of transmission electron microscope images of biological specimens.

PubMed

Rullgård, H; Ofverstedt, L-G; Masich, S; Daneholt, B; Oktem, O

2011-09-01

We present a new approach to simulate electron cryo-microscope images of biological specimens. The framework for simulation consists of two parts; the first is a phantom generator that generates a model of a specimen suitable for simulation, the second is a transmission electron microscope simulator. The phantom generator calculates the scattering potential of an atomic structure in aqueous buffer and allows the user to define the distribution of molecules in the simulated image. The simulator includes a well defined electron-specimen interaction model based on the scalar Schrödinger equation, the contrast transfer function for optics, and a noise model that includes shot noise as well as detector noise including detector blurring. To enable optimal performance, the simulation framework also includes a calibration protocol for setting simulation parameters. To test the accuracy of the new framework for simulation, we compare simulated images to experimental images recorded of the Tobacco Mosaic Virus (TMV) in vitreous ice. The simulated and experimental images show good agreement with respect to contrast variations depending on dose and defocus. Furthermore, random fluctuations present in experimental and simulated images exhibit similar statistical properties. The simulator has been designed to provide a platform for development of new instrumentation and image processing procedures in single particle electron microscopy, two-dimensional crystallography and electron tomography with well documented protocols and an open source code into which new improvements and extensions are easily incorporated. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

A Novel, Real-Time, In Vivo Mouse Retinal Imaging System

PubMed Central

Butler, Mark C.; Sullivan, Jack M.

2015-01-01

Purpose To develop an efficient, low-cost instrument for robust real-time imaging of the mouse retina in vivo, and assess system capabilities by evaluating various animal models. Methods Following multiple disappointing attempts to visualize the mouse retina during a subretinal injection using commercially available systems, we identified the key limitation to be inadequate illumination due to off axis illumination and poor optical train optimization. Therefore, we designed a paraxial illumination system for Greenough-type stereo dissecting microscope incorporating an optimized optical launch and an efficiently coupled fiber optic delivery system. Excitation and emission filters control spectral bandwidth. A color coupled-charged device (CCD) camera is coupled to the microscope for image capture. Although, field of view (FOV) is constrained by the small pupil aperture, the high optical power of the mouse eye, and the long working distance (needed for surgical manipulations), these limitations can be compensated by eye positioning in order to observe the entire retina. Results The retinal imaging system delivers an adjustable narrow beam to the dilated pupil with minimal vignetting. The optic nerve, vasculature, and posterior pole are crisply visualized and the entire retina can be observed through eye positioning. Normal and degenerative retinal phenotypes can be followed over time. Subretinal or intraocular injection procedures are followed in real time. Real-time, intravenous fluorescein angiography for the live mouse has been achieved. Conclusions A novel device is established for real-time viewing and image capture of the small animal retina during subretinal injections for preclinical gene therapy studies. PMID:26551329

Optical scanning tests of complex CMOS microcircuits

NASA Technical Reports Server (NTRS)

Levy, M. E.; Erickson, J. J.

1977-01-01

The new test method was based on the use of a raster-scanned optical stimulus in combination with special electrical test procedures. The raster-scanned optical stimulus was provided by an optical spot scanner, an instrument that combines a scanning optical microscope with electronic instrumentation to process and display the electric photoresponse signal induced in a device that is being tested.

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.

Microscope-integrated optical coherence tomography: A new surgical tool in vitreoretinal surgery.

PubMed

Jayadev, Chaitra; Dabir, Supriya; Vinekar, Anand; Shah, Urmil; Vaid, Tania; Yadav, Naresh Kumar

2015-05-01

Optical coherence tomography (OCT) has revolutionized imaging of ocular structures and various disease conditions. Though it has been used in the clinic for some decades, the OCT has only recently found its way into the operating theater. Early attempts at intraoperative OCT, hand-held and microscope mounted, have already improved our understanding of the surgical pathology and the role it might play in surgical decision-making. The microscope-integrated OCT now allows seamless, high-resolution, real-time imaging of surgical maneuvers from the incision to wound closure. Visualization of instruments and intraoperative tissue manipulation are possible with this in vivo modality and, therefore, help improve the outcome of surgery. In this article, we describe the advantages it offers during various vitreoretinal procedures.

Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf

NASA Astrophysics Data System (ADS)

Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; An Nguyen, Thien; Alfano, Robert R.

2014-06-01

Two-photon (2P) excitation of the second singlet (S) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S2 state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf.

PubMed

Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; Nguyen, Thien An; Alfano, Robert R

2014-06-01

Two-photon (2P) excitation of the second singlet (S₂) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S₂ state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S₂ state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

Superhydrophobic, Biomimetic Surfaces with High and Low Adhesion, Optical Transmittance, and Nanoscale Mechanical Wear Resistance

NASA Astrophysics Data System (ADS)

Ebert, Daniel R.

Superhydrophobic surfaces (defined as surfaces having water contact angle greater than 150°) show great promise for use in a rapidly growing number of engineering applications, ranging from biomedical devices to fluid drag reduction in pipelines. In nature, the surfaces of many organisms, such as certain plant leaves, are known to exhibit superhydrophobicity. In some cases, droplet adhesion is very low (droplet rolls away easily), while in other cases adhesion is high (droplet remains adhered when surface is inverted). The recent advent and development of microscopes with resolution down to a few nanometers (such as atomic force microscopes and scanning electron microscopes) has allowed for in-depth understanding of the micro- and nanoscale mechanisms employed by these plant leaves and other natural surfaces to achieve their particular wetting properties. Biomimetics (or "mimicking nature") is therefore a very promising approach for the development of engineering surfaces with desired wetting characteristics. However, research in creating biomimetic surfaces is still in its early stages, and many of the surfaces created thus far are not mechanically robust, which is required for many potential real-world applications. In addition, for applications such as self-cleaning windows and solar panels, optical transparency is required. In this thesis, a set of original studies are presented in which superhydrophobic surfaces were designed based on biomimetics and fabricated using a wide of variety of techniques. The surfaces were characterized with regard to wetting characteristics such as water contact angle and contact angle hysteresis, micro- and nanoscale mechanical durability, and in some cases optical transmittance. Theoretical wetting models served as guides both in the design and in the understanding of experimental results, especially in regard to different wetting regime and regime transition. This work provides important conclusions and valuable insight for identifying materials, techniques, and designs for mechanically durable, optically transparent superhydrophobic surfaces.

Coherent anti-Stokes Raman scattering spectroscope/microscope based on a widely tunable laser source

NASA Astrophysics Data System (ADS)

Dementjev, A.; Gulbinas, V.; Serbenta, A.; Kaucikas, M.; Niaura, G.

2010-03-01

We present a coherent anti-Stokes Raman scattering (CARS) microscope based on a robust and simple laser source. A picosecond laser operating in a cavity dumping regime at the 1 MHz repetition rate was used to pump a traveling wave optical parametric generator, which serves as a two-color excitation light source for the CARS microscope. We demonstrate the ability of the presented CARS microscope to measure CARS spectra and images by using several detection schemes.

Preclinical evaluation and intraoperative human retinal imaging with a high-resolution microscope-integrated spectral domain optical coherence tomography device.

PubMed

Hahn, Paul; Migacz, Justin; O'Donnell, Rachelle; Day, Shelley; Lee, Annie; Lin, Phoebe; Vann, Robin; Kuo, Anthony; Fekrat, Sharon; Mruthyunjaya, Prithvi; Postel, Eric A; Izatt, Joseph A; Toth, Cynthia A

2013-01-01

The authors have recently developed a high-resolution microscope-integrated spectral domain optical coherence tomography (MIOCT) device designed to enable OCT acquisition simultaneous with surgical maneuvers. The purpose of this report is to describe translation of this device from preclinical testing into human intraoperative imaging. Before human imaging, surgical conditions were fully simulated for extensive preclinical MIOCT evaluation in a custom model eye system. Microscope-integrated spectral domain OCT images were then acquired in normal human volunteers and during vitreoretinal surgery in patients who consented to participate in a prospective institutional review board-approved study. Microscope-integrated spectral domain OCT images were obtained before and at pauses in surgical maneuvers and were compared based on predetermined diagnostic criteria to images obtained with a high-resolution spectral domain research handheld OCT system (HHOCT; Bioptigen, Inc) at the same time point. Cohorts of five consecutive patients were imaged. Successful end points were predefined, including ≥80% correlation in identification of pathology between MIOCT and HHOCT in ≥80% of the patients. Microscope-integrated spectral domain OCT was favorably evaluated by study surgeons and scrub nurses, all of whom responded that they would consider participating in human intraoperative imaging trials. The preclinical evaluation identified significant improvements that were made before MIOCT use during human surgery. The MIOCT transition into clinical human research was smooth. Microscope-integrated spectral domain OCT imaging in normal human volunteers demonstrated high resolution comparable to tabletop scanners. In the operating room, after an initial learning curve, surgeons successfully acquired human macular MIOCT images before and after surgical maneuvers. Microscope-integrated spectral domain OCT imaging confirmed preoperative diagnoses, such as full-thickness macular hole and vitreomacular traction, and demonstrated postsurgical changes in retinal morphology. Two cohorts of five patients were imaged. In the second cohort, the predefined end points were exceeded with ≥80% correlation between microscope-mounted OCT and HHOCT imaging in 100% of the patients. This report describes high-resolution MIOCT imaging using the prototype device in human eyes during vitreoretinal surgery, with successful achievement of predefined end points for imaging. Further refinements and investigations will be directed toward fully integrating MIOCT with vitreoretinal and other ocular surgery to image surgical maneuvers in real time.

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.

Stimulated penetrating keratoplasty using real-time virtual intraoperative surgical optical coherence tomography

PubMed Central

Lee, Changho; Kim, Kyungun; Han, Seunghoon; Kim, Sehui; Lee, Jun Hoon; Kim, Hong kyun; Kim, Chulhong; Jung, Woonggyu; Kim, Jeehyun

2014-01-01

Abstract. An intraoperative surgical microscope is an essential tool in a neuro- or ophthalmological surgical environment. Yet, it has an inherent limitation to classify subsurface information because it only provides the surface images. To compensate for and assist in this problem, combining the surgical microscope with optical coherence tomography (OCT) has been adapted. We developed a real-time virtual intraoperative surgical OCT (VISOCT) system by adapting a spectral-domain OCT scanner with a commercial surgical microscope. Thanks to our custom-made beam splitting and image display subsystems, the OCT images and microscopic images are simultaneously visualized through an ocular lens or the eyepiece of the microscope. This improvement helps surgeons to focus on the operation without distraction to view OCT images on another separate display. Moreover, displaying the OCT live images on the eyepiece helps surgeon’s depth perception during the surgeries. Finally, we successfully processed stimulated penetrating keratoplasty in live rabbits. We believe that these technical achievements are crucial to enhance the usability of the VISOCT system in a real surgical operating condition. PMID:24604471

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.

Effect of drug loading method against drug dissolution mechanism of encapsulated amoxicillin trihydrate in matrix of semi-IPN chitosan-poly(N-vinylpyrrolidone) hydrogel with KHCO3 as pore forming agent in floating drug delivery system

NASA Astrophysics Data System (ADS)

Fimantari, Khansa; Budianto, Emil

2018-04-01

Helicobacterpylori infection can be treated using trihydrate amoxicillin. However, this treatment is not effective enough, as the conventional dosage treatment has a relatively short retention time in the human stomach. In the present study, the amoxicillin trihydrate drug will be encapsulated into a semi-IPN K-PNVP hydrogel matrix with 7,5% KHCO3 as a pore-forming agent. The encapsulated drug is tested with in vitro method to see the efficiency of its encapsulation and dissolution. The hydrogel in situ loading produces an encapsulation efficiency value. The values of the encapsulation efficiency are 95% and 98%, while post loading hydrogel yields an encapsulation efficiency value is 77% and the dissolution is 84%. The study of drug dissolution mechanism was done by using mathematical equation model to know its kinetics and its mechanism of dissolution. The post loading hydrogel was done by using thefirst-order model, while hydrogel in situ loading used Higuchi model. The Korsmeyer-Peppas model shows that post loading hydrogel dissolution mechanism is a mixture of diffusion and erosion, and in situ loading hydrogel in the form of diffusion. It is supported by the results of hydrogel characterization, before and after dissolution test with an optical microscope. The results of the optical microscope show that the hydrogel surface before and after the dissolution tested for both methods shows the change becomes rougher.

Simple and cost-effective hardware and software for functional brain mapping using intrinsic optical signal imaging.

PubMed

Harrison, Thomas C; Sigler, Albrecht; Murphy, Timothy H

2009-09-15

We describe a simple and low-cost system for intrinsic optical signal (IOS) imaging using stable LED light sources, basic microscopes, and commonly available CCD cameras. IOS imaging measures activity-dependent changes in the light reflectance of brain tissue, and can be performed with a minimum of specialized equipment. Our system uses LED ring lights that can be mounted on standard microscope objectives or video lenses to provide a homogeneous and stable light source, with less than 0.003% fluctuation across images averaged from 40 trials. We describe the equipment and surgical techniques necessary for both acute and chronic mouse preparations, and provide software that can create maps of sensory representations from images captured by inexpensive 8-bit cameras or by 12-bit cameras. The IOS imaging system can be adapted to commercial upright microscopes or custom macroscopes, eliminating the need for dedicated equipment or complex optical paths. This method can be combined with parallel high resolution imaging techniques such as two-photon microscopy.

Combined use of optical and electron microscopic techniques for the measurement of hygroscopic property, chemical composition, and morphology of individual aerosol particles.

PubMed

Ahn, Kang-Ho; Kim, Sun-Man; Jung, Hae-Jin; Lee, Mi-Jung; Eom, Hyo-Jin; Maskey, Shila; Ro, Chul-Un

2010-10-01

In this work, an analytical method for the characterization of the hygroscopic property, chemical composition, and morphology of individual aerosol particles is introduced. The method, which is based on the combined use of optical and electron microscopic techniques, is simple and easy to apply. An optical microscopic technique was used to perform the visual observation of the phase transformation and hygroscopic growth of aerosol particles on a single particle level. A quantitative energy-dispersive electron probe X-ray microanalysis, named low-Z particle EPMA, was used to perform a quantitative chemical speciation of the same individual particles after the measurement of the hygroscopic property. To validate the analytical methodology, the hygroscopic properties of artificially generated NaCl, KCl, (NH(4))(2)SO(4), and Na(2)SO(4) aerosol particles of micrometer size were investigated. The practical applicability of the analytical method for studying the hygroscopic property, chemical composition, and morphology of ambient aerosol particles is demonstrated.

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

Enhanced optical coupling and Raman scattering via microscopic interface engineering

NASA Astrophysics Data System (ADS)

Thompson, Jonathan V.; Hokr, Brett H.; Kim, Wihan; Ballmann, Charles W.; Applegate, Brian E.; Jo, Javier A.; Yamilov, Alexey; Cao, Hui; Scully, Marlan O.; Yakovlev, Vladislav V.

2017-11-01

Spontaneous Raman scattering is an extremely powerful tool for the remote detection and identification of various chemical materials. However, when those materials are contained within strongly scattering or turbid media, as is the case in many biological and security related systems, the sensitivity and range of Raman signal generation and detection is severely limited. Here, we demonstrate that through microscopic engineering of the optical interface, the optical coupling of light into a turbid material can be substantially enhanced. This improved coupling facilitates the enhancement of the Raman scattering signal generated by molecules within the medium. In particular, we detect at least two-orders of magnitude more spontaneous Raman scattering from a sample when the pump laser light is focused into a microscopic hole in the surface of the sample. Because this approach enhances both the interaction time and interaction region of the laser light within the material, its use will greatly improve the range and sensitivity of many spectroscopic techniques, including Raman scattering and fluorescence emission detection, inside highly scattering environments.

Shear Stress Sensing with Elastic Microfence Structures

NASA Technical Reports Server (NTRS)

Cisotto, Alexxandra; Palmieri, Frank L.; Saini, Aditya; Lin, Yi; Thurman, Christopher S; Kim, Jinwook; Kim, Taeyang; Connell, John W.; Zhu, Yong; Gopalarathnam, Ashok;

Nonlinear network model analysis of vibrational energy transfer and localisation in the Fenna-Matthews-Olson complex

NASA Astrophysics Data System (ADS)

Morgan, Sarah E.; Cole, Daniel J.; Chin, Alex W.

2016-11-01

Collective protein modes are expected to be important for facilitating energy transfer in the Fenna-Matthews-Olson (FMO) complex of photosynthetic green sulphur bacteria, however to date little work has focussed on the microscopic details of these vibrations. The nonlinear network model (NNM) provides a computationally inexpensive approach to studying vibrational modes at the microscopic level in large protein structures, whilst incorporating anharmonicity in the inter-residue interactions which can influence protein dynamics. We apply the NNM to the entire trimeric FMO complex and find evidence for the existence of nonlinear discrete breather modes. These modes tend to transfer energy to the highly connected core pigments, potentially opening up alternative excitation energy transfer routes through their influence on pigment properties. Incorporating localised modes based on these discrete breathers in the optical spectra calculations for FMO using ab initio site energies and excitonic couplings can substantially improve their agreement with experimental results.

Optical and Acoustical Techniques for Non-viral Gene Delivery to Mammalian Cells and In-situ Study of Cytoskeletal Mechanics

NASA Astrophysics Data System (ADS)

Ma, Zili

Since the first optical microscope invented by Anton van Leeuwenhoek in 1674, the great development of laser technique and its applications in biophotonics have helped us reveal the mechanisms underlying numerous biological activities gradually. The introduction of fs lasers to the studies of biology has emerged as a fast developing area calling for the efforts and skills both from optics and electric engineering and biology and medicine. Due to the fast update of laser source techniques, there has been an increasing number of commercialized fs lasers available for this growing market of biophotonics. To better utilize the potential offered by fs lasers, we studied the technique of optical gene delivery and tried to narrow the gap between laboratorial research and industrial/clinical applications, in that the strict experimental conditions of specific optical laboratorial studies are generally not appropriate for the practical biological applications. To carry out our experiments, we built a two-stage amplifier fs laser system to generate the desired pulse train. The laser pulse train was coupled into an invert fluorescence microscope for the imaging and manipulation of each cell. To overcome limitations brought by the tight focus of laser beam due to high NA objective, we introduced gold nanorods (GNRs), a metallic nanomaterial, with tunable optical property. With these additional membrane for membrane permeabilization, which could significantly improve the manipulation speed than that based on the tightly focused laser. We used GFP plasmid to demonstrate the applications of this technique in gene delivery, and successfully transfected and GFP-expressed cells were observed one day after the optical transfection. Additionally, as an important trend of biophotonics, the integration of optics with microfluidic chips has become the new frontier of both biology and engineering. Here we firstly demonstrated a technique of gene delivery by an on-chip device generating surface acoustic waves, which not only achieved a high efficiency of cells permeabilization in a quick speed, but also allowed us to observe the permeabilization process in real time by microscope. This device is also compatible with biophotonics studies based on fs laser, which can be further developed as a powerful tool for optical gene delivery with the capability of precisely controlling the fluid on-chip by SAW. SAW devices could also achieve exogenous gene delivery through the cell membrane without the need of adding chemical agents. Our results showed that the membrane of mammalian adherent cells could be effectively perforated transiently by applying a SAW. The transfection of pEGFP plasmids into endothelial cells was carried out successfully via this SAW-induced cell perforation. The expression of GFP was observed after 24-hour incubation subsequent to the SAW treatment. In regard to the application of fs lasers in cellular and subcellular level studies, we applied the optical nanoscissoring technique based on fs lasers in biomechanical studies to study the mechanical properties of single SF in-situ. Integrated into a confocal microscope, the fs laser showed great power in manipulating targeted in-situ subcellular structures under real-time imaging without damaging nearby regions. Here, how oxidative challenges would alter the mechanical properties of SFs in myoblasts was firstly investigated using the optical nanoscissoring technique to comprehend the whole picture of muscle tissue injury and repair from the basics. The prestress of stress fibers after the oxidative challenges was found through our modified viscoelastic retraction model and experiment result.

Quantitative orientation-independent differential interference contrast (DIC) microscopy

NASA Astrophysics Data System (ADS)

Shribak, Michael; LaFountain, James; Biggs, David; Inoué, Shinya

2007-02-01

We describe a new DIC technique, which records phase gradients within microscopic specimens independently of their orientation. The proposed system allows the generation of images representing the distribution of dry mass (optical path difference) in the specimen. Unlike in other forms of interference microscopes, this approach does not require a narrow illuminating cone. The orientation-independent differential interference contrast (OI-DIC) system can also be combined with orientation-independent polarization (OI-Pol) measurements to yield two complementary images: one showing dry mass distribution (which is proportional to refractive index) and the other showing distribution of birefringence (due to structural or internal anisotropy). With a model specimen used for this work -- living spermatocytes from the crane fly, Nephrotoma suturalis --- the OI-DIC image clearly reveals the detailed shape of the chromosomes while the polarization image quantitatively depicts the distribution of the birefringent microtubules in the spindle, both without any need for staining or other modifications of the cell. We present examples of a pseudo-color combined image incorporating both orientation-independent DIC and polarization images of a spermatocyte at diakinesis and metaphase of meiosis I. Those images provide clear evidence that the proposed technique can reveal fine architecture and molecular organization in live cells without perturbation associated with staining or fluorescent labeling. The phase image was obtained using optics having a numerical aperture 1.4, thus achieving a level of resolution never before achieved with any interference microscope.

First-principles approach to the dynamic magnetoelectric couplings for the non-reciprocal directional dichroism in BiFeO 3

DOE PAGES

Kezsmarki, I.; Fishman, Randy Scott

2016-04-18

Due to the complicated magnetic and crystallographic structures of BiFeO 3, its magnetoelectric (ME) couplings and microscopic model Hamiltonian remain poorly understood. By employing a firstprinciples approach, we uncover all possibleMEcouplings associated with the spin-current (SC) and exchange-striction (ES) polarizations, and construct an appropriate Hamiltonian for the long-range spin-cycloid in BiFeO 3. First-principles calculations are used to understand the microscopic origins of theMEcouplings.Wefind that inversion symmetries broken by ferroelectric and antiferroelectric distortions induce the SC and the ES polarizations, which cooperatively produce the dynamicME effects in BiFeO 3. A model motivated by first principles reproduces the absorption difference of counter-propagatingmore » light beams called non-reciprocal directional dichroism. The current paper focuses on the spin-driven (SD) polarizations produced by a dynamic electric field, i.e. the dynamic MEcouplings. Due to the inertial properties of Fe, the dynamic SD polarizations differ significantly from the static SD polarizations. Our systematic approach can be generally applied to any multiferroic material, laying the foundation for revealing hiddenMEcouplings on the atomic scale and for exploiting opticalMEeffects in the next generation of technological devices such as optical diodes.« less

Analytical modeling of electron energy loss spectroscopy of graphene: Ab initio study versus extended hydrodynamic model.

PubMed

Djordjević, Tijana; Radović, Ivan; Despoja, Vito; Lyon, Keenan; Borka, Duško; Mišković, Zoran L

2018-01-01

We present an analytical modeling of the electron energy loss (EEL) spectroscopy data for free-standing graphene obtained by scanning transmission electron microscope. The probability density for energy loss of fast electrons traversing graphene under normal incidence is evaluated using an optical approximation based on the conductivity of graphene given in the local, i.e., frequency-dependent form derived by both a two-dimensional, two-fluid extended hydrodynamic (eHD) model and an ab initio method. We compare the results for the real and imaginary parts of the optical conductivity in graphene obtained by these two methods. The calculated probability density is directly compared with the EEL spectra from three independent experiments and we find very good agreement, especially in the case of the eHD model. Furthermore, we point out that the subtraction of the zero-loss peak from the experimental EEL spectra has a strong influence on the analytical model for the EEL spectroscopy data. Copyright © 2017 Elsevier B.V. All rights reserved.

Elastic and inelastic scattering of neutrons on 238U nucleus

NASA Astrophysics Data System (ADS)

Capote, R.; Trkov, A.; Sin, M.; Herman, M. W.; Soukhovitskiĩ, E. Sh.

2014-04-01

Advanced modelling of neutron induced reactions on the 238U nucleus is aimed at improving our knowledge of neutron scattering. Capture and fission channels are well constrained by available experimental data and neutron standard evaluation. A focus of this contribution is on elastic and inelastic scattering cross sections. The employed nuclear reaction model includes - a new rotational-vibrational dispersive optical model potential coupling the low-lying collective bands of vibrational character observed in even-even actinides; - the Engelbrecht-Weidenmüller transformation allowing for inclusion of compound-direct interference effects; - and a multi-humped fission barrier with absorption in the secondary well described within the optical model for fission. Impact of the advanced modelling on elastic and inelastic scattering cross sections including angular distributions and emission spectra is assessed both by comparison with selected microscopic experimental data and integral criticality benchmarks including measured reaction rates (e.g. JEMIMA, FLAPTOP and BIG TEN). Benchmark calculations provided feedback to improve the reaction modelling. Improvement of existing libraries will be discussed.

Suspension and simple optical characterization of two-dimensional membranes

NASA Astrophysics Data System (ADS)

Northeast, David B.; Knobel, Robert G.

2018-03-01

We report on a method for suspending two-dimensional crystal materials in an electronic circuit using an only photoresists and solvents. Graphene and NbSe2 are suspended tens of nanometers above metal electrodes with clamping diameters of several microns. The optical cavity formed from the membrane/air/metal structures enables a quick method to measure the number of layers and the gap separation using comparisons between the expected colour and optical microscope images. This characterization technique can be used with just an illuminated microscope with a digital camera which makes it adaptable to environments where other means of characterization are not possible, such as inside nitrogen glove boxes used in handling oxygen-sensitive materials.

Enhancement of graphene visibility on transparent substrates by refractive index optimization.

PubMed

Gonçalves, Hugo; Alves, Luís; Moura, Cacilda; Belsley, Michael; Stauber, Tobias; Schellenberg, Peter

2013-05-20

Optical reflection microscopy is one of the main imaging tools to visualize graphene microstructures. Here is reported a novel method that employs refractive index optimization in an optical reflection microscope, which greatly improves the visibility of graphene flakes. To this end, an immersion liquid with a refractive index that is close to that of the glass support is used in-between the microscope lens and the support improving the contrast and resolution of the sample image. Results show that the contrast of single and few layer graphene crystals and structures can be enhanced by a factor of 4 compared to values commonly achieved with transparent substrates using optical reflection microscopy lacking refractive index optimization.

Optical characterization and bandgap engineering of flat and wrinkle-textured FA0.83Cs0.17Pb(I1-xBrx)3 perovskite thin films

NASA Astrophysics Data System (ADS)

Tejada, A.; Braunger, S.; Korte, L.; Albrecht, S.; Rech, B.; Guerra, J. A.

2018-05-01

The complex refractive indices of formamidinium cesium lead mixed-halide [FA0.83Cs0.17Pb(I1- xBrx)3] perovskite thin films of compositions ranging from x = 0 to 0.4, with both flat and wrinkle-textured surface topographies, are reported. The films are characterized using a combination of variable angle spectroscopic ellipsometry and spectral transmittance in the wavelength range of 190 nm to 850 nm. Optical constants, film thicknesses and roughness layers are obtained point-by-point by minimizing a global error function, without using optical dispersion models, and including topographical information supplied by a laser confocal microscope. To evaluate the bandgap engineering potential of the material, the optical bandgaps and Urbach energies are then accurately determined by applying a band fluctuation model for direct semiconductors, which considers both the Urbach tail and the fundamental band-to-band absorption region in a single equation. With this information, the composition yielding the optimum bandgap of 1.75 eV for a Si-perovskite tandem solar cell is determined.

Quantum Simulation

NASA Astrophysics Data System (ADS)

Orzel, Chad

2017-06-01

One of the most active areas in atomic, molecular and optical physics is the use of ultracold atomic gases in optical lattices to simulate the behaviour of electrons in condensed matter systems. The larger mass, longer length scale, and tuneable interactions in these systems allow the dynamics of atoms moving in these systems to be followed in real time, and resonant light scattering by the atoms allows this motion to be probed on a microscopic scale using site-resolved imaging. This book reviews the physics of Hubbard-type models for both bosons and fermions in an optical lattice, which give rise to a rich variety of insulating and conducting phases depending on the lattice properties and interparticle interactions. It also discusses the effect of disorder on the transport of atoms in these models, and the recently discovered phenomenon of many-body localization. It presents several examples of experiments using both density and momentum imaging and quantum gas microscopy to study the motion of atoms in optical lattices. These illustrate the power and flexibility of ultracold-lattice analogues for exploring exotic states of matter at an unprecedented level of precision.

Modeling of Yb3+/Er3+-codoped microring resonators

NASA Astrophysics Data System (ADS)

Vallés, Juan A.; Gălătuş, Ramona

2015-03-01

The performance of a highly Yb3+/Er3+-codoped phosphate glass add-drop microring resonator is numerically analyzed. The model assumes resonant behaviour of both pump and signal powers and the dependences of pump intensity build-up inside the microring resonator and of the signal transfer functions to the device through and drop ports are evaluated. Detailed equations for the evolution of the rare-earth ions levels population densities and the propagation of the optical powers inside the microring resonator are included in the model. Moreover, due to the high dopant concentrations considered, the microscopic statistical formalism based on the statistical average of the excitation probability of the Er3+ ion in a microscopic level has been used to describe energy-transfer inter-atomic mechanisms. Realistic parameters and working conditions are used for the calculations. Requirements to achieve amplification and laser oscillation within these devices are obtainable as a function of rare earth ions concentration and coupling losses.

Dynamic-focusing microscope objective for optical coherence tomography

NASA Astrophysics Data System (ADS)

Murali, Supraja; Rolland, Jannick

2007-01-01

Optical Coherence Tomography (OCT) is a novel optical imaging technique that has assumed significant importance in bio-medical imaging in the last two decades because it is non-invasive and provides accurate, high resolution images of three dimensional cross-sections of body tissue, exceeding the capabilities of the current predominant imaging technique - ultrasound. In this paper, the application of high resolution OCT, known as optical coherence microscopy (OCM) is investigated for in vivo detection of abnormal skin pathology for the early diagnosis of cancer. A main challenge in OCM is maintaining invariant resolution throughout the sample. The technology presented is based on a dynamic focusing microscope imaging probe conceived for skin imaging and the detection of abnormalities in the epithelium. A novel method for dynamic focusing in the biological sample is presented using variable-focus lens technology to obtain three dimensional images with invariant resolution throughout the cross-section and depth of the sample is presented and discussed. A low coherence broadband source centered at near IR wavelengths is used to illuminate the sample. The design, analysis and predicted performance of the dynamic focusing microscope objective designed for dynamic three dimensional imaging at 5μm resolution for the chosen broadband spectrum is presented.

Atomic force-multi-optical imaging integrated microscope for monitoring molecular dynamics in live cells.

PubMed

Trache, Andreea; Meininger, Gerald A

2005-01-01

A novel hybrid imaging system is constructed integrating atomic force microscopy (AFM) with a combination of optical imaging techniques that offer high spatial resolution. The main application of this instrument (the NanoFluor microscope) is the study of mechanotransduction with an emphasis on extracellular matrix-integrin-cytoskeletal interactions and their role in the cellular responses to changes in external chemical and mechanical factors. The AFM allows the quantitative assessment of cytoskeletal changes, binding probability, adhesion forces, and micromechanical properties of the cells, while the optical imaging applications allow thin sectioning of the cell body at the coverslip-cell interface, permitting the study of focal adhesions using total internal reflection fluorescence (TIRF) and internal reflection microscopy (IRM). Combined AFM-optical imaging experiments show that mechanical stimulation at the apical surface of cells induces a force-generating cytoskeletal response, resulting in focal contact reorganization on the basal surface that can be monitored in real time. The NanoFluor system is also equipped with a novel mechanically aligned dual camera acquisition system for synthesized Forster resonance energy transfer (FRET). The integrated NanoFluor microscope system is described, including its characteristics, applications, and limitations.

Structural and optical analysis of 60Co gamma-irradiated thin films of polycrystalline Ga10Se85Sn5

NASA Astrophysics Data System (ADS)

Ahmad, Shabir; Asokan, K.; Shahid Khan, Mohd.; Zulfequar, M.

2015-12-01

The present study focuses on the effects of gamma irradiation on structural and optical properties of polycrystalline Ga10Se85Sn5 thin films with a thickness of ∼300 nm deposited by the thermal evaporation technique on cleaned glass substrates. X-ray diffraction patterns of the investigated thin films show that crystallite growth occurs in the orthorhombic phase structure. The surface study carried out by using the scanning electron microscope (SEM) confirms that the grain size increases with gamma irradiation. The optical parameters were estimated from optical transmission spectra data measured from a UV-vis-spectrophotometer in the wavelength range of 200-1100 nm. The refractive index dispersion data of the investigated thin films follow the single oscillator model. The estimated values of static refractive index n0, oscillator strength Ed, zero frequency dielectric constant ε0, optical conductivity σoptical and the dissipation factor increases after irradiation, while the single oscillator energy Eo decreases after irradiation. It was found that the value of the optical band gap of the investigated thin films decreases and the corresponding absorption coefficient increases continuously with an increase in the dose of gamma irradiation. This post irradiation changes in the values of optical band gap and absorption coefficient were interpreted in terms of the bond distribution model.

Multimodal hyperspectral optical microscopy

DOE PAGES

Novikova, Irina V.; Smallwood, Chuck R.; Gong, Yu; ...

2017-09-02

We describe a unique and convenient approach to multimodal hyperspectral optical microscopy, herein achieved by coupling a portable and transferable hyperspectral imager to various optical microscopes. The experimental and data analysis schemes involved in recording spectrally and spatially resolved fluorescence, dark field, and optical absorption micrographs are illustrated through prototypical measurements targeting selected model systems. Namely, hyperspectral fluorescence micrographs of isolated fluorescent beads are employed to ensure spectral calibration of our detector and to gauge the attainable spatial resolution of our measurements; the recorded images are diffraction-limited. Moreover, spatially over-sampled absorption spectroscopy of a single lipid (18:1 Liss Rhod PE)more » layer reveals that optical densities on the order of 10-3 may be resolved by spatially averaging the recorded optical signatures. We also briefly illustrate two applications of our setup in the general areas of plasmonics and cell biology. Most notably, we deploy hyperspectral optical absorption microscopy to identify and image algal pigments within a single live Tisochrysis lutea cell. Overall, this work paves the way for multimodal multidimensional spectral imaging measurements spanning the realms of several scientific disciples.« less

Multimodal hyperspectral optical microscopy

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

Novikova, Irina V.; Smallwood, Chuck R.; Gong, Yu

We describe a unique and convenient approach to multimodal hyperspectral optical microscopy, herein achieved by coupling a portable and transferable hyperspectral imager to various optical microscopes. The experimental and data analysis schemes involved in recording spectrally and spatially resolved fluorescence, dark field, and optical absorption micrographs are illustrated through prototypical measurements targeting selected model systems. Namely, hyperspectral fluorescence micrographs of isolated fluorescent beads are employed to ensure spectral calibration of our detector and to gauge the attainable spatial resolution of our measurements; the recorded images are diffraction-limited. Moreover, spatially over-sampled absorption spectroscopy of a single lipid (18:1 Liss Rhod PE)more » layer reveals that optical densities on the order of 10-3 may be resolved by spatially averaging the recorded optical signatures. We also briefly illustrate two applications of our setup in the general areas of plasmonics and cell biology. Most notably, we deploy hyperspectral optical absorption microscopy to identify and image algal pigments within a single live Tisochrysis lutea cell. Overall, this work paves the way for multimodal multidimensional spectral imaging measurements spanning the realms of several scientific disciples.« less

Assessing the Penetrating Abilities of Experimental Preparation with Dental Infiltrant Features Using Optical Microscope: Preliminary Study.

PubMed

Skucha-Nowak, Małgorzata; Machorowska-Pieniążek, Agnieszka; Tanasiewicz, Marta

2016-01-01

The aim of the infiltration technique is to penetrate demineralized enamel with a low viscosity resin. Icon® (DMG) is the first ever and so far the only dental infiltrant. Bacteriostaticity is one of the properties that should be inherent in dental infiltrants, but Icon lacks this feature. The aim of the preliminary study was to properly choose a dye which would allow us to assess the penetrating abilities of our own, experimental preparation with features of a dental infiltrant with bacteriostatic properties and to compare using an optical microscope the depth of infiltration of the designed experimental preparation with the infiltrant available on the market. The preparation is supposed to infiltrate decalcified human enamel and be assessed with an optical microscope. Eosin, neutral fuchsine and methylene blue were added to experimental preparation with dental infiltrant features and to Icon® (DMG) in order to assess the depth of penetration of the experimental solution into the decalcified layers of enamel. The experimental solution mixes well with eosin, neutral fuchsine, and methylene blue. During the preliminary study, the authors concluded that the experimental solution mixes well with methylene blue, neutral fuchsine, and eosin. An addition of eosin to a preparation which infiltrates inner, demineralized enamel layers, facilitates the assessment of such a preparation with an optical microscope. A designed experimental solution with the main ingredients, i.e., 2-hydroxyethyl methacrylate (HEMA) and tetraethylene glycol dimethacrylate (TEGDMA) with a ratio of 75% to 25% penetrates the demineralized (decalcified) inner parts of the enamel and polymerizes when exposed to light. In order to assess the infiltration of the experimental solution into the demineralized enamel layers, it is required to improve the measurement techniques that utilize optical microscopy.

Comparative analysis of imaging configurations and objectives for Fourier microscopy.

PubMed

Kurvits, Jonathan A; Jiang, Mingming; Zia, Rashid

2015-11-01

Fourier microscopy is becoming an increasingly important tool for the analysis of optical nanostructures and quantum emitters. However, achieving quantitative Fourier space measurements requires a thorough understanding of the impact of aberrations introduced by optical microscopes that have been optimized for conventional real-space imaging. Here we present a detailed framework for analyzing the performance of microscope objectives for several common Fourier imaging configurations. To this end, we model objectives from Nikon, Olympus, and Zeiss using parameters that were inferred from patent literature and confirmed, where possible, by physical disassembly. We then examine the aberrations most relevant to Fourier microscopy, including the alignment tolerances of apodization factors for different objective classes, the effect of magnification on the modulation transfer function, and vignetting-induced reductions of the effective numerical aperture for wide-field measurements. Based on this analysis, we identify an optimal objective class and imaging configuration for Fourier microscopy. In addition, the Zemax files for the objectives and setups used in this analysis have been made publicly available as a resource for future studies.

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.

Environmental Integrity of Coating/Metal Interface

DTIC Science & Technology

1989-03-01

occurred. For this explanation to be correct, microblisters would be optically detectable using a microscope with Nomarsky contrast or by examining the...polymer surface with an interferometric microscope. Both of these techniques have been tried, but neither of them was able to detect any surface

A simple optical tweezers for trapping polystyrene particles

NASA Astrophysics Data System (ADS)

Shiddiq, Minarni; Nasir, Zulfa; Yogasari, Dwiyana

2013-09-01

Optical tweezers is an optical trap. For decades, it has become an optical tool that can trap and manipulate any particle from the very small size like DNA to the big one like bacteria. The trapping force comes from the radiation pressure of laser light which is focused to a group of particles. Optical tweezers has been used in many research areas such as atomic physics, medical physics, biophysics, and chemistry. Here, a simple optical tweezers has been constructed using a modified Leybold laboratory optical microscope. The ocular lens of the microscope has been removed for laser light and digital camera accesses. A laser light from a Coherent diode laser with wavelength λ = 830 nm and power 50 mW is sent through an immersion oil objective lens with magnification 100 × and NA 1.25 to a cell made from microscope slides containing polystyrene particles. Polystyrene particles with size 3 μm and 10 μm are used. A CMOS Thorlabs camera type DCC1545M with USB Interface and Thorlabs camera lens 35 mm are connected to a desktop and used to monitor the trapping and measure the stiffness of the trap. The camera is accompanied by camera software which makes able for the user to capture and save images. The images are analyzed using ImageJ and Scion macro. The polystyrene particles have been trapped successfully. The stiffness of the trap depends on the size of the particles and the power of the laser. The stiffness increases linearly with power and decreases as the particle size larger.

An Improved Optical Tweezers Assay for Measuring the Force Generation of Single Kinesin Molecules

PubMed Central

Nicholas, Matthew P.; Rao, Lu; Gennerich, Arne

2014-01-01

Numerous microtubule-associated molecular motors, including several kinesins and cytoplasmic dynein, produce opposing forces that regulate spindle and chromosome positioning during mitosis. The motility and force generation of these motors are therefore critical to normal cell division, and dysfunction of these processes may contribute to human disease. Optical tweezers provide a powerful method for studying the nanometer motility and piconewton force generation of single motor proteins in vitro. Using kinesin-1 as a prototype, we present a set of step-by-step, optimized protocols for expressing a kinesin construct (K560-GFP) in Escherichia coli, purifying it, and studying its force generation in an optical tweezers microscope. We also provide detailed instructions on proper alignment and calibration of an optical trapping microscope. These methods provide a foundation for a variety of similar experiments. PMID:24633799

Simulation study on compressive laminar optical tomography for cardiac action potential propagation

PubMed Central

Harada, Takumi; Tomii, Naoki; Manago, Shota; Kobayashi, Etsuko; Sakuma, Ichiro

2017-01-01

To measure the activity of tissue at the microscopic level, laminar optical tomography (LOT), which is a microscopic form of diffuse optical tomography, has been developed. However, obtaining sufficient recording speed to determine rapidly changing dynamic activity remains major challenges. For a high frame rate of the reconstructed data, we here propose a new LOT method using compressed sensing theory, called compressive laminar optical tomography (CLOT), in which novel digital micromirror device-based illumination and data reduction in a single reconstruction are applied. In the simulation experiments, the reconstructed volumetric images of the action potentials that were acquired from 5 measured images with random pattern featured a wave border at least to a depth of 2.5 mm. Consequently, it was shown that CLOT has potential for over 200 fps required for the cardiac electrophysiological phenomena. PMID:28736675

Imaging arrangement and microscope

DOEpatents

Pertsinidis, Alexandros; Chu, Steven

2015-12-15

An embodiment of the present invention is an imaging arrangement that includes imaging optics, a fiducial light source, and a control system. In operation, the imaging optics separate light into first and second tight by wavelength and project the first and second light onto first and second areas within first and second detector regions, respectively. The imaging optics separate fiducial light from the fiducial light source into first and second fiducial light and project the first and second fiducial light onto third and fourth areas within the first and second detector regions, respectively. The control system adjusts alignment of the imaging optics so that the first and second fiducial light projected onto the first and second detector regions maintain relatively constant positions within the first and second detector regions, respectively. Another embodiment of the present invention is a microscope that includes the imaging arrangement.

3D printing of optical materials: an investigation of the microscopic properties

NASA Astrophysics Data System (ADS)

Persano, Luana; Cardarelli, Francesco; Arinstein, Arkadii; Uttiya, Sureeporn; Zussman, Eyal; Pisignano, Dario; Camposeo, Andrea

2018-02-01

3D printing technologies are currently enabling the fabrication of objects with complex architectures and tailored properties. In such framework, the production of 3D optical structures, which are typically based on optical transparent matrices, optionally doped with active molecular compounds and nanoparticles, is still limited by the poor uniformity of the printed structures. Both bulk inhomogeneities and surface roughness of the printed structures can negatively affect the propagation of light in 3D printed optical components. Here we investigate photopolymerization-based printing processes by laser confocal microscopy. The experimental method we developed allows the printing process to be investigated in-situ, with microscale spatial resolution, and in real-time. The modelling of the photo-polymerization kinetics allows the different polymerization regimes to be investigated and the influence of process variables to be rationalized. In addition, the origin of the factors limiting light propagation in printed materials are rationalized, with the aim of envisaging effective experimental strategies to improve optical properties of printed materials.

Optical-sectioning microscopy of protoporphyrin IX fluorescence in human gliomas: standardization and quantitative comparison with histology

NASA Astrophysics Data System (ADS)

Wei, Linpeng; Chen, Ye; Yin, Chengbo; Borwege, Sabine; Sanai, Nader; Liu, Jonathan T. C.

2017-04-01

Systemic delivery of 5-aminolevulinic acid leads to enhanced fluorescence image contrast in many tumors due to the increased accumulation of protoporphyrin IX (PpIX), a fluorescent porphyrin that is associated with tumor burden and proliferation. The value of PpIX-guided resection of malignant gliomas has been demonstrated in prospective randomized clinical studies in which a twofold greater extent of resection and improved progression-free survival have been observed. In low-grade gliomas and at the diffuse infiltrative margins of all gliomas, PpIX fluorescence is often too weak to be detected with current low-resolution surgical microscopes that are used in operating rooms. However, it has been demonstrated that high-resolution optical-sectioning microscopes are capable of detecting the sparse and punctate accumulations of PpIX that are undetectable via conventional low-power surgical fluorescence microscopes. To standardize the performance of high-resolution optical-sectioning devices for future clinical use, we have developed an imaging phantom and methods to ensure that the imaging of PpIX-expressing brain tissues can be performed reproducibly. Ex vivo imaging studies with a dual-axis confocal microscope demonstrate that these methods enable the acquisition of images from unsectioned human brain tissues that quantitatively and consistently correlate with images of histologically processed tissue sections.

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.

[Intraoperative Optical Coherence Tomography (MI-OCT) for the Treatment of Corneal Dystrophies].

PubMed

Siebelmann, Sebastian; Matthaei, Mario; Heindl, Ludwig M; Bachmann, Björn O; Cursiefen, Claus

2018-06-01

The surgical treatment of corneal dystrophies develops rapidly as the use of lamellar corneal grafting techniques continue. While penetrating keratoplasty was the gold standard for treating a variety of dystrophies a few years ago, the affected layers of the cornea can, nowadays, be selectively replaced or ablated using laser technology. Of particular importance for these methods is optical coherence tomography, which has recently been integrated into surgical microscopes (MI-OCT). Literature overview from PubMed and Google.scholar.de supplemented with own imaging data. The MI-OCT enables the intraoperative real-time monitoring of different ophthalmic surgical procedures, such as deep anterior lamellar keratoplasty, Descemet's membrane endothelial keratoplasty, as well as minimally-invasive procedures, such as phototherapeutic keratectomy. In addition, it enables an evaluation of the cornea, but also of structures of the anterior chamber, in situations in which the cornea, for example, is clouded by an edema. Microscope-integrated, intraoperative optical coherence tomography (MI-OCT) represents a useful supplement to the normal surgical microscope. It is superior to the sole surgical microscope, especially in already severely clouded corneas, and represents a sensible supplement, especially for novel lamellar transplantation procedures. Prospective randomized trials are necessary to increase safety and efficacy when using MI-OCT for different indications. Georg Thieme Verlag KG Stuttgart · New York.

Light field creating and imaging with different order intensity derivatives

NASA Astrophysics Data System (ADS)

Wang, Yu; Jiang, Huan

2014-10-01

Microscopic image restoration and reconstruction is a challenging topic in the image processing and computer vision, which can be widely applied to life science, biology and medicine etc. A microscopic light field creating and three dimensional (3D) reconstruction method is proposed for transparent or partially transparent microscopic samples, which is based on the Taylor expansion theorem and polynomial fitting. Firstly the image stack of the specimen is divided into several groups in an overlapping or non-overlapping way along the optical axis, and the first image of every group is regarded as reference image. Then different order intensity derivatives are calculated using all the images of every group and polynomial fitting method based on the assumption that the structure of the specimen contained by the image stack in a small range along the optical axis are possessed of smooth and linear property. Subsequently, new images located any position from which to reference image the distance is Δz along the optical axis can be generated by means of Taylor expansion theorem and the calculated different order intensity derivatives. Finally, the microscopic specimen can be reconstructed in 3D form using deconvolution technology and all the images including both the observed images and the generated images. The experimental results show the effectiveness and feasibility of our method.

Model-based aberration correction in a closed-loop wavefront-sensor-less adaptive optics system.

PubMed

Song, H; Fraanje, R; Schitter, G; Kroese, H; Vdovin, G; Verhaegen, M

2010-11-08

In many scientific and medical applications, such as laser systems and microscopes, wavefront-sensor-less (WFSless) adaptive optics (AO) systems are used to improve the laser beam quality or the image resolution by correcting the wavefront aberration in the optical path. The lack of direct wavefront measurement in WFSless AO systems imposes a challenge to achieve efficient aberration correction. This paper presents an aberration correction approach for WFSlss AO systems based on the model of the WFSless AO system and a small number of intensity measurements, where the model is identified from the input-output data of the WFSless AO system by black-box identification. This approach is validated in an experimental setup with 20 static aberrations having Kolmogorov spatial distributions. By correcting N=9 Zernike modes (N is the number of aberration modes), an intensity improvement from 49% of the maximum value to 89% has been achieved in average based on N+5=14 intensity measurements. With the worst initial intensity, an improvement from 17% of the maximum value to 86% has been achieved based on N+4=13 intensity measurements.

Compact whole-body fluorescent imaging of nude mice bearing EGFP expressing tumor

NASA Astrophysics Data System (ADS)

Chen, Yanping; Xiong, Tao; Chu, Jun; Yu, Li; Zeng, Shaoqun; Luo, Qingming

2005-01-01

Issue of tumor has been a hotspot of current medicine. It is important for tumor research to detect tumors bearing in animal models easily, fast, repetitively and noninvasivly. Many researchers have paid their increasing interests on the detecting. Some contrast agents, such as green fluorescent protein (GFP) and Discosoma red fluorescent protein (Dsred) were applied to enhance image quality. Three main kinds of imaging scheme were adopted to visualize fluorescent protein expressing tumors in vivo. These schemes based on fluorescence stereo microscope, cooled charge-coupled-device (CCD) or camera as imaging set, and laser or mercury lamp as excitation light source. Fluorescence stereo microscope, laser and cooled CCD are expensive to many institutes. The authors set up an inexpensive compact whole-body fluorescent imaging tool, which consisted of a Kodak digital camera (model DC290), fluorescence filters(B and G2;HB Optical, Shenyang, Liaoning, P.R. China) and a mercury 50-W lamp power supply (U-LH50HG;Olympus Optical, Japan) as excitation light source. The EGFP was excited directly by mercury lamp with D455/70 nm band-pass filter and fluorescence was recorded by digital camera with 520nm long-pass filter. By this easy operation tool, the authors imaged, in real time, fluorescent tumors growing in live mice. The imaging system is external and noninvasive. For half a year our experiments suggested the imaging scheme was feasible. Whole-body fluorescence optical imaging for fluorescent expressing tumors in nude mouse is an ideal tool for antitumor, antimetastatic, and antiangiogenesis drug screening.

A nano grating tunable MEMS optical filter for high-speed on-chip multispectral fluorescent detection.

PubMed

Truxal, Steven C; Huang, Nien-Tsu; Kurabayashi, Katsuo

2009-01-01

We report a microelectromechanical (MEMS) tunable optical filter and its integration in a fluorescence microscope for high speed on-chip spectral measurements. This integration allows for measurements of any fluorescence sample placed onto the microscope stage. We demonstrate the system capabilities by taking spectral measurements of multicolor fluorescent beads and fluorescently labeled cells passing through a microfluidic cytometer. The system has applications in biological studies where the measurement of multiple fluorescent peaks is restricted by the detection method's speed and sensitivity.

Electron microscopic and optical studies of prism faces of synthetic quartz

NASA Technical Reports Server (NTRS)

Buzek, B. C.; Vagh, A. S.

1977-01-01

Application of electron and optical microscopic techniques to the study of growth spirals on quartz crystal faces is described. Attention is centered on the centers of the spirals and on screw ledges; overhanging kinks are revealed on one side of the spiral centers. The possibility that these special features may have developed after growth of the crystals went to completion is explored. The conjecture is raised that such structures might result from adsorption of growth-inhibiting impurities at the center of the growth spiral on the quartz habit faces.

Laboratory-size three-dimensional water-window x-ray microscope with Wolter type I mirror optics

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

Ohsuka, Shinji; The Graduate School for the Creation of New Photonics Industries, 1955-1 Kurematsu-cho, Nishi-ku, Hamamatsu-City, 431-1202; Ohba, Akira

2016-01-28

We constructed a laboratory-size three-dimensional water-window x-ray microscope that combines wide-field transmission x-ray microscopy with tomographic reconstruction techniques. It consists of an electron-impact x-ray source emitting oxygen Kα x-rays, Wolter type I grazing incidence mirror optics, and a back-illuminated CCD for x-ray imaging. A spatial resolution limit better than 1.0 line pairs per micrometer was obtained for two-dimensional transmission images, and 1-μm-scale three-dimensional fine structures were resolved.

Artificial testing targets with controllable blur for adaptive optics microscopes

NASA Astrophysics Data System (ADS)

Hattori, Masayuki; Tamada, Yosuke; Murata, Takashi; Oya, Shin; Hasebe, Mitsuyasu; Hayano, Yutaka; Kamei, Yasuhiro

2017-08-01

This letter proposes a method of configuring a testing target to evaluate the performance of adaptive optics microscopes. In this method, a testing slide with fluorescent beads is used to simultaneously determine the point spread function and the field of view. The point spread function is reproduced to simulate actual biological samples by etching a microstructure on the cover glass. The fabrication process is simplified to facilitate an onsite preparation. The artificial tissue consists of solid materials and silicone oil and is stable for use in repetitive experiments.

Fast and Accurate Cell Tracking by a Novel Optical-Digital Hybrid Method

NASA Astrophysics Data System (ADS)

Torres-Cisneros, M.; Aviña-Cervantes, J. G.; Pérez-Careta, E.; Ambriz-Colín, F.; Tinoco, Verónica; Ibarra-Manzano, O. G.; Plascencia-Mora, H.; Aguilera-Gómez, E.; Ibarra-Manzano, M. A.; Guzman-Cabrera, R.; Debeir, Olivier; Sánchez-Mondragón, J. J.

2013-09-01

An innovative methodology to detect and track cells using microscope images enhanced by optical cross-correlation techniques is proposed in this paper. In order to increase the tracking sensibility, image pre-processing has been implemented as a morphological operator on the microscope image. Results show that the pre-processing process allows for additional frames of cell tracking, therefore increasing its robustness. The proposed methodology can be used in analyzing different problems such as mitosis, cell collisions, and cell overlapping, ultimately designed to identify and treat illnesses and malignancies.

Demonstration of magnetic domain boundary movement using an easily assembled videocam-microscope system

NASA Technical Reports Server (NTRS)

Patterson, John W.

1992-01-01

The objectives are to build and demonstrate a low cost and highly flexible TV microscope facility and then use it to view the motion of magnetic domain boundaries as the local magnetic field is varied. The expense of an optical microscope and the videocam adapters sold for them is largely avoided by using the facility described below. The equipment, supplies, and procedure 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.

Microscope-integrated optical coherence tomography for image-aided positioning of glaucoma surgery

NASA Astrophysics Data System (ADS)

Li, Xiqi; Wei, Ling; Dong, Xuechuan; Huang, Ping; Zhang, Chun; He, Yi; Shi, Guohua; Zhang, Yudong

2015-07-01

Most glaucoma surgeries involve creating new aqueous outflow pathways with the use of a small surgical instrument. This article reported a microscope-integrated, real-time, high-speed, swept-source optical coherence tomography system (SS-OCT) with a 1310-nm light source for glaucoma surgery. A special mechanism was designed to produce an adjustable system suitable for use in surgery. A two-graphic processing unit architecture was used to speed up the data processing and real-time volumetric rendering. The position of the surgical instrument can be monitored and measured using the microscope and a grid-inserted image of the SS-OCT. Finally, experiments were simulated to assess the effectiveness of this integrated system. Experimental results show that this system is a suitable positioning tool for glaucoma surgery.

A versatile optical microscope for time-dependent single-molecule and single-particle spectroscopy

NASA Astrophysics Data System (ADS)

Li, Hao; Yang, Haw

2018-03-01

This work reports the design and implementation of a multi-function optical microscope for time-dependent spectroscopy on single molecules and single nanoparticles. It integrates the now-routine single-object measurements into one standalone platform so that no reconfiguration is needed when switching between different types of sample or spectroscopy modes. The illumination modes include evanescent field through total internal reflection, dark-field illumination, and epi-excitation onto a diffraction-limited spot suitable for confocal detection. The detection modes include spectrally resolved line imaging, wide-field imaging with dual-color capability, and two-color single-element photon-counting detection. The switch between different spectroscopy and data acquisition modes is fully automated and executed through computer programming. The capability of this microscope is demonstrated through selected proof-of-principle experiments.

Hyperspectral stimulated emission depletion microscopy and methods of use thereof

DOEpatents

Timlin, Jerilyn A; Aaron, Jesse S

2014-04-01

A hyperspectral stimulated emission depletion ("STED") microscope system for high-resolution imaging of samples labeled with multiple fluorophores (e.g., two to ten fluorophores). The hyperspectral STED microscope includes a light source, optical systems configured for generating an excitation light beam and a depletion light beam, optical systems configured for focusing the excitation and depletion light beams on a sample, and systems for collecting and processing data generated by interaction of the excitation and depletion light beams with the sample. Hyperspectral STED data may be analyzed using multivariate curve resolution analysis techniques to deconvolute emission from the multiple fluorophores. The hyperspectral STED microscope described herein can be used for multi-color, subdiffraction imaging of samples (e.g., materials and biological materials) and for analyzing a tissue by Forster Resonance Energy Transfer ("FRET").

A versatile optical microscope for time-dependent single-molecule and single-particle spectroscopy.

PubMed

Li, Hao; Yang, Haw

2018-03-28

This work reports the design and implementation of a multi-function optical microscope for time-dependent spectroscopy on single molecules and single nanoparticles. It integrates the now-routine single-object measurements into one standalone platform so that no reconfiguration is needed when switching between different types of sample or spectroscopy modes. The illumination modes include evanescent field through total internal reflection, dark-field illumination, and epi-excitation onto a diffraction-limited spot suitable for confocal detection. The detection modes include spectrally resolved line imaging, wide-field imaging with dual-color capability, and two-color single-element photon-counting detection. The switch between different spectroscopy and data acquisition modes is fully automated and executed through computer programming. The capability of this microscope is demonstrated through selected proof-of-principle experiments.

Asbestos Testing: Is the EPA Misleading You?

ERIC Educational Resources Information Center

Levins, Hoag

1983-01-01

Experts warn that only electron microscopes can see the smaller fibers of asbestos that are known to cause the most cancers, though the Environmental Protection Agency still endorses optical microscopes for asbestos removal verification. Asbestos testing methods are explained and sources of information are provided. (MLF)

Design and Construction of a Multi-wavelength, Micromirror Total Internal Reflectance Fluorescence Microscope

PubMed Central

Larson, Joshua; Kirk, Matt; Drier, Eric A.; O’Brien, William; MacKay, James F.; Friedman, Larry; Hoskins, Aaron

2015-01-01

Colocalization Single Molecule Spectroscopy (CoSMoS) has proven to be a useful method for studying the composition, kinetics, and mechanisms of complex cellular machines. Key to the technique is the ability to simultaneously monitor multiple proteins and/or nucleic acids as they interact with one another. Here we describe a protocol for constructing a CoSMoS micromirror Total Internal Reflection Fluorescence Microscope (mmTIRFM). Design and construction of a scientific microscope often requires a number of custom components and a significant time commitment. In our protocol, we have streamlined this process by implementation of a commercially available microscopy platform designed to accommodate the optical components necessary for a mmTIRFM. The mmTIRF system eliminates the need for machining custom parts by the end-user and facilitates optical alignment. Depending on the experience-level of the microscope builder, these time-savings and the following protocol can enable mmTIRF construction to be completed within two months. PMID:25188633

Design and construction of a multiwavelength, micromirror total internal reflectance fluorescence microscope.

PubMed

Larson, Joshua; Kirk, Matt; Drier, Eric A; O'Brien, William; MacKay, James F; Friedman, Larry J; Hoskins, Aaron A

2014-10-01

Colocalization single-molecule spectroscopy (CoSMoS) has proven to be a useful method for studying the composition, kinetics and mechanisms of complex cellular machines. Key to the technique is the ability to simultaneously monitor multiple proteins and/or nucleic acids as they interact with one another. Here we describe a protocol for constructing a CoSMoS micromirror total internal reflection fluorescence microscope (mmTIRFM). Design and construction of a scientific microscope often requires a number of custom components and a substantial time commitment. In our protocol, we have streamlined this process by implementation of a commercially available microscopy platform designed to accommodate the optical components necessary for an mmTIRFM. The mmTIRF system eliminates the need for machining custom parts by the end user and facilitates optical alignment. Depending on the experience level of the microscope builder, these time savings and the following protocol can enable mmTIRF construction to be completed within 2 months.

Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning.

PubMed

Cheng, Li-Chung; Chang, Chia-Yuan; Lin, Chun-Yu; Cho, Keng-Chi; Yen, Wei-Chung; Chang, Nan-Shan; Xu, Chris; Dong, Chen Yuan; Chen, Shean-Jen

2012-04-09

In this study, a microscope based on spatiotemporal focusing offering widefield multiphoton excitation has been developed to provide fast optical sectioning images. Key features of this microscope are the integrations of a 10 kHz repetition rate ultrafast amplifier featuring high instantaneous peak power (maximum 400 μJ/pulse at a 90 fs pulse width) and a TE-cooled, ultra-sensitive photon detecting, electron multiplying charge-coupled camera into a spatiotemporal focusing microscope. This configuration can produce multiphoton images with an excitation area larger than 200 × 100 μm² at a frame rate greater than 100 Hz (current maximum of 200 Hz). Brownian motions of fluorescent microbeads as small as 0.5 μm were observed in real-time with a lateral spatial resolution of less than 0.5 μm and an axial resolution of approximately 3.5 μm. Furthermore, second harmonic images of chicken tendons demonstrate that the developed widefield multiphoton microscope can provide high resolution z-sectioning for bioimaging.

Multifocal Fluorescence Microscope for Fast Optical Recordings of Neuronal Action Potentials

PubMed Central

Shtrahman, Matthew; Aharoni, Daniel B.; Hardy, Nicholas F.; Buonomano, Dean V.; Arisaka, Katsushi; Otis, Thomas S.

2015-01-01

In recent years, optical sensors for tracking neural activity have been developed and offer great utility. However, developing microscopy techniques that have several kHz bandwidth necessary to reliably capture optically reported action potentials (APs) at multiple locations in parallel remains a significant challenge. To our knowledge, we describe a novel microscope optimized to measure spatially distributed optical signals with submillisecond and near diffraction-limit resolution. Our design uses a spatial light modulator to generate patterned illumination to simultaneously excite multiple user-defined targets. A galvanometer driven mirror in the emission path streaks the fluorescence emanating from each excitation point during the camera exposure, using unused camera pixels to capture time varying fluorescence at rates that are ∼1000 times faster than the camera’s native frame rate. We demonstrate that this approach is capable of recording Ca2+ transients resulting from APs in neurons labeled with the Ca2+ sensor Oregon Green Bapta-1 (OGB-1), and can localize the timing of these events with millisecond resolution. Furthermore, optically reported APs can be detected with the voltage sensitive dye DiO-DPA in multiple locations within a neuron with a signal/noise ratio up to ∼40, resolving delays in arrival time along dendrites. Thus, the microscope provides a powerful tool for photometric measurements of dynamics requiring submillisecond sampling at multiple locations. PMID:25650920

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.

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.

Electron Optics for Biologists: Physical Origins of Spherical Aberrations

ERIC Educational Resources Information Center

Geissler, Peter; Zadunaisky, Jose

1974-01-01

Reports on the physical origins of spherical aberrations in axially symmetric electrostatic lenses to convey the essentials of electon optics to those who must think critically about the resolution of the electron microscope. (GS)

Applied physics: Optical trapping for space mirrors.

PubMed

McGloin, David

2014-02-27

Might it be possible to create mirrors for space telescopes, using nothing but microscopic particles held in place by light? A study that exploits a technique called optical binding provides a step towards this goal.

Wavelet analysis of polarization maps of the myocardium tissue microscopic images in the diagnosis of the causes of death

NASA Astrophysics Data System (ADS)

Ushenko, V. O.; Boichuk, T. M.; Bachinskiy, V. T.; Vanchuliak, O. Ya.; Minzer, O. P.; Dubolazov, O. V.; Marchuk, Yu. F.; Olar, O. I.

2015-08-01

The results of optical modeling of biological tissues polycrystalline multilayer networks have been presented. Algorithms of reconstruction of parameter distributions were determined that describe the linear and circular birefringence. For the separation of the manifestations of these mechanisms we propose a method of space-frequency filtering. Criteria for differentiation of benign and malignant tissues of the women reproductive sphere were found.

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

Microscopic theory of optical absorption in graphene enhanced by lattices of plasmonic nanoparticles

NASA Astrophysics Data System (ADS)

Mueller, Niclas S.; Reich, Stephanie

2018-06-01

We present a microscopic description of plasmon-enhanced optical absorption in graphene, which is based on perturbation theory. We consider the interaction of graphene with a lattice of plasmonic nanoparticles, as was previously realized experimentally. By using tight-binding wave functions for the electronic states of graphene and the dipole approximation for the plasmon, we obtain analytic expressions for the coupling matrix element and enhanced optical absorption. The plasmonic nanostructure induces nonvertical optical transitions in the band structure of graphene with selection rules for the momentum transfer that depend on the periodicity of the plasmonic lattice. The plasmon-mediated optical absorption leads to an anisotropic carrier population around the K point in phase space, which depends on the polarization pattern of the plasmonic near field in the graphene plane. Using Fourier optics, we draw a connection to a macroscopic approach, which is independent from graphene-specific parameters. Each Fourier component of the plasmonic near field corresponds to the momentum transfer of an optical transition. Both approaches lead to the same expression for the integrated optical absorption enhancement, which is relevant for the photocurrent enhancement in graphene-based optoelectronic devices.

Optical alignment using a CGH and an autostigmatic microscope

NASA Astrophysics Data System (ADS)

Parks, Robert E.

2017-08-01

We show how custom computer generated holograms (CGH) are used along with an autostigmatic microscope (ASM) to align both optical and mechanical components relative to the CGH. The patterns in the CGHs define points and lines in space when interrogated with the focus of the ASM. Once the ASM is aligned to the CGH, an optical or mechanical component such as a lens, a well-polished ball or a cylinder can be aligned to the ASM in 3 or 4 degrees of freedom and thus to the CGH. In this case we show how a CGH is used to make a fixture for cementing a doublet lens without the need for a rotary table or a precision vertical stage.

Reflective type objective based spectral-domain phase-sensitive optical coherence tomography for high-sensitive structural and functional imaging of cochlear microstructures through intact bone of an excised guinea pig cochlea

NASA Astrophysics Data System (ADS)

Subhash, Hrebesh M.; Wang, Ruikang K.; Chen, Fangyi; Nuttall, Alfred L.

2013-03-01

Most of the optical coherence tomographic (OCT) systems for high resolution imaging of biological specimens are based on refractive type microscope objectives, which are optimized for specific wave length of the optical source. In this study, we present the feasibility of using commercially available reflective type objective for high sensitive and high resolution structural and functional imaging of cochlear microstructures of an excised guinea pig through intact temporal bone. Unlike conventional refractive type microscopic objective, reflective objective are free from chromatic aberrations due to their all-reflecting nature and can support a broadband of spectrum with very high light collection efficiency.

Aligning Arrays of Lenses and Single-Mode Optical Fibers

NASA Technical Reports Server (NTRS)

Liu, Duncan

2004-01-01

A procedure now under development is intended to enable the precise alignment of sheet arrays of microscopic lenses with the end faces of a coherent bundle of as many as 1,000 single-mode optical fibers packed closely in a regular array (see Figure 1). In the original application that prompted this development, the precise assembly of lenses and optical fibers serves as a single-mode spatial filter for a visible-light nulling interferometer. The precision of alignment must be sufficient to limit any remaining wavefront error to a root-mean-square value of less than 1/10 of a wavelength of light. This wavefront-error limit translates to requirements to (1) ensure uniformity of both the lens and fiber arrays, (2) ensure that the lateral distance from the central axis of each lens and the corresponding optical fiber is no more than a fraction of a micron, (3) angularly align the lens-sheet planes and the fiber-bundle end faces to within a few arc seconds, and (4) axially align the lenses and the fiber-bundle end faces to within tens of microns of the focal distance. Figure 2 depicts the apparatus used in the alignment procedure. The beam of light from a Zygo (or equivalent) interferometer is first compressed by a ratio of 20:1 so that upon its return to the interferometer, the beam will be magnified enough to enable measurement of wavefront quality. The apparatus includes relay lenses that enable imaging of the arrays of microscopic lenses in a charge-coupled-device (CCD) camera that is part of the interferometer. One of the arrays of microscopic lenses is mounted on a 6-axis stage, in proximity to the front face of the bundle of optical fibers. The bundle is mounted on a separate stage. A mirror is attached to the back face of the bundle of optical fibers for retroreflection of light. When a microscopic lens and a fiber are aligned with each other, the affected portion of the light is reflected back by the mirror, recollimated by the microscopic lens, transmitted through the relay lenses and the beam compressor/expander, then split so that half goes to a detector and half to the interferometer. The output of the detector is used as a feedback control signal for the six-axis stage to effect alignment.

Classification of gram-positive and gram-negative foodborne pathogenic bacteria with hyperspectral microscope imaging

USDA-ARS?s Scientific Manuscript database

Optical method with hyperspectral microscope imaging (HMI) has potential for identification of foodborne pathogenic bacteria from microcolonies rapidly with a cell level. A HMI system that provides both spatial and spectral information could be an effective tool for analyzing spectral characteristic...

Development of an upconverting chelate assay

NASA Astrophysics Data System (ADS)

Xiao, Xudong; Haushalter, Jeanne P.; Kotz, Kenneth T.; Faris, Gregory W.

2005-04-01

We report progress on performing a cell-based assay for the detection of EGFR on cell surfaces by using upconverting chelates. An upconversion microscope has been developed for performing assays and testing optical response. A431 cells are labeled with europium DOTA and imaged using this upconverting microscope.

The dynamic process and microscopic mechanism of extraordinary terahertz transmission through perforated superconducting films

PubMed Central

Wu, J. B.; Zhang, X.; Jin, B. B.; Liu, H. T.; Chen, Y. H.; Li, Z. Y.; Zhang, C. H.; Kang, L.; Xu, W. W.; Chen, J.; Wang, H. B.; Tonouchi, M.; Wu, P. H.

2015-01-01

Superconductor is a compelling plasmonic medium at terahertz frequencies owing to its intrinsic low Ohmic loss and good tuning property. However, the microscopic physics of the interaction between terahertz wave and superconducting plasmonic structures is still unknown. In this paper, we conducted experiments of the enhanced terahertz transmission through a series of superconducting NbN subwavelength hole arrays, and employed microscopic hybrid wave model in theoretical analysis of the role of hybrid waves in the enhanced transmission. The theoretical calculation provided a good match of experimental data. In particular, we obtained the following results. When the width of the holes is far below wavelength, the enhanced transmission is mainly caused by localized resonance around individual holes. On the contrary, when the holes are large, hybrid waves scattered by the array of holes dominate the extraordinary transmission. The surface plasmon polaritions are proved to be launched on the surface of superconducting film and the excitation efficiency increases when the temperature approaches critical temperature and the working frequency goes near energy gap frequency. This work will enrich our knowledge on the microscopic physics of extraordinary optical transmission at terahertz frequencies and contribute to developing terahertz plasmonic devices. PMID:26498994

Modeling of Optoelectronic Devices

NASA Technical Reports Server (NTRS)

Li, Jian-Zhong; Woo, Alex C. (Technical Monitor)

2000-01-01

Ultrafast modulation of semiconductor quantum well (QW) laser is of technological importance for information technology. Improvement by order(s) of magnitude in data transfer rate is possible as terahertz (THz) radiation is available for heating the laser at picosecond time scale. Optical gain modulation in the QW is achieved via temperature modulation of electron-hole plasma (EHP). Applications include free-space THz communication, optical switching, and pulse generation. The EHP in the semiconductor QW is described with a two-band model. Semiconductor Bloch equations with many-body effects are used to derive a hydrodynamical model for the active QW region. Because of ultrafast carrier-carrier scatterings in the order of 50 fs, EHP follows quasiequilibrium Fermi-Dirac distributions and THz field interacts incoherently with it. Carrier-longitudinal optical (LO) phonon scatterings and coherent laser-EHP interaction are treated microscopically in our physical model. A set of hydrodynamical equations for plasma density, temperature, and laser envelop amplitude are derived and Runge-Kutta method is adopted for numerical simulation. A typical 8 nm GaAs/Al(0.3)Ga(0.7) As single QW at 300 K is used. Additional information is contained in the original extended abstract.

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

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.

Photon path distribution and optical responses of turbid media: theoretical analysis based on the microscopic Beer-Lambert law.

PubMed

Tsuchiya, Y

2001-08-01

A concise theoretical treatment has been developed to describe the optical responses of a highly scattering inhomogeneous medium using functions of the photon path distribution (PPD). The treatment is based on the microscopic Beer-Lambert law and has been found to yield a complete set of optical responses by time- and frequency-domain measurements. The PPD is defined for possible photons having a total zigzag pathlength of l between the points of light input and detection. Such a distribution is independent of the absorption properties of the medium and can be uniquely determined for the medium under quantification. Therefore, the PPD can be calculated with an imaginary reference medium having the same optical properties as the medium under quantification except for the absence of absorption. One of the advantages of this method is that the optical responses, the total attenuation, the mean pathlength, etc are expressed by functions of the PPD and the absorption distribution.

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.

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.

Microscope sterility during spine surgery.

PubMed

Bible, Jesse E; O'Neill, Kevin R; Crosby, Colin G; Schoenecker, Jonathan G; McGirt, Matthew J; Devin, Clinton J

2012-04-01

Prospective study. Assess the contamination rates of sterile microscope drapes after spine surgery. The use of the operating microscope has become more prevalent in certain spine procedures, providing superior magnification, visualization, and illumination of the operative field. However, it may represent an additional source of bacterial contamination and increase the risk of developing a postoperative infection. This study included 25 surgical spine cases performed by a single spine surgeon that required the use of the operative microscope. Sterile culture swabs were used to obtain samples from 7 defined locations on the microscope drape after its use during the operation. The undraped technician's console was sampled in each case as a positive control, and an additional 25 microscope drapes were swabbed immediately after they were applied to the microscope to obtain negative controls. Swab samples were assessed for bacterial growth on 5% sheep blood Columbia agar plates using a semiquantitative technique. No growth was observed on any of the 25 negative control drapes. In contrast, 100% of preoperative and 96% of postoperative positive controls demonstrated obvious contamination. In the postoperative group, all 7 sites of evaluation were found to be contaminated with rates of 12% to 44%. Four of the 7 evaluated locations were found to have significant contamination rates compared with negative controls, including the shafts of the optic eyepieces on the main surgeon side (24%, P = 0.022), "forehead" portion on both the main surgeon (24%, P = 0.022) and assistant sides (28%, P = 0.010), and "overhead" portion of the drape (44%, P = 0.0002). Bacterial contamination of the operative microscope was found to be significant after spine surgery. Contamination was more common around the optic eyepieces, likely due to inadvertent touching of unsterile portions. Similarly, all regions above the eyepieces also have a propensity for contamination because of unknown contact with unsterile parts of the surgeon. Therefore, we believe that changing gloves after making adjustments to the optic eyepieces and avoid handling any portion of the drape above the eyepieces may decrease the risks of intraoperative contamination and possibly postoperative infection as well.

Ultra-high resolution water window x ray microscope optics design and analysis

NASA Technical Reports Server (NTRS)

Shealy, David L.; Wang, C.

1993-01-01

This project has been focused on the design and analysis of an ultra-high resolution water window soft-x-ray microscope. These activities have been accomplished by completing two tasks contained in the statement of work of this contract. The new results from this work confirm: (1) that in order to achieve resolutions greater than three times the wavelength of the incident radiation, it will be necessary to use spherical mirror surfaces and to use graded multilayer coatings on the secondary in order to accommodate the large variations of the angle of incidence over the secondary when operating the microscope at numerical apertures of 0.35 or greater; (2) that surface contour errors will have a significant effect on the optical performance of the microscope and must be controlled to a peak-to-valley variation of 50-100 A and a frequency of 8 periods over the surface of a mirror; and (3) that tolerance analysis of the spherical Schwarzschild microscope has been shown that the water window operations will require 2-3 times tighter tolerances to achieve a similar performance of operations with 130 A radiation. These results have been included in a manuscript included in the appendix.

Theoretical model of the helium zone plate microscope

NASA Astrophysics Data System (ADS)

Salvador Palau, Adrià; Bracco, Gianangelo; Holst, Bodil

2017-01-01

Neutral helium microscopy is a new technique currently under development. Its advantages are the low energy, charge neutrality, and inertness of the helium atoms, a potential large depth of field, and the fact that at thermal energies the helium atoms do not penetrate into any solid material. This opens the possibility, among others, for the creation of an instrument that can measure surface topology on the nanoscale, even on surfaces with high aspect ratios. One of the most promising designs for helium microscopy is the zone plate microscope. It consists of a supersonic expansion helium beam collimated by an aperture (skimmer) focused by a Fresnel zone plate onto a sample. The resolution is determined by the focal spot size, which depends on the size of the skimmer, the optics of the system, and the velocity spread of the beam through the chromatic aberrations of the zone plate. An important factor for the optics of the zone plate is the width of the outermost zone, corresponding to the smallest opening in the zone plate. The width of the outermost zone is fabrication limited to around 10 nm with present-day state-of-the-art technology. Due to the high ionization potential of neutral helium atoms, it is difficult to build efficient helium detectors. Therefore, it is crucial to optimize the microscope design to maximize the intensity for a given resolution and width of the outermost zone. Here we present an optimization model for the helium zone plate microscope. Assuming constant resolution and width of the outermost zone, we are able to reduce the problem to a two-variable problem (zone plate radius and object distance) and we show that for a given beam temperature and pressure, there is always a single intensity maximum. We compare our model with the highest-resolution zone plate focusing images published and show that the intensity can be increased seven times. Reducing the width of the outermost zone to 10 nm leads to an increase in intensity of more than 8000 times. Finally, we show that with present-day state-of-the-art detector technology (ionization efficiency 1 ×10-3 ), a resolution of the order of 10 nm is possible. In order to make this quantification, we have assumed a Lambertian reflecting surface and calculated the beam spot size that gives a signal 100 cts/s within a solid angle of 0.02 sr, following an existing helium microscope design.

When the fl# Is Not the fl#.

ERIC Educational Resources Information Center

Biermann, Mark L.; Biermann, Lois A. A.

1996-01-01

Discusses descriptions of the way in which an optical system controls the quantity of light that reaches a point on the image plane, a basic feature of optical imaging systems such as cameras, telescopes, and microscopes. (JRH)

Field of view advantage of conjugate adaptive optics in microscopy applications

PubMed Central

Mertz, Jerome; Paudel, Hari; Bifano, Thomas G.

2015-01-01

The imaging performance of an optical microscope can be degraded by sample-induced aberrations. A general strategy to undo the effect of these aberrations is to apply wavefront correction with a deformable mirror (DM). In most cases the DM is placed conjugate to the microscope pupil, called pupil adaptive optics (AO). When the aberrations are spatially variant an alternative configuration involves placing the DM conjugate to the main source of aberrations, called conjugate AO. We provide a theoretical and experimental comparison of both configurations for the simplified case where spatially variant aberrations are produced by a well defined phase screen. We pay particular attention to the resulting correction field of view (FOV). Conjugate AO is found to provide a significant FOV advantage. While this result is well known in the astronomy community, our goal here is to recast it specifically for the optical microscopy community. PMID:25967343

Vapor-condensation-assisted optical microscopy for ultralong carbon nanotubes and other nanostructures.

PubMed

Wang, Jiangtao; Li, Tianyi; Xia, Bingyu; Jin, Xiang; Wei, Haoming; Wu, Wenyun; Wei, Yang; Wang, Jiaping; Liu, Peng; Zhang, Lina; Li, Qunqing; Fan, Shoushan; Jiang, Kaili

2014-06-11

Here we present a simple yet powerful approach for the imaging of nanostructures under an optical microscope with the help of vapor condensation on their surfaces. Supersaturated water vapor will first form a nanometer-sized water droplet on the condensation nuclei on the surface of nanostructures, and then the water droplet will grow bigger and scatter more light to make the outline of the nanostructure be visible under dark-field optical microscope. This vapor-condensation-assisted (VCA) optical microscopy is applicable to a variety of nanostructures from ultralong carbon nanotubes to functional groups, generating images with contrast coming from the difference in density of the condensation sites, and does not induce any impurities to the specimens. Moreover, this low-cost and efficient technique can be conveniently integrated with other facilities, such as Raman spectroscope and so forth, which will pave the way for widespread applications.

Fabrication of bright and thin Zn₂SiO₄ luminescent film for electron beam excitation-assisted optical microscope.

PubMed

Furukawa, Taichi; Kanamori, Satoshi; Fukuta, Masahiro; Nawa, Yasunori; Kominami, Hiroko; Nakanishi, Yoichiro; Sugita, Atsushi; Inami, Wataru; Kawata, Yoshimasa

2015-07-13

We fabricated a bright and thin Zn₂SiO₄ luminescent film to serve as a nanometric light source for high-spatial-resolution optical microscopy based on electron beam excitation. The Zn₂SiO₄ luminescent thin film was fabricated by annealing a ZnO film on a Si₃N₄ substrate at 1000 °C in N₂. The annealed film emitted bright cathodoluminescence compared with the as-deposited film. The film is promising for nano-imaging with electron beam excitation-assisted optical microscopy. We evaluated the spatial resolution of a microscope developed using this Zn₂SiO₄ luminescent thin film. This is the first report of the investigation and application of ZnO/Si₃N₄ annealed at a high temperature (1000 °C). The fabricated Zn₂SiO₄ film is expected to enable high-frame-rate dynamic observation with ultra-high resolution using our electron beam excitation-assisted optical microscopy.

Generation and spectroscopic signatures of a fractional quantum Hall liquid of photons in an incoherently pumped optical cavity

NASA Astrophysics Data System (ADS)

Umucalılar, R. O.; Carusotto, I.

2017-11-01

We investigate theoretically a driven dissipative model of strongly interacting photons in a nonlinear optical cavity in the presence of a synthetic magnetic field. We show the possibility of using a frequency-dependent incoherent pump to create a strongly correlated ν =1 /2 bosonic Laughlin state of light: Due to the incompressibility of the Laughlin state, fluctuations in the total particle number and excitation of edge modes can be tamed by imposing a suitable external potential profile for photons. We further propose angular-momentum-selective spectroscopy of the emitted light as a tool to obtain unambiguous signatures of the microscopic physics of the quantum Hall liquid of light.

Detection of vesicant-induced upper airway mucosa damage in the hamster cheek pouch model using optical coherence tomography.

PubMed

Hammer-Wilson, Marie J; Nguyen, Vi; Jung, Woong-Gyu; Ahn, Yehchen; Chen, Zhongping; Wilder-Smith, Petra

2010-01-01

Hamster cheek pouches were exposed to 2-chloroethyl ethyl sulfide [CEES, half-mustard gas (HMG)] at a concentration of 0.4, 2.0, or 5.0 mg/ml for 1 or 5 min. Twenty-four hours post-HMG exposure, tissue damage was assessed by both stereomicrography and optical coherence tomography (OCT). Damage that was not visible on gross visual examination was apparent in the OCT images. Tissue changes were found to be dependent on both HMG concentration and exposure time. The submucosal and muscle layers of the cheek pouch tissue showed the greatest amount of structural alteration. Routine light microscope histology was performed to confirm the OCT observations.

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

PubMed

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

2015-09-01

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

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.

Highest Resolution Image of Dust and Sand Yet Acquired on Mars

NASA Technical Reports Server (NTRS)

2008-01-01

[figure removed for brevity, see original site] [figure removed for brevity, see original site] [figure removed for brevity, see original site] Click on image for Figure 1Click on image for Figure 2Click on image for Figure 3

This mosaic of four side-by-side microscope images (one a color composite) was acquired by the Optical Microscope, a part of the Microscopy, Electrochemistry, and Conductivity Analyzer (MECA) instrument suite on NASA's Phoenix Mars Lander. Taken on the ninth Martian day of the mission, or Sol 9 (June 3, 2008), the image shows a 3 millimeter (0.12 inch) diameter silicone target after it has been exposed to dust kicked up by the landing. It is the highest resolution image of dust and sand ever acquired on Mars. The silicone substrate provides a sticky surface for holding the particles to be examined by the microscope.

Martian Particles on Microscope's Silicone Substrate In figure 1, the particles are on a silcone substrate target 3 millimeters (0.12 inch) in diameter, which provides a sticky surface for holding the particles while the microscope images them. Blow-ups of four of the larger particles are shown in the center. These particles range in size from about 30 microns to 150 microns (from about one one-thousandth of an inch to six one-thousandths of an inch).

Possible Nature of Particles Viewed by Mars Lander's Optical Microscope In figure 2, the color composite on the right was acquired to examine dust that had fallen onto an exposed surface. The translucent particle highlighted at bottom center is of comparable size to white particles in a Martian soil sample (upper pictures) seen two sols earlier inside the scoop of Phoenix's Robotic Arm as imaged by the lander's Robotic Arm Camera. The white particles may be examples of the abundant salts that have been found in the Martian soil by previous missions. Further investigations will be needed to determine the white material's composition and whether translucent particles like the one in this microscopic image are found in Martian soil samples.

Scale of Phoenix Optical Microscope Images This set of pictures in figure 3 gives context for the size of individual images from the Optical Microscope on NASA's Mars Phoenix Lander.

The picture in the upper left was taken on Mars by the Surface Stereo Imager on Phoenix. It shows a portion of the microscope's sample stage exposed to accept a sample. In this case, the sample was of dust kicked up by the spacecraft thrusters during landers. Later samples will include soil delivered by the Robotic Arm.

The other pictures were taken on Earth. They show close-ups of circular substrates on which the microscopic samples rest when the microscope images them. Each circular substrate target is 3 millimeters (about one-tenth of an inch) in diameter. Each image taken by the microscope covers and area 2 millimeters by 1 millimeter (0.08 inch by 0.04 inch), the size of a large grain of sand.

The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

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.

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.

Miniature fiber optic spectrometer-based quantitative fluorescence resonance energy transfer measurement in single living cells.

PubMed

Chai, Liuying; Zhang, Jianwei; Zhang, Lili; Chen, Tongsheng

2015-03-01

Spectral measurement of fluorescence resonance energy transfer (FRET), spFRET, is a widely used FRET quantification method in living cells today. We set up a spectrometer-microscope platform that consists of a miniature fiber optic spectrometer and a widefield fluorescence microscope for the spectral measurement of absolute FRET efficiency (E) and acceptor-to-donor concentration ratio (R(C)) in single living cells. The microscope was used for guiding cells and the spectra were simultaneously detected by the miniature fiber optic spectrometer. Moreover, our platform has independent excitation and emission controllers, so different excitations can share the same emission channel. In addition, we developed a modified spectral FRET quantification method (mlux-FRET) for the multiple donors and multiple acceptors FRET construct (mD∼nA) sample, and we also developed a spectra-based 2-channel acceptor-sensitized FRET quantification method (spE-FRET). We implemented these modified FRET quantification methods on our platform to measure the absolute E and R(C) values of tandem constructs with different acceptor/donor stoichiometries in single living Huh-7 cells.

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

Miranda, Adelaide; De Beule, Pieter A. A., E-mail: pieter.de-beule@inl.int; Martins, Marco

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

Chronic monitoring of cortical hemodynamics in behaving, freely-moving rats using a miniaturized head-mounted optical microscope

NASA Astrophysics Data System (ADS)

Sigal, Iliya; Gad, Raanan; Koletar, Margaret; Ringuette, Dene; Stefanovic, Bojana; Levi, Ofer

2016-03-01

Growing interest within the neurophysiology community in assessing healthy and pathological brain activity in animals that are awake and freely-behaving has triggered the need for optical systems that are suitable for such longitudinal studies. In this work we report label-free multi-modal imaging of cortical hemodynamics in the somatosensory cortex of awake, freely-behaving rats, using a novel head-mounted miniature optical microscope. The microscope employs vertical cavity surface emitting lasers (VCSELs) at three distinct wavelengths (680 nm, 795 nm, and 850 nm) to provide measurements of four hemodynamic markers: blood flow speeds, HbO, HbR, and total Hb concentration, across a > 2 mm field of view. Blood flow speeds are extracted using Laser Speckle Contrast Imaging (LSCI), while oxygenation measurements are performed using Intrinsic Optical Signal Imaging (IOSI). Longitudinal measurements on the same animal are made possible over the course of > 6 weeks using a chronic window that is surgically implanted into the skull. We use the device to examine changes in blood flow and blood oxygenation in superficial cortical blood vessels and tissue in response to drug-induced absence-like seizures, correlating motor behavior with changes in blood flow and blood oxygenation in the brain.

Quantitative high dynamic range beam profiling for fluorescence microscopy

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

Mitchell, T. J., E-mail: t.j.mitchell@dur.ac.uk; Saunter, C. D.; O’Nions, W.

2014-10-15

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

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

NASA Astrophysics Data System (ADS)

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

2016-03-01

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

Real-time restoration of white-light confocal microscope optical sections

PubMed Central

Balasubramanian, Madhusudhanan; Iyengar, S. Sitharama; Beuerman, Roger W.; Reynaud, Juan; Wolenski, Peter

2009-01-01

Confocal microscopes (CM) are routinely used for building 3-D images of microscopic structures. Nonideal imaging conditions in a white-light CM introduce additive noise and blur. The optical section images need to be restored prior to quantitative analysis. We present an adaptive noise filtering technique using Karhunen–Loéve expansion (KLE) by the method of snapshots and a ringing metric to quantify the ringing artifacts introduced in the images restored at various iterations of iterative Lucy–Richardson deconvolution algorithm. The KLE provides a set of basis functions that comprise the optimal linear basis for an ensemble of empirical observations. We show that most of the noise in the scene can be removed by reconstructing the images using the KLE basis vector with the largest eigenvalue. The prefiltering scheme presented is faster and does not require prior knowledge about image noise. Optical sections processed using the KLE prefilter can be restored using a simple inverse restoration algorithm; thus, the methodology is suitable for real-time image restoration applications. The KLE image prefilter outperforms the temporal-average prefilter in restoring CM optical sections. The ringing metric developed uses simple binary morphological operations to quantify the ringing artifacts and confirms with the visual observation of ringing artifacts in the restored images. PMID:20186290

Maximizing fluorescence collection efficiency in multiphoton microscopy

PubMed Central

Zinter, Joseph P.; Levene, Michael J.

2011-01-01

Understanding fluorescence propagation through a multiphoton microscope is of critical importance in designing high performance systems capable of deep tissue imaging. Optical models of a scattering tissue sample and the Olympus 20X 0.95NA microscope objective were used to simulate fluorescence propagation as a function of imaging depth for physiologically relevant scattering parameters. The spatio-angular distribution of fluorescence at the objective back aperture derived from these simulations was used to design a simple, maximally efficient post-objective fluorescence collection system. Monte Carlo simulations corroborated by data from experimental tissue phantoms demonstrate collection efficiency improvements of 50% – 90% over conventional, non-optimized fluorescence collection geometries at large imaging depths. Imaging performance was verified by imaging layer V neurons in mouse cortex to a depth of 850 μm. PMID:21934897

Multiscale Imaging of the Mouse Cortex Using Two-Photon Microscopy and Wide-Field Illumination

NASA Astrophysics Data System (ADS)

Bumstead, Jonathan R.

The mouse brain can be studied over vast spatial scales ranging from microscopic imaging of single neurons to macroscopic measurements of hemodynamics acquired over the majority of the mouse cortex. However, most neuroimaging modalities are limited by a fundamental trade-off between the spatial resolution and the field-of-view (FOV) over which the brain can be imaged, making it difficult to fully understand the functional and structural architecture of the healthy mouse brain and its disruption in disease. My dissertation has focused on developing multiscale optical systems capable of imaging the mouse brain at both microscopic and mesoscopic spatial scales, specifically addressing the difference in spatial scales imaged with two-photon microscopy (TPM) and optical intrinsic signal imaging (OISI). Central to this work has been the formulation of a principled design strategy for extending the FOV of the two-photon microscope. Using this design approach, we constructed a TPM system with subcellular resolution and a FOV area 100 times greater than a conventional two-photon microscope. To image the ellipsoidal shape of the mouse cortex, we also developed the microscope to image arbitrary surfaces within a single frame using an electrically tunable lens. Finally, to address the speed limitations of the TPM systems developed during my dissertation, I also conducted research in large-scale neural phenomena occurring in the mouse brain imaged with high-speed OISI. The work conducted during my dissertation addresses some of the fundamental principles in designing and applying optical systems for multiscale imaging of the mouse brain.

Images from Phoenix's MECA Instruments

NASA Technical Reports Server (NTRS)

2008-01-01

The image on the upper left is from NASA's Phoenix Mars Lander's Optical Microscope after a sample informally called 'Sorceress' was delivered to its silicon substrate on the 38th Martian day, or sol, of the mission (July 2, 2008).

A 3D representation of the same sample is on the right, as seen by Phoenix's Atomic Force Microscope. This is 200 times greater magnification than the view from the Optical Microscope, and the most highly magnified image ever seen from another world.

The image shows four round pits, only 5 microns in depth, that were micromachined into the silicon substrate, which is the background plane shown in red. This image has been processed to reflect the levelness of the substrate.

A Martian particle only one micrometer, or one millionth of a meter, across is held in the upper left pit.

The rounded particle shown at the highest magnification ever seen from another world is a particle of the dust that cloaks Mars. Such dust particles color the Martian sky pink, feed storms that regularly envelop the planet and produce Mars' distinctive red soil.

The Optical Microscope and the Atomic Force Microscope are part of Phoenix's Microscopy, Electrochemistry and Conductivity Analyzer instrument.

The AFM was developed by a Swiss-led consortium, with Imperial College London producing the silicon substrate that holds sampled particles.

The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

Making a Microscope with Readily Available Materials

ERIC Educational Resources Information Center

Vannoni, Maurizio; Buah-Bassuah, Paul K.; Molesini, Giuseppe

2007-01-01

The making of microscope devices using inexpensive or recovered materials is demonstrated. Examples of images illustrating the performance of such devices are presented. As a project at the undergraduate level, the task is effective in acquiring familiarity with optical imaging concepts and their practical implementation in the laboratory.…

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.

Elastic scattering and breakup reactions of the exotic nucleus 8B on nuclear targets

NASA Astrophysics Data System (ADS)

Lukyanov, V. K.; Kadrev, D. N.; Antonov, A. N.; Zemlyanaya, E. V.; Lukyanov, K. V.; Gaidarov, M. K.; Spasova, K.

2018-05-01

Microscopic calculations of the optical potentials (OPs) and elastic scattering cross sections of the proton-rich nucleus 8B on 12C, 58Ni and 208Pb targets are presented. The density distributions of 8B obtained within the variational Monte Carlo (VMC) model and the three-cluster model (3CM) are used to construct the optical potentials (OP). The real part of the hybrid OP (ReOP) is calculated using the folding model with the direct and exchange terms included, while the imaginary part (ImOP) is obtained on the base of the high energy approximation (HEA). In addition, the cluster model, in which 8B consists of a proton halo and a 7Be core is applied to calculate the breakup cross sections of 8B on 9Be, 12C and 197Au targets, as well as the momentum distributions of 7Be fragments. A comparison with the available experimental data is made and a good agreement is obtained.

A laboratory 8 keV transmission full-field x-ray microscope with a polycapillary as condenser for bright and dark field imaging

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

Baumbach, S., E-mail: baumbach@rheinahrcampus.de; Wilhein, T.; Kanngießer, B.

2015-08-15

This article introduces a laboratory setup of a transmission full-field x-ray microscope at 8 keV photon energy. The microscope operates in bright and dark field imaging mode with a maximum field of view of 50 μm. Since the illumination geometry determines whether the sample is illuminated homogeneously and moreover, if different imaging methods can be applied, the condenser optic is one of the most significant parts. With a new type of x-ray condenser, a polycapillary optic, we realized bright field imaging and for the first time dark field imaging at 8 keV photon energy in a laboratory setup. A detectormore » limited spatial resolution of 210 nm is measured on x-ray images of Siemens star test patterns.« less

Fabrication and characterization of novel microsphere-embedded optical devices for enhancing microscopy resolution

NASA Astrophysics Data System (ADS)

Darafsheh, Arash

2018-02-01

Microsphere-assisted imaging can be incorporated onto conventional light microscopes allowing wide-field and flourescence imaging with enhanced resolution. We demonstrated that imaging of specimens containing subdiffraction-limited features is achievable through high-index microspheres embedded in a transparent thin film placed over the specimen. We fabricated novel microsphere-embedded microscope slides composed of barium titanate glass microspheres (with diameter 10-100 μm and refractive index 1.9-2.2) embedded in a transparent polydimethylsiloxane (PDMS) elastomer layer with controllable thickness. We characterized the imaging performance of such microsphere-embedded devices in white-light microscopies, by measuring the imaging resolution, field-of-view, and magnification as a function of microsphere size. Our results inform on the design of novel optical devices, such as microsphere-embedded microscope slides for imaging applications.

Optomechanical design and tolerance of a microscope objective at 121.6 nm

NASA Astrophysics Data System (ADS)

Keyes, Derek S.; Jota, Thiago S.; Gao, Weichuan; Luepke, Dakota; Densmore, Victor; Kim, Young-Sik; Kim, Gun-Hee; Milster, Thomas D.

2015-08-01

By utilizing the Hydrogen-Lyman-α (HLA) source at 121.6 nm, we hope to achieve an intrinsic resolution of 247 nm at 0.3 numerical aperture (NA) and 92 nm at 0.8 NA. The motivation for 121.6 nm microscopy is the existence of a transparent window in the air absorption spectrum at that wavelength, which allows for the sample to be in air while the microscope is in an enclosed nitrogen environment. The microscope objective consists of two reflective optics and a LiF window, and it has been designed to demonstrate diffraction limited performance over a 160μm full field at 121.6 nm. The optomechanical design consists of mechanical subcells for each optical component, precision spacers and a barrel bore, which allow for submicron control of tolerance parameters.

A laboratory 8 keV transmission full-field x-ray microscope with a polycapillary as condenser for bright and dark field imaging.

PubMed

Baumbach, S; Kanngießer, B; Malzer, W; Stiel, H; Wilhein, T

2015-08-01

This article introduces a laboratory setup of a transmission full-field x-ray microscope at 8 keV photon energy. The microscope operates in bright and dark field imaging mode with a maximum field of view of 50 μm. Since the illumination geometry determines whether the sample is illuminated homogeneously and moreover, if different imaging methods can be applied, the condenser optic is one of the most significant parts. With a new type of x-ray condenser, a polycapillary optic, we realized bright field imaging and for the first time dark field imaging at 8 keV photon energy in a laboratory setup. A detector limited spatial resolution of 210 nm is measured on x-ray images of Siemens star test patterns.

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.

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.

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.

Foucault imaging by using non-dedicated transmission electron microscope

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

Taniguchi, Yoshifumi; Matsumoto, Hiroaki; Harada, Ken

2012-08-27

An electron optical system for observing Foucault images was constructed using a conventional transmission electron microscope without any special equipment for Lorentz microscopy. The objective lens was switched off and an electron beam was converged by a condenser optical system to the crossover on the selected area aperture plane. The selected area aperture was used as an objective aperture to select the deflected beam for Foucault mode, and the successive image-forming lenses were controlled for observation of the specimen images. The irradiation area on the specimen was controlled by selecting the appropriate diameter of the condenser aperture.

Modulus design multiwavelength polarization microscope for transmission Mueller matrix imaging.

PubMed

Zhou, Jialing; He, Honghui; Chen, Zhenhua; Wang, Ye; Ma, Hui

2018-01-01

We have developed a polarization microscope based on a commercial transmission microscope. We replace the halogen light source by a collimated LED light source module of six different colors. We use achromatic polarized optical elements that can cover the six different wavelength ranges in the polarization state generator (PSG) and polarization state analyzer (PSA) modules. The dual-rotating wave plate method is used to measure the Mueller matrix of samples, which requires the simultaneous rotation of the two quarter-wave plates in both PSG and PSA at certain angular steps. A scientific CCD detector is used as the image receiving module. A LabView-based software is developed to control the rotation angels of the wave plates and the exposure time of the detector to allow the system to run fully automatically in preprogrammed schedules. Standard samples, such as air, polarizers, and quarter-wave plates, are used to calibrate the intrinsic Mueller matrix of optical components, such as the objectives, using the eigenvalue calibration method. Errors due to the images walk-off in the PSA are studied. Errors in the Mueller matrices are below 0.01 using air and polarizer as standard samples. Data analysis based on Mueller matrix transformation and Mueller matrix polarization decomposition is used to demonstrate the potential application of this microscope in pathological diagnosis. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

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

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

Gofron, K. J., E-mail: kgofron@bnl.gov; Cai, Y. Q.; Coburn, D. S.

A novel on-axis X-ray microscope with 3 µm resolution, 3x magnification, and a working distance of 600 mm for in-situ sample alignment and X-ray beam visualization for the Inelastic X-ray Scattering (IXS) beamline at NSLS-II is presented. The microscope uses reflective optics, which minimizes dispersion, and allows imaging from Ultraviolet (UV) to Infrared (IR) with specifically chosen objective components (coatings, etc.). Additionally, a portable high resolution X-ray microscope for KB mirror alignment and X-ray beam characterization was developed.

The microscopes of Antoni van Leeuwenhoek.

PubMed

van Zuylen, J

1981-03-01

The seventeenth-century Dutch microscopist, Antoni van Leeuwenhoek, was the first man to make a protracted study of microscopical objects, and, unlike his contemporary Robert Hooke, he viewed by transmitted light. Leeuwenhoek made over 500 of his own, curious, simple microscopes, but now only nine are known to exist. The exact nature of the lenses Leeuwenhoek made, has for long been a puzzle. The existing microscopes have now been examined in detail, and their optical characteristics measured and tabulated. It is proposed that the lens of highest magnification, x 266, was made using a special blown bubble technique.

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.

Electron tomography simulator with realistic 3D phantom for evaluation of acquisition, alignment and reconstruction methods.

PubMed

Wan, Xiaohua; Katchalski, Tsvi; Churas, Christopher; Ghosh, Sreya; Phan, Sebastien; Lawrence, Albert; Hao, Yu; Zhou, Ziying; Chen, Ruijuan; Chen, Yu; Zhang, Fa; Ellisman, Mark H

2017-05-01

Because of the significance of electron microscope tomography in the investigation of biological structure at nanometer scales, ongoing improvement efforts have been continuous over recent years. This is particularly true in the case of software developments. Nevertheless, verification of improvements delivered by new algorithms and software remains difficult. Current analysis tools do not provide adaptable and consistent methods for quality assessment. This is particularly true with images of biological samples, due to image complexity, variability, low contrast and noise. We report an electron tomography (ET) simulator with accurate ray optics modeling of image formation that includes curvilinear trajectories through the sample, warping of the sample and noise. As a demonstration of the utility of our approach, we have concentrated on providing verification of the class of reconstruction methods applicable to wide field images of stained plastic-embedded samples. Accordingly, we have also constructed digital phantoms derived from serial block face scanning electron microscope images. These phantoms are also easily modified to include alignment features to test alignment algorithms. The combination of more realistic phantoms with more faithful simulations facilitates objective comparison of acquisition parameters, alignment and reconstruction algorithms and their range of applicability. With proper phantoms, this approach can also be modified to include more complex optical models, including distance-dependent blurring and phase contrast functions, such as may occur in cryotomography. Copyright © 2017 Elsevier Inc. All rights reserved.

X-ray microbeam stand-alone facility for cultured cells irradiation

NASA Astrophysics Data System (ADS)

Bożek, Sebastian; Bielecki, Jakub; Wiecheć, Anna; Lekki, Janusz; Stachura, Zbigniew; Pogoda, Katarzyna; Lipiec, Ewelina; Tkocz, Konrad; Kwiatek, Wojciech M.

2017-03-01

The article describes an X-ray microbeam standalone facility dedicated for irradiation of living cultured cells. The article can serve as an advice for such facilities construction, as it begins from engineering details, through mathematical modeling and experimental procedures, ending up with preliminary experimental results and conclusions. The presented system consists of an open type X-ray tube with microfocusing down to about 2 μm, an X-ray focusing system with optical elements arranged in the nested Kirckpatrick-Baez (or Montel) geometry, a sample stand and an optical microscope with a scientific digital CCD camera. For the beam visualisation an X-ray sensitive CCD camera and a spectral detector are used, as well as a scintillator screen combined with the microscope. A method of precise one by one irradiation of previously chosen cells is presented, as well as a fast method of uniform irradiation of a chosen sample area. Mathematical models of beam and cell with calculations of kerma and dose are presented. The experiments on dose-effect relationship, kinetics of DNA double strand breaks repair, as well as micronuclei observation were performed on PC-3 (Prostate Cancer) cultured cells. The cells were seeded and irradiated on Mylar foil, which covered a hole drilled in the Petri dish. DNA lesions were visualised with γ-H2AX marker combined with Alexa Fluor 488 fluorescent dye.

[Effect of electroacupuncture on cellular structure of hippocampus in splenic asthenia pedo-rats].

PubMed

Yang, Zhuo-xin; Zhuo, Yuan-yuan; Yu, Hai-bo; Wang, Ning

2010-02-01

To observe the effect of electroacupuncture (EA) on hippocampal structure in splenic asthenia pedo-rats. A total of 15 SD male rats were randomly assigned to normal control group (n=5), model group (n=5) and EA group (n=5). Splenic asthenic syndrome model was established by intragastric administration of rhubarb and intraperitoneal injection of Reserpine for 14 d. EA (1 mA, 3 Hz/iS Hz) was applied to bilateral "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) for 20 mm, once a day for 14 days. The cellular structure of hippocampus was observed by light microscope and transmission electron microscope. Optical microscopic observation showed that in normal control group, the cellular nucleus was distinct, and the granular cell layer well-arranged and tight. In model group, the intracellular space was widened, and the granular cell layer was out of order in the arrangement. In EA group, the celluldr nucleus and the granular cell layer were nearly normal. Results of the electronic microscope showed that cells in model group had a karyopyknosis with irregular appearance and clear incisure, and some of them presented dissolving and necrotic phenomena; and those in EA group were milder in injury, had nearly-normal nucleus with visible nucleoli and relatively-intact nuclear membrane. Regarding the cellular plasma, in comparison with rich normal organelles of control group, the mitochondria in model group were swelling, with vague, dissolved and broken cristae, while in EA group, majority of the organelles were well-kept, and slightly dissolved mitochondrial cristae found. In regard to the synaptic structure, in comparison with control group, synaptic apomorphosis and swelling mitochondria were found in model group While in EA group, milder swelling and hydropic degeneration were seen. Different from the distinct pre- and post-synaptic membrane and synaptic vesicles of control group, while those in EA group were nearly-normal. electroacupunture can effectively relieve splenasthenic syndrome induced pathohistological changes of neurons of the hippocampus in the rat.

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.

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.

Microscopic morphology evolution during ion beam smoothing of Zerodur® surfaces.

PubMed

Liao, Wenlin; Dai, Yifan; Xie, Xuhui; Zhou, Lin

2014-01-13

Ion sputtering of Zerodur material often results in the formation of nanoscale microstructures on the surfaces, which seriously influences optical surface quality. In this paper, we describe the microscopic morphology evolution during ion sputtering of Zerodur surfaces through experimental researches and theoretical analysis, which shows that preferential sputtering together with curvature-dependent sputtering overcomes ion-induced smoothing mechanisms leading to granular nanopatterns formation in morphology and the coarsening of the surface. Consequently, we propose a new method for ion beam smoothing (IBS) of Zerodur optics assisted by deterministic ion beam material adding (IBA) technology. With this method, Zerodur optics with surface roughness down to 0.15 nm root mean square (RMS) level is obtained through the experimental investigation, which demonstrates the feasibility of our proposed method.

Modeling and Observations of Phase-Mask Trapezoidal Profiles with Grating-Fiber Image Reproduction

NASA Technical Reports Server (NTRS)

Lyons, Donald R.; Lindesay, James V.; Lee, Hyung R.; Ndlela, Zolili U.; Thompso, Erica J.

2000-01-01

We report on an investigation of the trapezoidal design and fabrication defects in phase masks used to produce Bragg reflection gratings in optical fibers. We used a direct visualization technique to examine the nonuniformity of the interference patterns generated by several phase masks. Fringe patterns from the phase masks are compared with the analogous patterns resulting from two-beam interference. Atomic force microscope imaging of the actual phase gratings that give rise to anomalous fringe patterns is used to determine input parameters for a general theoretical model. Phase masks with pitches of 0.566 and 1.059 microns are modeled and investigated.

Grid Gap Measurement for an NSTAR Ion Thruster

NASA Technical Reports Server (NTRS)

Diaz, Esther M.; Soulas, George C.

2006-01-01

The change in gap between the screen and accelerator grids of an engineering model NSTAR ion optics assembly was measured during thruster operation with beam extraction. The molybdenum ion optics assembly was mounted onto an engineering model NSTAR ion thruster. The measurement technique consisted of measuring the difference in height of an alumina pin relative to the downstream accelerator grid surface. The alumina pin was mechanically attached to the center aperture of the screen grid and protruded through the center aperture of the accelerator grid. The change in pin height was monitored using a long distance microscope coupled to a digital imaging system. Transient and steady-state hot grid gaps were measured at three power levels: 0.5, 1.5 and 2.3 kW. Also, the change in grid gap was measured during the transition between power levels, and during the startup with high voltage applied just prior to discharge ignition. Performance measurements, such as perveance, electron backstreaming limit and screen grid ion transparency, were also made to confirm that this ion optics assembly performed similarly to past testing. Results are compared to a prior test of 30 cm titanium ion optics.

Developing a trend prediction model of subsurface damage for fixed-abrasive grinding of optics by cup wheels.

PubMed

Dong, Zhichao; Cheng, Haobo

2016-11-10

Fixed-abrasive grinding by cup wheels plays an important role in the production of precision optics. During cup wheel grinding, we strive for a large removal rate while maintaining fine integrity on the surface and subsurface layers (academically recognized as surface roughness and subsurface damage, respectively). This study develops a theoretical model used to predict the trend of subsurface damage of optics (with respect to various grinding parameters) in fixed-abrasive grinding by cup wheels. It is derived from the maximum undeformed chip thickness model, and it successfully correlates the pivotal parameters of cup wheel grinding with the subsurface damage depth. The efficiency of this model is then demonstrated by a set of experiments performed on a cup wheel grinding machine. In these experiments, the characteristics of subsurface damage are inspected by a wedge-polishing plus microscopic inspection method, revealing that the subsurface damage induced in cup wheel grinding is composed of craterlike morphologies and slender cracks, with depth ranging from ∼6.2 to ∼13.2  μm under the specified grinding parameters. With the help of the proposed model, an optimized grinding strategy is suggested for realizing fine subsurface integrity as well as high removal rate, which can alleviate the workload of subsequent lapping and polishing.

EDITORIAL: Recent developments in biomedical optics

NASA Astrophysics Data System (ADS)

Wang, Ruikang K.; Hebden, Jeremy C.; Tuchin, Valery V.

2004-04-01

The rapid growth in laser and photonic technology has resulted in new tools being proposed and developed for use in the medical and biological sciences. Specifically, a discipline known as biomedical optics has emerged which is providing a broad variety of optical techniques and instruments for diagnostic, therapeutic and basic science applications. New laser sources, detectors and measurement techniques are yielding powerful new methods for the study of diseases on all scales, from single molecules, to specific tissues and whole organs. For example, novel laser microscopes permit spectroscopic and force measurements to be performed on single protein molecules; new optical devices provide information on molecular dynamics and structure to perform `optical biopsy' non-invasively and almost instantaneously; and optical coherence tomography and diffuse optical tomography allow visualization of specific tissues and organs. Using genetic promoters to derive luciferase expression, bioluminescence methods can generate molecular light switches, which serve as functional indicator lights reporting cellular conditions and responses in living animals. This technique could allow rapid assessment of and response to the effects of anti-tumour drugs, antibiotics, or antiviral drugs. This issue of Physics in Medicine and Biology highlights recent research in biomedical optics, and is based on invited contributions to the International Conference on Advanced Laser Technology (Focused on Biomedical Optics) held at Cranfield University at Silsoe on 19--23 September 2003. This meeting included sessions devoted to: diffuse optical imaging and spectroscopy; optical coherence tomography and coherent domain techniques; optical sensing and applications in life science; microscopic, spectroscopic and opto-acoustic imaging; therapeutic and diagnostic applications; and laser interaction with organic and inorganic materials. Twenty-one papers are included in this special issue. The first paper gives an overview on the current status of scanning laser ophthalmoscopy and its role in bioscience and medicine, while the second paper describes the current problems in tissue engineering and the potential role for optical coherence tomography. The following seven papers present and discuss latest developments in infrared spectroscopy and diffuse optical tomography for medical diagnostics. Eight further papers report recent advances in optical coherence tomography, covering new and evolving methods and instrumentation, theoretical and numerical modelling, and its clinical applications. The remaining papers cover miscellaneous topics in biomedical optics, including new developments in opto-acoustic imaging techniques, laser speckle imaging of blood flow in microcirculations, and potential of hollow-core photonic-crystal fibres for laser dentistry. We thank all the authors for their valuable contributions and their prompt responses to reviewers' comments. We are also very grateful to the reviewers for their hard work and their considerable efforts to meet tight deadlines.

Near-field microscopy with a microfabricated solid immersion lens

NASA Astrophysics Data System (ADS)

Fletcher, Daniel Alden

2001-07-01

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

Power spectrum analysis with least-squares fitting: amplitude bias and its elimination, with application to optical tweezers and atomic force microscope cantilevers.

PubMed

Nørrelykke, Simon F; Flyvbjerg, Henrik

2010-07-01

Optical tweezers and atomic force microscope (AFM) cantilevers are often calibrated by fitting their experimental power spectra of Brownian motion. We demonstrate here that if this is done with typical weighted least-squares methods, the result is a bias of relative size between -2/n and +1/n on the value of the fitted diffusion coefficient. Here, n is the number of power spectra averaged over, so typical calibrations contain 10%-20% bias. Both the sign and the size of the bias depend on the weighting scheme applied. Hence, so do length-scale calibrations based on the diffusion coefficient. The fitted value for the characteristic frequency is not affected by this bias. For the AFM then, force measurements are not affected provided an independent length-scale calibration is available. For optical tweezers there is no such luck, since the spring constant is found as the ratio of the characteristic frequency and the diffusion coefficient. We give analytical results for the weight-dependent bias for the wide class of systems whose dynamics is described by a linear (integro)differential equation with additive noise, white or colored. Examples are optical tweezers with hydrodynamic self-interaction and aliasing, calibration of Ornstein-Uhlenbeck models in finance, models for cell migration in biology, etc. Because the bias takes the form of a simple multiplicative factor on the fitted amplitude (e.g. the diffusion coefficient), it is straightforward to remove and the user will need minimal modifications to his or her favorite least-squares fitting programs. Results are demonstrated and illustrated using synthetic data, so we can compare fits with known true values. We also fit some commonly occurring power spectra once-and-for-all in the sense that we give their parameter values and associated error bars as explicit functions of experimental power-spectral values.

Camera, handlens, and microscope optical system for imaging and coupled optical spectroscopy

NASA Technical Reports Server (NTRS)

Mungas, Greg S. (Inventor); Boynton, John (Inventor); Sepulveda, Cesar A. (Inventor); Nunes de Sepulveda, legal representative, Alicia (Inventor); Gursel, Yekta (Inventor)

2012-01-01

An optical system comprising two lens cells, each lens cell comprising multiple lens elements, to provide imaging over a very wide image distance and within a wide range of magnification by changing the distance between the two lens cells. An embodiment also provides scannable laser spectroscopic measurements within the field-of-view of the instrument.

Camera, handlens, and microscope optical system for imaging and coupled optical spectroscopy

NASA Technical Reports Server (NTRS)

Mungas, Greg S. (Inventor); Boynton, John (Inventor); Sepulveda, Cesar A. (Inventor); Nunes de Sepulveda, Alicia (Inventor); Gursel, Yekta (Inventor)

2011-01-01

An optical system comprising two lens cells, each lens cell comprising multiple lens elements, to provide imaging over a very wide image distance and within a wide range of magnification by changing the distance between the two lens cells. An embodiment also provides scannable laser spectroscopic measurements within the field-of-view of the instrument.

Resolving the optical anisotropy of low-symmetry 2D materials.

PubMed

Shen, Wanfu; Hu, Chunguang; Tao, Jin; Liu, Jun; Fan, Shuangqing; Wei, Yaxu; An, Chunhua; Chen, Jiancui; Wu, Sen; Li, Yanning; Liu, Jing; Zhang, Daihua; Sun, Lidong; Hu, Xiaotang

2018-05-03

Optical anisotropy is one of the most fundamental physical characteristics of emerging low-symmetry two-dimensional (2D) materials. It provides abundant structural information and is crucial for creating diverse nanoscale devices. Here, we have proposed an azimuth-resolved microscopic approach to directly resolve the normalized optical difference along two orthogonal directions at normal incidence. The differential principle ensures that the approach is only sensitive to anisotropic samples and immune to isotropic materials. We studied the optical anisotropy of bare and encapsulated black phosphorus (BP) and unveiled the interference effect on optical anisotropy, which is critical for practical applications in optical and optoelectronic devices. A multi-phase model based on the scattering matrix method was developed to account for the interference effect and then the crystallographic directions were unambiguously determined. Our result also suggests that the optical anisotropy is a probe to measure the thickness with monolayer resolution. Furthermore, the optical anisotropy of rhenium disulfide (ReS2), another class of anisotropic 2D materials, with a 1T distorted crystal structure, was investigated, which demonstrates that our approach is suitable for other anisotropic 2D materials. This technique is ideal for optical anisotropy characterization and will inspire future efforts in BP and related anisotropic 2D nanomaterials for engineering new conceptual nanodevices.

Computational imaging through a fiber-optic bundle

NASA Astrophysics Data System (ADS)

Lodhi, Muhammad A.; Dumas, John Paul; Pierce, Mark C.; Bajwa, Waheed U.

2017-05-01

Compressive sensing (CS) has proven to be a viable method for reconstructing high-resolution signals using low-resolution measurements. Integrating CS principles into an optical system allows for higher-resolution imaging using lower-resolution sensor arrays. In contrast to prior works on CS-based imaging, our focus in this paper is on imaging through fiber-optic bundles, in which manufacturing constraints limit individual fiber spacing to around 2 μm. This limitation essentially renders fiber-optic bundles as low-resolution sensors with relatively few resolvable points per unit area. These fiber bundles are often used in minimally invasive medical instruments for viewing tissue at macro and microscopic levels. While the compact nature and flexibility of fiber bundles allow for excellent tissue access in-vivo, imaging through fiber bundles does not provide the fine details of tissue features that is demanded in some medical situations. Our hypothesis is that adapting existing CS principles to fiber bundle-based optical systems will overcome the resolution limitation inherent in fiber-bundle imaging. In a previous paper we examined the practical challenges involved in implementing a highly parallel version of the single-pixel camera while focusing on synthetic objects. This paper extends the same architecture for fiber-bundle imaging under incoherent illumination and addresses some practical issues associated with imaging physical objects. Additionally, we model the optical non-idealities in the system to get lower modelling errors.

Refractive-index profiling of embedded microstructures in optical materials

NASA Astrophysics Data System (ADS)

Dave, Digant P.; Milner, Thomas E.

2002-04-01

We describe use of a phase-sensitive low-coherence reflectometer to measure spatial variation of refractive index in optical materials. The described interferometric technique is demonstrated to be a valuable tool to profile the refractive index of optical elements such as integrated waveguides and photowritten optical microstructures. As an example, a refractive-index profile is mapped of a microstructure written in a microscope glass slide with an ultrashort-pulse laser.

All-optical materials design of chiral edge modes in transition-metal dichalcogenides

DOE PAGES

Claassen, Martin; Jia, Chunjing; Moritz, Brian; ...

2016-10-10

Monolayer transition-metal dichalcogenides are novel materials which at low energies constitute a condensed-matter realization of massive relativistic fermions in two dimensions. Here, we show that this picture breaks for optical pumping—instead, the added complexity of a realistic materials description leads to a new mechanism to optically induce topologically protected chiral edge modes, facilitating optically switchable conduction channels that are insensitive to disorder. In contrast to graphene and previously discussed toy models, the underlying mechanism relies on the intrinsic three-band nature of transition-metal dichalcogenide monolayers near the band edges. Photo-induced band inversions scale linearly in applied pump field and exhibit transitionsmore » from one to two chiral edge modes on sweeping from red to blue detuning. As a result, we develop an ab initio strategy to understand non-equilibrium Floquet–Bloch bands and topological transitions, and illustrate for WS 2 that control of chiral edge modes can be dictated solely from symmetry principles and is not qualitatively sensitive to microscopic materials details.« less

All-optical materials design of chiral edge modes in transition-metal dichalcogenides

PubMed Central

Claassen, Martin; Jia, Chunjing; Moritz, Brian; Devereaux, Thomas P.

2016-01-01

Monolayer transition-metal dichalcogenides are novel materials which at low energies constitute a condensed-matter realization of massive relativistic fermions in two dimensions. Here, we show that this picture breaks for optical pumping—instead, the added complexity of a realistic materials description leads to a new mechanism to optically induce topologically protected chiral edge modes, facilitating optically switchable conduction channels that are insensitive to disorder. In contrast to graphene and previously discussed toy models, the underlying mechanism relies on the intrinsic three-band nature of transition-metal dichalcogenide monolayers near the band edges. Photo-induced band inversions scale linearly in applied pump field and exhibit transitions from one to two chiral edge modes on sweeping from red to blue detuning. We develop an ab initio strategy to understand non-equilibrium Floquet–Bloch bands and topological transitions, and illustrate for WS2 that control of chiral edge modes can be dictated solely from symmetry principles and is not qualitatively sensitive to microscopic materials details. PMID:27721504

Scene-based Shack-Hartmann wavefront sensor for light-sheet microscopy

NASA Astrophysics Data System (ADS)

Lawrence, Keelan; Liu, Yang; Dale, Savannah; Ball, Rebecca; VanLeuven, Ariel J.; Sornborger, Andrew; Lauderdale, James D.; Kner, Peter

2018-02-01

Light-sheet microscopy is an ideal imaging modality for long-term live imaging in model organisms. However, significant optical aberrations can be present when imaging into an organism that is hundreds of microns or greater in size. To measure and correct optical aberrations, an adaptive optics system must be incorporated into the microscope. Many biological samples lack point sources that can be used as guide stars with conventional Shack-Hartmann wavefront sensors. We have developed a scene-based Shack-Hartmann wavefront sensor for measuring the optical aberrations in a light-sheet microscopy system that does not require a point-source and can measure the aberrations for different parts of the image. The sensor has 280 lenslets inside the pupil, creates an image from each lenslet with a 500 micron field of view and a resolution of 8 microns, and has a resolution for the wavefront gradient of 75 milliradians per lenslet. We demonstrate the system on both fluorescent bead samples and zebrafish embryos.

Self-propagated combustion synthesis of few-layered graphene: an optical properties perspective.

PubMed

Mohandoss, Manonmani; Sen Gupta, Soujit; Kumar, Ramesh; Islam, Md Rabiul; Som, Anirban; Mohd, Azhardin Ganayee; Pradeep, T; Maliyekkal, Shihabudheen M

2018-04-26

This paper describes a labour efficient and cost-effective strategy to prepare few-layered of reduced graphene oxide like (RGOL) sheets from graphite. The self-propagated combustion route enables the bulk production of RGOL sheets. Microscopic and spectroscopic analyses confirmed the formation of few-layer graphene sheets of an average thickness of ∼3 nm and the presence of some oxygen functional groups with a C/O ratio of 8.74. A possible mechanistic pathway for the formation of RGOL sheets is proposed. The optical properties of the RGOL sample were studied in detail by means of Spectroscopic Ellipsometry (SE). The experimental abilities of SE in relating the optical properties with the number of oxygen functionalities present in the samples are explored. The data were analysed by a double-layered optical model along with the Drude-Lorentz oscillatory dispersion relation. The refractive index (n = 2.24), extinction coefficient (k = 2.03), and dielectric functions are obtained using point-by-point analysis and are also checked for Kramers-Kronig (KK) consistency.

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

Ilnytskyi, Jaroslav M.; Neher, Dieter; Saphiannikova, Marina

Photo-induced deformations in azobenzene-containing polymers (azo-polymers) are central to a number of applications, such as optical storage and fabrication of diffractive elements. The microscopic nature of the underlying opto-mechanical coupling is yet not clear. In this study, we address the experimental finding that the scenario of the effects depends on molecular architecture of the used azo-polymer. Typically, opposite deformations in respect to the direction of light polarization are observed for liquid crystalline and amorphous azo-polymers. In this study, we undertake molecular dynamics simulations of two different models that mimic these two types of azo-polymers. We employ hybrid force field modelingmore » and consider only trans-isomers of azobenzene, represented as Gay-Berne sites. The effect of illumination on the orientation of the chromophores is considered on the level of orientational hole burning and emphasis is given to the resulting deformation of the polymer matrix. We reproduce deformations of opposite sign for the two models being considered here and discuss the relevant microscopic mechanisms in both cases.« less

A quantitative and spatially resolved analysis of the performance-bottleneck in high efficiency, planar hybrid perovskite solar cells

DOE PAGES

Draguta, Sergiu; Christians, Jeffrey A.; Morozov, Yurii V.; ...

2018-01-01

Hybrid perovskites represent a potential paradigm shift for the creation of low-cost solar cells. Current power conversion efficiencies (PCEs) exceed 22%. However, despite this, record PCEs are still far from their theoretical Shockley–Queisser limit of 31%. To increase these PCE values, there is a pressing need to understand, quantify and microscopically model charge recombination processes in full working devices. Here, we present a complete microscopic account of charge recombination processes in high efficiency (18–19% PCE) hybrid perovskite (mixed cation and methylammonium lead iodide) solar cells. We employ diffraction-limited optical measurements along with relevant kinetic modeling to establish, for the firstmore » time, local photoluminescence quantum yields, trap densities, trapping efficiencies, charge extraction efficiencies, quasi-Fermi-level splitting, and effective PCE estimates. Correlations between these spatially resolved parameters, in turn, allow us to conclude that intrinsic electron traps in the perovskite active layers limit the performance of these state-of-the-art hybrid perovskite solar cells.« less

A quantitative and spatially resolved analysis of the performance-bottleneck in high efficiency, planar hybrid perovskite solar cells

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

Draguta, Sergiu; Christians, Jeffrey A.; Morozov, Yurii V.

Hybrid perovskites represent a potential paradigm shift for the creation of low-cost solar cells. Current power conversion efficiencies (PCEs) exceed 22%. However, despite this, record PCEs are still far from their theoretical Shockley–Queisser limit of 31%. To increase these PCE values, there is a pressing need to understand, quantify and microscopically model charge recombination processes in full working devices. Here, we present a complete microscopic account of charge recombination processes in high efficiency (18–19% PCE) hybrid perovskite (mixed cation and methylammonium lead iodide) solar cells. We employ diffraction-limited optical measurements along with relevant kinetic modeling to establish, for the firstmore » time, local photoluminescence quantum yields, trap densities, trapping efficiencies, charge extraction efficiencies, quasi-Fermi-level splitting, and effective PCE estimates. Correlations between these spatially resolved parameters, in turn, allow us to conclude that intrinsic electron traps in the perovskite active layers limit the performance of these state-of-the-art hybrid perovskite solar cells.« less

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.

Rapid and early detection of salmonella serotypes with hyperspectral microscope and multivariate data analysis

USDA-ARS?s Scientific Manuscript database

This study was designed to evaluate hyperspectral microscope images for early and rapid detection of Salmonella serotypes: S. Enteritidis, S. Heidelberg, S. Infantis, S. Kentucky, and S. Typhimurium at incubation times of 6, 8, 10, 12, and 24 hours. Images were collected by an acousto-optical tunab...

Real-time spectral imaging in three spatial dimensions

NASA Astrophysics Data System (ADS)

Liu, Wenhai; Psaltis, Demetri; Barbastathis, George

2002-05-01

We report what is to our knowledge the first volume-holographic optical imaging instrument with the capability to return three-dimensional spatial as well as spectral information about semitranslucent microscopic objects in a single measurement. The four-dimensional volume-holographic microscope is characterized theoretically and experimentally by use of fluorescent microspheres as objects.

Visualizing Fluorescence: Using a Homemade Fluorescence "Microscope" to View Latent Fingerprints on Paper

ERIC Educational Resources Information Center

LaFratta, Christopher N.; Huh, Sun Phill; Mallillin, Allistair C.; Riviello, Peter J.; Walt, David R.

2010-01-01

We describe an inexpensive hand-held fluorescence imager (low-magnification microscope), constructed from poly(vinyl chloride) pipe and other inexpensive components for use as a teaching tool to understand the principles of fluorescence detection. Optical filters are used to select the excitation and emission wavelengths and can be easily…

A submersible digital in-line holographic microscope

NASA Astrophysics Data System (ADS)

Jericho, Manfred; Jericho, Stefan; Kreuzer, Hans Juergen; Garcia, Jeorge; Klages, Peter

Few instruments exist that can image microscopic marine organisms in their natural environment so that their locomotion mechanisms, feeding habits and interactions with surfaces, such as bio-fouling, can be investigated in situ. In conventional optical microscopy under conditions of high magnification, only objects confined to the narrow focal plane can be imaged and processes that involve translation of the object perpendicular to this plane are not accessible. To overcome this severe limitation of optical microscopy, we developed digital in-line holographic microscopy (DIHM) as a high-resolution tool for the tracking of organisms in three dimensions. We describe here the design and performance of a very simple submersible digital in-line holographic microscope (SDIHM) that can image organisms and their motion with micron resolution and that can be deployed from small vessels. Holograms and reconstructed images of several microscopic marine organisms were successfully obtained down to a depth of 20 m. The maximum depth was limited by the length of data transmission cables available at the time and operating depth in excess of 100 m are easily possible for the instrument.

Is localized infrared spectroscopy now possible in the electron microscope?

PubMed

Rez, Peter

2014-06-01

The recently developed in-column monochromators make it possible to record energy-c spectra with resolutions better than 30 meV from nanometer-sized regions. It should therefore in principle be possible to detect localized vibrational excitations. The scattering geometry in the electron microscope means that bond stretching in the specimen plane or longitudinal optic phonons dominate the scattering. Most promising for initial studies are vibrations with energies between 300 and 400 meV from hydrogen bonded to other atoms. Estimates of the scattering cross-sections on the basis of a simple model show that they are about the same as inner shell scattering cross-sections. Cross-sections also increase with charge transfer between the atoms, and theory incorporating realistic charge distributions shows that signal/noise is the only limitation to high-resolution imaging. Given the magnitude of the scattering cross-sections, minimizing the tail of the zero-loss peak is just as important as achieving a small-width at half-maximum. Improvements in both resolution and controlling the zero-loss tail will be necessary before it is practical to detect optic phonons in solids between 40 and 60 meV.

Boundary-artifact-free phase retrieval with the transport of intensity equation II: applications to microlens characterization.

PubMed

Zuo, Chao; Chen, Qian; Li, Hongru; Qu, Weijuan; Asundi, Anand

2014-07-28

Boundary conditions play a crucial role in the solution of the transport of intensity equation (TIE). If not appropriately handled, they can create significant boundary artifacts across the reconstruction result. In a previous paper [Opt. Express 22, 9220 (2014)], we presented a new boundary-artifact-free TIE phase retrieval method with use of discrete cosine transform (DCT). Here we report its experimental investigations with applications to the micro-optics characterization. The experimental setup is based on a tunable lens based 4f system attached to a non-modified inverted bright-field microscope. We establish inhomogeneous Neumann boundary values by placing a rectangular aperture in the intermediate image plane of the microscope. Then the boundary values are applied to solve the TIE with our DCT-based TIE solver. Experimental results on microlenses highlight the importance of boundary conditions that often overlooked in simplified models, and confirm that our approach effectively avoid the boundary error even when objects are located at the image borders. It is further demonstrated that our technique is non-interferometric, accurate, fast, full-field, and flexible, rendering it a promising metrological tool for the micro-optics inspection.

Motion mechanics of non-adherent giant liposomes with a combined optical and atomic force microscope

NASA Astrophysics Data System (ADS)

Moreno-Flores, Susana; Ortíz, Rocío

2017-11-01

Herein we present an investigation of the motional dynamics of single mesoscopic bodies of biological relevance with an AFM-based macromanipulation tool and an optical microscope. Giant liposomes are prominent case examples as minimal cell models; studying their mechanics provides a means to address the influence of structural components in the mechanical behaviour of living cells. However, they also pose an experimental challenge due to their lightness, fragility, and high mobility. Their entrapment in wells in a fluid of lower density allows their study under conditions of constrained motion, which enables the synchronous measurement of nanoforces with motion tracking. The procedure enables to estimate sliding friction coefficients and masses of vesicles, and sheds light upon the region between the vesicle and the underlying substrate. The present study paves the way for the investigation of motion and deformation mechanics with one combined technique and a single type of experiment traditionally vetoed to objects that can move as well as deform. Such an approach can be directly applied to cells in suspension, adherent cells or cellular 3D-assemblies so as to assess substrate biocompatibility, monitor adhesion, detachment, motility as well as deformability.

Optimization of a Widefield Structured Illumination Microscope for Non-Destructive Assessment and Quantification of Nuclear Features in Tumor Margins of a Primary Mouse Model of Sarcoma

PubMed Central

Fu, Henry L.; Mueller, Jenna L.; Javid, Melodi P.; Mito, Jeffrey K.; Kirsch, David G.; Ramanujam, Nimmi; Brown, J. Quincy

2013-01-01

Cancer is associated with specific cellular morphological changes, such as increased nuclear size and crowding from rapidly proliferating cells. In situ tissue imaging using fluorescent stains may be useful for intraoperative detection of residual cancer in surgical tumor margins. We developed a widefield fluorescence structured illumination microscope (SIM) system with a single-shot FOV of 2.1×1.6 mm (3.4 mm2) and sub-cellular resolution (4.4 µm). The objectives of this work were to measure the relationship between illumination pattern frequency and optical sectioning strength and signal-to-noise ratio in turbid (i.e. thick) samples for selection of the optimum frequency, and to determine feasibility for detecting residual cancer on tumor resection margins, using a genetically engineered primary mouse model of sarcoma. The SIM system was tested in tissue mimicking solid phantoms with various scattering levels to determine impact of both turbidity and illumination frequency on two SIM metrics, optical section thickness and modulation depth. To demonstrate preclinical feasibility, ex vivo 50 µm frozen sections and fresh intact thick tissue samples excised from a primary mouse model of sarcoma were stained with acridine orange, which stains cell nuclei, skeletal muscle, and collagenous stroma. The cell nuclei were segmented using a high-pass filter algorithm, which allowed quantification of nuclear density. The results showed that the optimal illumination frequency was 31.7 µm−1 used in conjunction with a 4×0.1 NA objective ( = 0.165). This yielded an optical section thickness of 128 µm and an 8.9×contrast enhancement over uniform illumination. We successfully demonstrated the ability to resolve cell nuclei in situ achieved via SIM, which allowed segmentation of nuclei from heterogeneous tissues in the presence of considerable background fluorescence. Specifically, we demonstrate that optical sectioning of fresh intact thick tissues performed equivalently in regards to nuclear density quantification, to physical frozen sectioning and standard microscopy. PMID:23894357

Cryo X-ray microscope with flat sample geometry for correlative fluorescence and nanoscale tomographic imaging.

PubMed

Schneider, Gerd; Guttmann, Peter; Rehbein, Stefan; Werner, Stephan; Follath, Rolf

2012-02-01

X-ray imaging offers a new 3-D view into cells. With its ability to penetrate whole hydrated cells it is ideally suited for pairing fluorescence light microscopy and nanoscale X-ray tomography. In this paper, we describe the X-ray optical set-up and the design of the cryo full-field transmission X-ray microscope (TXM) at the electron storage ring BESSY II. Compared to previous TXM set-ups with zone plate condenser monochromator, the new X-ray optical layout employs an undulator source, a spherical grating monochromator and an elliptically shaped glass capillary mirror as condenser. This set-up improves the spectral resolution by an order of magnitude. Furthermore, the partially coherent object illumination improves the contrast transfer of the microscope compared to incoherent conditions. With the new TXM, cells grown on flat support grids can be tilted perpendicular to the optical axis without any geometrical restrictions by the previously required pinhole for the zone plate monochromator close to the sample plane. We also developed an incorporated fluorescence light microscope which permits to record fluorescence, bright field and DIC images of cryogenic cells inside the TXM. For TXM tomography, imaging with multi-keV X-rays is a straightforward approach to increase the depth of focus. Under these conditions phase contrast imaging is necessary. For soft X-rays with shrinking depth of focus towards 10nm spatial resolution, thin optical sections through a thick specimen might be obtained by deconvolution X-ray microscopy. As alternative 3-D X-ray imaging techniques, the confocal cryo-STXM and the dual beam cryo-FIB/STXM with photoelectron detection are proposed. Copyright © 2012 Elsevier Inc. All rights reserved.

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.

Measuring the complex optical conductivity of graphene by Fabry-Pérot reflectance spectroscopy [Determination of the optical index for few-layer graphene by reflectivity spectroscopy

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

Ghamsari, Behnood G.; Tosado, Jacob; Yamamoto, Mahito

Here, we have experimentally studied the optical refractive index of few-layer graphene through reflection spectroscopy at visible wavelengths. A laser scanning microscope (LSM) with a coherent supercontinuum laser source measured the reflectivity of an exfoliated graphene flake on a Si/SiO 2 substrate, containing monolayer, bilayer and trilayer areas, as the wavelength of the laser was varied from 545nm to 710nm. The complex refractive index of few-layer graphene, n-ik, was extracted from the reflectivity contrast to the bare substrate and the Fresnel reflection theory. Since the SiO 2 thickness enters to the modeling as a parameter, it was precisely measured atmore » the location of the sample. It was found that a common constant optical index cannot explain the wavelength-dependent reflectivity data for single-, double- and three-layer graphene simultaneously, but rather each individual few-layer graphene possesses a unique optical index whose complex values were precisely and accurately determined from the experimental data.« less

Measuring the complex optical conductivity of graphene by Fabry-Pérot reflectance spectroscopy [Determination of the optical index for few-layer graphene by reflectivity spectroscopy

DOE PAGES

Ghamsari, Behnood G.; Tosado, Jacob; Yamamoto, Mahito; ...

2016-09-29

Here, we have experimentally studied the optical refractive index of few-layer graphene through reflection spectroscopy at visible wavelengths. A laser scanning microscope (LSM) with a coherent supercontinuum laser source measured the reflectivity of an exfoliated graphene flake on a Si/SiO 2 substrate, containing monolayer, bilayer and trilayer areas, as the wavelength of the laser was varied from 545nm to 710nm. The complex refractive index of few-layer graphene, n-ik, was extracted from the reflectivity contrast to the bare substrate and the Fresnel reflection theory. Since the SiO 2 thickness enters to the modeling as a parameter, it was precisely measured atmore » the location of the sample. It was found that a common constant optical index cannot explain the wavelength-dependent reflectivity data for single-, double- and three-layer graphene simultaneously, but rather each individual few-layer graphene possesses a unique optical index whose complex values were precisely and accurately determined from the experimental data.« less

Superluminescence from an optically pumped molecular tunneling junction by injection of plasmon induced hot electrons

PubMed Central

Braun, Kai; Wang, Xiao; Kern, Andreas M; Adler, Hilmar; Peisert, Heiko; Chassé, Thomas; Zhang, Dai

2015-01-01

Summary Here, we demonstrate a bias-driven superluminescent point light-source based on an optically pumped molecular junction (gold substrate/self-assembled molecular monolayer/gold tip) of a scanning tunneling microscope, operating at ambient conditions and providing almost three orders of magnitude higher electron-to-photon conversion efficiency than electroluminescence induced by inelastic tunneling without optical pumping. A positive, steadily increasing bias voltage induces a step-like rise of the Stokes shifted optical signal emitted from the junction. This emission is strongly attenuated by reversing the applied bias voltage. At high bias voltage, the emission intensity depends non-linearly on the optical pump power. The enhanced emission can be modelled by rate equations taking into account hole injection from the tip (anode) into the highest occupied orbital of the closest substrate-bound molecule (lower level) and radiative recombination with an electron from above the Fermi level (upper level), hence feeding photons back by stimulated emission resonant with the gap mode. The system reflects many essential features of a superluminescent light emitting diode. PMID:26171286

Microscopic Materials on a Magnet

NASA Technical Reports Server (NTRS)

2008-01-01

These images show a comparison of the weak magnet OM7 from the Optical Microscope on NASA's Phoenix Mars Lander before (left) and after (right) soil deposition.

The microscope took the left image during Phoenix's Sol 15 (June 10, 2008) and the right image during Sol 21 (Jun 16, 2008).

The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

Closed loop adaptive optics for microscopy without a wavefront sensor.

PubMed

Kner, Peter; Winoto, Lukman; Agard, David A; Sedat, John W

2010-02-24

A three-dimensional wide-field image of a small fluorescent bead contains more than enough information to accurately calculate the wavefront in the microscope objective back pupil plane using the phase retrieval technique. The phase-retrieved wavefront can then be used to set a deformable mirror to correct the point-spread function (PSF) of the microscope without the use of a wavefront sensor. This technique will be useful for aligning the deformable mirror in a widefield microscope with adaptive optics and could potentially be used to correct aberrations in samples where small fluorescent beads or other point sources are used as reference beacons. Another advantage is the high resolution of the retrieved wavefont as compared with current Shack-Hartmann wavefront sensors. Here we demonstrate effective correction of the PSF in 3 iterations. Starting from a severely aberrated system, we achieve a Strehl ratio of 0.78 and a greater than 10-fold increase in maximum intensity.

A novel 3D deformation measurement method under optical microscope for micro-scale bulge-test

NASA Astrophysics Data System (ADS)

Wu, Dan; Xie, Huimin

2017-11-01

A micro-scale 3D deformation measurement method combined with optical microscope is proposed in this paper. The method is based on gratings and phase shifting algorithm. By recording the grating images before and after deformation from two symmetrical angles and calculating the phases of the grating patterns, the 3D deformation field of the specimen can be extracted from the phases of the grating patterns. The proposed method was applied to the micro-scale bulge test. A micro-scale thermal/mechanical coupling bulge-test apparatus matched with the super-depth microscope was exploited. With the gratings fabricated onto the film, the deformed morphology of the bulged film was measured reliably. The experimental results show that the proposed method and the exploited bulge-test apparatus can be used to characterize the thermal/mechanical properties of the films at micro-scale successfully.

PHOTONICS AND NANOTECHNOLOGY Microscopic theory of optical properties of composite media with chaotically distributed nanoparticles

NASA Astrophysics Data System (ADS)

Shalin, A. S.

2010-12-01

The boundary problem of light reflection and transmission by a film with chaotically distributed nanoinclusions is considered. Based on the proposed microscopic approach, analytic expressions are derived for distributions inside and outside the nanocomposite medium. Good agreement of the results with exact calculations and (at low concentrations of nanoparticles) with the integral Maxwell-Garnett effective-medium theory is demonstrated. It is shown that at high nanoparticle concentrations, averaging the dielectric constant in volume as is done within the framework of the effective-medium theory yields overestimated values of the optical film density compared to the values yielded by the proposed microscopic approach. We also studied the dependence of the reflectivity of a system of gold nanoparticles on their size, the size dependence of the plasmon resonance position along the wavelength scale, and demonstrated a good agreement with experimental data.