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

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

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.

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

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.

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.

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.

Generic distortion model for metrology under optical microscopes

NASA Astrophysics Data System (ADS)

Liu, Xingjian; Li, Zhongwei; Zhong, Kai; Chao, YuhJin; Miraldo, Pedro; Shi, Yusheng

2018-04-01

For metrology under optical microscopes, lens distortion is the dominant source of error. Previous distortion models and correction methods mostly rely on the assumption that parametric distortion models require a priori knowledge of the microscopes' lens systems. However, because of the numerous optical elements in a microscope, distortions can be hardly represented by a simple parametric model. In this paper, a generic distortion model considering both symmetric and asymmetric distortions is developed. Such a model is obtained by using radial basis functions (RBFs) to interpolate the radius and distortion values of symmetric distortions (image coordinates and distortion rays for asymmetric distortions). An accurate and easy to implement distortion correction method is presented. With the proposed approach, quantitative measurement with better accuracy can be achieved, such as in Digital Image Correlation for deformation measurement when used with an optical microscope. The proposed technique is verified by both synthetic and real data experiments.

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

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.

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.

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.

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.

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.

Multimodal optical workstation for simultaneous linear, nonlinear microscopy and nanomanipulation: upgrading a commercial confocal inverted microscope.

PubMed

Mathew, Manoj; Santos, Susana I C O; Zalvidea, Dobryna; Loza-Alvarez, Pablo

2009-07-01

In this work we propose and build a multimodal optical workstation that extends a commercially available confocal microscope (Nikon Confocal C1-Si) to include nonlinear/multiphoton microscopy and optical manipulation/stimulation tools such as nanosurgery. The setup allows both subsystems (confocal and nonlinear) to work independently and simultaneously. The workstation enables, for instance, nanosurgery along with simultaneous confocal and brightfield imaging. The nonlinear microscopy capabilities are added around the commercial confocal microscope by exploiting all the flexibility offered by this microscope and without need for any mechanical or electronic modification of the confocal microscope systems. As an example, the standard differential interference contrast condenser and diascopic detector in the confocal microscope are readily used as a forward detection mount for second harmonic generation imaging. The various capabilities of this workstation, as applied directly to biology, are demonstrated using the model organism Caenorhabditis elegans.

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.

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

Analytical model of the optical vortex microscope.

PubMed

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

2016-04-20

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

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.

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 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.

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.

Modeling off-resonant nonlinear-optical cascading in mesoscopic thin films and guest-host molecular systems

NASA Astrophysics Data System (ADS)

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

2013-12-01

A model for off-resonant microscopic cascading of (hyper)polarizabilities is developed using a self-consistent field approach to study mesoscopic systems of nonlinear polarizable atoms and molecules. We find enhancements in the higher-order susceptibilities resulting from geometrical and boundary orientation effects. We include an example of the dependence on excitation beam cross sectional structure and a simplified derivation of the microscopic cascading of the nonlinear-optical response in guest-host systems.

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.

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.

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

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

Diamond Quantum Nanoemitters: Cross Discipline Research on Hyperbolic Optical Systems for Control of Quantum Nanoemitters

DTIC Science & Technology

2017-05-05

results of this project there are: (1) the investigation of the effect of phonons on the optical properties of solid state emitters. A microscopic ...In  what  follows  we  list  the  main  results  and  undergoing  research.   2. Results 2.1   Microscopic  modeling...fluorescent  markers   for   biological   measurements.   Here,   we   present   a   first-­‐principles   microscopic   description

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.

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.

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.

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").

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.

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.

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

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)

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.

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.

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

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.

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.

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.

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.

Correlative super-resolution fluorescence microscopy combined with optical coherence microscopy

NASA Astrophysics Data System (ADS)

Kim, Sungho; Kim, Gyeong Tae; Jang, Soohyun; Shim, Sang-Hee; Bae, Sung Chul

2015-03-01

Recent development of super-resolution fluorescence imaging technique such as stochastic optical reconstruction microscopy (STORM) and photoactived localization microscope (PALM) has brought us beyond the diffraction limits. It allows numerous opportunities in biology because vast amount of formerly obscured molecular structures, due to lack of spatial resolution, now can be directly observed. A drawback of fluorescence imaging, however, is that it lacks complete structural information. For this reason, we have developed a super-resolution multimodal imaging system based on STORM and full-field optical coherence microscopy (FF-OCM). FF-OCM is a type of interferometry systems based on a broadband light source and a bulk Michelson interferometer, which provides label-free and non-invasive visualization of biological samples. The integration between the two systems is simple because both systems use a wide-field illumination scheme and a conventional microscope. This combined imaging system gives us both functional information at a molecular level (~20nm) and structural information at the sub-cellular level (~1μm). For thick samples such as tissue slices, while FF-OCM is readily capable of imaging the 3D architecture, STORM suffer from aberrations and high background fluorescence that substantially degrade the resolution. In order to correct the aberrations in thick tissues, we employed an adaptive optics system in the detection path of the STORM microscope. We used our multimodal system to obtain images on brain tissue samples with structural and functional information.

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.

Laser-induced fluorescence microscopic system using an optical parametric oscillator for tunable detection in microchip analysis.

PubMed

Kumemura, Momoko; Odake, Tamao; Korenaga, Takashi

2005-06-01

A laser-induced fluorescence microscopic system based on optical parametric oscillation has been constructed as a tunable detector for microchip analysis. The detection limit of sulforhodamine B (Ex. 520 nm, Em. 570 nm) was 0.2 mumol, which was approximately eight orders of magnitude better than with a conventional fluorophotometer. The system was applied to the determination of fluorescence-labeled DNA (Ex. 494 nm, Em. 519 nm) in a microchannel and the detection limit reached a single molecule. These results showed the feasibility of this system as a highly sensitive and tunable fluorescence detector for microchip analysis.

A light field microscope imaging spectrometer based on the microlens array

NASA Astrophysics Data System (ADS)

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

2017-10-01

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

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.

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.

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.

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.

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.

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

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.

A Compact "Water Window" Microscope with 60 nm Spatial Resolution for Applications in Biology and Nanotechnology.

PubMed

Wachulak, Przemyslaw; Torrisi, Alfio; Nawaz, Muhammad F; Bartnik, Andrzej; Adjei, Daniel; Vondrová, Šárka; Turňová, Jana; Jančarek, Alexandr; Limpouch, Jiří; Vrbová, Miroslava; Fiedorowicz, Henryk

2015-10-01

Short illumination wavelength allows an extension of the diffraction limit toward nanometer scale; thus, improving spatial resolution in optical systems. Soft X-ray (SXR) radiation, from "water window" spectral range, λ=2.3-4.4 nm wavelength, which is particularly suitable for biological imaging due to natural optical contrast provides better spatial resolution than one obtained with visible light microscopes. The high contrast in the "water window" is obtained because of selective radiation absorption by carbon and water, which are constituents of the biological samples. The development of SXR microscopes permits the visualization of features on the nanometer scale, but often with a tradeoff, which can be seen between the exposure time and the size and complexity of the microscopes. Thus, herein, we present a desk-top system, which overcomes the already mentioned limitations and is capable of resolving 60 nm features with very short exposure time. Even though the system is in its initial stage of development, we present different applications of the system for biology and nanotechnology. Construction of the microscope with recently acquired images of various samples will be presented and discussed. Such a high resolution imaging system represents an interesting solution for biomedical, material science, and nanotechnology applications.

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.

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 (

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

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.

Three-dimensional motion-picture imaging of dynamic object by parallel-phase-shifting digital holographic microscopy using an inverted magnification optical system

NASA Astrophysics Data System (ADS)

Fukuda, Takahito; Shinomura, Masato; Xia, Peng; Awatsuji, Yasuhiro; Nishio, Kenzo; Matoba, Osamu

2017-04-01

We constructed a parallel-phase-shifting digital holographic microscopy (PPSDHM) system using an inverted magnification optical system, and succeeded in three-dimensional (3D) motion-picture imaging for 3D displacement of a microscopic object. In the PPSDHM system, the inverted and afocal magnification optical system consisted of a microscope objective (16.56 mm focal length and 0.25 numerical aperture) and a convex lens (300 mm focal length and 82 mm aperture diameter). A polarization-imaging camera was used to record multiple phase-shifted holograms with a single-shot exposure. We recorded an alum crystal, sinking down in aqueous solution of alum, by the constructed PPSDHM system at 60 frames/s for about 20 s and reconstructed high-quality 3D motion-picture image of the crystal. Then, we calculated amounts of displacement of the crystal from the amounts in the focus plane and the magnifications of the magnification optical system, and obtained the 3D trajectory of the crystal by that amounts.

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.

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.

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.

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.

Mechanical vibration compensation method for 3D+t multi-particle tracking in microscopic volumes.

PubMed

Pimentel, A; Corkidi, G

2009-01-01

The acquisition and analysis of data in microscopic systems with spatiotemporal evolution is a very relevant topic. In this work, we describe a method to optimize an experimental setup for acquiring and processing spatiotemporal (3D+t) data in microscopic systems. The method is applied to a three-dimensional multi-tracking and analysis system of free-swimming sperm trajectories previously developed. The experimental set uses a piezoelectric device making oscillate a large focal-distance objective mounted on an inverted microscope (over its optical axis) to acquire stacks of images at a high frame rate over a depth on the order of 250 microns. A problem arise when the piezoelectric device oscillates, in such a way that a vibration is transmitted to the whole microscope, inducing undesirable 3D vibrations to the whole set. For this reason, as a first step, the biological preparation was isolated from the body of the microscope to avoid modifying the free swimming pattern of the microorganism due to the transmission of these vibrations. Nevertheless, as the image capturing device is mechanically attached to the "vibrating" microscope, the resulting acquired data are contaminated with an undesirable 3D movement that biases the original trajectory of these high speed moving cells. The proposed optimization method determines the functional form of these 3D oscillations to neutralize them from the original acquired data set. Given the spatial scale of the system, the added correction increases significantly the data accuracy. The optimized system may be very useful in a wide variety of 3D+t applications using moving optical devices.

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.

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.

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.

Development of Nomarski microscopy for quantitative determination of surface topography

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

Hartman, J. S.; Gordon, R. L.; Lessor, D. L.

1979-01-01

The use of Nomarski differential interference contrast (DIC) microscopy has been extended to provide nondestructive, quantitative analysis of a sample's surface topography. Theoretical modeling has determined the dependence of the image intensity on the microscope's optical components, the sample's optical properties, and the sample's surface orientation relative to the microscope. Results include expressions to allow the inversion of image intensity data to determine sample surface slopes. A commercial Nomarski system has been modified and characterized to allow the evaluation of the optical model. Data have been recorded with smooth, planar samples that verify the theoretical predictions.

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

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.

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.

Transfer function characteristics of super resolving systems

NASA Technical Reports Server (NTRS)

Milster, Tom D.; Curtis, Craig H.

1992-01-01

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

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.

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.

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.

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.

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.

Optical tweezers based force measurement system for quantitating binding interactions: system design and application for the study of bacterial adhesion.

PubMed

Fällman, Erik; Schedin, Staffan; Jass, Jana; Andersson, Magnus; Uhlin, Bernt Eric; Axner, Ove

2004-06-15

An optical force measurement system for quantitating forces in the pN range between micrometer-sized objects has been developed. The system was based upon optical tweezers in combination with a sensitive position detection system and constructed around an inverted microscope. A trapped particle in the focus of the high numerical aperture microscope-objective behaves like an omnidirectional mechanical spring in response to an external force. The particle's displacement from the equilibrium position is therefore a direct measure of the exerted force. A weak probe laser beam, focused directly below the trapping focus, was used for position detection of the trapped particle (a polystyrene bead). The bead and the condenser focus the light to a distinct spot in the far field, monitored by a position sensitive detector. Various calibration procedures were implemented in order to provide absolute force measurements. The system has been used to measure the binding forces between Escherichia coli bacterial adhesins and galabiose-functionalized beads.

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.

A fibre optic fluorescence sensor to measure redox level in tissues

NASA Astrophysics Data System (ADS)

Zhang, Wen Qi; Morrison, Janna L.; Darby, Jack R. T.; Plush, Sally; Sorvina, Alexandra; Brooks, Doug; Monro, Tanya M.; Afshar Vahid, Shahraam

2018-01-01

We report the design of a fibre optic-based redox detection system for investigating differences in metabolic activities of tissues. Our system shows qualitative agreement with the results collected from a commercial two- photon microscope system. Thus, demonstrating the feasibility of building an ex vivo and in vivo redox detection system that is low cost and portable.

Spectral ophthalmoscopy based on supercontinuum

NASA Astrophysics Data System (ADS)

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

2010-02-01

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

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.

Enabling the detection of UV signal in multimodal nonlinear microscopy with catalogue lens components.

PubMed

Vogel, Martin; Wingert, Axel; Fink, Rainer H A; Hagl, Christian; Ganikhanov, Feruz; Pfeffer, Christian P

2015-10-01

Using an optical system made from fused silica catalogue optical components, third-order nonlinear microscopy has been enabled on conventional Ti:sapphire laser-based multiphoton microscopy setups. The optical system is designed using two lens groups with straightforward adaptation to other microscope stands when one of the lens groups is exchanged. Within the theoretical design, the optical system collects and transmits light with wavelengths between the near ultraviolet and the near infrared from an object field of at least 1 mm in diameter within a resulting numerical aperture of up to 0.56. The numerical aperture can be controlled with a variable aperture stop between the two lens groups of the condenser. We demonstrate this new detection capability in third harmonic generation imaging experiments at the harmonic wavelength of ∼300 nm and in multimodal nonlinear optical imaging experiments using third-order sum frequency generation and coherent anti-Stokes Raman scattering microscopy so that the wavelengths of the detected signals range from ∼300 nm to ∼660 nm. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

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.

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...

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.

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.

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.

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.

Low cost, high performance, self-aligning miniature optical systems

PubMed Central

Kester, Robert T.; Christenson, Todd; Kortum, Rebecca Richards; Tkaczyk, Tomasz S.

2009-01-01

The most expensive aspects in producing high quality miniature optical systems are the component costs and long assembly process. A new approach for fabricating these systems that reduces both aspects through the implementation of self-aligning LIGA (German acronym for lithographie, galvanoformung, abformung, or x-ray lithography, electroplating, and molding) optomechanics with high volume plastic injection molded and off-the-shelf glass optics is presented. This zero alignment strategy has been incorporated into a miniature high numerical aperture (NA = 1.0W) microscope objective for a fiber confocal reflectance microscope. Tight alignment tolerances of less than 10 μm are maintained for all components that reside inside of a small 9 gauge diameter hypodermic tubing. A prototype system has been tested using the slanted edge modulation transfer function technique and demonstrated to have a Strehl ratio of 0.71. This universal technology is now being developed for smaller, needle-sized imaging systems and other portable point-of-care diagnostic instruments. PMID:19543344

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

NASA Astrophysics Data System (ADS)

Joshi, Narahari V.; Medina, Honorio

2000-05-01

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

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.

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.

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.

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.

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

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.

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.

Successfully using optical components and systems in novel ways during educational outreach programs for K-12 grades

NASA Astrophysics Data System (ADS)

Silberman, Donn

2006-08-01

Much work has been done in efforts to reach students in the K-12 grades to encourage them to learn about optics and related sciences and technologies. One goal of these efforts is to develop the future optical scientists and engineers to carry on the work of this and related societies. One main obstacle is to create low costs novel and effective hands-on optical components and systems for these students to use and from which to get excited. Students at different grade levels and abilities are receptive to different kinds of components and systems and this must be taken into account when preparing for outreach programs. There are, however, some guiding principles which can be used throughout the various levels, including making sure the components and systems are good examples and not marginal. Small telescopes or microscopes that use poor quality optics which provide poor quality images do more to discourage young students from going into the sciences than if they never had the experience at all. Some examples of both poor and good quality optical components and systems that will be described and demonstrated include: lenses, telescopes, microscopes, diffraction gratings, Kaleidoscopes, Fresnel Lenses, polarization filters and liquid crystals. The figures in this paper are in color and best viewed on-line or printed with a good color printer.

Optical tweezers for the measurement of binding forces: system description and application for the study of E. coli adhesion

NASA Astrophysics Data System (ADS)

Fallman, Erik G.; Schedin, Staffan; Andersson, Magnus J.; Jass, Jana; Axner, Ove

2003-06-01

Optical tweezers together with a position sensitive detection system allows measurements of forces in the pN range between micro-sized biological objects. A prototype force measurement system has been constructed around in inverted microscope with an argon-ion pumped Ti:sapphire laser as light source for optical trapping. A trapped particle in the focus of the high numerical aperture microscope-objective behaves like an omni-directional mechanical spring if an external force displaces it. The displacement from the equilibrium position is a measure of the exerted force. For position detection of the trapped particle (polystyrene beads), a He-Ne laser beam is focused a small distance below the trapping focus. An image of the bead appears as a distinct spot in the far field, monitored by a photosensitive detector. The position data is converted to a force measurement by a calibration procedure. The system has been used for measuring the binding forces between E-coli bacterial adhesin and their receptor sugars.

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

PubMed

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

1997-06-01

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

RMB identification based on polarization parameters inversion imaging

NASA Astrophysics Data System (ADS)

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

2016-10-01

Social order is threatened by counterfeit money. Conventional anti-counterfeit technology is much too old to identify its authenticity or not. The intrinsic difference between genuine notes and counterfeit notes is its paper tissue. In this paper a new technology of detecting RMB is introduced, the polarization parameter indirect microscopic imaging technique. A conventional reflection microscopic system is used as the basic optical system, and inserting into it with polarization-modulation mechanics. The near-field structural characteristics can be delivered by optical wave and material coupling. According to coupling and conduction physics, calculate the changes of optical wave parameters, then get the curves of the intensity of the image. By analyzing near-field polarization parameters in nanoscale, finally calculate indirect polarization parameter imaging of the fiber of the paper tissue in order to identify its authenticity.

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.

Low-power noncontact photoacoustic microscope for bioimaging applications

NASA Astrophysics Data System (ADS)

Sathiyamoorthy, Krishnan; Strohm, Eric M.; Kolios, Michael C.

2017-04-01

An inexpensive noncontact photoacoustic (PA) imaging system using a low-power continuous wave laser and a kilohertz-range microphone has been developed. The system operates in both optical and PA imaging modes and is designed to be compatible with conventional optical microscopes. Aqueous coupling fluids are not required for the detection of the PA signals; air is used as the coupling medium. The main component of the PA system is a custom designed PA imaging sensor that consists of an air-filled sample chamber and a resonator chamber that isolates a standard kilohertz frequency microphone from the input laser. A sample to be examined is placed on the glass substrate inside the chamber. A laser focused to a small spot by a 40× objective onto the substrate enables generation of PA signals from the sample. Raster scanning the laser over the sample with micrometer-sized steps enables high-resolution PA images to be generated. A lateral resolution of 1.37 μm was achieved in this proof of concept study, which can be further improved using a higher numerical aperture objective. The application of the system was investigated on a red blood cell, with a noise-equivalent detection sensitivity of 43,887 hemoglobin molecules (72.88×10-21 mol or 72.88 zeptomol). The minimum pressure detectable limit of the system was 19.1 μPa. This inexpensive, compact noncontact PA sensor is easily integrated with existing commercial optical microscopes, enabling optical and PA imaging of the same sample. Applications include forensic measurements, blood coagulation tests, and monitoring the penetration of drugs into human membrane.

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.

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

Real-time processing of interferograms for monitoring protein crystal growth on the Space Station

NASA Technical Reports Server (NTRS)

Choudry, A.; Dupuis, N.

1988-01-01

The possibility of using microscopic interferometric techniques to monitor the growth of protein crystals on the Space Station is studied. Digital image processing techniques are used to develop a system for the real-time analysis of microscopic interferograms of nucleation sites during protein crystal growth. Features of the optical setup and the image processing system are discussed and experimental results are presented.

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

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.

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

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.

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.

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.

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.

Near real-time measurement of forces applied by an optical trap to a rigid cylindrical object

NASA Astrophysics Data System (ADS)

Glaser, Joseph; Hoeprich, David; Resnick, Andrew

2014-07-01

An automated data acquisition and processing system is established to measure the force applied by an optical trap to an object of unknown composition in real time. Optical traps have been in use for the past 40 years to manipulate microscopic particles, but the magnitude of applied force is often unknown and requires extensive instrument characterization. Measuring or calculating the force applied by an optical trap to nonspherical particles presents additional difficulties which are also overcome with our system. Extensive experiments and measurements using well-characterized objects were performed to verify the system performance.

Design of a self-aligned, wide temperature range (300 mK-300 K) atomic force microscope/magnetic force microscope with 10 nm magnetic force microscope resolution

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

Karcı, Özgür; Department of Nanotechnology and Nanomedicine, Hacettepe University, Beytepe, 06800 Ankara; Dede, Münir

We describe the design of a wide temperature range (300 mK-300 K) atomic force microscope/magnetic force microscope with a self-aligned fibre-cantilever mechanism. An alignment chip with alignment groves and a special mechanical design are used to eliminate tedious and time consuming fibre-cantilever alignment procedure for the entire temperature range. A low noise, Michelson fibre interferometer was integrated into the system for measuring deflection of the cantilever. The spectral noise density of the system was measured to be ~12 fm/√Hz at 4.2 K at 3 mW incident optical power. Abrikosov vortices in BSCCO(2212) single crystal sample and a high density hardmore » disk sample were imaged at 10 nm resolution to demonstrate the performance of the system.« less

Optical filters for wavelength selection in fluorescence instrumentation.

PubMed

Erdogan, Turan

2011-04-01

Fluorescence imaging and analysis techniques have become ubiquitous in life science research, and they are poised to play an equally vital role in in vitro diagnostics (IVD) in the future. Optical filters are crucial for nearly all fluorescence microscopes and instruments, not only to provide the obvious function of spectral control, but also to ensure the highest possible detection sensitivity and imaging resolution. Filters make it possible for the sample to "see" light within only the absorption band, and the detector to "see" light within only the emission band. Without filters, the detector would not be able to distinguish the desired fluorescence from scattered excitation light and autofluorescence from the sample, substrate, and other optics in the system. Today the vast majority of fluorescence instruments, including the widely popular fluorescence microscope, use thin-film interference filters to control the spectra of the excitation and emission light. Hence, this unit emphasizes thin-film filters. After briefly introducing different types of thin-film filters and how they are made, the unit describes in detail different optical filter configurations in fluorescence instruments, including both single-color and multicolor imaging systems. Several key properties of thin-film filters, which can significantly affect optical system performance, are then described. In the final section, tunable optical filters are also addressed in a relative comparison.

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…

Adaptive optical microscope for brain imaging in vivo

NASA Astrophysics Data System (ADS)

Wang, Kai

2017-04-01

The optical heterogeneity of biological tissue imposes a major limitation to acquire detailed structural and functional information deep in the biological specimens using conventional microscopes. To restore optimal imaging performance, we developed an adaptive optical microscope based on direct wavefront sensing technique. This microscope can reliably measure and correct biological samples induced aberration. We demonstrated its performance and application in structural and functional brain imaging in various animal models, including fruit fly, zebrafish and mouse.

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

Optics of wide-angle panoramic viewing system-assisted vitreous surgery.

PubMed

Chalam, Kakarla V; Shah, Vinay A

2004-01-01

The purpose of the article is to describe the optics of the contact wide-angle lens system with stereo-reinverter for vitreous surgery. A panoramic viewing system is made up of two components; an indirect ophthalmoscopy lens system for fundus image viewing, which is placed on the patient's cornea as a contact lens, and a separate removable prism system for reinversion of the image mounted on the microscope above the zooming system. The system provides a 104 degrees field of view in a phakic emmetropic eye with minification, which can be magnified by the operating microscope. It permits a binocular stereoptic view even through a small pupil (3 mm) or larger. In an air-filled phakic eye, field of view increases to approximately 130 degrees. The obtained image of the patient's fundus is reinverted to form true, erect, stereoscopic image by the reinversion system. In conclusion, this system permits wide-angle panoramic view of the surgical field. The contact lens neutralizes the optical irregularities of the corneal surface and allows improved visualization in eyes with irregular astigmatism induced by corneal scars. Excellent visualization is achieved in complex clinical situations such as miotic pupils, lenticular opacities, and in air-filled phakic eyes.

Hand-held photomicroscopy system

NASA Technical Reports Server (NTRS)

Zabower, H. R.

1972-01-01

Photomicroscopy system, with simple optics and any standard microscope objective, is used with any type of motion picture, still, or television camera system. Device performs well under difficult environmental conditions and applies to work in ecological studies, field hospitals, and geological surveys.

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

PubMed

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

2015-08-01

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

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.

Laser beam shaping for biomedical microscopy techniques

NASA Astrophysics Data System (ADS)

Laskin, Alexander; Kaiser, Peter; Laskin, Vadim; Ostrun, Aleksei

2016-04-01

Uniform illumination of a working field is very important in optical systems of confocal microscopy and various implementations of fluorescence microscopy like TIR, SSIM, STORM, PALM to enhance performance of these laser-based research techniques. Widely used TEM00 laser sources are characterized by essentially non-uniform Gaussian intensity profile which leads usually to non-uniform intensity distribution in a microscope working field or in a field of microlenses array of a confocal microscope optical system, this non-uniform illumination results in instability of measuring procedure and reducing precision of quantitative measurements. Therefore transformation of typical Gaussian distribution of a TEM00 laser to flat-top (top hat) profile is an actual technical task, it is solved by applying beam shaping optics. Due to high demands to optical image quality the mentioned techniques have specific requirements to a uniform laser beam: flatness of phase front and extended depth of field, - from this point of view the microscopy techniques are similar to holography and interferometry. There are different refractive and diffractive beam shaping approaches used in laser industrial and scientific applications, but only few of them are capable to fulfil the optimum conditions for beam quality required in discussed microscopy techniques. We suggest applying refractive field mapping beam shapers πShaper, which operational principle presumes almost lossless transformation of Gaussian to flat-top beam with flatness of output wavefront, conserving of beam consistency, providing collimated low divergent output beam, high transmittance, extended depth of field, negligible wave aberration, and achromatic design provides capability to work with several lasers with different wavelengths simultaneously. The main function of a beam shaper is transformation of laser intensity profile, further beam transformation to provide optimum for a particular technique spot size and shape has to be realized by an imaging optical system which can include microscope objectives and tube lenses. This paper will describe design basics of refractive beam shapers and optical layouts of their applying in microscopy systems. Examples of real implementations and experimental results will be presented as well.

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

NASA Astrophysics Data System (ADS)

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

2017-06-01

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

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).

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.

Three-dimensional polarization algebra for all polarization sensitive optical systems.

PubMed

Li, Yahong; Fu, Yuegang; Liu, Zhiying; Zhou, Jianhong; Bryanston-Cross, P J; Li, Yan; He, Wenjun

2018-05-28

Using three-dimensional (3D) coherency vector (9 × 1), we develop a new 3D polarization algebra to calculate the polarization properties of all polarization sensitive optical systems, especially when the incident optical field is partially polarized or un-polarized. The polarization properties of a high numerical aperture (NA) microscope objective (NA = 1.25 immersed in oil) are analyzed based on the proposed 3D polarization algebra. Correspondingly, the polarization simulation of this high NA optical system is performed by the commercial software VirtualLAB Fusion. By comparing the theoretical calculations with polarization simulations, a perfect matching relation is obtained, which demonstrates that this 3D polarization algebra is valid to quantify the 3D polarization properties for all polarization sensitive optical systems.

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.

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

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-22

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

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.

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.

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.

On-Chip Biomedical Imaging

PubMed Central

Göröcs, Zoltán; Ozcan, Aydogan

2012-01-01

Lab-on-a-chip systems have been rapidly emerging to pave the way toward ultra-compact, efficient, mass producible and cost-effective biomedical research and diagnostic tools. Although such microfluidic and micro electromechanical systems achieved high levels of integration, and are capable of performing various important tasks on the same chip, such as cell culturing, sorting and staining, they still rely on conventional microscopes for their imaging needs. Recently several alternative on-chip optical imaging techniques have been introduced, which have the potential to substitute conventional microscopes for various lab-on-a-chip applications. Here we present a critical review of these recently emerging on-chip biomedical imaging modalities, including contact shadow imaging, lensfree holographic microscopy, fluorescent on-chip microscopy and lensfree optical tomography. PMID:23558399

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

PubMed Central

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

2017-01-01

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

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.

On-axis programmable microscope using liquid crystal spatial light modulator

NASA Astrophysics Data System (ADS)

García-Martínez, Pascuala; Martínez, José Luís.; Moreno, Ignacio

2017-06-01

Spatial light modulators (SLM) are currently used in many applications in optical microscopy and imaging. One of the most promising methods is the use of liquid crystal displays (LCD) as programmable phase diffractive optical elements (DOE) placed in the Fourier plane giving access to the spatial frequencies which can be phased shifted individually, allowing to emulate a wealth of contrast enhancing methods for both amplitude and phase samples. We use phase and polarization modulation of LCD to implement an on-axis microscope optical system. The LCD used are Hamamatsu liquid crystal on silicon (LCOS) SLM free of flicker, thus showing a full profit of the SLM space bandwidth, as opposed to optical systems in the literature forced to work off-axis due to the strong zero-order component. Taking benefits of the phase modulation of the LCOS we have implemented different microscopic imaging operations, such as high-pass and low-pass filtering in parallel using programmable blazed gratings. Moreover, we are able to control polarization modulation to display two orthogonal linear state of polarization images than can be subtracted or added by changing the period of the blazed grating. In that sense, Differential Interference Contrast (DIC) microscopy can be easily done by generating two images exploiting the polarization splitting properties when a blazed grating is displayed in the SLM. Biological microscopy samples are also used.

A review on optical actuators for microfluidic systems

NASA Astrophysics Data System (ADS)

Yang, Tie; Chen, Yue; Minzioni, Paolo

2017-12-01

During the last few decades microfluidic systems have become more and more popular and their relevance in different fields is continually growing. In fact, the use of microchannels allows a significant reduction of the required sample-volume and opens the way to a completely new set of possible investigations, including the study of the properties of cells, the development of new cells’ separation techniques and the analysis of single-cell proteins. One of the main differences between microscopic and macroscopic systems is obviously dictated by the need for suitable actuation mechanisms, which should allow precise control of microscopic fluid volumes and of micro-samples inside the fluid. Even if both syringe-pump and pneumatic-pump technologies significantly evolved and they currently enable sub-μL samples control, completely new approaches were recently developed for the manipulation of samples inside the microchannel. This review is dedicated to describing different kinds of optical actuators that can be applied in microfluidic systems for sample manipulation as well as for pumping. The basic principles underlying the optical actuation mechanisms will be described first, and then several experimental demonstrations will be reviewed and compared.

High-resolution retinal imaging using adaptive optics and Fourier-domain optical coherence tomography

DOEpatents

Olivier, Scot S.; Werner, John S.; Zawadzki, Robert J.; Laut, Sophie P.; Jones, Steven M.

2010-09-07

This invention permits retinal images to be acquired at high speed and with unprecedented resolution in three dimensions (4.times.4.times.6 .mu.m). The instrument achieves high lateral resolution by using adaptive optics to correct optical aberrations of the human eye in real time. High axial resolution and high speed are made possible by the use of Fourier-domain optical coherence tomography. Using this system, we have demonstrated the ability to image microscopic blood vessels and the cone photoreceptor mosaic.

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.

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

PubMed

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

2012-01-01

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

Surface plasmon resonance microscopy: achieving a quantitative optical response

PubMed Central

Peterson, Alexander W.; Halter, Michael; Plant, Anne L.; Elliott, John T.

2016-01-01

Surface plasmon resonance (SPR) imaging allows real-time label-free imaging based on index of refraction, and changes in index of refraction at an interface. Optical parameter analysis is achieved by application of the Fresnel model to SPR data typically taken by an instrument in a prism based configuration. We carry out SPR imaging on a microscope by launching light into a sample, and collecting reflected light through a high numerical aperture microscope objective. The SPR microscope enables spatial resolution that approaches the diffraction limit, and has a dynamic range that allows detection of subnanometer to submicrometer changes in thickness of biological material at a surface. However, unambiguous quantitative interpretation of SPR changes using the microscope system could not be achieved using the Fresnel model because of polarization dependent attenuation and optical aberration that occurs in the high numerical aperture objective. To overcome this problem, we demonstrate a model to correct for polarization diattenuation and optical aberrations in the SPR data, and develop a procedure to calibrate reflectivity to index of refraction values. The calibration and correction strategy for quantitative analysis was validated by comparing the known indices of refraction of bulk materials with corrected SPR data interpreted with the Fresnel model. Subsequently, we applied our SPR microscopy method to evaluate the index of refraction for a series of polymer microspheres in aqueous media and validated the quality of the measurement with quantitative phase microscopy. PMID:27782542

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

Mirror-based broadband scanner with minimized aberration

NASA Astrophysics Data System (ADS)

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

2009-02-01

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

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.

Microscopic optical path length difference and polarization measurement system for cell analysis

NASA Astrophysics Data System (ADS)

Satake, H.; Ikeda, K.; Kowa, H.; Hoshiba, T.; Watanabe, E.

2018-03-01

In recent years, noninvasive, nonstaining, and nondestructive quantitative cell measurement techniques have become increasingly important in the medical field. These cell measurement techniques enable the quantitative analysis of living cells, and are therefore applied to various cell identification processes, such as those determining the passage number limit during cell culturing in regenerative medicine. To enable cell measurement, we developed a quantitative microscopic phase imaging system based on a Mach-Zehnder interferometer that measures the optical path length difference distribution without phase unwrapping using optical phase locking. The applicability of our phase imaging system was demonstrated by successful identification of breast cancer cells amongst normal cells. However, the cell identification method using this phase imaging system exhibited a false identification rate of approximately 7%. In this study, we implemented a polarimetric imaging system by introducing a polarimetric module to one arm of the Mach-Zehnder interferometer of our conventional phase imaging system. This module was comprised of a quarter wave plate and a rotational polarizer on the illumination side of the sample, and a linear polarizer on the optical detector side. In addition, we developed correction methods for the measurement errors of the optical path length and birefringence phase differences that arose through the influence of elements other than cells, such as the Petri dish. As the Petri dish holding the fluid specimens was transparent, it did not affect the amplitude information; however, the optical path length and birefringence phase differences were affected. Therefore, we proposed correction of the optical path length and birefringence phase for the influence of elements other than cells, as a prerequisite for obtaining highly precise phase and polarimetric images.

Portable fiber-optic taper coupled optical microscopy platform

NASA Astrophysics Data System (ADS)

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

2017-04-01

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

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.

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.

A frameless stereotaxic operating microscope for neurosurgery.

PubMed

Friets, E M; Strohbehn, J W; Hatch, J F; Roberts, D W

1989-06-01

A new system, which we call the frameless stereotaxic operating microscope, is discussed. Its purpose is to display CT or other image data in the operating microscope in the correct scale, orientation, and position without the use of a stereotaxic frame. A nonimaging ultrasonic rangefinder allows the position of the operating microscope and the position of the patient to be determined. Discrete fiducial points on the patient's external anatomy are located in both image space and operating room space, linking the image data and the operating room. Physician-selected image information, e.g., tumor contours or guidance to predetermined targets, is projected through the optics of the operating microscope using a miniature cathode ray tube and a beam splitter. Projected images superpose the surgical field, reconstructed from image data to match the focal plane of the operating microscope. The algorithms on which the system is based are described, and the sources and effects of errors are discussed. The system's performance is simulated, providing an estimate of accuracy. Two phantoms are used to measure accuracy experimentally. Clinical results and observations are given.

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.

Characteristics of the annular beam using a single axicon and a pair of lens

NASA Astrophysics Data System (ADS)

Ji, Ke; Lei, Ming; Yao, Baoli; Yan, Shaohui; Yang, Yanlong; Li, Ze; Dan, Dan; Menke, Neimule

2012-10-01

In optical trapping, annular beam as a kind of hollow beam is used to increase the axial trapping efficiency as well as the trapping stability. In this paper, a method for producing an annular beam by a system consisting of a single axicon and a pair of lens is proposed. The generated beam was also used as the optical tweezers. We use the geometrical optics to describe the propagation of light in the system. The calculated intensity distribution in three-dimensional space after the system shows a good agreement with the experimental results. The advantages of this method are simplicity of operation, good stability, and high transmittance, having possible applications in fields like optical microscopic, optical manipulation and electronic acceleration, etc.

Confocal Microscopy Imaging with an Optical Transition Edge Sensor

NASA Astrophysics Data System (ADS)

Fukuda, D.; Niwa, K.; Hattori, K.; Inoue, S.; Kobayashi, R.; Numata, T.

2018-05-01

Fluorescence color imaging at an extremely low excitation intensity was performed using an optical transition edge sensor (TES) embedded in a confocal microscope for the first time. Optical TES has the ability to resolve incident single photon energy; therefore, the wavelength of each photon can be measured without spectroscopic elements such as diffraction gratings. As target objects, animal cells labeled with two fluorescent dyes were irradiated with an excitation laser at an intensity below 1 μW. In our confocal system, an optical fiber-coupled TES device is used to detect photons instead of the pinhole and photomultiplier tube used in typical confocal microscopes. Photons emitted from the dyes were collected by the objective lens, and sent to the optical TES via the fiber. The TES measures the wavelength of each photon arriving in an exposure time of 70 ms, and a fluorescent photon spectrum is constructed. This measurement is repeated by scanning the target sample, and finally a two-dimensional RGB-color image is obtained. The obtained image showed that the photons emitted from the dyes of mitochondria and cytoskeletons were clearly resolved at a detection intensity level of tens of photons. TES exhibits ideal performance as a photon detector with a low dark count rate (< 1 Hz) and wavelength resolving power. In the single-mode fiber-coupled system, the confocal microscope can be operated in the super-resolution mode. These features are very promising to realize high-sensitivity and high-resolution photon spectral imaging, and would help avoid cell damage and photobleaching of fluorescence dyes.

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

PubMed

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

2007-05-01

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

Optical analysis of electro-optical systems by MTF calculus

NASA Astrophysics Data System (ADS)

Barbarini, Elisa Signoreto; Dos Santos, Daniel, Jr.; Stefani, Mário Antonio; Yasuoka, Fátima Maria Mitsue; Castro Neto, Jarbas C.; Rodrigues, Evandro Luís Linhari

2011-08-01

One of the widely used methods for performance analysis of an optical system is the determination of the Modulation Transfer Function (MTF). The MTF represents a quantitative and direct measure of image quality, and, besides being an objective test, it can be used on concatenated optical system. This paper presents the application of software called SMTF (software modulation transfer function), built in C++ and Open CV platforms for MTF calculation on electro-optical system. Through this technique, it is possible to develop specific method to measure the real time performance of a digital fundus camera, an infrared sensor and an ophthalmological surgery microscope. Each optical instrument mentioned has a particular device to measure the MTF response, which is being developed. Then the MTF information assists the analysis of the optical system alignment, and also defines its resolution limit by the MTF graphic. The result obtained from the implemented software is compared with the theoretical MTF curve from the analyzed systems.

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

PubMed Central

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

2012-01-01

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

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.

An intraoperative spectroscopic imaging system for quantification of Protoporphyrin IX during glioma surgery (Conference Presentation)

NASA Astrophysics Data System (ADS)

Angulo-Rodríguez, Leticia M.; Laurence, Audrey; Jermyn, Michael; Sheehy, Guillaume; Sibai, Mira; Petrecca, Kevin; Roberts, David W.; Paulsen, Keith D.; Wilson, Brian C.; Leblond, Frédéric

2016-03-01

Cancer tissue often remains after brain tumor resection due to the inability to detect the full extent of cancer during surgery, particularly near tumor boundaries. Commercial systems are available for intra-operative real-time aminolevulenic acid (ALA)-induced protoporphyrin IX (PpIX) fluorescence imaging. These are standard white-light neurosurgical microscopes adapted with optical components for fluorescence excitation and detection. However, these instruments lack sensitivity and specificity, which limits the ability to detect low levels of PpIX and distinguish it from tissue auto-fluorescence. Current systems also cannot provide repeatable and un-biased quantitative fluorophore concentration values because of the unknown and highly variable light attenuation by tissue. We present a highly sensitive spectroscopic fluorescence imaging system that is seamlessly integrated onto a neurosurgical microscope. Hardware and software were developed to achieve through-microscope spatially-modulated illumination for 3D profilometry and to use this information to extract tissue optical properties to correct for the effects of tissue light attenuation. This gives pixel-by-pixel quantified fluorescence values and improves detection of low PpIX concentrations. This is achieved using a high-sensitivity Electron Multiplying Charge Coupled Device (EMCCD) with a Liquid Crystal Tunable Filter (LCTF) whereby spectral bands are acquired sequentially; and a snapshot camera system with simultaneous acquisition of all bands is used for profilometry and optical property recovery. Sensitivity and specificity to PpIX is demonstrated using brain tissue phantoms and intraoperative human data acquired in an on-going clinical study using PpIX fluorescence to guide glioma resection.

Toolkit for the Automated Characterization of Optical Trapping Forces on Microscopic Particles

NASA Astrophysics Data System (ADS)

Glaser, Joseph; Hoeprich, David; Resnick, Andrew

2014-03-01

Optical traps have been in use in microbiological studies for the past 40 years to obtain noninvasive control of microscopic particles. However, the magnitude of the applied forces is often unknown. Therefore, we have developed an automated data acquisition and processing system which characterizes trap properties for known particle geometries. Extensive experiments and measurements utilizing well-characterized objects were performed and compared to literature to confirm the system's performance. This system will enable the future analysis of a trapped primary cilium, a slender rod-shaped organelle with aspect ratio L/R >30, where `L' is the cilium length and `R' the cilium diameter. The trapping of cilia is of primary importance, as it will lead to the precise measurements of mechanical properties of the organelle and its significance to the epithelial cell. Support from the National Institutes of Health, 1R15DK092716 is gratefully acknowledged.

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.

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.

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.

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

Micro-optics technology and sensor systems applications

NASA Technical Reports Server (NTRS)

Gal, George; Herman, B.; Anderson, W.; Whitney, R.; Morrow, H.

1993-01-01

The current generation of electro-optical sensors utilizing refractive and reflective optical elements require sophisticated, complex, and expensive designs. Advanced-technology-based electro-optical sensors of minimum size and weight require miniaturization of optical, electrical, and mechanical devices with an increasing trend toward integration of various components. Micro-optics technology has the potential in a number of areas to simplify optical design with improved performance. This includes internally cooled apertures, hybrid optical design, microlenses, dispersive multicolor microlenses, active dither, electronically controlled optical beam steer, and microscopic integration of micro-optics, detectors, and signal processing layers. This paper describes our approach to the development of micro-optics technology with our main emphasis for sensors applications.

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 tracking of nanoscale particles in microscale environments

NASA Astrophysics Data System (ADS)

Mathai, P. P.; Liddle, J. A.; Stavis, S. M.

2016-03-01

The trajectories of nanoscale particles through microscale environments record useful information about both the particles and the environments. Optical microscopes provide efficient access to this information through measurements of light in the far field from nanoparticles. Such measurements necessarily involve trade-offs in tracking capabilities. This article presents a measurement framework, based on information theory, that facilitates a more systematic understanding of such trade-offs to rationally design tracking systems for diverse applications. This framework includes the degrees of freedom of optical microscopes, which determine the limitations of tracking measurements in theory. In the laboratory, tracking systems are assemblies of sources and sensors, optics and stages, and nanoparticle emitters. The combined characteristics of such systems determine the limitations of tracking measurements in practice. This article reviews this tracking hardware with a focus on the essential functions of nanoparticles as optical emitters and microenvironmental probes. Within these theoretical and practical limitations, experimentalists have implemented a variety of tracking systems with different capabilities. This article reviews a selection of apparatuses and techniques for tracking multiple and single particles by tuning illumination and detection, and by using feedback and confinement to improve the measurements. Prior information is also useful in many tracking systems and measurements, which apply across a broad spectrum of science and technology. In the context of the framework and review of apparatuses and techniques, this article reviews a selection of applications, with particle diffusion serving as a prelude to tracking measurements in biological, fluid, and material systems, fabrication and assembly processes, and engineered devices. In so doing, this review identifies trends and gaps in particle tracking that might influence future research.

Handheld optical-resolution photoacoustic microscopy

NASA Astrophysics Data System (ADS)

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

2017-04-01

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

Inspection and characterization of flexo-printing plates

NASA Astrophysics Data System (ADS)

Hahlweg, Cornelius; Pescoller, Lukas; Zhao, Wenjing

2013-09-01

In continuation of last year's paper on distorting optics for inspection of 2 1/2D surfaces with convex or elevated elements - like braille paper and other special printing products - the present paper is dedicated to the quality control and characterization of flexo-printing plates. The need for high optical resolution contradicts the need for depth of field. A rugged optical system for gathering a series of microscopic images at various planes of focus is discussed.

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.

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.

Standoff detection of explosives: a challenging approach for optical technologies

NASA Astrophysics Data System (ADS)

Désilets, S.; Hô, N.; Mathieu, P.; Simard, J. R.; Puckrin, E.; Thériault, J. M.; Lavoie, H.; Théberge, F.; Babin, F.; Gay, D.; Forest, R.; Maheux, J.; Roy, G.; Châteauneuf, M.

2011-06-01

Standoff detection of explosives residues on surfaces at few meters was made using optical technologies based on Raman scattering, Laser-Induced Breakdown Spectroscopy (LIBS) and passive standoff FTIR radiometry. By comparison, detection and analysis of nanogram samples of different explosives was made with a microscope system where Raman scattering from a micron-size single point illuminated crystal of explosive was observed. Results from standoff detection experiments using a telescope were compared to experiments using a microscope to find out important parameters leading to the detection. While detection and spectral identification of the micron-size explosive particles was possible with a microscope, standoff detection of these particles was very challenging due to undesired light reflected and produced by the background surface or light coming from other contaminants. Results illustrated the challenging approach of detecting at a standoff distance the presence of low amount of micron or submicron explosive particles.

Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo.

PubMed

Engelbrecht, Christoph J; Johnston, Richard S; Seibel, Eric J; Helmchen, Fritjof

2008-04-14

We present a small, lightweight two-photon fiberscope and demonstrate its suitability for functional imaging in the intact brain. Our device consists of a hollow-core photonic crystal fiber for efficient delivery of near-IR femtosecond laser pulses, a spiral fiber-scanner for resonant beam steering, and a gradient-index lens system for fluorescence excitation, dichroic beam splitting, and signal collection. Fluorescence light is remotely detected using a standard photomultiplier tube. All optical components have 1 mm dimensions and the microscope's headpiece weighs only 0.6 grams. The instrument achieves micrometer resolution at frame rates of typically 25 Hz with a field-of-view of up to 200 microns. We demonstrate functional imaging of calcium signals in Purkinje cell dendrites in the cerebellum of anesthetized rats. The microscope will be easily portable by a rat or mouse and thus should enable functional imaging in freely behaving animals.

Shack-Hartmann reflective micro profilometer

NASA Astrophysics Data System (ADS)

Gong, Hai; Soloviev, Oleg; Verhaegen, Michel; Vdovin, Gleb

2018-01-01

We present a quantitative phase imaging microscope based on a Shack-Hartmann sensor, that directly reconstructs the optical path difference (OPD) in reflective mode. Comparing with the holographic or interferometric methods, the SH technique needs no reference beam in the setup, which simplifies the system. With a preregistered reference, the OPD image can be reconstructed from a single shot. Also, the method has a rather relaxed requirement on the illumination coherence, thus a cheap light source such as a LED is feasible in the setup. In our previous research, we have successfully verified that a conventional transmissive microscope can be transformed into an optical path difference microscope by using a Shack-Hartmann wavefront sensor under incoherent illumination. The key condition is that the numerical aperture of illumination should be smaller than the numerical aperture of imaging lens. This approach is also applicable to characterization of reflective and slightly scattering surfaces.

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.

In vivo study of rat cortical hemodynamics using a stereotaxic-apparatus-compatible photoacoustic microscope.

PubMed

Guo, Heng; Chen, Qian; Qi, Weizhi; Chen, Xingxing; Xi, Lei

2018-04-19

Brain imaging is an important technique in cognitive neuroscience. In this article, we designed a stereotaxic-apparatus-compatible photoacoustic microscope for the studies of rat cortical hemodynamics. Compared with existing optical resolution photoacoustic microscopy (ORPAM) systems, the probe owns feature of fast, light and miniature. In this microscope, we integrated a miniaturized ultrasound transducer with a center frequency of 10 MHz to detect photoacoustic signals and a 2-dimensional (2D) microelectromechanical system (MEMS) scanner to achieve raster scanning of the optical focus. Based on phantom evaluation, this imaging probe has a high lateral resolution of 3.8 μm and an effective imaging domain of 2 × 2 mm 2 . Different from conventional ORPAMs, combining with standard stereotaxic apparatus enables broad studies of rodent brains without any motion artifact. To show its capability, we successfully captured red blood cell flow in the capillary, monitored the vascular changes during bleeding and blood infusion and visualized cortical hemodynamics induced by middle cerebral artery occlusion. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Sub-cell turning to accomplish micron-level alignment of precision assemblies

NASA Astrophysics Data System (ADS)

Kumler, James J.; Buss, Christian

2017-08-01

Higher performance expectations for complex optical systems demand tighter alignment requirements for lens assembly alignment. In order to meet diffraction limited imaging performance over wide spectral bands across the UV and visible wavebands, new manufacturing approaches and tools must be developed if the optical systems will be produced consistently in volume production. This is especially applicable in the field of precision microscope objectives for life science, semiconductor inspection and laser material processing systems. We observe a rising need for the improvement in the optical imaging performance of objective lenses. The key challenge lies in the micron-level decentration and tilt of each lens element. One solution for the production of high quality lens systems is sub-cell assembly with alignment turning. This process relies on an automatic alignment chuck to align the optical axis of a mounted lens to the spindle axis of the machine. Subsequently, the mount is cut with diamond tools on a lathe with respect to the optical axis of the mount. Software controlled integrated measurement technology ensures highest precision. In addition to traditional production processes, further dimensions can be controlled in a very precise manner, e.g. the air gaps between the lenses. Using alignment turning simplifies further alignment steps and reduces the risk of errors. This paper describes new challenges in microscope objective design and manufacturing, and addresses difficulties with standard production processes. A new measurement and alignment technique is described, and strengths and limitations are outlined.

Microscope-Integrated OCT Feasibility and Utility With the EnFocus System in the DISCOVER Study.

PubMed

Runkle, Anne; Srivastava, Sunil K; Ehlers, Justis P

2017-03-01

To evaluate the feasibility and utility of a novel microscope-integrated intraoperative optical coherence tomography (OCT) system. The DISCOVER study is an investigational device study evaluating microscope-integrated intraoperative OCT systems for ophthalmic surgery. This report focuses on subjects imaged with the EnFocus prototype system (Leica Microsystems/Bioptigen, Morrisville, NC). OCT was performed at surgeon-directed milestones. Surgeons completed a questionnaire after each case to evaluate the impact of OCT on intraoperative management. Fifty eyes underwent imaging with the EnFocus system. Successful imaging was obtained in 46 of 50 eyes (92%). In eight cases (16%), surgical management was changed based on intraoperative OCT findings. In membrane peeling procedures, intraoperative OCT findings were discordant from the surgeon's initial impression in seven of 20 cases (35%). This study demonstrates the feasibility of microscope-integrated intraoperative OCT using the Bioptigen EnFocus system. Intraoperative OCT may provide surgeons with additional information that may influence surgical decision-making. [Ophthalmic Surg Lasers Imaging Retina. 2017;48:216-222.]. Copyright 2017, SLACK Incorporated.

Observation of development of breast cancer cell lines in real time by fluorescence microscopy under simulated microgravity

NASA Astrophysics Data System (ADS)

Lavan, David; Valdivia-Silva, Julio E.; Sanabria, Gabriela; Orihuela, Diego; Suarez, Juan; Quispe, Marco; Chuchon, Mariano; Martin, David; Maroto, Marcos; Egea, Javier

2016-07-01

This project consist in the implementation of a fluorescence microscope for the in real time monitoring of biological labeled samples by several fluorophores in microgravity conditions keeping the temperature, humidity, and (CO)2 controlled by an electronic platform. The system (fluorescence microscope and incubator) is integrated to a microgravity simulator machine which was presented on the "30th Annual American Society for Gravitation and Space Research Meeting" October 2014 in Pasadena, CA, USA. Currently, we have the microgravity machine biologically validated by genetic expression studies in pupal stage of Drosophila melanogaster. The fluorescence microscope has a platform designed to hold a culture flask, and a fluorescence camera (Leica DFC3000 G) connected to an optical system (Fluorescence Light source Leica EL6000, optic fiber, fiber adapter, and fluorescence filter) in order to take images in real time. The mechanical system of the fluorescence microsc ope is designed to allow the displacement of the fluorescence camera through a parallel plane to the culture flask's plane and also the movement of the platform through a perpendicular axis to the culture flask in order to focus the samples to the optical system. The mechanical system is propelled by four DC moto-reductors with encoder (A-max 26 Maxon motor, GP 32S screw and MR encoder) that generate displacements in the order of micrometers. The angular position control of the DC motoreductor's shaft of all the DC moto-reductors is done by PWM signals based on the interpretation of the signals provided by the encoders during the movement. The system is remotely operated by a graphic interface installed on a personal computer or any mobile device (smartphone, laptop or tablet) by using the internet. Acknowledgments: Grant of INNOVATE PERU (Formerly FINCYT)

Polarization anisotropy in fiber-optic second harmonic generation microscopy.

PubMed

Fu, Ling; Gu, Min

2008-03-31

We report the investigation and implementation of a compact second harmonic generation microscope that uses a single-mode fiber coupler and a double-clad photonic crystal fiber. Second harmonic polarization anisotropy through the fiber-optic microscope systems is quantitatively measured with KTP microcrystals, fish scale and rat tail tendon. It is demonstrated that the polarized second harmonic signals can be excited and collected through the single-mode fiber coupler to analyze the molecular orientations of structural proteins. It has been discovered that a double-clad photonic crystal fiber can preserve the linear polarization in the core, although a depolarization effect is observed in the inner cladding region. The feasibility of polarization anisotropy measurements in fiber-optic second harmonic generation microscopy will benefit the in vivo study of collagen-related diseases with a compact imaging probe.

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.

Malaria Diagnosis Using a Mobile Phone Polarized Microscope

NASA Astrophysics Data System (ADS)

Pirnstill, Casey W.; Coté, Gerard L.

2015-08-01

Malaria remains a major global health burden, and new methods for low-cost, high-sensitivity, diagnosis are essential, particularly in remote areas with low-resource around the world. In this paper, a cost effective, optical cell-phone based transmission polarized light microscope system is presented for imaging the malaria pigment known as hemozoin. It can be difficult to determine the presence of the pigment from background and other artifacts, even for skilled microscopy technicians. The pigment is much easier to observe using polarized light microscopy. However, implementation of polarized light microscopy lacks widespread adoption because the existing commercial devices have complicated designs, require sophisticated maintenance, tend to be bulky, can be expensive, and would require re-training for existing microscopy technicians. To this end, a high fidelity and high optical resolution cell-phone based polarized light microscopy system is presented which is comparable to larger bench-top polarized microscopy systems but at much lower cost and complexity. The detection of malaria in fixed and stained blood smears is presented using both, a conventional polarized microscope and our cell-phone based system. The cell-phone based polarimetric microscopy design shows the potential to have both the resolution and specificity to detect malaria in a low-cost, easy-to-use, modular platform.

Malaria Diagnosis Using a Mobile Phone Polarized Microscope

PubMed Central

Pirnstill, Casey W.; Coté, Gerard L.

2015-01-01

Malaria remains a major global health burden, and new methods for low-cost, high-sensitivity, diagnosis are essential, particularly in remote areas with low-resource around the world. In this paper, a cost effective, optical cell-phone based transmission polarized light microscope system is presented for imaging the malaria pigment known as hemozoin. It can be difficult to determine the presence of the pigment from background and other artifacts, even for skilled microscopy technicians. The pigment is much easier to observe using polarized light microscopy. However, implementation of polarized light microscopy lacks widespread adoption because the existing commercial devices have complicated designs, require sophisticated maintenance, tend to be bulky, can be expensive, and would require re-training for existing microscopy technicians. To this end, a high fidelity and high optical resolution cell-phone based polarized light microscopy system is presented which is comparable to larger bench-top polarized microscopy systems but at much lower cost and complexity. The detection of malaria in fixed and stained blood smears is presented using both, a conventional polarized microscope and our cell-phone based system. The cell-phone based polarimetric microscopy design shows the potential to have both the resolution and specificity to detect malaria in a low-cost, easy-to-use, modular platform. PMID:26303238

A Multiscale Material Testing System for In Situ Optical and Electron Microscopes and Its Application

PubMed Central

Ye, Xuan; Cui, Zhiguo; Fang, Huajun; Li, Xide

2017-01-01

We report a novel material testing system (MTS) that uses hierarchical designs for in-situ mechanical characterization of multiscale materials. This MTS is adaptable for use in optical microscopes (OMs) and scanning electron microscopes (SEMs). The system consists of a microscale material testing module (m-MTM) and a nanoscale material testing module (n-MTM). The MTS can measure mechanical properties of materials with characteristic lengths ranging from millimeters to tens of nanometers, while load capacity can vary from several hundred micronewtons to several nanonewtons. The m-MTM is integrated using piezoelectric motors and piezoelectric stacks/tubes to form coarse and fine testing modules, with specimen length from millimeters to several micrometers, and displacement distances of 12 mm with 0.2 µm resolution for coarse level and 8 µm with 1 nm resolution for fine level. The n-MTM is fabricated using microelectromechanical system technology to form active and passive components and realizes material testing for specimen lengths ranging from several hundred micrometers to tens of nanometers. The system’s capabilities are demonstrated by in-situ OM and SEM testing of the system’s performance and mechanical properties measurements of carbon fibers and metallic microwires. In-situ multiscale deformation tests of Bacillus subtilis filaments are also presented. PMID:28777341

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

PubMed Central

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

2007-01-01

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

Preparing and probing many-body correlated systems in a Quantum Gas Microscope by engineering arbitrary landscape potentials

NASA Astrophysics Data System (ADS)

Rispoli, Matthew; Lukin, Alexander; Ma, Ruichao; Preiss, Philipp; Tai, M. Eric; Islam, Rajibul; Greiner, Markus

2015-05-01

Ultracold atoms in optical lattices provide a versatile tool box for observing the emergence of strongly correlated physics in quantum systems. Dynamic control of optical potentials on the single-site level allows us to prepare and probe many-body quantum states through local Hamiltonian engineering. We achieve these high precision levels of optical control through spatial light modulation with a DMD (digital micro-mirror device). This allows for both arbitrary beam shaping and aberration compensation in our imaging system to produce high fidelity optical potentials. We use these techniques to control state initialization, Hamiltonian dynamics, and measurement in experiments investigating low-dimensional many-body physics - from one-dimensional correlated quantum walks to characterizing entanglement.

Parallel and patterned optogenetic manipulation of neurons in the brain slice using a DMD-based projector.

PubMed

Sakai, Seiichiro; Ueno, Kenichi; Ishizuka, Toru; Yawo, Hiromu

2013-01-01

Optical manipulation technologies greatly advanced the understanding of the neuronal network and its dysfunctions. To achieve patterned and parallel optical switching, we developed a microscopic illumination system using a commercial DMD-based projector and a software program. The spatiotemporal patterning of the system was evaluated using acute slices of the hippocampus. The neural activity was optically manipulated, positively by the combination of channelrhodopsin-2 (ChR2) and blue light, and negatively by the combination of archaerhodopsin-T (ArchT) and green light. It is suggested that our projector-managing optical system (PMOS) would effectively facilitate the optogenetic analyses of neurons and their circuits. Copyright © 2012 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

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.

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.

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

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

NASA Astrophysics Data System (ADS)

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

2017-12-01

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

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

PubMed

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

2013-06-01

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

Animated Optical Microscope Zoom in from Phoenix Launch to Martian Surface

NASA Technical Reports Server (NTRS)

2008-01-01

[figure removed for brevity, see original site] Click on image for animation

This animated camera view zooms in from NASA's Phoenix Mars Lander launch site all the way to Phoenix's Microscopy and Electrochemistry and C Eonductivity Analyzer (MECA) aboard the spacecraft on the Martian surface. The final frame shows the soil sample delivered to MECA as viewed through the Optical Microscope (OM) on Sol 17 (June 11, 2008), or the 17th Martian day.

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.

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.

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

NASA Astrophysics Data System (ADS)

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

2017-02-01

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

Purple sea urchin Strongylocentrotus purpuratus gamete manipulation using optical trapping and microfluidics

NASA Astrophysics Data System (ADS)

Chandsawangbhuwana, Charlie; Shi, Linda Z.; Zhu, Qingyuan; Berns, Michael W.

2013-04-01

A system has been developed that allows for optical and fluidic manipulation of gametes. The optical manipulation is performed by using a single-point gradient trap with a 40× oil immersion PH3 1.3 NA objective on a Zeiss inverted microscope. The fluidic manipulation is performed by using a custom microfluidic chamber designed to fit into the short working distance between the condenser and objective. The system is validated using purple sea urchin Strongylocentrotus purpuratus gametes and has the potential to be used for mammalian in vitro fertilization and animal husbandry.

Investigation of the potential of optical coherence tomography (OCT) as a non-invasive diagnostic tool in reproductive medicine

NASA Astrophysics Data System (ADS)

Trottmann, Matthias; Homann, Christian; Leeb, R.; Doering, D.; Kuznetsova, J.; Reese, S.; Stief, C. G.; Koelle, S.; Sroka, R.

2015-02-01

Introduction and objective: In Europe, nearly every sixth couple in the reproductive age is involuntarily childless. In about 30%, both male and female reveal fertility problems. In about 10% of infertile men, azoospermia is the underlying cause. As conventional therapeutic options are limited, surgical testicular sperm extraction (TESE) is necessary to obtain sperms for assisted reproductive techniques. Regarding the females, up to 30% of all idiopathic infertilities are due to alterations of the uterine tube So far, no imaging technique, which does not require any labelling, is available to evaluate the male and female genital tract at a microscopic level under in vivo conditions. Thus, the aim of this study was to investigate the potential of optical coherence tomography (OCT) as a non-invasive diagnostic tool in gynaecology and andrology. Material and Methods: Tissues samples from the bovine testis, epididymis, vas deferens, ovary, oviduct (ampulla and isthmus) and uterus were obtained immediately after slaughter (14 cows aged 3 to 8 years and 14 bulls aged 3 to 6 years; breeds: Holstein- Friesian, and Deutsches Fleckvieh). Imaging was done by using the US Food and Drug Administration (FDA) approved probe-based Niris Imaging System (Imalux, Cleveland, Ohio, USA) and the Telesto 1325 nm OCT System and Ganymede 930 nm OCT System (Thorlabs Inc., Dachau, Germany). All images obtained were compared to histological images after paraffin embedding and HE staining. Results: OCT imaging visualized the microarchitecture of the testis, epididymis, spermatic duct and the ovary, oviduct and uterus. Using the Thorlabs systems a axial resolution of approx. 5μm and lateral resolution of 8- 15μm could be achieved. Different optical tissue volumes could be visualized, which depends on the optical penetration depth of the wavelength of the system used. While the tissue volume observed by probe based Imalux-OCT is similar to the used Thorlabs systems, the optical resolution is reduced. By means of the microscopic OCT-system differentiation of testical tissue structures like content and diameter of seminiferous tubules and the epididymal duct was possible. Structures of the female oviduct, like the primary, secondary and tertiary folds including the typical epithelium consisting of secretory and ciliated cells were identified. Ampulla and isthmus were clearly differentiated by the height of the folds and the thickness of the smooth muscle layer. Imaging was successful both from the outside wall and from the inner lumen. After experience with microscopic OCT-structure identification such structures could also be identified by means of probe based OCT. Conclusions: Technical improvement of probe-based OCT up to a high-resolution level of nowadays-available OCT microscopic systems could open up new ways of in vivo imaging in the reproductive tract. Potential applications could be an OCT-guided testicular biopsy for improving sperm retrieval or microscopic evaluation of the oviduct by OCT-assisted fertiloscopy. The latter would provide a valuable tool to facilitate the decision of which type of assisted reproductive techniques might be preferred.

Ion photon emission microscope

DOEpatents

Doyle, Barney L.

2003-04-22

An ion beam analysis system that creates microscopic multidimensional image maps of the effects of high energy ions from an unfocussed source upon a sample by correlating the exact entry point of an ion into a sample by projection imaging of the ion-induced photons emitted at that point with a signal from a detector that measures the interaction of that ion within the sample. The emitted photons are collected in the lens system of a conventional optical microscope, and projected on the image plane of a high resolution single photon position sensitive detector. Position signals from this photon detector are then correlated in time with electrical effects, including the malfunction of digital circuits, detected within the sample that were caused by the individual ion that created these photons initially.

Miniaturized video-rate epi-third-harmonic-generation fiber-microscope.

PubMed

Chia, Shih-Hsuan; Yu, Che-Hang; Lin, Chih-Han; Cheng, Nai-Chia; Liu, Tzu-Ming; Chan, Ming-Che; Chen, I-Hsiu; Sun, Chi-Kuang

2010-08-02

With a micro-electro-mechanical system (MEMS) mirror, we successfully developed a miniaturized epi-third-harmonic-generation (epi-THG) fiber-microscope with a video frame rate (31 Hz), which was designed for in vivo optical biopsy of human skin. With a large-mode-area (LMA) photonic crystal fiber (PCF) and a regular microscopic objective, the nonlinear distortion of the ultrafast pulses delivery could be much reduced while still achieving a 0.4 microm lateral resolution for epi-THG signals. In vivo real time virtual biopsy of the Asian skin with a video rate (31 Hz) and a sub-micron resolution was obtained. The result indicates that this miniaturized system was compact enough for the least invasive hand-held clinical use.

A Low-Cost Digital Microscope with Real-Time Fluorescent Imaging Capability.

PubMed

Hasan, Md Mehedi; Alam, Mohammad Wajih; Wahid, Khan A; Miah, Sayem; Lukong, Kiven Erique

2016-01-01

This paper describes the development of a prototype of a low-cost digital fluorescent microscope built from commercial off-the-shelf (COTS) components. The prototype was tested to detect malignant tumor cells taken from a living organism in a preclinical setting. This experiment was accomplished by using Alexa Fluor 488 conjugate dye attached to the cancer cells. Our prototype utilizes a torch along with an excitation filter as a light source for fluorophore excitation, a dichroic mirror to reflect the excitation and pass the emitted green light from the sample under test and a barrier filter to permit only appropriate wavelength. The system is designed out of a microscope using its optical zooming property and an assembly of exciter filter, dichroic mirror and transmitter filter. The microscope is connected to a computer or laptop through universal serial bus (USB) that allows real-time transmission of captured florescence images; this also offers real-time control of the microscope. The designed system has comparable features of high-end commercial fluorescent microscopes while reducing cost, power, weight and size.

A Low-Cost Digital Microscope with Real-Time Fluorescent Imaging Capability

PubMed Central

Hasan, Md. Mehedi; Wahid, Khan A.; Miah, Sayem; Lukong, Kiven Erique

2016-01-01

This paper describes the development of a prototype of a low-cost digital fluorescent microscope built from commercial off-the-shelf (COTS) components. The prototype was tested to detect malignant tumor cells taken from a living organism in a preclinical setting. This experiment was accomplished by using Alexa Fluor 488 conjugate dye attached to the cancer cells. Our prototype utilizes a torch along with an excitation filter as a light source for fluorophore excitation, a dichroic mirror to reflect the excitation and pass the emitted green light from the sample under test and a barrier filter to permit only appropriate wavelength. The system is designed out of a microscope using its optical zooming property and an assembly of exciter filter, dichroic mirror and transmitter filter. The microscope is connected to a computer or laptop through universal serial bus (USB) that allows real-time transmission of captured florescence images; this also offers real-time control of the microscope. The designed system has comparable features of high-end commercial fluorescent microscopes while reducing cost, power, weight and size. PMID:27977709

Correction of cell-induced optical aberrations in a fluorescence fluctuation microscope

PubMed Central

Leroux, Charles-Edouard; Grichine, Alexei; Wang, Irène; Delon, Antoine

2013-01-01

We describe the effect of optical aberrations on fluorescence fluctuations microscopy (FFM), when focusing through a single living cell. FFM measurements are performed in an aqueous fluorescent solution, and prove to be a highly sensitive tool to assess the optical aberrations introduced by the cell. We demonstrate an adaptive optics (AO) system to remove the aberration-related bias in the FFM measurements. Our data show that AO is not only useful when imaging deep in tissues, but also when performing FFM measurements through a single cellular layer. PMID:23939061

Scientific Applications of Optical Instruments to Materials Research

NASA Technical Reports Server (NTRS)

Witherow, William K.

1997-01-01

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

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

PubMed

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

2008-07-01

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

Compact Video Microscope Imaging System Implemented in Colloid Studies

NASA Technical Reports Server (NTRS)

McDowell, Mark

2002-01-01

Long description Photographs showing fiber-optic light source, microscope and charge-coupled discharge (CCD) camera head connected to camera body, CCD camera body feeding data to image acquisition board in PC, and Cartesian robot controlled via PC board. The Compact Microscope Imaging System (CMIS) is a diagnostic tool with intelligent controls for use in space, industrial, medical, and security applications. CMIS can be used in situ with a minimum amount of user intervention. This system can scan, find areas of interest in, focus on, and acquire images automatically. Many multiple-cell experiments require microscopy for in situ observations; this is feasible only with compact microscope systems. CMIS is a miniature machine vision system that combines intelligent image processing with remote control. The software also has a user-friendly interface, which can be used independently of the hardware for further post-experiment analysis. CMIS has been successfully developed in the SML Laboratory at the NASA Glenn Research Center and adapted for use for colloid studies and is available for telescience experiments. The main innovations this year are an improved interface, optimized algorithms, and the ability to control conventional full-sized microscopes in addition to compact microscopes. The CMIS software-hardware interface is being integrated into our SML Analysis package, which will be a robust general-purpose image-processing package that can handle over 100 space and industrial applications.

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).

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.

Optical tracking of nanoscale particles in microscale environments

PubMed Central

Mathai, P. P.; Liddle, J. A.; Stavis, S. M.

2016-01-01

The trajectories of nanoscale particles through microscale environments record useful information about both the particles and the environments. Optical microscopes provide efficient access to this information through measurements of light in the far field from nanoparticles. Such measurements necessarily involve trade-offs in tracking capabilities. This article presents a measurement framework, based on information theory, that facilitates a more systematic understanding of such trade-offs to rationally design tracking systems for diverse applications. This framework includes the degrees of freedom of optical microscopes, which determine the limitations of tracking measurements in theory. In the laboratory, tracking systems are assemblies of sources and sensors, optics and stages, and nanoparticle emitters. The combined characteristics of such systems determine the limitations of tracking measurements in practice. This article reviews this tracking hardware with a focus on the essential functions of nanoparticles as optical emitters and microenvironmental probes. Within these theoretical and practical limitations, experimentalists have implemented a variety of tracking systems with different capabilities. This article reviews a selection of apparatuses and techniques for tracking multiple and single particles by tuning illumination and detection, and by using feedback and confinement to improve the measurements. Prior information is also useful in many tracking systems and measurements, which apply across a broad spectrum of science and technology. In the context of the framework and review of apparatuses and techniques, this article reviews a selection of applications, with particle diffusion serving as a prelude to tracking measurements in biological, fluid, and material systems, fabrication and assembly processes, and engineered devices. In so doing, this review identifies trends and gaps in particle tracking that might influence future research. PMID:27213022

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.

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.

Excitonic linewidth and coherence lifetime in monolayer transition metal dichalcogenides

DOE PAGES

Selig, Malte; Berghäuser, Gunnar; Raja, Archana; ...

2016-11-07

Atomically thin transition metal dichalcogenides are direct-gap semiconductors with strong light–matter and Coulomb interactions. The latter accounts for tightly bound excitons, which dominate their optical properties. Besides the optically accessible bright excitons, these systems exhibit a variety of dark excitonic states. They are not visible in the optical spectra, but can strongly influence the coherence lifetime and the linewidth of the emission from bright exciton states. We investigate the microscopic origin of the excitonic coherence lifetime in two representative materials (WS 2 and MoSe 2) through a study combining microscopic theory with spectroscopic measurements. We also show that the excitonicmore » coherence lifetime is determined by phonon-induced intravalley scattering and intervalley scattering into dark excitonic states. Particularly, we identify exciton relaxation processes involving phonon emission into lower-lying dark states that are operative at all temperatures, in WS 2.« less

Optical method for high magnification imaging and video recording of live cells at sub-micron resolution

NASA Astrophysics Data System (ADS)

Romo, Jaime E., Jr.

Optical microscopy, the most common technique for viewing living microorganisms, is limited in resolution by Abbe's criterion. Recent microscopy techniques focus on circumnavigating the light diffraction limit by using different methods to obtain the topography of the sample. Systems like the AFM and SEM provide images with fields of view in the nanometer range with high resolvable detail, however these techniques are expensive, and limited in their ability to document live cells. The Dino-Lite digital microscope coupled with the Zeiss Axiovert 25 CFL microscope delivers a cost-effective method for recording live cells. Fields of view ranging from 8 microns to 300 microns with fair resolution provide a reliable method for discovering native cell structures at the nanoscale. In this report, cultured HeLa cells are recorded using different optical configurations resulting in documentation of cell dynamics at high magnification and resolution.

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

NASA Technical Reports Server (NTRS)

Lauer, James L.; Abel, Phillip B.

1988-01-01

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

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

PubMed

Majima, K

1998-01-01

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

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.

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.

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.

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

PubMed

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

2018-01-01

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

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

NASA Astrophysics Data System (ADS)

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

2018-01-01

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

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.

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.

A fully automated calibration method for an optical see-through head-mounted operating microscope with variable zoom and focus.

PubMed

Figl, Michael; Ede, Christopher; Hummel, Johann; Wanschitz, Felix; Ewers, Rolf; Bergmann, Helmar; Birkfellner, Wolfgang

2005-11-01

Ever since the development of the first applications in image-guided therapy (IGT), the use of head-mounted displays (HMDs) was considered an important extension of existing IGT technologies. Several approaches to utilizing HMDs and modified medical devices for augmented reality (AR) visualization were implemented. These approaches include video-see through systems, semitransparent mirrors, modified endoscopes, and modified operating microscopes. Common to all these devices is the fact that a precise calibration between the display and three-dimensional coordinates in the patient's frame of reference is compulsory. In optical see-through devices based on complex optical systems such as operating microscopes or operating binoculars-as in the case of the system presented in this paper-this procedure can become increasingly difficult since precise camera calibration for every focus and zoom position is required. We present a method for fully automatic calibration of the operating binocular Varioscope M5 AR for the full range of zoom and focus settings available. Our method uses a special calibration pattern, a linear guide driven by a stepping motor, and special calibration software. The overlay error in the calibration plane was found to be 0.14-0.91 mm, which is less than 1% of the field of view. Using the motorized calibration rig as presented in the paper, we were also able to assess the dynamic latency when viewing augmentation graphics on a mobile target; spatial displacement due to latency was found to be in the range of 1.1-2.8 mm maximum, the disparity between the true object and its computed overlay represented latency of 0.1 s. We conclude that the automatic calibration method presented in this paper is sufficient in terms of accuracy and time requirements for standard uses of optical see-through systems in a clinical environment.

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.

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.

Wide-field and high-resolution optical imaging for early detection of oral neoplasia

NASA Astrophysics Data System (ADS)

Pierce, Mark C.; Schwarz, Richard A.; Rosbach, Kelsey; Roblyer, Darren; Muldoon, Tim; Williams, Michelle D.; El-Naggar, Adel K.; Gillenwater, Ann M.; Richards-Kortum, Rebecca

2010-02-01

Current procedures for oral cancer screening typically involve visual inspection of the entire tissue surface at risk under white light illumination. However, pre-cancerous lesions can be difficult to distinguish from many benign conditions when viewed under these conditions. We have developed wide-field (macroscopic) imaging system which additionally images in cross-polarized white light, narrowband reflectance, and fluorescence imaging modes to reduce specular glare, enhance vascular contrast, and detect disease-related alterations in tissue autofluorescence. We have also developed a portable system to enable high-resolution (microscopic) evaluation of cellular features within the oral mucosa in situ. This system is a wide-field epi-fluorescence microscope coupled to a 1 mm diameter, flexible fiber-optic imaging bundle. Proflavine solution was used to specifically label cell nuclei, enabling the characteristic differences in N/C ratio and nuclear distribution between normal, dysplastic, and cancerous oral mucosa to be quantified. This paper discusses the technical design and performance characteristics of these complementary imaging systems. We will also present data from ongoing clinical studies aimed at evaluating diagnostic performance of these systems for detection of oral neoplasia.

High-Bandwidth Dynamic Full-Field Profilometry for Nano-Scale Characterization of MEMS

NASA Astrophysics Data System (ADS)

Chen, Liang-Chia; Huang, Yao-Ting; Chang, Pi-Bai

2006-10-01

The article describes an innovative optical interferometric methodology to delivery dynamic surface profilometry with a measurement bandwidth up to 10MHz or higher and a vertical resolution up to 1 nm. Previous work using stroboscopic microscopic interferometry for dynamic characterization of micro (opto)electromechanical systems (M(O)EMS) has been limited in measurement bandwidth mainly within a couple of MHz. For high resonant mode analysis, the stroboscopic light pulse is insufficiently short to capture the moving fringes from dynamic motion of the detected structure. In view of this need, a microscopic prototype based on white-light stroboscopic interferometry with an innovative light superposition strategy was developed to achieve dynamic full-field profilometry with a high measurement bandwidth up to 10MHz or higher. The system primarily consists of an optical microscope, on which a Mirau interferometric objective embedded with a piezoelectric vertical translator, a high-power LED light module with dual operation modes and light synchronizing electronics unit are integrated. A micro cantilever beam used in AFM was measured to verify the system capability in accurate characterisation of dynamic behaviours of the device. The full-field seventh-mode vibration at a vibratory frequency of 3.7MHz can be fully characterized and nano-scale vertical measurement resolution as well as tens micrometers of vertical measurement range can be performed.

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

NASA Astrophysics Data System (ADS)

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

2001-12-01

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

M3: Microscope-based maskless micropatterning with dry film photoresist

PubMed Central

Leigh, Steven Y.; Tattu, Aashay; Mitchell, Joseph S. B.

2011-01-01

We present a maskless micropatterning system that utilizes a fluorescence microscope with programmable X-Y stage and dry film photoresist to realize feature sizes in the sub-millimeter range (40–700 μm). The method allows for flexible in-house maskless photolithography without a dedicated microfabrication facility and is well-suited for rapid prototyping of microfluidic channels, scaffold templates for protein/cell patterning or optically-guided cell encapsulation for biomedical applications. PMID:21190086

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.

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.

Single-photon counting multicolor multiphoton fluorescence microscope.

PubMed

Buehler, Christof; Kim, Ki H; Greuter, Urs; Schlumpf, Nick; So, Peter T C

2005-01-01

We present a multicolor multiphoton fluorescence microscope with single-photon counting sensitivity. The system integrates a standard multiphoton fluorescence microscope, an optical grating spectrograph operating in the UV-Vis wavelength region, and a 16-anode photomultiplier tube (PMT). The major technical innovation is in the development of a multichannel photon counting card (mC-PhCC) for direct signal collection from multi-anode PMTs. The electronic design of the mC-PhCC employs a high-throughput, fully-parallel, single-photon counting scheme along with a high-speed electrical or fiber-optical link interface to the data acquisition computer. There is no electronic crosstalk among the detection channels of the mC-PhCC. The collected signal remains linear up to an incident photon rate of 10(8) counts per second. The high-speed data interface offers ample bandwidth for real-time readout: 2 MByte lambda-stacks composed of 16 spectral channels, 256 x 256 pixel image with 12-bit dynamic range can be transferred at 30 frames per second. The modular design of the mC-PhCC can be readily extended to accommodate PMTs of more anodes. Data acquisition from a 64-anode PMT has been verified. As a demonstration of system performance, spectrally resolved images of fluorescent latex spheres and ex-vivo human skin are reported. The multicolor multiphoton microscope is suitable for highly sensitive, real-time, spectrally-resolved three-dimensional imaging in biomedical applications.

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.

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.

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

Quantitative luminescence imaging system

DOEpatents

Erwin, D.N.; Kiel, J.L.; Batishko, C.R.; Stahl, K.A.

1990-08-14

The QLIS images and quantifies low-level chemiluminescent reactions in an electromagnetic field. It is capable of real time nonperturbing measurement and simultaneous recording of many biochemical and chemical reactions such as luminescent immunoassays or enzyme assays. The system comprises image transfer optics, a low-light level digitizing camera with image intensifying microchannel plates, an image process or, and a control computer. The image transfer optics may be a fiber image guide with a bend, or a microscope, to take the light outside of the RF field. Output of the camera is transformed into a localized rate of cumulative digitalized data or enhanced video display or hard-copy images. The system may be used as a luminescent microdosimetry device for radiofrequency or microwave radiation, as a thermal dosimeter, or in the dosimetry of ultra-sound (sonoluminescence) or ionizing radiation. It provides a near-real-time system capable of measuring the extremely low light levels from luminescent reactions in electromagnetic fields in the areas of chemiluminescence assays and thermal microdosimetry, and is capable of near-real-time imaging of the sample to allow spatial distribution analysis of the reaction. It can be used to instrument three distinctly different irradiation configurations, comprising (1) RF waveguide irradiation of a small Petri-dish-shaped sample cell, (2) RF irradiation of samples in a microscope for the microscopic imaging and measurement, and (3) RF irradiation of small to human body-sized samples in an anechoic chamber. 22 figs.

Depth of focus extended microscope configuration for imaging of incorporated groups of molecules, DNA constructs and clusters inside bacterial cells

NASA Astrophysics Data System (ADS)

Fessl, Tomas; Ben-Yaish, Shai; Vacha, Frantisek; Adamec, Frantisek; Zalevsky, Zeev

2009-07-01

Imaging of small objects such as single molecules, DNA clusters and single bacterial cells is problematic not only due to the lateral resolution that is obtainable in currently existing microscopy but also, and as much fundamentally limiting, due to the lack of sufficient axial depth of focus to have the full object focused simultaneously. Extension in depth of focus is helpful also for single molecule steady state FRET measurements. In this technique it is crucial to obtain data from many well focused molecules, which are often located in different axial depths. In this paper we present the implementation of an all-optical and a real time technique of extension in the depth of focus that may be incorporated in any high NA microscope system and to be used for the above mentioned applications. We demonstrate experimentally how after the integration of special optical element in high NA 100× objective lens of a single molecule imaging microscope system, the depth of focus is significantly improved while maintaining the same lateral resolution in imaging applications of incorporated groups of molecules, DNA constructs and clusters inside bacterial cells.

Color calibration of an RGB camera mounted in front of a microscope with strong color distortion.

PubMed

Charrière, Renée; Hébert, Mathieu; Trémeau, Alain; Destouches, Nathalie

2013-07-20

This paper aims at showing that performing color calibration of an RGB camera can be achieved even in the case where the optical system before the camera introduces strong color distortion. In the present case, the optical system is a microscope containing a halogen lamp, with a nonuniform irradiance on the viewed surface. The calibration method proposed in this work is based on an existing method, but it is preceded by a three-step preprocessing of the RGB images aiming at extracting relevant color information from the strongly distorted images, taking especially into account the nonuniform irradiance map and the perturbing texture due to the surface topology of the standard color calibration charts when observed at micrometric scale. The proposed color calibration process consists first in computing the average color of the color-chart patches viewed under the microscope; then computing white balance, gamma correction, and saturation enhancement; and finally applying a third-order polynomial regression color calibration transform. Despite the nonusual conditions for color calibration, fairly good performance is achieved from a 48 patch Lambertian color chart, since an average CIE-94 color difference on the color-chart colors lower than 2.5 units is obtained.

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.

Combined laser heating and tandem acousto-optical filter for two-dimensional temperature distribution on the surface of the heated microobject

NASA Astrophysics Data System (ADS)

Bykov, A. A.; Kutuza, I. B.; Zinin, P. V.; Machikhin, A. S.; Troyan, I. A.; Bulatov, K. M.; Batshev, V. I.; Mantrova, Y. V.; Gaponov, M. I.; Prakapenka, V. B.; Sharma, S. K.

2018-01-01

Recently it has been shown that it is possible to measure the two-dimensional distribution of the surface temperature of microscopic specimens. The main component of the system is a tandem imaging acousto-optical tunable filter synchronized with a video camera. In this report, we demonstrate that combining the laser heating system with a tandem imaging acousto-optical tunable filter allows measurement of the temperature distribution under laser heating of the platinum plates as well as a visualization of the infrared laser beam, that is widely used for laser heating in diamond anvil cells.

Closed loop adaptive optics for microscopy without a wavefront sensor

PubMed Central

Kner, Peter; Winoto, Lukman; Agard, David A.; Sedat, John W.

2013-01-01

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. PMID:24392198

Design and fabrication of x-ray Kirkpatrick-Baez microscope for ICF

NASA Astrophysics Data System (ADS)

Mu, Baozhong; Wang, Zhanshan; Huang, Shengling; Yi, Shengzhen; Shen, Zhengxiang

2007-12-01

A hard x-ray (8 keV, Kα line of Cu) Kirkpatrick-Baez (KB) microscope was designed for the diagnostics of inertial confinement fusion (ICF). Three main parts including optical design, fabrication of multilayers, and alignment method were discussed in this paper. According to the deduced equation of aberration in whole field, an optical system was designed, which gives attention to not only spatial resolution but also the collection efficiency. Tungsten (W) and boron carbide (B4C) were chosen as multilayer materials and the non-periodic multilayer with 40 layers was deposited. The measured reflectivity by XRD is better than 18% in the bandwidth range of about 0.3%. Super accurately alignment is another difficulty in the application of KB microscope. To meet the requirements of pointing and co-focusing, a binocular laser pointer which is flexible enough was designed. Finally, an 8keV x-ray tube was used as source in x-ray imaging experiment and images with magnification of 2× were obtained.

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

NASA Astrophysics Data System (ADS)

Yacoot, Andrew; Koenders, Ludger; Wolff, Helmut

2007-02-01

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

Resolution of 90 nm (lambda/5) in an optical transmission microscope with an annular condenser.

PubMed

Vainrub, Arnold; Pustovyy, Oleg; Vodyanoy, Vitaly

2006-10-01

Resolution of 90 nm was achieved with a research microscope simply by replacing the standard bright-field condenser with a homebuilt illumination system with a cardioid annular condenser. Diffraction gratings with 100 nm width lines as well as less than 100 nm size features of different-shaped objects were clearly visible on a calibrated microscope test slide. The resolution increase results from a known narrower diffraction pattern in coherent illumination for the annular aperture compared with the circular aperture. This explanation is supported by an excellent accord of calculated and measured diffraction patterns for a 50 nm radius disk.

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.

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

PubMed

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

2018-01-01

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

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.

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.

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.

Fiber-optic fluorescence imaging

PubMed Central

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

2010-01-01

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

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.

A Practical Guide to Experimental Geometrical Optics

NASA Astrophysics Data System (ADS)

Garbovskiy, Yuriy A.; Glushchenko, Anatoliy V.

2017-12-01

Preface; 1. Markets of optical materials, components, accessories, light sources and detectors; 2. Introduction to optical experiments: light producing, light managing, light detection and measuring; 3. Light detectors based on semiconductors: photoresistors, photodiodes in a photo-galvanic regime. Principles of operation and measurements; 4. Linear light detectors based on photodiodes; 5. Basic laws of geometrical optics: experimental verification; 6. Converging and diverging thin lenses; 7. Thick lenses; 8. Lens systems; 9. Simple optical instruments I: the eye and the magnifier, eyepieces and telescopes; 10. Simple optical instruments II: light illuminators and microscope; 11. Spherical mirrors; 12. Introduction to optical aberrations; 13. Elements of optical radiometry; 14. Cylindrical lenses and vials; 15. Methods of geometrical optics to measure refractive index; 16. Dispersion of light and prism spectroscope; 17. Elements of computer aided optical design; Index.

Fiber laser-microscope system for femtosecond photodisruption of biological samples

PubMed Central

Yavaş, Seydi; Erdogan, Mutlu; Gürel, Kutan; Ilday, F. Ömer; Eldeniz, Y. Burak; Tazebay, Uygar H.

2012-01-01

We report on the development of a ultrafast fiber laser-microscope system for femtosecond photodisruption of biological targets. A mode-locked Yb-fiber laser oscillator generates few-nJ pulses at 32.7 MHz repetition rate, amplified up to ∼125 nJ at 1030 nm. Following dechirping in a grating compressor, ∼240 fs-long pulses are delivered to the sample through a diffraction-limited microscope, which allows real-time imaging and control. The laser can generate arbitrary pulse patterns, formed by two acousto-optic modulators (AOM) controlled by a custom-developed field-programmable gate array (FPGA) controller. This capability opens the route to fine optimization of the ablation processes and management of thermal effects. Sample position, exposure time and imaging are all computerized. The capability of the system to perform femtosecond photodisruption is demonstrated through experiments on tissue and individual cells. PMID:22435105

Phase Sensitive Demodulation in Multiphoton Microscopy

NASA Astrophysics Data System (ADS)

Fisher, Walt G.; Piston, David W.; Wachter, Eric A.

2002-06-01

Multiphoton laser scanning microscopy offers advantages in depth of penetration into intact samples over other optical sectioning techniques. To achieve these advantages it is necessary to detect the emitted light without spatial filtering. In this nondescanned (nonconfocal) approach, ambient room light can easily contaminate the signal, forcing experiments to be performed in absolute darkness. For multiphoton microscope systems employing mode-locked lasers, signal processing can be used to reduce such problems by taking advantage of the pulsed characteristics of such lasers. Specifically, by recovering fluorescence generated at the mode-locked frequency, interference from stray light and other ambient noise sources can be significantly reduced. This technology can be adapted to existing microscopes by inserting demodulation circuitry between the detector and data collection system. The improvement in signal-to-noise ratio afforded by this approach yields a more robust microscope system and opens the possibility of moving multiphoton microscopy from the research lab to more demanding settings, such as the clinic.

Digital micromirror devices: principles and applications in imaging.

PubMed

Bansal, Vivek; Saggau, Peter

2013-05-01

A digital micromirror device (DMD) is an array of individually switchable mirrors that can be used in many advanced optical systems as a rapid spatial light modulator. With a DMD, several implementations of confocal microscopy, hyperspectral imaging, and fluorescence lifetime imaging can be realized. The DMD can also be used as a real-time optical processor for applications such as the programmable array microscope and compressive sensing. Advantages and disadvantages of the DMD for these applications as well as methods to overcome some of the limitations will be discussed in this article. Practical considerations when designing with the DMD and sample optical layouts of a completely DMD-based imaging system and one in which acousto-optic deflectors (AODs) are used in the illumination pathway are also provided.

Tying Knots in DNA with Holographic Optical Tweezers

NASA Astrophysics Data System (ADS)

Miles, Mervyn; Foster, David; Seddon, Annela; Phillips, David; Carberry, David; Padgett, Miles; Dennis, Mark

It has been demonstrated that holographic optical tweezers can be used to tie a trefoil knot in double-stranded DNA. We have developed an advanced holographic optical tweezers system with several types of intuitive control interfaces. It has been used in a range of research projects including the characterization and assembly of structures. Here the process of tying increasingly complex knots with holographic tweezers will be described. The DNA is of the order of 50 μ m in length and is fluorescently labeled, in order that it can be visualized in the optical microscope of the tweezers system. With a knot was tied, the effect of increasing the persistence length of the DNA by partial methylation of the DNA molecule was investigated. Leverhulme Trust.

Focus and perspective adaptive digital surgical microscope: optomechanical design and experimental implementation

NASA Astrophysics Data System (ADS)

Claus, Daniel; Reichert, Carsten; Herkommer, Alois

2017-05-01

This paper relates to the improvement of conventional surgical stereo microscopy via the application of digital recording devices and adaptive optics. The research is aimed at improving the working conditions of the surgeon during the operation, such that free head movement is possible. The depth clues known from conventional stereo microscopy in interaction with the human eye's functionality, such as convergence, disparity, angular elevation, parallax, and accommodation, are implemented in a digital recording system via adaptive optomechanical components. Two laterally moving pupil apertures have been used mimicking the digital implementation of the eye's vergence and head motion. The natural eye's accommodation is mimicked via the application of a tunable lens. Additionally, another system has been built, which enables tracking the surgeon's eye pupil through a digital displaying stereoscopic microscope to supply the necessary information for steering the recording system. The optomechanical design and experimental results for both systems, digital recording stereoscopic microscope and pupil tracking system, are shown.

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.

Microscope Image of a Martian Soil Surface Sample

NASA Technical Reports Server (NTRS)

2008-01-01

This is the closest view of the material underneath NASA's Phoenix Mars Lander. This sample was taken from the top centimeter of the Martian soil, and this image from the lander's Optical Microscope demonstrates its overall composition.

The soil is mostly composed of fine orange particles, and also contains larger grains, about a tenth of a millimeter in diameter, and of various colors. The soil is sticky, keeping together as a slab of material on the supporting substrate even though the substrate is tilted to the vertical.

The fine orange grains are at or below the resolution of the Optical Microscope. Mixed into the soil is a small amount&mdashabout 0.5 percent&mdashof white grains, possibly of a salt. The larger grains range from black to almost transparent in appearance. At the bottom of the image, the shadows of the Atomic Force Microscope (AFM) beams are visible. This image is 1 millimeter x 2 millimeters.

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 compact "water-window" microscope with 60-nm spatial resolution based on a double stream gas-puff target and Fresnel zone plate optics

NASA Astrophysics Data System (ADS)

Wachulak, Przemyslaw; Torrisi, Alfio; Nawaz, Muhammad F.; Adjei, Daniel; Bartnik, Andrzej; Kostecki, Jerzy; Wegrzynski, Łukasz; Vondrová, Šárka; Turňová, Jana; Fok, Tomasz; Jančarek, Alexandr; Fiedorowicz, Henryk

2015-05-01

Radiation with shorter illumination wavelength allows for extension of the diffraction limit towards nanometer scale, which is a straightforward way to significantly improve a spatial resolution in photon based microscopes. Soft X-ray (SXR) radiation, from the so called "water window" spectral range, λ=2.3-4.4 nm, which is particularly suitable for biological imaging due to natural optical contrast, providing much better spatial resolution than one obtained with visible light microscopes. The high contrast is obtained because of selective absorption of radiation by carbon and water, being constituents of the biological samples. We present a desk-top system, capable of resolving 60 nm features in few seconds exposure time. We exploit the advantages of a compact, laser-plasma SXR source, based on a double stream nitrogen gas puff target, developed at the Institute of Optoelectronics, Military University of Technology. The source, emitting quasi-monochromatic, incoherent radiation, in the "water widow" spectral range at λ = 2.88 nm, is coupled with ellipsoidal, grazing incidence condenser and Fresnel zone plate objective. The construction of the microscope with some recent images of test and real samples will be presented and discussed.

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...

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.

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.

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

NASA Technical Reports Server (NTRS)

Germani, Mark S.

1992-01-01

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

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.

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

NASA Astrophysics Data System (ADS)

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

2017-06-01

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

Increasing Student Understanding of Microscope Optics by Building and Testing the Limits of Simple, Hand-Made Model Microscopes†

PubMed Central

Drace, Kevin; Couch, Brett; Keeling, Patrick J.

2012-01-01

The ability to effectively use a microscope to observe microorganisms is a crucial skill required for many disciplines within biology, especially general microbiology and cell biology. A basic understanding of the optical properties of light microscopes is required for students to use microscopes effectively, but this subject can also be a challenge to make personally interesting to students. To explore basic optical principles of magnification and resolving power in a more engaging and hands-on fashion, students constructed handmade lenses and microscopes based on Antony van Leeuwenhoek’s design using simple materials—paper, staples, glass, and adhesive putty. Students determined the power of their lenses using a green laser pointer to magnify a copper grid of known size, which also allowed students to examine variables affecting the power and resolution of a lens such as diameter, working distance, and wavelength of light. To assess the effectiveness of the laboratory’s learning objectives, four sections of a general microbiology course were given a brief pre-activity assessment quiz to determine their background knowledge on the subject. One week after the laboratory activity, students were given the same quiz (unannounced) under similar conditions. Students showed significant gains in their understanding of microscope optics. PMID:23653781

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.

Simultaneous dual-color fluorescence microscope: a characterization study.

PubMed

Li, Zheng; Chen, Xiaodong; Ren, Liqiang; Song, Jie; Li, Yuhua; Zheng, Bin; Liu, Hong

2013-01-01

High spatial resolution and geometric accuracy is crucial for chromosomal analysis of clinical cytogenetic applications. High resolution and rapid simultaneous acquisition of multiple fluorescent wavelengths can be achieved by utilizing concurrent imaging with multiple detectors. However, such class of microscopic systems functions differently from traditional fluorescence microscopes. To develop a practical characterization framework to assess and optimize the performance of a high resolution and dual-color fluorescence microscope designed for clinical chromosomal analysis. A dual-band microscopic imaging system utilizes a dichroic mirror, two sets of specially selected optical filters, and two detectors to simultaneously acquire two fluorescent wavelengths. The system's geometric distortion, linearity, the modulation transfer function, and the dual detectors' alignment were characterized. Experiment results show that the geometric distortion at lens periphery is less than 1%. Both fluorescent channels show linear signal responses, but there exists discrepancy between the two due to the detectors' non-uniform response ratio to different wavelengths. In terms of the spatial resolution, the two contrast transfer function curves trend agreeably with the spatial frequency. The alignment measurement allows quantitatively assessing the cameras' alignment. A result image of adjusted alignment is demonstrated to show the reduced discrepancy by using the alignment measurement method. In this paper, we present a system characterization study and its methods for a specially designed imaging system for clinical cytogenetic applications. The presented characterization methods are not only unique to this dual-color imaging system but also applicable to evaluation and optimization of other similar multi-color microscopic image systems for improving their clinical utilities for future cytogenetic applications.

Rapid constructions of microstructures for optical fiber sensors using a commercial CO2 laser system.

PubMed

Irawan, Rudi; Chuan, Tjin Swee; Meng, Tay Chia; Ming, Tan Khay

2008-06-27

Exposing an optical fiber core to the measurand surrounding the fiber is often used to enhance the sensitivity of an optical fiber sensor. This paper reports on the rapid fabrication of microstructures in an optical fiber using a CO₂ laser system which help exposing the optical fiber core to the measurand. The direct-write CO₂ laser system used is originally designed for engraving the polymeric material. Fabrications of microstructures such as in-fiber microhole, D-shaped fiber, in-fiber microchannel, side-sliced fiber and tapered fiber were attempted. The microstructures in the fibers were examined using a SEM and an optical microscope. Quality of microstructures shown by the SEM images and promising results from fluorescence sensor tests using in-fiber microchannels of 100μm width, 210μm depth and 10mm length show the prospect of this method for use in optical fiber sensor development. The direct-write CO₂ laser system is a flexible and fast machining tool for fabricating microstructures in an optical fiber, and can possibly be a replacement of the time consuming chemical etching and polishing methods used for microstructure fabrications of optical the fiber sensors reported in other literatures.

Rapid Constructions of Microstructures for Optical Fiber Sensors Using a Commercial CO2 Laser System

PubMed Central

Irawan, Rudi; Chuan, Tjin Swee; Meng, Tay Chia; Ming, Tan Khay

2008-01-01

Exposing an optical fiber core to the measurand surrounding the fiber is often used to enhance the sensitivity of an optical fiber sensor. This paper reports on the rapid fabrication of microstructures in an optical fiber using a CO2 laser system which help exposing the optical fiber core to the measurand. The direct-write CO2 laser system used is originally designed for engraving the polymeric material. Fabrications of microstructures such as in-fiber microhole, D-shaped fiber, in-fiber microchannel, side-sliced fiber and tapered fiber were attempted. The microstructures in the fibers were examined using a SEM and an optical microscope. Quality of microstructures shown by the SEM images and promising results from fluorescence sensor tests using in-fiber microchannels of 100μm width, 210μm depth and 10mm length show the prospect of this method for use in optical fiber sensor development. The direct-write CO2 laser system is a flexible and fast machining tool for fabricating microstructures in an optical fiber, and can possibly be a replacement of the time consuming chemical etching and polishing methods used for microstructure fabrications of optical the fiber sensors reported in other literatures. PMID:19662114

Martian Dust Collected by Phoenix's Arm

NASA Technical Reports Server (NTRS)

2008-01-01

This image from NASA's Phoenix Lander's Optical Microscope shows particles of Martian dust lying on the microscope's silicon substrate. The Robotic Arm sprinkled a sample of the soil from the Snow White trench onto the microscope on July 2, 2008, the 38th Martian day, or sol, of the mission after landing.

Subsequently, the Atomic Force Microscope, or AFM, zoomed in one of the fine particles, creating the first-ever image of a particle of Mars' ubiquitous fine dust, the most highly magnified image ever seen from another world.

The Atomic Force Microscope was developed by a Swiss-led consortium in collaboration with Imperial College London. The AFM is part of Phoenix's Microscopy, Electrochemistry and Conductivity Analyzer instrument.

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.

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

PubMed Central

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

1999-01-01

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

Optical microscope using an interferometric source of two-color, two-beam entangled photons

DOEpatents

Dress, William B.; Kisner, Roger A.; Richards, Roger K.

2004-07-13

Systems and methods are described for an optical microscope using an interferometric source of multi-color, multi-beam entangled photons. A method includes: downconverting a beam of coherent energy to provide a beam of multi-color entangled photons; converging two spatially resolved portions of the beam of multi-color entangled photons into a converged multi-color entangled photon beam; transforming at least a portion of the converged multi-color entangled photon beam by interaction with a sample to generate an entangled photon specimen beam; and combining the entangled photon specimen beam with an entangled photon reference beam within a single beamsplitter. An apparatus includes: a multi-refringent device providing a beam of multi-color entangled photons; a condenser device optically coupled to the multi-refringent device, the condenser device converging two spatially resolved portions of the beam of multi-color entangled photons into a converged multi-color entangled photon beam; a beam probe director and specimen assembly optically coupled to the condenser device; and a beam splitter optically coupled to the beam probe director and specimen assembly, the beam splitter combining an entangled photon specimen beam from the beam probe director and specimen assembly with an entangled photon reference beam.

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

PubMed

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

2018-06-22

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

Epi-detected quadruple-modal nonlinear optical microscopy for label-free imaging of the tooth

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

Wang, Zi; Zheng, Wei; Huang, Zhiwei, E-mail: biehzw@nus.edu.sg

2015-01-19

We present an epi-detected quadruple-modal nonlinear optical microscopic imaging technique (i.e., coherent anti-Stokes Raman scattering (CARS), second-harmonic generation (SHG), third-harmonic generation (THG), and two-photon excited fluorescence (TPEF)) based on a picosecond (ps) laser-pumped optical parametric oscillator system for label-free imaging of the tooth. We demonstrate that high contrast ps-CARS images covering both the fingerprint (500–1800 cm{sup −1}) and high-wavenumber (2500–3800 cm{sup −1}) regions can be acquired to uncover the distributions of mineral and organic biomaterials in the tooth, while high quality TPEF, SHG, and THG images of the tooth can also be acquired under ps laser excitation without damaging the samples. Themore » quadruple-modal nonlinear microscopic images (CARS/SHG/THG/TPEF) acquired provide better understanding of morphological structures and biochemical/biomolecular distributions in the dentin, enamel, and the dentin-enamel junction of the tooth without labeling, facilitating optical diagnosis and characterization of the tooth in dentistry.« less

Optical workstation with concurrent, independent multiphoton imaging and experimental laser microbeam capabilities

PubMed Central

Wokosin, David L.; Squirrell, Jayne M.; Eliceiri, Kevin W.; White, John G.

2008-01-01

Experimental laser microbeam techniques have become established tools for studying living specimens. A steerable, focused laser beam may be used for a variety of experimental manipulations such as laser microsurgery, optical trapping, localized photolysis of caged bioactive probes, and patterned photobleaching. Typically, purpose-designed experimental systems have been constructed for each of these applications. In order to assess the consequences of such experimental optical interventions, long-term, microscopic observation of the specimen is often required. Multiphoton excitation, because of its ability to obtain high-contrast images from deep within a specimen with minimal phototoxic effects, is a preferred technique for in vivo imaging. An optical workstation is described that combines the functionality of an experimental optical microbeam apparatus with a sensitive multiphoton imaging system designed for use with living specimens. Design considerations are discussed and examples of ongoing biological applications are presented. The integrated optical workstation concept offers advantages in terms of flexibility and versatility relative to systems implemented with separate imaging and experimental components. PMID:18607511

Optical workstation with concurrent, independent multiphoton imaging and experimental laser microbeam capabilities

NASA Astrophysics Data System (ADS)

Wokosin, David L.; Squirrell, Jayne M.; Eliceiri, Kevin W.; White, John G.

2003-01-01

Experimental laser microbeam techniques have become established tools for studying living specimens. A steerable, focused laser beam may be used for a variety of experimental manipulations such as laser microsurgery, optical trapping, localized photolysis of caged bioactive probes, and patterned photobleaching. Typically, purpose-designed experimental systems have been constructed for each of these applications. In order to assess the consequences of such experimental optical interventions, long-term, microscopic observation of the specimen is often required. Multiphoton excitation, because of its ability to obtain high-contrast images from deep within a specimen with minimal phototoxic effects, is a preferred technique for in vivo imaging. An optical workstation is described that combines the functionality of an experimental optical microbeam apparatus with a sensitive multiphoton imaging system designed for use with living specimens. Design considerations are discussed and examples of ongoing biological applications are presented. The integrated optical workstation concept offers advantages in terms of flexibility and versatility relative to systems implemented with separate imaging and experimental components.

In situ synthesis of metal nanoparticles in polymer matrix and their optical limiting applications.

PubMed

Porel, S; Venkatram, N; Rao, D Narayana; Radhakrishnan, T P

2007-06-01

We present an overview of the simple and environmentally benign protocol we have developed recently, for the in situ generation of metal nanoparticles inside polymer films by mild thermal annealing, leading to free-standing as well as supported thin films of nanoparticle-embedded polymer. The fabrication chemistry is discussed and spectroscopic/microscopic characterizations of silver and gold nanoparticles in poly(vinyl alcohol) film are presented. Optical limiting characteristics of the silver-polymer system are investigated in detail and preliminary results for the gold-polymer system are reported.

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.

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

NASA Technical Reports Server (NTRS)

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

1989-01-01

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

Nano-biosilica from marine diatoms: A brand new material for photonic applications

NASA Astrophysics Data System (ADS)

De Stefano, L.; Maddalena, P.; Moretti, L.; Rea, I.; Rendina, I.; De Tommasi, E.; Mocella, V.; De Stefano, M.

2009-07-01

Several biological organisms, from some sea shells to butterflies, exhibit sophisticated optical systems, which have been developed during the evolution of each species. The diatoms are microscopic algae enclosed between two valves of hydrated amorphous silica. These intricate structures, called frustules, show quite symmetric patterns of micrometric and nanometric pores. Their strong similarity with man-made objects suggests to exploit the optical properties of the frustules in light guiding and optical transducing. We have found very interesting results, both from the experimental and numerical points of view.

Upconversion fiber-optic confocal microscopy under near-infrared pumping.

PubMed

Kim, Do-Hyun; Kang, Jin U; Ilev, Ilko K

2008-03-01

We present a simple upconversion fiber-optic confocal microscope design using a near-infrared laser for pumping of a rare-earth-doped glass powder. The nonlinear optical frequency conversion process is highly efficient with more than 2% upconversion fluorescence efficiency at a near-infrared pumping wavelength of 1.55 microm. The upconversion confocal design allows the use of conventional Si detectors and 1.55 microm near-infrared pump light. The lateral and axial resolutions of the system were equal to or better than 1.10 and 13.11 microm, respectively.

Zoom microscope objective using electrowetting lenses.

PubMed

Li, Lei; Wang, Di; Liu, Chao; Wang, Qiong-Hua

2016-02-08

We report a zoom microscope objective which can achieve continuous zoom change and correct the aberrations dynamically. The objective consists of three electrowetting liquid lenses and two glass lenses. The magnification is changed by applying voltages on the three electrowetting lenses. Besides, the three electrowetting liquid lenses can play a role to correct the aberrations. A digital microscope based on the proposed objective is demonstrated. We analyzed the properties of the proposed objective. In contrast to the conventional objectives, the proposed objective can be tuned from ~7.8 × to ~13.2 × continuously. For our objective, the working distance is fixed, which means no movement parts are needed to refocus or change its magnification. Moreover, the zoom objective can be dynamically optimized for a wide range of wavelength. Using such an objective, the fabrication tolerance of the optical system is larger than that of a conventional system, which can decrease the fabrication cost. The proposed zoom microscope objective cannot only take place of the conventional objective, but also has potential application in the 3D microscopy.

Rotary-scanning optical resolution photoacoustic microscopy

NASA Astrophysics Data System (ADS)

Qi, Weizhi; Xi, Lei

2016-10-01

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

(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)

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.

Site-Resolved Imaging with the Fermi Gas Microscope

NASA Astrophysics Data System (ADS)

Huber, Florian Gerhard

The recent development of quantum gas microscopy for bosonic rubidium atoms trapped in optical lattices has made it possible to study local structure and correlations in quantum many-body systems. Quantum gas microscopes are a perfect platform to perform quantum simulation of condensed matter systems, offering unprecedented control over both internal and external degrees of freedom at a single-site level. In this thesis, this technique is extended to fermionic particles, paving the way to fermionic quantum simulation, which emulate electrons in real solids. Our implementation uses lithium, the lightest atom amenable to laser cooling. The absolute timescales of dynamics in optical lattices are inversely proportional to the mass. Therefore, experiments are more than six times faster than for the only other fermionic alkali atom, potassium, and more then fourteen times faster than an equivalent rubidium experiment. Scattering and collecting a sufficient number of photons with our high-resolution imaging system requires continuous cooling of the atoms during the fluorescence imaging. The lack of a resolved excited hyperfine structure on the D2 line of lithium prevents efficient conventional sub-Doppler cooling. To address this challenge we have applied a Raman sideband cooling scheme and achieved the first site-resolved imaging of ultracold fermions in an optical lattice.

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.

A Compact Soft X-Ray Microscope using an Electrode-less Z-Pinch Source.

PubMed

Horne, S F; Silterra, J; Holber, W

2009-01-01

Soft X-rays (< 1Kev) are of medical interest both for imaging and microdosimetry applications. X-ray sources at this low energy present a technological challenge. Synchrotrons, while very powerful and flexible, are enormously expensive national research facilities. Conventional X-ray sources based on electron bombardment can be compact and inexpensive, but low x-ray production efficiencies at low electron energies restrict this approach to very low power applications. Laser-based sources tend to be expensive and unreliable. Energetiq Technology, Inc. (Woburn, MA, USA) markets a 92 eV, 10W(2pi sr) electrode-less Z-pinch source developed for advanced semiconductor lithography. A modified version of this commercial product has produced 400 mW at 430 eV (2pi sr), appropriate for water window soft X-ray microscopy. The US NIH has funded Energetiq to design and construct a demonstration microscope using this source, coupled to a condenser optic, as the illumination system. The design of the condenser optic matches the unique characteristics of the source to the illumination requirements of the microscope, which is otherwise a conventional design. A separate program is underway to develop a microbeam system, in conjunction with the RARAF facility at Columbia University, NY, USA. The objective is to develop a focused, sub-micron beam capable of delivering > 1 Gy/second to the nucleus of a living cell. While most facilities of this type are coupled to a large and expensive particle accelerator, the Z-pinch X-ray source enables a compact, stand-alone design suitable to a small laboratory. The major technical issues in this system involve development of suitable focusing X-ray optics. Current status of these programs will be reported.

A Compact Soft X-Ray Microscope using an Electrode-less Z-Pinch Source

PubMed Central

Silterra, J; Holber, W

2009-01-01

Soft X-rays (< 1Kev) are of medical interest both for imaging and microdosimetry applications. X-ray sources at this low energy present a technological challenge. Synchrotrons, while very powerful and flexible, are enormously expensive national research facilities. Conventional X-ray sources based on electron bombardment can be compact and inexpensive, but low x-ray production efficiencies at low electron energies restrict this approach to very low power applications. Laser-based sources tend to be expensive and unreliable. Energetiq Technology, Inc. (Woburn, MA, USA) markets a 92 eV, 10W(2pi sr) electrode-less Z-pinch source developed for advanced semiconductor lithography. A modified version of this commercial product has produced 400 mW at 430 eV (2pi sr), appropriate for water window soft X-ray microscopy. The US NIH has funded Energetiq to design and construct a demonstration microscope using this source, coupled to a condenser optic, as the illumination system. The design of the condenser optic matches the unique characteristics of the source to the illumination requirements of the microscope, which is otherwise a conventional design. A separate program is underway to develop a microbeam system, in conjunction with the RARAF facility at Columbia University, NY, USA. The objective is to develop a focused, sub-micron beam capable of delivering > 1 Gy/second to the nucleus of a living cell. While most facilities of this type are coupled to a large and expensive particle accelerator, the Z-pinch X-ray source enables a compact, stand-alone design suitable to a small laboratory. The major technical issues in this system involve development of suitable focusing X-ray optics. Current status of these programs will be reported. PMID:20198115

Waveguide detection of right-angle-scattered light in flow cytometry

DOEpatents

Mariella, Jr., Raymond P.

2000-01-01

A transparent flow cell is used as an index-guided optical waveguide. A detector for the flow cell but not the liquid stream detects the Right-Angle-Scattered (RAS) Light exiting from one end of the flow cell. The detector(s) could view the trapped RAS light from the flow cell either directly or through intermediate optical light guides. If the light exits one end of the flow cell, then the other end of the flow cell can be given a high-reflectivity coating to approximately double the amount of light collected. This system is more robust in its alignment than the traditional flow cytometry systems which use imaging optics, such as microscope objectives.

Optical polymers for laser medical applications

NASA Astrophysics Data System (ADS)

Sultanova, Nina G.; Kasarova, Stefka N.; Nikolov, Ivan D.

2016-01-01

In medicine, optical polymers are used not only in ophthalmology but in many laser surgical, diagnostic and therapeutic systems. The application in lens design is determined by their refractive and dispersive properties in the considered spectral region. We have used different measuring techniques to obtain precise refractometric data in the visible and near-infrared spectral regions. Dispersive, thermal and other important optical characteristics of polymers have been studied. Design of a plastic achromatic objective, used in a surgical stereo-microscope at 1064 nm laser wavelength, is accomplished. Geometrical and wavefront aberrations are calculated. Another example of application of polymers is the designed all-mirror apochromatic micro-lens, intended for superluminescent diode fiber coupling in medical systems.

Microtextured metals for stray-light suppression in the Clementine startracker

NASA Technical Reports Server (NTRS)

Johnson, E. A.

1993-01-01

Anodized blacks for suppressing stray light in optical systems can now be replaced by microscopically textured metal surfaces. An application of these black surfaces to the Clementine star-tracker navigational system, which will be launched in early 1994 to examine the Moon, en route to intercept an asteroid, is detailed. Rugged black surfaces with Lambertian BRDF less than 10(exp -2) srad(sup -1) are critical for suppressing stray light in the star-tracker optical train. Previously available materials spall under launch vibrations to contaminate mirrors and lenses. Microtextured aluminum is nearly as dark, but much less fragile. It is made by differential ion beam sputtering, which generates light-trapping pores and cones slightly smaller than the wavelength to be absorbed. This leaves a sturdy but light-absorbing surface that can survive challenging conditions without generating debris or contaminants. Both seeded ion beams and plasma immersion (from ECR plasmas) extraction can produce these microscopic textures without fragile interfaces. Process parameters control feature size, spacing, and optical effects (THR, BRDF). Both broad and narrow absorption bands can be engineered with tuning for specific wavelengths and applications. Examples are presented characterized by FTIR in reflection librators (0.95 normal emissivity), heat rejection, and enhanced nucleate boiling.

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

PubMed

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

2003-05-01

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

In situ TEM Raman spectroscopy and laser-based materials modification.

PubMed

Allen, F I; Kim, E; Andresen, N C; Grigoropoulos, C P; Minor, A M

2017-07-01

We present a modular assembly that enables both in situ Raman spectroscopy and laser-based materials processing to be performed in a transmission electron microscope. The system comprises a lensed Raman probe mounted inside the microscope column in the specimen plane and a custom specimen holder with a vacuum feedthrough for a tapered optical fiber. The Raman probe incorporates both excitation and collection optics, and localized laser processing is performed using pulsed laser light delivered to the specimen via the tapered optical fiber. Precise positioning of the fiber is achieved using a nanomanipulation stage in combination with simultaneous electron-beam imaging of the tip-to-sample distance. Materials modification is monitored in real time by transmission electron microscopy. First results obtained using the assembly are presented for in situ pulsed laser ablation of MoS 2 combined with Raman spectroscopy, complimented by electron-beam diffraction and electron energy-loss spectroscopy. Copyright © 2016 Elsevier B.V. All rights reserved.

Communication Applications for Deformable Mirror Devices.

DTIC Science & Technology

1997-06-01

is mean deflection [after Rhoadarmer. 1994] 4.5 Improved interference microscope system for micromirror characterization [after Michalicek. et...identical hexagonal micromirrors [after Michalicek. et al.. 1995] 4.7 (a) Optical system design for micromirror array (or DMD ) interfacing...constructive and destructive interference between the reflective and nonreflective portions of the element (about 75% of the element is reflective

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.

A portable liquid crystal-based polarized light system for the detection of organophosphorus nerve gas.

PubMed

He, Feng Jie; Liu, Hui Long; Chen, Long Cong; Xiong, Xing Liang

2018-03-01

Liquid crystal (LC)-based sensors have the advantageous properties of being fast, sensitive, and label-free, the results of which can be accessed directly only through the naked eye. However, the inherent disadvantages possessed by LC sensors, such as relying heavily on polarizing microscopes and the difficulty to quantify, have limited the possibility of field applications. Herein, we have addressed these issues by constructing a portable polarized detection system with constant temperature control. This system is mainly composed of four parts: the LC cell, the optics unit, the automatic temperature control unit, and the image processing unit. The LC cell was based on the ordering transitions of LCs in the presence of analytes. The optics unit based on the imaging principle of LCs was designed to substitute the polarizing microscope for the real-time observation. The image processing unit is expected to quantify the concentration of analytes. The results have shown that the presented system can detect dimethyl methyl phosphonate (a stimulant for organophosphorus nerve gas) within 25 s, and the limit of detection is about 10 ppb. In all, our portable system has potential in field applications.

A portable liquid crystal-based polarized light system for the detection of organophosphorus nerve gas

NASA Astrophysics Data System (ADS)

He, Feng Jie; Liu, Hui Long; Chen, Long Cong; Xiong, Xing Liang

2018-03-01

Liquid crystal (LC)-based sensors have the advantageous properties of being fast, sensitive, and label-free, the results of which can be accessed directly only through the naked eye. However, the inherent disadvantages possessed by LC sensors, such as relying heavily on polarizing microscopes and the difficulty to quantify, have limited the possibility of field applications. Herein, we have addressed these issues by constructing a portable polarized detection system with constant temperature control. This system is mainly composed of four parts: the LC cell, the optics unit, the automatic temperature control unit, and the image processing unit. The LC cell was based on the ordering transitions of LCs in the presence of analytes. The optics unit based on the imaging principle of LCs was designed to substitute the polarizing microscope for the real-time observation. The image processing unit is expected to quantify the concentration of analytes. The results have shown that the presented system can detect dimethyl methyl phosphonate (a stimulant for organophosphorus nerve gas) within 25 s, and the limit of detection is about 10 ppb. In all, our portable system has potential in field applications.

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.

Automatic real time evaluation of red blood cell elasticity by optical tweezers

NASA Astrophysics Data System (ADS)

Moura, Diógenes S.; Silva, Diego C. N.; Williams, Ajoke J.; Bezerra, Marcos A. C.; Fontes, Adriana; de Araujo, Renato E.

2015-05-01

Optical tweezers have been used to trap, manipulate, and measure individual cell properties. In this work, we show that the association of a computer controlled optical tweezers system with image processing techniques allows rapid and reproducible evaluation of cell deformability. In particular, the deformability of red blood cells (RBCs) plays a key role in the transport of oxygen through the blood microcirculation. The automatic measurement processes consisted of three steps: acquisition, segmentation of images, and measurement of the elasticity of the cells. An optical tweezers system was setup on an upright microscope equipped with a CCD camera and a motorized XYZ stage, computer controlled by a Labview platform. On the optical tweezers setup, the deformation of the captured RBC was obtained by moving the motorized stage. The automatic real-time homemade system was evaluated by measuring RBCs elasticity from normal donors and patients with sickle cell anemia. Approximately 150 erythrocytes were examined, and the elasticity values obtained by using the developed system were compared to the values measured by two experts. With the automatic system, there was a significant time reduction (60 × ) of the erythrocytes elasticity evaluation. Automated system can help to expand the applications of optical tweezers in hematology and hemotherapy.

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

NASA Astrophysics Data System (ADS)

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

2016-08-01

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

Computational-optical microscopy for 3D biological imaging beyond the diffraction limit

NASA Astrophysics Data System (ADS)

Grover, Ginni

In recent years, super-resolution imaging has become an important fluorescent microscopy tool. It has enabled imaging of structures smaller than the optical diffraction limit with resolution less than 50 nm. Extension to high-resolution volume imaging has been achieved by integration with various optical techniques. In this thesis, development of a fluorescent microscope to enable high resolution, extended depth, three dimensional (3D) imaging is discussed; which is achieved by integration of computational methods with optical systems. In the first part of the thesis, point spread function (PSF) engineering for volume imaging is discussed. A class of PSFs, referred to as double-helix (DH) PSFs, is generated. The PSFs exhibit two focused spots in the image plane which rotate about the optical axis, encoding depth in rotation of the image. These PSFs extend the depth-of-field up to a factor of ˜5. Precision performance of the DH-PSFs, based on an information theoretical analysis, is compared with other 3D methods with conclusion that the DH-PSFs provide the best precision and the longest depth-of-field. Out of various possible DH-PSFs, a suitable PSF is obtained for super-resolution microscopy. The DH-PSFs are implemented in imaging systems, such as a microscope, with a special phase modulation at the pupil plane. Surface-relief elements which are polarization-insensitive and ˜90% light efficient are developed for phase modulation. The photon-efficient DH-PSF microscopes thus developed are used, along with optimal position estimation algorithms, for tracking and super-resolution imaging in 3D. Imaging at depths-of-field of up to 2.5 microm is achieved without focus scanning. Microtubules were imaged with 3D resolution of (6, 9, 39) nm, which is in close agreement with the theoretical limit. A quantitative study of co-localization of two proteins in volume was conducted in live bacteria. In the last part of the thesis practical aspects of the DH-PSF microscope are discussed. A method to stabilize it, for extended periods of time, with 3-4 nm precision in 3D is developed. 3D Super-resolution is demonstrated without drift. A PSF correction algorithm is demonstrated to improve characteristics of the DH-PSF in an experiment, where it is implemented with a polarization-insensitive liquid crystal spatial light modulator.

Particle detection, number estimation, and feature measurement in gene transfer studies: optical fractionator stereology integrated with digital image processing and analysis.

PubMed

King, Michael A; Scotty, Nicole; Klein, Ronald L; Meyer, Edwin M

2002-10-01

Assessing the efficacy of in vivo gene transfer often requires a quantitative determination of the number, size, shape, or histological visualization characteristics of biological objects. The optical fractionator has become a choice stereological method for estimating the number of objects, such as neurons, in a structure, such as a brain subregion. Digital image processing and analytic methods can increase detection sensitivity and quantify structural and/or spectral features located in histological specimens. We describe a hardware and software system that we have developed for conducting the optical fractionator process. A microscope equipped with a video camera and motorized stage and focus controls is interfaced with a desktop computer. The computer contains a combination live video/computer graphics adapter with a video frame grabber and controls the stage, focus, and video via a commercial imaging software package. Specialized macro programs have been constructed with this software to execute command sequences requisite to the optical fractionator method: defining regions of interest, positioning specimens in a systematic uniform random manner, and stepping through known volumes of tissue for interactive object identification (optical dissectors). The system affords the flexibility to work with count regions that exceed the microscope image field size at low magnifications and to adjust the parameters of the fractionator sampling to best match the demands of particular specimens and object types. Digital image processing can be used to facilitate object detection and identification, and objects that meet criteria for counting can be analyzed for a variety of morphometric and optical properties. Copyright 2002 Elsevier Science (USA)

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

NASA Astrophysics Data System (ADS)

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

2016-03-01

It is difficult to achieve sub-micron resolution in backward mode OA microscopy using conventional piezoelectric detectors, because of wavefront distortions caused by components placed in the optical path, between the sample and the objective lens, that are required to separate the acoustic wave from the optical beam. As an alternate approach, an optoacoustic microscope (OAM) was constructed using the probe beam deflection technique (PBDT) to detect laserinduced acoustic signals. The all-optical OAM detects laser-generated pressure waves using a probe beam passing through a coupling medium, such as water, filling the space between the microscope objective lens and sample. The acoustic waves generated in the sample propagate through the coupling medium, causing transient changes in the refractive index that deflect the probe beam. These deflections are measured with a high-speed, balanced photodiode position detector. The deflection amplitude is directly proportional to the magnitude of the acoustic pressure wave, and provides the data required for image reconstruction. The sensitivity of the PBDT detector expressed as noise equivalent pressure was 12 Pa, comparable to that of existing high-performance ultrasound detectors. Because of the unimpeded working distance, a high numerical aperture objective lens, i.e. NA = 1, was employed in the OAM to achieve near diffraction-limited lateral resolution of 0.5 μm at 532nm. The all-optical OAM provides several benefits over current piezoelectric detector-based systems, such as increased lateral and axial resolution, higher sensitivity, robustness, and potentially more compatibility with multimodal instruments.

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...

A high-resolution optical imaging system for obtaining the serial transverse section images of biologic tissue

NASA Astrophysics Data System (ADS)

Wu, Li; Zhang, Bin; Wu, Ping; Liu, Qian; Gong, Hui

2007-05-01

A high-resolution optical imaging system was designed and developed to obtain the serial transverse section images of the biologic tissue, such as the mouse brain, in which new knife-edge imaging technology, high-speed and high-sensitive line-scan CCD and linear air bearing stages were adopted and incorporated with an OLYMPUS microscope. The section images on the tip of the knife-edge were synchronously captured by the reflection imaging in the microscope while cutting the biologic tissue. The biologic tissue can be sectioned at interval of 250 nm with the same resolution of the transverse section images obtained in x and y plane. And the cutting job can be automatically finished based on the control program wrote specially in advance, so we save the mass labor of the registration of the vast images data. In addition, by using this system a larger sample can be cut than conventional ultramicrotome so as to avoid the loss of the tissue structure information because of splitting the tissue sample to meet the size request of the ultramicrotome.

Optical Scatter Imaging with a digital micromirror device.

PubMed

Zheng, Jing-Yi; Pasternack, Robert M; Boustany, Nada N

2009-10-26

We had developed Optical Scatter Imaging (OSI) as a method which combines light scattering spectroscopy with microscopic imaging to probe local particle size in situ. Using a variable diameter iris as a Fourier spatial filter, the technique consisted of collecting images that encoded the intensity ratio of wide-to-narrow angle scatter at each pixel in the full field of view. In this paper, we replace the variable diameter Fourier filter with a digital micromirror device (DMD) to extend our assessment of morphology to the characterization of particle shape and orientation. We describe our setup in detail and demonstrate how to eliminate aberrations associated with the placement of the DMD in a conjugate Fourier plane of our microscopic imaging system. Using bacteria and polystyrene spheres, we show how this system can be used to assess particle aspect ratio even when imaged at low resolution. We also show the feasibility of detecting alterations in organelle aspect ratio in situ within living cells. This improved OSI system could be further developed to automate morphological quantification and sorting of non-spherical particles in situ.

Test target for characterizing 3D resolution of optical coherence tomography

NASA Astrophysics Data System (ADS)

Hu, Zhixiong; Hao, Bingtao; Liu, Wenli; Hong, Baoyu; Li, Jiao

2014-12-01

Optical coherence tomography (OCT) is a non-invasive 3D imaging technology which has been applied or investigated in many diagnostic fields including ophthalmology, dermatology, dentistry, cardiovasology, endoscopy, brain imaging and so on. Optical resolution is an important characteristic that can describe the quality and utility of an image acquiring system. We employ 3D printing technology to design and fabricate a test target for characterizing 3D resolution of optical coherence tomography. The test target which mimics USAF 1951 test chart was produced with photopolymer. By measuring the 3D test target, axial resolution as well as lateral resolution of a spectral domain OCT system was evaluated. For comparison, conventional microscope and surface profiler were employed to characterize the 3D test targets. The results demonstrate that the 3D resolution test targets have the potential of qualitatively and quantitatively validating the performance of OCT systems.

Multiplexed fluorescence detector system for capillary electrophoresis

DOEpatents

Yeung, E.S.; Taylor, J.A.

1996-03-12

A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis. 14 figs.

Multiplexed fluorescence detector system for capillary electrophoresis

DOEpatents

Yeung, E.S.; Taylor, J.A.

1994-06-28

A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis. 14 figures.

Multiplexed fluorescence detector system for capillary electrophoresis

DOEpatents

Yeung, Edward S.; Taylor, John A.

1996-03-12

A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

Multiplexed fluorescence detector system for capillary electrophoresis

DOEpatents

Yeung, Edward S.; Taylor, John A.

1994-06-28

A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

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.

Optical characterization of a low solubility organic compound.

PubMed

Huber, H E

1981-10-01

The X, Y, and Z principal vibration directions along with the principal refractive indexes, optic angle, optical sign, birefringence, optical orientation, and crystal system for the low solubility compound 5-(tetradecyloxy)-2-furancarboxylic acid were determined with a polarizing microscope and spindle stage. The X and Z principal vibration directions are not coincident with the a and c crystallographic axes; however, the Y direction is considered to be coincident with the b axis. Therefore, the crystal is assigned to the monoclinic crystal system. The bladed/lath-shaped crystals rest on one of the two large orthopinacoid (100) faces and present the microscopist with a single plane of optical symmetry. A beta refractive index of 1.555 is observed with the crystal axis of elongation parallel to the polarizer, and a gamma of 1.600-1.660 is observed in the contiguous extinction position. Determination of the optic angle principal vibration directions, and principal refractive indexes was facilitated by mounting the crystals on a spindle stage for rotation about the b crystallographic axis (optic normal).

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

PubMed Central

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

2016-01-01

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

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

PubMed

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

2016-09-01

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

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.

Visualizing substructure of Ca2+ waves by total internal reflection fluorescence microscopy

NASA Astrophysics Data System (ADS)

Bai, Yongqiang; Tang, Aihui; Wang, Shiqiang; Zhu, Xing

2005-02-01

Total internal reflection fluorescence microscope is a new optical microscopic system based on near-field optical theory. Its character of illumination by evanescent wave, together with the great signal-to-noise ratio and temporal resolution achieved by high quality CCD, allows us to analyze the spatiotemporal details of local Ca2+ dynamics within the nanoscale microdomain surrounding different Ca2+ channels. We have recently constructed a versatile objective TIRFM equipped with a high numerical aperture (NA=1.45) objective. Using fluo-4 as the Ca2+ indicator, we visualized the near-membrane profiles of Ca2+ waves and elementary Ca2+ sparks generated by Ca2+ release channels in rat ventricular myocytes. Different from those detected using conventional and confocal microscopy, Ca2+ waves observed with TIRFM exhibited fine inhomogenous substructures composed of fluctuating Ca2+ sparks. The anfractuous routes of spark recruitment suggested that the propagation of Ca2+ waves is much more complicated than previously imagined. We believe that TIRFM will provide a unique tool for dissecting the microscopic mechanisms of intracellular Ca2+ signaling.

An estimate of biofilm properties using an acoustic microscope.

PubMed

Good, Morris S; Wend, Christopher F; Bond, Leonard J; Mclean, Jeffrey S; Panetta, Paul D; Ahmed, Salahuddin; Crawford, Susan L; Daly, Don S

2006-09-01

Noninvasive measurements over a biofilm, a three-dimensional (3-D) community of microorganisms immobilized at a substratum, were made using an acoustic microscope operating at frequencies up to 70 MHz. The microscope scanned a 2.5-mm by 2.5-mm region of a living biofilm having a nominal thickness of 100 microm. Spatial variation of surface heterogeneity, thickness, interior structure, and biomass were estimated. Thickness was estimated as the product of the speed of sound of the medium and the interim between the highest signal peak and that of the substratum plane without biofilm. The thickest portions of biofilm were 145 microm; however, slender structures attributed as streamers extended above, with one obtaining a 274-microm height above the substratum. Three-dimensional iso-contours of amplitude were used to estimate the internal structure of the biofilm. Backscatter amplitude was examined at five zones of increasing height from the substratum to examine biomass distribution. Ultrasound-based estimates of thickness were corroborated with optical microscopy. The experimental acoustic and optical systems, methods used to estimate biofilm properties, and potential applications for the resulting data are discussed.

Modulus design multiwavelength polarization microscope for transmission Mueller matrix imaging

NASA Astrophysics Data System (ADS)

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.

Characterization of Akiyama probe applied to dual-probes atomic force microscope

NASA Astrophysics Data System (ADS)

Wang, Hequn; Gao, Sitian; Li, Wei; Shi, Yushu; Li, Qi; Li, Shi; Zhu, Zhendong

2016-10-01

The measurement of nano-scale line-width has always been important and difficult in the field of nanometer measurements, while the rapid development of integrated circuit greatly raises the demand again. As one kind of scanning probe microscope (SPM), atomic force microscope (AFM) can realize quasi three-dimensional measurement, which is widely used in nanometer scale line-width measurement. Our team researched a dual-probes atomic force microscope, which can eliminate the prevalent effect of probe width on measurement results. In dual-probes AFM system, a novel head are newly designed. A kind of self-sensing and self-exciting probes which is Nanosensors cooperation's patented probe—Akiyama probe, is used in this novel head. The Akiyama probe applied to dual-probe atomic force microscope is one of the most important issues. The characterization of Akiyama probe would affect performance and accuracy of the whole system. The fundamental features of the Akiyama probe are electrically and optically characterized in "approach-withdraw" experiments. Further investigations include the frequency response of an Akiyama probe to small mechanical vibrations externally applied to the tip and the effective loading force yielding between the tip and the sample during the periodic contact. We hope that the characterization of the Akiyama probe described in this paper will guide application for dual-probe atomic force microscope.

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

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

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

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

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.

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

High-Speed, Three Dimensional Object Composition Mapping Technology

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

Ishikawa, M Y

2001-02-14

This document overviews an entirely new approach to determining the composition--the chemical-elemental, isotopic and molecular make-up--of complex, highly structured objects, moreover with microscopic spatial resolution in all 3 dimensions. The front cover depicts the new type of pulsed laser system at the heart of this novel technology under adjustment by Alexis Wynne, and schematically indicates two of its early uses: swiftly analyzing the 3-D composition governed structure of a transistor circuit with both optical and mass-spectrometric detectors, and of fossilized dinosaur and turtle bones high-speed probed by optical detection means. Studying the composition-cued 3-D micro-structures of advanced composite materials andmore » the microscopic scale composition-texture of biological tissues are two near-term examples of the rich spectrum of novel applications enabled by this field-opening analytic tool-set.« less

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.

A fiber-optic-based imaging system for nondestructive assessment of cell-seeded tissue-engineered scaffolds.

PubMed

Hofmann, Matthias C; Whited, Bryce M; Criswell, Tracy; Rylander, Marissa Nichole; Rylander, Christopher G; Soker, Shay; Wang, Ge; Xu, Yong

2012-09-01

A major limitation in tissue engineering is the lack of nondestructive methods that assess the development of tissue scaffolds undergoing preconditioning in bioreactors. Due to significant optical scattering in most scaffolding materials, current microscope-based imaging methods cannot "see" through thick and optically opaque tissue constructs. To address this deficiency, we developed a fiber-optic-based imaging method that is capable of nondestructive imaging of fluorescently labeled cells through a thick and optically opaque scaffold, contained in a bioreactor. This imaging modality is based on the local excitation of fluorescent cells, the acquisition of fluorescence through the scaffold, and fluorescence mapping based on the position of the excitation light. To evaluate the capability and accuracy of the imaging system, human endothelial cells (ECs), stably expressing green fluorescent protein (GFP), were imaged through a fibrous scaffold. Without sacrificing the scaffolds, we nondestructively visualized the distribution of GFP-labeled cells through a ~500 μm thick scaffold with cell-level resolution and distinct localization. These results were similar to control images obtained using an optical microscope with direct line-of-sight access. Through a detailed quantitative analysis, we demonstrated that this method achieved a resolution on the order of 20-30 μm, with 10% or less deviation from standard optical microscopy. Furthermore, we demonstrated that the penetration depth of the imaging method exceeded that of confocal laser scanning microscopy by more than a factor of 2. Our imaging method also possesses a working distance (up to 8 cm) much longer than that of a standard confocal microscopy system, which can significantly facilitate bioreactor integration. This method will enable the nondestructive monitoring of ECs seeded on the lumen of a tissue-engineered vascular graft during preconditioning in vitro, as well as for other tissue-engineered constructs in the future.

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

PubMed

Boruah, B R; Neil, M A A

2009-01-01

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

[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)

Development of a dual joystick-controlled laser trapping and cutting system for optical micromanipulation of chromosomes inside living cells.

PubMed

Harsono, Marcellinus S; Zhu, Qingyuan; Shi, Linda Z; Duquette, Michelle; Berns, Michael W

2013-02-01

A multi-joystick robotic laser microscope system used to control two optical traps (tweezers) and one laser scissors has been developed for subcellular organelle manipulation. The use of joysticks has provided a "user-friendly" method for both trapping and cutting of organelles such as chromosomes in live cells. This innovative design has enabled the clean severing of chromosome arms using the laser scissors as well as the ability to easily hold and pull the severed arm using the laser tweezers. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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.

Endoscopic Optical Coherence Tomography for Clinical Gastroenterology

PubMed Central

Tsai, Tsung-Han; Fujimoto, James G.; Mashimo, Hiroshi

2014-01-01

Optical coherence tomography (OCT) is a real-time optical imaging technique that is similar in principle to ultrasonography, but employs light instead of sound waves and allows depth-resolved images with near-microscopic resolution. Endoscopic OCT allows the evaluation of broad-field and subsurface areas and can be used ancillary to standard endoscopy, narrow band imaging, chromoendoscopy, magnification endoscopy, and confocal endomicroscopy. This review article will provide an overview of the clinical utility of endoscopic OCT in the gastrointestinal tract and of recent achievements using state-of-the-art endoscopic 3D-OCT imaging systems. PMID:26852678

Application of principal component analysis to multispectral-multimodal optical image analysis for malaria diagnostics.

PubMed

Omucheni, Dickson L; Kaduki, Kenneth A; Bulimo, Wallace D; Angeyo, Hudson K

2014-12-11

Multispectral imaging microscopy is a novel microscopic technique that integrates spectroscopy with optical imaging to record both spectral and spatial information of a specimen. This enables acquisition of a large and more informative dataset than is achievable in conventional optical microscopy. However, such data are characterized by high signal correlation and are difficult to interpret using univariate data analysis techniques. In this work, the development and application of a novel method which uses principal component analysis (PCA) in the processing of spectral images obtained from a simple multispectral-multimodal imaging microscope to detect Plasmodium parasites in unstained thin blood smear for malaria diagnostics is reported. The optical microscope used in this work has been modified by replacing the broadband light source (tungsten halogen lamp) with a set of light emitting diodes (LEDs) emitting thirteen different wavelengths of monochromatic light in the UV-vis-NIR range. The LEDs are activated sequentially to illuminate same spot of the unstained thin blood smears on glass slides, and grey level images are recorded at each wavelength. PCA was used to perform data dimensionality reduction and to enhance score images for visualization as well as for feature extraction through clusters in score space. Using this approach, haemozoin was uniquely distinguished from haemoglobin in unstained thin blood smears on glass slides and the 590-700 spectral range identified as an important band for optical imaging of haemozoin as a biomarker for malaria diagnosis. This work is of great significance in reducing the time spent on staining malaria specimens and thus drastically reducing diagnosis time duration. The approach has the potential of replacing a trained human eye with a trained computerized vision system for malaria parasite blood screening.

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

PubMed Central

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

2017-01-01

High-throughput sectioning and optical imaging of tissue samples using traditional immunohistochemical techniques can be costly and inaccessible in resource-limited areas. We demonstrate three-dimensional (3D) imaging and phenotyping in optically transparent tissue using lens-free holographic on-chip microscopy as a low-cost, simple, and high-throughput alternative to conventional approaches. The tissue sample is passively cleared using a simplified CLARITY method and stained using 3,3′-diaminobenzidine to target cells of interest, enabling bright-field optical imaging and 3D sectioning of thick samples. The lens-free computational microscope uses pixel super-resolution and multi-height phase recovery algorithms to digitally refocus throughout the cleared tissue and obtain a 3D stack of complex-valued images of the sample, containing both phase and amplitude information. We optimized the tissue-clearing and imaging system by finding the optimal illumination wavelength, tissue thickness, sample preparation parameters, and the number of heights of the lens-free image acquisition and implemented a sparsity-based denoising algorithm to maximize the imaging volume and minimize the amount of the acquired data while also preserving the contrast-to-noise ratio of the reconstructed images. As a proof of concept, we achieved 3D imaging of neurons in a 200-μm-thick cleared mouse brain tissue over a wide field of view of 20.5 mm2. The lens-free microscope also achieved more than an order-of-magnitude reduction in raw data compared to a conventional scanning optical microscope imaging the same sample volume. Being low cost, simple, high-throughput, and data-efficient, we believe that this CLARITY-enabled computational tissue imaging technique could find numerous applications in biomedical diagnosis and research in low-resource settings. PMID:28819645

A laminar optical tomography system for the early cervical cancer diagnosis

NASA Astrophysics Data System (ADS)

Cui, Shanshan; Jia, Mengyu; Chen, Xueying; Meng, Wei; Gao, Feng; Zhao, Huijuan

2014-03-01

Laminar optical tomography (LOT) is a new mesoscopic functional optical imaging technique, which is an extension of a confocal microscope and diffuse optical tomography to acquire both the coaxial and off-axis scattered light at the same time. In this paper, a LOT system with a larger detection area aiming at the in vivo detection of early cervical cancer is developed. The field of view of our system is 10 mm x 10 mm. In order to improve the image quality of the system, two methods were performed: the correction of image distortion and the restriction of returning light. The performance of the system with aperture stop was assessed by liquid phantom experiments. Comparing with the Monte Carlo simulation, the measurement results show that the average relative errors of eight different source-detector distances corresponding to 4 source points are lower than the errors of the system taking the frame of objective lens as the aperture stop by 5.7%, 4.8%, 6.1%, 6.1% respectively. Moreover, the experiment based on the phantom with specified structure and optical parameters to simulate the cervix demonstrates that the system perform well for the cervix measurement.

Laser tweezer actuated microphotonic array devices for high resolution imaging and analysis in chip-based biosystems

NASA Astrophysics Data System (ADS)

Birkbeck, Aaron L.

A new technology is developed that functionally integrates arrays of lasers and micro-optics into microfluidic systems for the purpose of imaging, analyzing, and manipulating objects and biological cells. In general, the devices and technologies emerging from this area either lack functionality through the reliance on mechanical systems or provide a serial-based, time consuming approach. As compared to the current state of art, our all-optical design methodology has several distinguishing features, such as parallelism, high efficiency, low power, auto-alignment, and high yield fabrication methods, which all contribute to minimizing the cost of the integration process. The potential use of vertical cavity surface emitting lasers (VCSELs) for the creation of two-dimensional arrays of laser optical tweezers that perform independently controlled, parallel capture, and transport of large numbers of individual objects and biological cells is investigated. One of the primary biological applications for which VCSEL array sourced laser optical tweezers are considered is the formation of engineered tissues through the manipulation and spatial arrangement of different types of cells in a co-culture. Creating devices that combine laser optical tweezers with select micro-optical components permits optical imaging and analysis functions to take place inside the microfluidic channel. One such device is a micro-optical spatial filter whose motion and alignment is controlled using a laser optical tweezer. Unlike conventional spatial filter systems, our device utilizes a refractive optical element that is directly incorporated onto the lithographically patterned spatial filter. This allows the micro-optical spatial filter to automatically align itself in three-dimensions to the focal point of the microscope objective, where it then filters out the higher frequency additive noise components present in the laser beam. As a means of performing high resolution imaging in the microfluidic channel, we developed a novel technique that integrates the capacity of a laser tweezer to optically trap and manipulate objects in three-dimensions with the resolution-enhanced imaging capabilities of a solid immersion lens (SIL). In our design, the SIL is a free-floating device whose imaging beam, motion control and alignment is provided by a laser optical tweezer, which allows the microfluidic SIL to image in areas that are inaccessible to traditional solid immersion microscopes.

Fluorescence fibre-optic confocal microscopy of skin in vivo: microscope and fluorophores.

PubMed

Suihko, Christian; Swindle, Lucinda D; Thomas, Steven G; Serup, Jørgen

2005-11-01

Fibre-optic confocal imaging in vivo is a new approach in the assessment of human skin. The objective is to describe a novel instrument and its operation and use in combination with fluorophores. The Stratum is a fibre-optic fluorescence confocal microscope especially developed for the study of skin and mucous membranes. The system is flexible and any body site can be studied with a hand-held scanner. The light source is a 488 nm argon ion laser. Horizontal (en face) images of the epidermis and outer dermis are produced with cellular resolution. Magnification is approximately 1000 x . Fluorescein sodium is routinely used as fluorophore (intradermal injection or application to the skin surface). This fluorophore is safe for human use in vivo, but other substances (rhodamine B, Acridine Orange, green fluorescent protein, curcumin) have also been studied. The instrument produces sharp images of epidermal cell layers from the epidermal surface to the sub-papillary dermis, with sub-cellular resolution. The scanner is flexible in use. The technique of intradermal fluorophore injection requires some skill. We consider this fibre-optic instrument a potentially important tool in skin research for non-invasive optical biopsy of primarily the epidermis. Present use is focussed on research applications, where the fluorophore distribution in the skin may illustrate morphological changes in the epidermis.

Exploring the limits of optical microscopy: live cell and superresolution fluorescence microscopy of HIV-1 Transfer Between T lymphocytes Across the Virological Synapse

NASA Astrophysics Data System (ADS)

McNerney, Gregory Paul

Human immunodeficiency virus 1 (HIV-1) is a human retrovirus that efficiently, albeit gradually, overruns the immune system. An already infected T lymphocyte can latch onto another T lymphocyte whereby creating a virological synapse (VS); this junction drives viral assembly and transfer to the target cell in batches in an efficient, protective manor. My Ph.D. doctoral thesis focused on studying this transmission mechanism using advanced optical imaging modalities and the fully infectious fluorescent clone HIV Gag-iGFP. T lymphocytes are non-adherent cells (˜10 um thick) and the viral transmission process is fairly dynamic, hence we employed a custom spinning disk confocal microscope that revealed many interesting characteristics of this cooperative event. This methodology has low throughput as cell contact and transfer is at random. Optical tweezers was then added to the microscope to directly initiate cell contact at will. To assess when viral maturation occurs post-transfer, an optical assay based off of Forster resonance energy transfer was developed to monitor maturation. Structured illumination microscopy was further used to image the process at higher resolution and it showed that viral particles are not entering existing degradative compartments. Non-HIV-1 applications of the optical technologies are also reviewed.

Development of HiLo Microscope and its use in In-Vivo Applications

NASA Astrophysics Data System (ADS)

Patel, Shreyas J.

The functionality of achieving optical sectioning in biomedical research is invaluable as it allows for visualization of a biological sample at different depths while being free of background scattering. Most current microscopy techniques that offer optical sectioning, unfortunately, require complex instrumentation and thus are generally costly. HiLo microscopy, on the other hand, offers the same functionality and advantage at a relatively low cost. Hence, the work described in this thesis involves the design, build, and application of a HiLo microscope. More specifically, a standalone HiLo microscope was built in addition to implementing HiLo microscopy on a standard fluorescence microscope. In HiLo microscopy, optical sectioning is achieved by acquiring two different types of images per focal plane. One image is acquired under uniform illumination and the other is acquired under speckle illumination. These images are processed using an algorithm that extracts in-focus information and removes features and glare that occur as a result of background fluorescence. To show the benefits of the HiLo microscopy, several imaging experiments on various samples were performed under a HiLo microscope and compared against a traditional fluorescence microscope and a confocal microscope, which is considered the gold standard in optical imaging. In-vitro and ex-vivo imaging was performed on a set of pollen grains, and optically cleared mouse brain and heart slices. Each of these experiments showed great reduction in background scattering at different depths under HiLo microscopy. More importantly, HiLo imaging of optically cleared heart slice demonstrated emergence of different vasculature at different depths. Reduction of out-of-focus light increased the spatial resolution and allowed better visualization of capillary vessels. Furthermore, HiLo imaging was tested in an in-vivo model of a rodent dorsal window chamber model. When imaging the same sample under confocal microscope, the results were comparable between the two modalities. Additionally, a method of achieving blood flow maps at different depth using a combination of HiLo and LSI imaging is also discussed. The significance of this combined technique could help categorize blood flow to particular depths; this can help improve outcomes of medical treatments such pulse dye laser and photodynamic therapy treatments.

Single-shot measurement of phase and amplitude by using a heterodyne time-lens system and ultrafast digital time-holography

NASA Astrophysics Data System (ADS)

Tikan, Alexey; Bielawski, Serge; Szwaj, Christophe; Randoux, Stéphane; Suret, Pierre

2018-04-01

Temporal imaging systems are outstanding tools for single-shot observation of optical signals that have irregular and ultrafast dynamics. They allow long time windows to be recorded with femtosecond resolution, and do not rely on complex algorithms. However, simultaneous recording of amplitude and phase remains an open challenge for these systems. Here, we present a new heterodyne time-lens arrangement that efficiently records both the amplitude and phase of complex and random signals over large temporal windows (tens of picoseconds). Phase and time are encoded onto the two spatial dimensions of a camera. We implement this phase-sensitive time-lens system in two configurations: a time microscope and a digital temporal-holography device that enables single-shot measurement with a temporal resolution of 80 fs. We demonstrate direct application of our heterodyne time-lens to turbulent-like optical fields and optical rogue waves generated from nonlinear propagation of partially coherent waves inside optical fibres.

Optical track width measurements below 100 nm using artificial neural networks

NASA Astrophysics Data System (ADS)

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

2005-12-01

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

Calibration of the optical torque wrench.

PubMed

Pedaci, Francesco; Huang, Zhuangxiong; van Oene, Maarten; Dekker, Nynke H

2012-02-13

The optical torque wrench is a laser trapping technique that expands the capability of standard optical tweezers to torque manipulation and measurement, using the laser linear polarization to orient tailored microscopic birefringent particles. The ability to measure torque of the order of kBT (∼4 pN nm) is especially important in the study of biophysical systems at the molecular and cellular level. Quantitative torque measurements rely on an accurate calibration of the instrument. Here we describe and implement a set of calibration approaches for the optical torque wrench, including methods that have direct analogs in linear optical tweezers as well as introducing others that are specifically developed for the angular variables. We compare the different methods, analyze their differences, and make recommendations regarding their implementations.

Common path in-line holography using enhanced joint object reference digital interferometers

PubMed Central

Kelner, Roy; Katz, Barak; Rosen, Joseph

2014-01-01

Joint object reference digital interferometer (JORDI) is a recently developed system capable of recording holograms of various types [Opt. Lett. 38(22), 4719 (2013)24322115]. Presented here is a new enhanced system design that is based on the previous JORDI. While the previous JORDI has been based purely on diffractive optical elements, displayed on spatial light modulators, the present design incorporates an additional refractive objective lens, thus enabling hologram recording with improved resolution and increased system applicability. Experimental results demonstrate successful hologram recording for various types of objects, including transmissive, reflective, three-dimensional, phase and highly scattering objects. The resolution limit of the system is analyzed and experimentally validated. Finally, the suitability of JORDI for microscopic applications is verified as a microscope objective based configuration of the system is demonstrated. PMID:24663838

Intraoperative video-rate hemodynamic response assessment in human cortex using snapshot hyperspectral optical imaging

PubMed Central

Pichette, Julien; Laurence, Audrey; Angulo, Leticia; Lesage, Frederic; Bouthillier, Alain; Nguyen, Dang Khoa; Leblond, Frederic

2016-01-01

Abstract. Using light, we are able to visualize the hemodynamic behavior of the brain to better understand neurovascular coupling and cerebral metabolism. In vivo optical imaging of tissue using endogenous chromophores necessitates spectroscopic detection to ensure molecular specificity as well as sufficiently high imaging speed and signal-to-noise ratio, to allow dynamic physiological changes to be captured, isolated, and used as surrogate of pathophysiological processes. An optical imaging system is introduced using a 16-bands on-chip hyperspectral camera. Using this system, we show that up to three dyes can be imaged and quantified in a tissue phantom at video-rate through the optics of a surgical microscope. In vivo human patient data are presented demonstrating brain hemodynamic response can be measured intraoperatively with molecular specificity at high speed. PMID:27752519

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.

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

NASA Astrophysics Data System (ADS)

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

2008-03-01

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

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.

Superresolving dendritic spine morphology with STED microscopy under holographic photostimulation

PubMed Central

Lauterbach, Marcel Andreas; Guillon, Marc; Desnos, Claire; Khamsing, Dany; Jaffal, Zahra; Darchen, François; Emiliani, Valentina

2016-01-01

Abstract. Emerging all-optical methods provide unique possibilities for noninvasive studies of physiological processes at the cellular and subcellular scale. On the one hand, superresolution microscopy enables observation of living samples with nanometer resolution. On the other hand, light can be used to stimulate cells due to the advent of optogenetics and photolyzable neurotransmitters. To exploit the full potential of optical stimulation, light must be delivered to specific cells or even parts of cells such as dendritic spines. This can be achieved with computer generated holography (CGH), which shapes light to arbitrary patterns by phase-only modulation. We demonstrate here in detail how CGH can be incorporated into a stimulated emission depletion (STED) microscope for photostimulation of neurons and monitoring of nanoscale morphological changes. We implement an original optical system to allow simultaneous holographic photostimulation and superresolution STED imaging. We present how synapses can be clearly visualized in live cells using membrane stains either with lipophilic organic dyes or with fluorescent proteins. We demonstrate the capabilities of this microscope to precisely monitor morphological changes of dendritic spines after stimulation. These all-optical methods for cell stimulation and monitoring are expected to spread to various fields of biological research in neuroscience and beyond. PMID:27413766

4D microscope-integrated OCT improves accuracy of ophthalmic surgical maneuvers

NASA Astrophysics Data System (ADS)

Carrasco-Zevallos, Oscar; Keller, Brenton; Viehland, Christian; Shen, Liangbo; Todorich, Bozho; Shieh, Christine; Kuo, Anthony; Toth, Cynthia; Izatt, Joseph A.

2016-03-01

Ophthalmic surgeons manipulate micron-scale tissues using stereopsis through an operating microscope and instrument shadowing for depth perception. While ophthalmic microsurgery has benefitted from rapid advances in instrumentation and techniques, the basic principles of the stereo operating microscope have not changed since the 1930's. Optical Coherence Tomography (OCT) has revolutionized ophthalmic imaging and is now the gold standard for preoperative and postoperative evaluation of most retinal and many corneal procedures. We and others have developed initial microscope-integrated OCT (MIOCT) systems for concurrent OCT and operating microscope imaging, but these are limited to 2D real-time imaging and require offline post-processing for 3D rendering and visualization. Our previously presented 4D MIOCT system can record and display the 3D surgical field stereoscopically through the microscope oculars using a dual-channel heads-up display (HUD) at up to 10 micron-scale volumes per second. In this work, we show that 4D MIOCT guidance improves the accuracy of depth-based microsurgical maneuvers (with statistical significance) in mock surgery trials in a wet lab environment. Additionally, 4D MIOCT was successfully performed in 38/45 (84%) posterior and 14/14 (100%) anterior eye human surgeries, and revealed previously unrecognized lesions that were invisible through the operating microscope. These lesions, such as residual and potentially damaging retinal deformation during pathologic membrane peeling, were visualized in real-time by the surgeon. Our integrated system provides an enhanced 4D surgical visualization platform that can improve current ophthalmic surgical practice and may help develop and refine future microsurgical techniques.

3D refractive index measurements of special optical fibers

NASA Astrophysics Data System (ADS)

Yan, Cheng; Huang, Su-Juan; Miao, Zhuang; Chang, Zheng; Zeng, Jun-Zhang; Wang, Ting-Yun

2016-09-01

A digital holographic microscopic chromatography-based approach with considerably improved accuracy, simplified configuration and performance stability is proposed to measure three dimensional refractive index of special optical fibers. Based on the approach, a measurement system is established incorporating a modified Mach-Zehnder interferometer and lab-developed supporting software for data processing. In the system, a phase projection distribution of an optical fiber is utilized to obtain an optimal digital hologram recorded by a CCD, and then an angular spectrum theory-based algorithm is adopted to extract the phase distribution information of an object wave. The rotation of the optic fiber enables the experimental measurements of multi-angle phase information. Based on the filtered back projection algorithm, a 3D refraction index of the optical fiber is thus obtained at high accuracy. To evaluate the proposed approach, both PANDA fibers and special elliptical optical fiber are considered in the system. The results measured in PANDA fibers agree well with those measured using S14 Refractive Index Profiler, which is, however, not suitable for measuring the property of a special elliptical fiber.

Adaptive optics improves multiphoton super-resolution imaging

NASA Astrophysics Data System (ADS)

Zheng, Wei; Wu, Yicong; Winter, Peter; Shroff, Hari

2018-02-01

Three dimensional (3D) fluorescence microscopy has been essential for biological studies. It allows interrogation of structure and function at spatial scales spanning the macromolecular, cellular, and tissue levels. Critical factors to consider in 3D microscopy include spatial resolution, signal-to-noise (SNR), signal-to-background (SBR), and temporal resolution. Maintaining high quality imaging becomes progressively more difficult at increasing depth (where optical aberrations, induced by inhomogeneities of refractive index in the sample, degrade resolution and SNR), and in thick or densely labeled samples (where out-of-focus background can swamp the valuable, in-focus-signal from each plane). In this report, we introduce our new instrumentation to address these problems. A multiphoton structured illumination microscope was simply modified to integrate an adpative optics system for optical aberrations correction. Firstly, the optical aberrations are determined using direct wavefront sensing with a nonlinear guide star and subsequently corrected using a deformable mirror, restoring super-resolution information. We demonstrate the flexibility of our adaptive optics approach on a variety of semi-transparent samples, including bead phantoms, cultured cells in collagen gels and biological tissues. The performance of our super-resolution microscope is improved in all of these samples, as peak intensity is increased (up to 40-fold) and resolution recovered (up to 176+/-10 nm laterally and 729+/-39 nm axially) at depths up to 250 μm from the coverslip surface.

Nanoscale live cell optical imaging of the dynamics of intracellular microvesicles in neural cells.

PubMed

Lee, Sohee; Heo, Chaejeong; Suh, Minah; Lee, Young Hee

2013-11-01

Recent advances in biotechnology and imaging technology have provided great opportunities to investigate cellular dynamics. Conventional imaging methods such as transmission electron microscopy, scanning electron microscopy, and atomic force microscopy are powerful techniques for cellular imaging, even at the nanoscale level. However, these techniques have limitations applications in live cell imaging because of the experimental preparation required, namely cell fixation, and the innately small field of view. In this study, we developed a nanoscale optical imaging (NOI) system that combines a conventional optical microscope with a high resolution dark-field condenser (Cytoviva, Inc.) and halogen illuminator. The NOI system's maximum resolution for live cell imaging is around 100 nm. We utilized NOI to investigate the dynamics of intracellular microvesicles of neural cells without immunocytological analysis. In particular, we studied direct, active random, and moderate random dynamic motions of intracellular microvesicles and visualized lysosomal vesicle changes after treatment of cells with a lysosomal inhibitor (NH4Cl). Our results indicate that the NOI system is a feasible, high-resolution optical imaging system for live small organelles that does not require complicated optics or immunocytological staining processes.

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

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

NASA Astrophysics Data System (ADS)

Shen, Jin H.; Joos, Karen M.

2015-03-01

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

4D megahertz optical coherence tomography (OCT): imaging and live display beyond 1 gigavoxel/sec (Conference Presentation)

NASA Astrophysics Data System (ADS)

Huber, Robert A.; Draxinger, Wolfgang; Wieser, Wolfgang; Kolb, Jan Philip; Pfeiffer, Tom; Karpf, Sebastian N.; Eibl, Matthias; Klein, Thomas

2016-03-01

Over the last 20 years, optical coherence tomography (OCT) has become a valuable diagnostic tool in ophthalmology with several 10,000 devices sold today. Other applications, like intravascular OCT in cardiology and gastro-intestinal imaging will follow. OCT provides 3-dimensional image data with microscopic resolution of biological tissue in vivo. In most applications, off-line processing of the acquired OCT-data is sufficient. However, for OCT applications like OCT aided surgical microscopes, for functional OCT imaging of tissue after a stimulus, or for interactive endoscopy an OCT engine capable of acquiring, processing and displaying large and high quality 3D OCT data sets at video rate is highly desired. We developed such a prototype OCT engine and demonstrate live OCT with 25 volumes per second at a size of 320x320x320 pixels. The computer processing load of more than 1.5 TFLOPS was handled by a GTX 690 graphics processing unit with more than 3000 stream processors operating in parallel. In the talk, we will describe the optics and electronics hardware as well as the software of the system in detail and analyze current limitations. The talk also focuses on new OCT applications, where such a system improves diagnosis and monitoring of medical procedures. The additional acquisition of hyperspectral stimulated Raman signals with the system will be discussed.

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

DOE PAGES

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

2017-10-05

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

Advantages of using newly developed quartz contact lens with slit illumination from operating microscope.

PubMed

Kiyokawa, Masatoshi; Sakuma, Toshiro; Hatano, Noriko; Mizota, Atsushi; Tanaka, Minoru

2009-06-01

The purpose of this article is to report the characteristics and advantages of using a newly designed quartz contact lens with slit illumination from an operating microscope for intraocular surgery. The new contact lens is made of quartz. The lens is convex-concave and is used in combination with slit illumination from an operating microscope. The optical properties of quartz make this lens less reflective with greater transmittance. The combination of a quartz contact lens with slit illumination provided a brighter and wider field of view than conventional lenses. This system enabled us to perform bimanual vitrectomy and scleral buckling surgery without indirect ophthalmoscope. Small intraocular structures in the posterior pole or in the periphery were detected more easily. In conclusion, the newly designed quartz lens with slit beam illumination from an operating microscope provided a bright, clear and wide surgical field, and allowed intraocular surgery to be performed more easily.

Application of differential interference contrast with inverted microscopes to the in vitro perfused nephron.

PubMed

Horster, M; Gundlach, H

1979-12-01

The study of in vitro perfused individual nephron segments requires a microscope which provides: (1) easy access to the specimen for measurement of cellular solute flux and voltage; (2) an image with high resolution and contrast; (3) optical sectioning of the object at different levels; and (4) rapid recording of the morphological phenomena. This paper describes an example of commercially available apparatus meeting the above requirements, and illustrates its efficiency. The microscope is of the inverted type (Zeiss IM 35) equipped with differential-interference-contrast (DIC) with a long working distance, and an automatically controlled camera system. The microscopic image exhibits cellular and intercellular details in the unstained transporting mammalian nephron segments despite their tubular structure and great thickness and makes obvious function-structure correlations (e.g. cell volume changes); luminal and contraluminal cell borders are well resolved for controlled microelectrode impalement.

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

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

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

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

Screening unlabeled DNA targets with randomly ordered fiber-optic gene arrays.

PubMed

Steemers, F J; Ferguson, J A; Walt, D R

2000-01-01

We have developed a randomly ordered fiber-optic gene array for rapid, parallel detection of unlabeled DNA targets with surface immobilized molecular beacons (MB) that undergo a conformational change accompanied by a fluorescence change in the presence of a complementary DNA target. Microarrays are prepared by randomly distributing MB-functionalized 3-microm diameter microspheres in an array of wells etched in a 500-microm diameter optical imaging fiber. Using several MBs, each designed to recognize a different target, we demonstrate the selective detection of genomic cystic fibrosis related targets. Positional registration and fluorescence response monitoring of the microspheres was performed using an optical encoding scheme and an imaging fluorescence microscope system.

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.

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

NASA Astrophysics Data System (ADS)

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

2017-02-01

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

PC-based control unit for a head-mounted operating microscope for augmented-reality visualization in surgical navigation

NASA Astrophysics Data System (ADS)

Figl, Michael; Birkfellner, Wolfgang; Watzinger, Franz; Wanschitz, Felix; Hummel, Johann; Hanel, Rudolf A.; Ewers, Rolf; Bergmann, Helmar

2002-05-01

Two main concepts of Head Mounted Displays (HMD) for augmented reality (AR) visualization exist, the optical and video-see through type. Several research groups have pursued both approaches for utilizing HMDs for computer aided surgery. While the hardware requirements for a video see through HMD to achieve acceptable time delay and frame rate seem to be enormous the clinical acceptance of such a device is doubtful from a practical point of view. Starting from previous work in displaying additional computer-generated graphics in operating microscopes, we have adapted a miniature head mounted operating microscope for AR by integrating two very small computer displays. To calibrate the projection parameters of this so called Varioscope AR we have used Tsai's Algorithm for camera calibration. Connection to a surgical navigation system was performed by defining an open interface to the control unit of the Varioscope AR. The control unit consists of a standard PC with a dual head graphics adapter to render and display the desired augmentation of the scene. We connected this control unit to a computer aided surgery (CAS) system by the TCP/IP interface. In this paper we present the control unit for the HMD and its software design. We tested two different optical tracking systems, the Flashpoint (Image Guided Technologies, Boulder, CO), which provided about 10 frames per second, and the Polaris (Northern Digital, Ontario, Canada) which provided at least 30 frames per second, both with a time delay of one frame.

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.

Longitudinal spatial coherence gated high-resolution tomography and quantitative phase microscopy of biological cells and tissues with uniform illumination

NASA Astrophysics Data System (ADS)

Mehta, Dalip Singh; Ahmad, Azeem; Dubey, Vishesh; Singh, Veena; Butola, Ankit; Mohanty, Tonmoy; Nandi, Sreyankar

2018-02-01

We report longitudinal spatial coherence (LSC) gated high-resolution tomography and quantitative phase microscopy of biological cells and tissues with uniform illumination using laser as a light source. To accomplish this a pseudo thermal light source was synthesized by passing laser beams through an optical system, which is basically a speckle reduction system with combined effect of spatial, temporal, angular and polarisation diversity. The longitudinal spatial coherence length of such light was significantly reduced by synthesizing a pseudo thermal source with the combined effect of spatial, angular and temporal diversity. This results in a low spatially coherent (i.e., broad angular frequency spectrum) light source with narrow temporal frequency spectrum. Light from such a pseudo thermal light source was passed through an interference microscope with varying magnification, such as, 10X and 50X. The interference microscope was used for full-field OCT imaging of multilayer objects and topography of industrial objects. Experimental results of optical sectioning of multilayer biological objects with high axial-resolution less than 10μm was achieved which is comparable to broadband white light source. The synthesized light source with reduced speckles having uniform illumination on the sample, which can be very useful for fluorescence microscopy as well as quantitative phase microscopy with less phase noise. The present system does not require any dispersion compensation optical system for biological samples as a highly monochromatic light source is used.

Epifluorescence light collection for multiphoton microscopic endoscopy

NASA Astrophysics Data System (ADS)

Brown, Christopher M.; Rivera, David R.; Xu, Chris; Webb, Watt W.

2011-03-01

Multiphoton microscopic endoscopy (MPM-E) is a promising medical in vivo diagnostic imaging technique because it captures intrinsic fluorescence and second harmonic generation signals to reveal anatomical and histological information about disease states in tissue. However, maximizing light collection from multiphoton endoscopes remains a challenge: weak nonlinear emissions from endogenous structures, miniature optics, large imaging depths, and light scattering in tissue all hamper light collection. The quantity of light that may be collected using a dual-clad fiber system from scattering phantoms that mimic the properties of the in vivo environment is measured. In this experiment, 800nm excitation light from a Ti:Sapphire laser is dispersion compensated and focused through a SM800 optical fiber and lens system into the tissue phantom. Emission light from the phantom passes through the lens system, reflects off the dichroic and is then collected by a second optical fiber actuated by a micromanipulator. The lateral position of the collection fiber varies, measuring the distribution of emitted light 2000μm on either side of the focal point reimaged to the object plane. This spatial collection measurement is performed at depths up to 200μm from the phantom surface. The tissue phantoms are composed of a 15.8 μM fluorescein solution mixed with microspheres, approximating the scattering properties of human bladder and dermis tissue. Results show that commercially available dual-clad optical fibers collect more than 47% of the total emission returning to the object plane from both phantoms. Based on these results, initial MPM-E devices will image the surface of epithelial tissues.

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.

Benchtop and animal validation of a portable fluorescence microscopic imaging system for potential use in cholecystectomy.

PubMed

Ye, Jian; Liu, Guanghui; Liu, Peng; Zhang, Shiwu; Shao, Pengfei; Smith, Zachary J; Liu, Chenhai; Xu, Ronald X

2018-02-01

We propose a portable fluorescence microscopic imaging system (PFMS) for intraoperative display of biliary structure and prevention of iatrogenic injuries during cholecystectomy. The system consists of a light source module, a camera module, and a Raspberry Pi computer with an LCD. Indocyanine green (ICG) is used as a fluorescent contrast agent for experimental validation of the system. Fluorescence intensities of the ICG aqueous solution at different concentration levels are acquired by our PFMS and compared with those of a commercial Xenogen IVIS system. We study the fluorescence detection depth by superposing different thicknesses of chicken breast on an ICG-loaded agar phantom. We verify the technical feasibility for identifying potential iatrogenic injury in cholecystectomy using a rat model in vivo. The proposed PFMS system is portable, inexpensive, and suitable for deployment in resource-limited settings. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

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.

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.

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

NASA Astrophysics Data System (ADS)

Ekberg, Peter; Mattsson, Lars

2018-03-01

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

A precise method for adjusting the optical system of laser sub-aperture

NASA Astrophysics Data System (ADS)

Song, Xing; Zhang, Xue-min; Yang, Jianfeng; Xue, Li

2018-02-01

In order to adapt to the requirement of modern astronomical observation and warfare, the resolution of the space telescope is needed to improve, sub-aperture stitching imaging technique is one method to improve the resolution, which could be used not only the foundation and space-based large optical systems, also used in laser transmission and microscopic imaging. A large aperture main mirror of sub-aperture stitching imaging system is composed of multiple sub-mirrors distributed according to certain laws. All sub-mirrors are off-axis mirror, so the alignment of sub-aperture stitching imaging system is more complicated than a single off-axis optical system. An alignment method based on auto-collimation imaging and interferometric imaging is introduced in this paper, by using this alignment method, a sub-aperture stitching imaging system which is composed of 12 sub-mirrors was assembled with high resolution, the beam coincidence precision is better than 0.01mm, and the system wave aberration is better than 0.05λ.

An Efficient Referencing And Sample Positioning System To Investigate Heterogeneous Substances With Combined Microfocused Synchrotron X-ray Techniques

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

Spangenberg, Thomas; Goettlicher, Joerg; Steininger, Ralph

2009-01-29

A referencing and sample positioning system has been developed to transfer object positions measured with an offline microscope to a synchrotron experimental station. The accuracy should be sufficient to deal with heterogeneous samples on micrometer scale. Together with an online fluorescence mapping visualisation the optical alignment helps to optimize measuring procedures for combined microfocused X-ray techniques.

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.

Aberration correction in wide-field fluorescence microscopy by segmented-pupil image interferometry.

PubMed

Scrimgeour, Jan; Curtis, Jennifer E

2012-06-18

We present a new technique for the correction of optical aberrations in wide-field fluorescence microscopy. Segmented-Pupil Image Interferometry (SPII) uses a liquid crystal spatial light modulator placed in the microscope's pupil plane to split the wavefront originating from a fluorescent object into an array of individual beams. Distortion of the wavefront arising from either system or sample aberrations results in displacement of the images formed from the individual pupil segments. Analysis of image registration allows for the local tilt in the wavefront at each segment to be corrected with respect to a central reference. A second correction step optimizes the image intensity by adjusting the relative phase of each pupil segment through image interferometry. This ensures that constructive interference between all segments is achieved at the image plane. Improvements in image quality are observed when Segmented-Pupil Image Interferometry is applied to correct aberrations arising from the microscope's optical path.

Review of intraoperative optical coherence tomography: technology and applications [Invited

PubMed Central

Carrasco-Zevallos, Oscar M.; Viehland, Christian; Keller, Brenton; Draelos, Mark; Kuo, Anthony N.; Toth, Cynthia A.; Izatt, Joseph A.

2017-01-01

During microsurgery, en face imaging of the surgical field through the operating microscope limits the surgeon’s depth perception and visualization of instruments and sub-surface anatomy. Surgical procedures outside microsurgery, such as breast tumor resections, may also benefit from visualization of the sub-surface tissue structures. The widespread clinical adoption of optical coherence tomography (OCT) in ophthalmology and its growing prominence in other fields, such as cancer imaging, has motivated the development of intraoperative OCT for real-time tomographic visualization of surgical interventions. This article reviews key technological developments in intraoperative OCT and their applications in human surgery. We focus on handheld OCT probes, microscope-integrated OCT systems, and OCT-guided laser treatment platforms designed for intraoperative use. Moreover, we discuss intraoperative OCT adjuncts and processing techniques currently under development to optimize the surgical feedback derivable from OCT data. Lastly, we survey salient clinical studies of intraoperative OCT for human surgery. PMID:28663853

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.

The rheo-Raman microscope: Simultaneous chemical, conformational, mechanical, and microstructural measures of soft materials

NASA Astrophysics Data System (ADS)

Kotula, Anthony P.; Meyer, Matthew W.; De Vito, Francesca; Plog, Jan; Hight Walker, Angela R.; Migler, Kalman B.

2016-10-01

The design and performance of an instrument capable of simultaneous Raman spectroscopy, rheology, and optical microscopy are described. The instrument couples a Raman spectrometer and optical microscope to a rotational rheometer through an optically transparent base, and the resulting simultaneous measurements are particularly advantageous in situations where flow properties vary due to either chemical or conformational changes in molecular structure, such as in crystallization, melting, gelation, or curing processes. Instrument performance is demonstrated on two material systems that show thermal transitions. First, we perform steady state rotational tests, Raman spectroscopy, and polarized reflection microscopy during a melting transition in a cosmetic emulsion. Second, we perform small amplitude oscillatory shear measurements along with Raman spectroscopy and polarized reflection microscopy during crystallization of a high density polyethylene. The instrument can be applied to study structure-property relationships in a variety of soft materials including thermoset resins, liquid crystalline materials, colloidal suspensions undergoing sol-gel processes, and biomacromolecules. Official contribution of the National Institute of Standards and Technology; not subject to copyright in the United States.

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.

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.

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

NASA Astrophysics Data System (ADS)

Guo, Kaikai; Zheng, Guoan

2017-03-01

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

Digital holographic microscope with low-frequency attenuation filter for position measurement of a nanoparticle.

PubMed

Pham, Quang Duc; Kusumi, Yuichi; Hasegawa, Satoshi; Hayasaki, Yoshio

2012-10-01

We propose a new method for three-dimensional (3D) position measurement of nanoparticles using an in-line digital holographic microscope. The method improves the signal-to-noise ratio of the amplitude of the interference fringes to achieve higher accuracy in the position measurement by increasing weak scattered light from a nanoparticle relative to the reference light by using a low spatial frequency attenuation filter. We demonstrated the improvements of signal-to-noise ratio of the optical system and contrast of the interference fringes, allowing the 3D positions of nanoparticles to be determined more precisely.

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

PubMed

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

2013-09-01

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

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

Refractive multiple optical tweezers for parallel biochemical analysis in micro-fluidics

NASA Astrophysics Data System (ADS)

Merenda, Fabrice; Rohner, Johann; Pascoal, Pedro; Fournier, Jean-Marc; Vogel, Horst; Salathé, René-Paul

2007-02-01

We present a multiple laser tweezers system based on refractive optics. The system produces an array of 100 optical traps thanks to a refractive microlens array, whose focal plane is imaged into the focal plane of a high-NA microscope objective. This refractive multi-tweezers system is combined to micro-fluidics, aiming at performing simultaneous biochemical reactions on ensembles of free floating objects. Micro-fluidics allows both transporting the particles to the trapping area, and conveying biochemical reagents to the trapped particles. Parallel trapping in micro-fluidics is achieved with polystyrene beads as well as with native vesicles produced from mammalian cells. The traps can hold objects against fluid flows exceeding 100 micrometers per second. Parallel fluorescence excitation and detection on the ensemble of trapped particles is also demonstrated. Additionally, the system is capable of selectively and individually releasing particles from the tweezers array using a complementary steerable laser beam. Strategies for high-yield particle capture and individual particle release in a micro-fluidic environment are discussed. A comparison with diffractive optical tweezers enhances the pros and cons of refractive systems.

Time lapse video recordings of highly purified human hematopoietic progenitor cells in culture.

PubMed

Denkers, I A; Dragowska, W; Jaggi, B; Palcic, B; Lansdorp, P M

1993-05-01

Major hurdles in studies of stem cell biology include the low frequency and heterogeneity of human hematopoietic precursor cells in bone marrow and the difficulty of directly studying the effect of various culture conditions and growth factors on such cells. We have adapted the cell analyzer imaging system for monitoring and recording the morphology of limited numbers of cells under various culture conditions. Hematopoietic progenitor cells with a CD34+ CD45RAlo CD71lo phenotype were purified from previously frozen organ donor bone marrow by fluorescence activated cell sorting. Cultures of such cells were analyzed with the imaging system composed of an inverted microscope contained in an incubator, a video camera, an optical memory disk recorder and a computer-controlled motorized microscope XYZ precision stage. Fully computer-controlled video images at defined XYZ positions were captured at selected time intervals and recorded at a predetermined sequence on an optical memory disk. In this study, the cell analyzer system was used to obtain descriptions and measurements of hematopoietic cell behavior, like cell motility, cell interactions, cell shape, cell division, cell cycle time and cell size changes under different culture conditions.

Portable fluorescence microendoscope system for smartphones and its applications

NASA Astrophysics Data System (ADS)

Gómez García, Pablo Aurelio; Teixeira Rosa, Ramon Gabriel; Pratavieira, Sebastião.; Kurachi, Cristina

2015-06-01

A portable microscope/microendoscope will be presented in this article. The system was specially designed for Smartphones and taking into account its simplicity, will be able to bring this technology to almost every doctor's office. It is worth mentioning its flexibility of use, that allows several modes since all the components are interchangeable (the illumination LED, the lens, the optic filters, etc) resulting in different applications, from medical applications until other areas (for example, the inspection of non-accessible pieces of plane engines). In addition, the system has a double platform, working as a conventional microscope or as a fiberoptic microendoscope. In situ and cell smear interrogation of oral mucosa, using a proflavine as dye will be presented. The price of the system does not exceed US 350, plus the price of the fiber bundle (around US 500) turning it onto a high resolution affordable system.

Potential Engineering of Fermi-Hubbard Systems using a Quantum Gas Microscope

NASA Astrophysics Data System (ADS)

Ji, Geoffrey; Mazurenko, Anton; Chiu, Christie; Parsons, Maxwell; Kanász-Nagy, Márton; Schmidt, Richard; Grusdt, Fabian; Demler, Eugene; Greif, Daniel; Greiner, Markus

2017-04-01

Arbitrary control of optical potentials has emerged as an important tool in manipulating ultracold atomic systems, especially when combined with the single-site addressing afforded by quantum gas microscopy. Already, experiments have used digital micromirror devices (DMDs) to initialize and control ultracold atomic systems in the context of studying quantum walks, quantum thermalization, and many-body localization. Here, we report on progress in using a DMD located in the image plane of a quantum gas microscope to explore static and dynamic properties of a 2D Fermi-Hubbard system. By projecting a large, ring-shaped anti-confining potential, we demonstrate entropy redistribution and controlled doping of the system. Moreover, we use the DMD to prepare localized holes, which upon release interact with and disrupt the surrounding spin environment. These techniques pave the way for controlled investigations of dynamics in the low-temperature phases of the Hubbard model.

Nondestructive measurement of an optical fiber refractive-index profile by a transmitted-light differential interference contact microscope.

PubMed

Liu, Zhongyao; Dong, Xiaoman; Chen, Qianghua; Yin, Chunyong; Xu, Yuxian; Zheng, Yingjun

2004-03-01

A novel transmitted-light differential interference contrast (DIC) system is used for nondestructive measurement of the refractive-index profile (RIP) of an optical fiber. By means of this system the phase of a measured light beam can be modulated with an analyzer, and the phase distribution of a fiber is obtained by calculation of the various interference patterns. The measurement theory and structure and some typical applications of this system are demonstrated. The results of measuring RIPs in graded-index fiber are presented. Both the experimental results and theoretical analysis show that the system takes the advantage of high index resolution and of sufficient measurement accuracy for measuring the refractive index of the optical fiber. The system has strong ability to overcome environmental disturbance because of its common-path design. Moreover, one can use the system to measure the RIP along the fiber axis and acquire an image of the three-dimensional RIP of the fiber.

Adaptive metalenses with simultaneous electrical control of focal length, astigmatism, and shift.

PubMed

She, Alan; Zhang, Shuyan; Shian, Samuel; Clarke, David R; Capasso, Federico

2018-02-01

Focal adjustment and zooming are universal features of cameras and advanced optical systems. Such tuning is usually performed longitudinally along the optical axis by mechanical or electrical control of focal length. However, the recent advent of ultrathin planar lenses based on metasurfaces (metalenses), which opens the door to future drastic miniaturization of mobile devices such as cell phones and wearable displays, mandates fundamentally different forms of tuning based on lateral motion rather than longitudinal motion. Theory shows that the strain field of a metalens substrate can be directly mapped into the outgoing optical wavefront to achieve large diffraction-limited focal length tuning and control of aberrations. We demonstrate electrically tunable large-area metalenses controlled by artificial muscles capable of simultaneously performing focal length tuning (>100%) as well as on-the-fly astigmatism and image shift corrections, which until now were only possible in electron optics. The device thickness is only 30 μm. Our results demonstrate the possibility of future optical microscopes that fully operate electronically, as well as compact optical systems that use the principles of adaptive optics to correct many orders of aberrations simultaneously.

Nondestructive defect detection in laser optical coatings

NASA Astrophysics Data System (ADS)

Marrs, C. D.; Porteus, J. O.; Palmer, J. R.

1985-03-01

Defects responsible for laser damage in visible-wavelength mirrors are observed at nondamaging intensities using a new video microscope system. Studies suggest that a defect scattering phenomenon combined with lag characteristics of video cameras makes this possible. Properties of the video-imaged light are described for multilayer dielectric coatings and diamond-turned metals.

Fibre optic microarrays.

PubMed

Walt, David R

2010-01-01

This tutorial review describes how fibre optic microarrays can be used to create a variety of sensing and measurement systems. This review covers the basics of optical fibres and arrays, the different microarray architectures, and describes a multitude of applications. Such arrays enable multiplexed sensing for a variety of analytes including nucleic acids, vapours, and biomolecules. Polymer-coated fibre arrays can be used for measuring microscopic chemical phenomena, such as corrosion and localized release of biochemicals from cells. In addition, these microarrays can serve as a substrate for fundamental studies of single molecules and single cells. The review covers topics of interest to chemists, biologists, materials scientists, and engineers.

Fabrication and Theoretical Evaluation of Microlens Arrays on Layered Polymers

NASA Astrophysics Data System (ADS)

Oder, Tom; McMaster, Michael; Merlo, Corey; Bagheri, Camron; Reakes, Clayton; Petrus, Joshua; Li, Dingqiang; Crescimanno, Michael; Andrews, James

2014-03-01

Arrays of microlens were fabricated on nano-layered polymers using reactive ion etching. Semi hemispherical patterns with diameters ranging from 20 to 80 micrometers were first formed on a thick photoresist film that was spin-coated on the layered polymers using standard photolithographic process employing a gray scale glass mask. These patterns were then transferred to the polymers using dry etching in a reactive ion etching system. The optimized etch condition included a mixture of sulfur hexafluoride and oxygen, which resulted in an etch depth of 5 micrometers and successfully exposed the individual sub-micron thick layers in the polymers. Physical characterization of the microlens arrays was done using atomic force microscope and scanning electron microscope. We combine basic physical optics theory with the transfer matrix analysis of optical transport in nano-layered polymers to address subtleties in the chromatic response of microlenses made from these materials. In particular this method explains the len's behavior in and around the reflection band of the materials. We wish to acknowledge support of funds from NSF through its Center for Layered Polymeric Systems (CLiPS) at Case Western Reserve University.

Mapping optical path length and image enhancement using quantitative orientation-independent differential interference contrast microscopy

PubMed Central

Shribak, Michael; Larkin, Kieran G.; Biggs, David

2017-01-01

Abstract. We describe the principles of using orientation-independent differential interference contrast (OI-DIC) microscopy for mapping optical path length (OPL). Computation of the scalar two-dimensional OPL map is based on an experimentally received map of the OPL gradient vector field. Two methods of contrast enhancement for the OPL image, which reveal hardly visible structures and organelles, are presented. The results obtained can be used for reconstruction of a volume image. We have confirmed that a standard research grade light microscope equipped with the OI-DIC and 100×/1.3 NA objective lens, which was not specially selected for minimum wavefront and polarization aberrations, provides OPL noise level of ∼0.5  nm and lateral resolution if ∼300  nm at a wavelength of 546 nm. The new technology is the next step in the development of the DIC microscopy. It can replace standard DIC prisms on existing commercial microscope systems without modification. This will allow biological researchers that already have microscopy setups to expand the performance of their systems. PMID:28060991

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

NASA Astrophysics Data System (ADS)

Egorov, D. I.

2017-06-01

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

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.

Acoustic imaging microscope

DOEpatents

Deason, Vance A.; Telschow, Kenneth L.

2006-10-17

An imaging system includes: an object wavefront source and an optical microscope objective all positioned to direct an object wavefront onto an area of a vibrating subject surface encompassed by a field of view of the microscope objective, and to direct a modulated object wavefront reflected from the encompassed surface area through a photorefractive material; and a reference wavefront source and at least one phase modulator all positioned to direct a reference wavefront through the phase modulator and to direct a modulated reference wavefront from the phase modulator through the photorefractive material to interfere with the modulated object wavefront. The photorefractive material has a composition and a position such that interference of the modulated object wavefront and modulated reference wavefront occurs within the photorefractive material, providing a full-field, real-time image signal of the encompassed surface area.

Complex Pupil Masks for Aberrated Imaging of Closely Spaced Objects

NASA Astrophysics Data System (ADS)

Reddy, A. N. K.; Sagar, D. K.; Khonina, S. N.

2017-12-01

Current approach demonstrates the suppression of optical side-lobes and the contraction of the main lobe in the composite image of two object points of the optical system under the influence of defocusing effect when an asymmetric phase edges are imposed over the apodized circular aperture. The resolution of two point sources having different intensity ratio is discussed in terms of the modified Sparrow criterion, functions of the degree of coherence of the illumination, the intensity difference and the degree of asymmetric phase masking. Here we have introduced and explored the effects of focus aberration (defect-of-focus) on the two-point resolution of the optical systems. Results on the aberrated composite image of closely spaced objects with amplitude mask and asymmetric phase masks forms a significant contribution in astronomical and microscopic observations.

Design and simulation of high resolution optical imaging system based on near-field using solid immersion lens with NA = 2.2

NASA Astrophysics Data System (ADS)

Abbasian, Karim; Sadeghi, Rasool; Sadeghi, Parvin

2014-03-01

In this work, by changing annular aperture zones transmittance, we could get a spot size smaller than any reported one by utilizing annular aperture. Where, by dividing the annular aperture to more than three zones and utilizing of Sony corporation Produced SIL that has NA higher than 2, we could improve imaging resolution for radial polarization (RP); also we could decrease the FWHM from around ? to near ?. Here, the FWHM variation, according to the refractive index changing, has decreased to zero for RP. After that, circular polarization (CP) has been introduced to get a spot size less than ?. This image resolution improving can be applied to enhance optical data storage, microscopes and lithographic and other high accurate optical systems.

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.

A fast low-power optical memory based on coupled micro-ring lasers

NASA Astrophysics Data System (ADS)

Hill, Martin T.; Dorren, Harmen J. S.; de Vries, Tjibbe; Leijtens, Xaveer J. M.; den Besten, Jan Hendrik; Smalbrugge, Barry; Oei, Yok-Siang; Binsma, Hans; Khoe, Giok-Djan; Smit, Meint K.

2004-11-01

The increasing speed of fibre-optic-based telecommunications has focused attention on high-speed optical processing of digital information. Complex optical processing requires a high-density, high-speed, low-power optical memory that can be integrated with planar semiconductor technology for buffering of decisions and telecommunication data. Recently, ring lasers with extremely small size and low operating power have been made, and we demonstrate here a memory element constructed by interconnecting these microscopic lasers. Our device occupies an area of 18 × 40µm2 on an InP/InGaAsP photonic integrated circuit, and switches within 20ps with 5.5fJ optical switching energy. Simulations show that the element has the potential for much smaller dimensions and switching times. Large numbers of such memory elements can be densely integrated and interconnected on a photonic integrated circuit: fast digital optical information processing systems employing large-scale integration should now be viable.

Compact Microscope Imaging System with Intelligent Controls

NASA Technical Reports Server (NTRS)

McDowell, Mark

2004-01-01

The figure presents selected views of a compact microscope imaging system (CMIS) that includes a miniature video microscope, a Cartesian robot (a computer- controlled three-dimensional translation stage), and machine-vision and control subsystems. The CMIS was built from commercial off-the-shelf instrumentation, computer hardware and software, and custom machine-vision software. The machine-vision and control subsystems include adaptive neural networks that afford a measure of artificial intelligence. The CMIS can perform several automated tasks with accuracy and repeatability . tasks that, heretofore, have required the full attention of human technicians using relatively bulky conventional microscopes. In addition, the automation and control capabilities of the system inherently include a capability for remote control. Unlike human technicians, the CMIS is not at risk of becoming fatigued or distracted: theoretically, it can perform continuously at the level of the best human technicians. In its capabilities for remote control and for relieving human technicians of tedious routine tasks, the CMIS is expected to be especially useful in biomedical research, materials science, inspection of parts on industrial production lines, and space science. The CMIS can automatically focus on and scan a microscope sample, find areas of interest, record the resulting images, and analyze images from multiple samples simultaneously. Automatic focusing is an iterative process: The translation stage is used to move the microscope along its optical axis in a succession of coarse, medium, and fine steps. A fast Fourier transform (FFT) of the image is computed at each step, and the FFT is analyzed for its spatial-frequency content. The microscope position that results in the greatest dispersal of FFT content toward high spatial frequencies (indicating that the image shows the greatest amount of detail) is deemed to be the focal position.

Detection of cervical intraepithelial neoplasia by using optical coherence tomography in combination with microscopy

NASA Astrophysics Data System (ADS)

Gallwas, Julia; Jalilova, Aydan; Ladurner, Roland; Kolben, Theresa Maria; Kolben, Thomas; Ditsch, Nina; Homann, Christian; Lankenau, Eva; Dannecker, Christian

2017-01-01

Optical coherence tomography (OCT) is a noninvasive high-resolution imaging technique that permits the detection of cancerous and precancerous lesions of the uterine cervix. The purpose of this study was to evaluate a new system that integrates an OCT device into a microscope. OCT images were taken from loop electrosurgical excision procedure (LEEP) specimens under microscopic guidance. The images were blinded with respect to their origin within the microscopic image and analyzed independently by two investigators using initially defined criteria and later compared to the corresponding histology. Sensitivity and specificity were calculated with respect to the correct identification of high-grade squamous intraepithelial lesions (HSIL). The interinvestigator agreement was assessed by using Cohen's kappa statistics. About 160 OCT images were obtained from 20 LEEP specimens. Sixty randomly chosen images were used to define reproducible criteria for evaluation. The assessment of the remaining 100 images showed a sensitivity of 88% (second investigator 84%) and a specificity of 69% (65%) in detecting HSIL. Surgical microscopy-guided OCT appears to be a promising technique for immediate assessment of microanatomical changes. In the gynecological setting, the combination of OCT with a colposcope may improve the detection of high-grade squamous intraepithelial lesions.

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

PubMed

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

2018-02-28

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

Enhancing the performance of the light field microscope using wavefront coding.

PubMed

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

2014-10-06

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.

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.

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

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

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.

Automated micromanipulation desktop station based on mobile piezoelectric microrobots

NASA Astrophysics Data System (ADS)

Fatikow, Sergej

1996-12-01

One of the main problems of present-day research on microsystem technology (MST) is to assemble a whole micro- system from different microcomponents. This paper presents a new concept of an automated micromanipulation desktop- station including piezoelectrically driven microrobots placed on a high-precise x-y-stage of a light microscope, a CCD-camera as a local sensor subsystem, a laser sensor unit as a global sensor subsystem, a parallel computer system with C167 microcontrollers, and a Pentium PC equipped additionally with an optical grabber. The microrobots can perform high-precise manipulations (with an accuracy of up to 10 nm) and a nondestructive transport (at a speed of about 3 cm/sec) of very small objects under the microscope. To control the desktop-station automatically, an advanced control system that includes a task planning level and a real-time execution level is being developed. The main function of the task planning sub-system is to interpret the implicit action plan and to generate a sequence of explicit operations which are sent to the execution level of the control system. The main functions of the execution control level are the object recognition, image processing and feedback position control of the microrobot and the microscope stage.

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.

Perspective: Electronic systems of knowledge in the world of virtual microscopy.

PubMed

Maybury, Terrence; Farah, Camile S

2009-09-01

Across a broad range of medical disciplines, learning how to use an optical or light microscope has been a mandatory inclusion in the undergraduate curriculum. The development of virtual microscopy (VM) technology during the past 10 years has called into question the use of the optical microscope in educational contexts. VM allows slide specimens to be digitized, which, in turn, allows the computer to mimic the workings of the light microscope. This move from analog technology (the light microscope) to digital technology (the computer as microscope) is part of the many significant changes going on in education, a singular manifestation of the broader move from print-literate traditions of knowledge (requiring literacy) to an electronics-literate, or "electrate," mode (requiring "electracy"). VM is here used as an exemplar of this broad transition from literacy to electracy, some components of which include data deluge, a multimodal structure, and modularity. Understandably, this transition is important to clarify educationally, especially in a global context mediated via digital means. A related aspect of these educational changes is the move from teacher-directed learning to student-centered learning, or "user-led education," which points to a redefinition of "pedagogy" as "andragogy." The dissemination of the specific value of VM, then, is critical to both learners and teachers and to a more coherent understanding of electracy. A practical consequence of this clarity might be a better application of this knowledge in the evolving fields of computer simulation and telemedicine, areas in which today's medical students will need future expertise.

Hollow fiber-optic Raman probes for small experimental animals

NASA Astrophysics Data System (ADS)

Katagiri, Takashi; Hattori, Yusuke; Suzuki, Toshiaki; Matsuura, Yuji; Sato, Hidetoshi

2007-02-01

Two types of hollow fiber-optic probes are developed to measure the in vivo Raman spectra of small animals. One is the minimized probe which is end-sealed with the micro-ball lens. The measured spectra reflect the information of the sample's sub-surface. This probe is used for the measurement of the esophagus and the stomach via an endoscope. The other probe is a confocal Raman probe which consists of a single fiber and a lens system. It is integrated into the handheld microscope. A simple and small multimodal probe is realized because the hollow optical fiber requires no optical filters. The performance of each probe is examined and the effectiveness of these probes for in vivo Raman spectroscopy is shown by animal tests.

Integrated optical motor.

PubMed

Kelemen, Lóránd; Valkai, Sándor; Ormos, Pál

2006-04-20

A light-driven micrometer-sized mechanical motor is created by laser-light-induced two-photon photopolymerization. All necessary components of the engine are built upon a glass surface by an identical procedure and include the following: a rigid mechanical framework, a rotor freely rotating on an axis, and an integrated optical waveguide carrying the actuating light to the rotor. The resulting product is a most practical stand-alone system. The light introduced into the integrated optical waveguide input of the motor provides the driving force: neither optical tweezers or even a microscope are needed for the function. The power and efficiency of the motor are evaluated. The independent unit is expected to become an important component of more complex integrated lab-on-a-chip devices.

Extreme ultraviolet patterned mask inspection performance of advanced projection electron microscope system for 11nm half-pitch generation

NASA Astrophysics Data System (ADS)

Hirano, Ryoichi; Iida, Susumu; Amano, Tsuyoshi; Watanabe, Hidehiro; Hatakeyama, Masahiro; Murakami, Takeshi; Suematsu, Kenichi; Terao, Kenji

2016-03-01

Novel projection electron microscope optics have been developed and integrated into a new inspection system named EBEYE-V30 ("Model EBEYE" is an EBARA's model code) , and the resulting system shows promise for application to half-pitch (hp) 16-nm node extreme ultraviolet lithography (EUVL) patterned mask inspection. To improve the system's inspection throughput for 11-nm hp generation defect detection, a new electron-sensitive area image sensor with a high-speed data processing unit, a bright and stable electron source, and an image capture area deflector that operates simultaneously with the mask scanning motion have been developed. A learning system has been used for the mask inspection tool to meet the requirements of hp 11-nm node EUV patterned mask inspection. Defects are identified by the projection electron microscope system using the "defectivity" from the characteristics of the acquired image. The learning system has been developed to reduce the labor and costs associated with adjustment of the detection capability to cope with newly-defined mask defects. We describe the integration of the developed elements into the inspection tool and the verification of the designed specification. We have also verified the effectiveness of the learning system, which shows enhanced detection capability for the hp 11-nm node.

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.

Lensless Imaging for Battlefield On-Chip Blood Diagnostics

DTIC Science & Technology

2010-12-06

Applications” 7th International Conference on Optics-Photonics Design and Fabrication, (April 19-21 2010) Yokohoma, Japan 16. A. Ozcan, “Photonics based...MicroTAS 2010 - The 14th International Conference on Miniaturized Systems for Chemistry and Life Sciences, October 3-7, 2010, Groningen, The...Chip Microscope,” MicroTAS 2010 - The 14th International Conference on Miniaturized Systems for Chemistry and Life Sciences, October 3-7, 2010

MultiFocus Polarization Microscope (MF-PolScope) for 3D polarization imaging of up to 25 focal planes simultaneously

PubMed Central

Abrahamsson, Sara; McQuilken, Molly; Mehta, Shalin B.; Verma, Amitabh; Larsch, Johannes; Ilic, Rob; Heintzmann, Rainer; Bargmann, Cornelia I.; Gladfelter, Amy S.; Oldenbourg, Rudolf

2015-01-01

We have developed an imaging system for 3D time-lapse polarization microscopy of living biological samples. Polarization imaging reveals the position, alignment and orientation of submicroscopic features in label-free as well as fluorescently labeled specimens. Optical anisotropies are calculated from a series of images where the sample is illuminated by light of different polarization states. Due to the number of images necessary to collect both multiple polarization states and multiple focal planes, 3D polarization imaging is most often prohibitively slow. Our MF-PolScope system employs multifocus optics to form an instantaneous 3D image of up to 25 simultaneous focal-planes. We describe this optical system and show examples of 3D multi-focus polarization imaging of biological samples, including a protein assembly study in budding yeast cells. PMID:25837112

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.

Novel approach to real-time flash photolysis and confocal [Ca2+] imaging

PubMed Central

Sobie, Eric A.; Kao, Joseph P.Y.; Lederer, W. J.

2008-01-01

Flash photolysis of “caged” compounds using ultraviolet light is a powerful experimental technique for producing rapid changes in concentrations of bioactive signaling molecules. Studies that employ this technique have used diverse strategies for controlling the spatial and temporal application of light to the specimen. Here we describe a new system for flash photolysis that delivers light from a pulsed, adjustable intensity laser through an optical fiber coupled into the epifluorescence port of a commercial confocal microscope. Photolysis is achieved with extremely brief (5 ns) pulses of ultraviolet light (355 nm) that can be synchronized with respect to confocal laser scanning. The system described also localizes the UV intensity spatially so that uncaging only occurs in defined sub-cellular regions; moreover, since the microscope optics are used in localization, the photolysis volume can be easily adjusted. Experiments performed on rat ventricular myocytes loaded with the Ca2+ indicator fluo-3 and the Ca2+ cage NP-EGTA demonstrate the system's capabilities. Localized intracellular increases in [Ca2+] can trigger sarcoplasmic reticular Ca2+ release events such as Ca2+ sparks and, under certain conditions, regenerative Ca2+ waves. This relatively simple and inexpensive system is therefore a useful tool for examining local signaling in heart and other tissues. PMID:17323075

Quantitative optical imaging and sensing by joint design of point spread functions and estimation algorithms

NASA Astrophysics Data System (ADS)

Quirin, Sean Albert

The joint application of tailored optical Point Spread Functions (PSF) and estimation methods is an important tool for designing quantitative imaging and sensing solutions. By enhancing the information transfer encoded by the optical waves into an image, matched post-processing algorithms are able to complete tasks with improved performance relative to conventional designs. In this thesis, new engineered PSF solutions with image processing algorithms are introduced and demonstrated for quantitative imaging using information-efficient signal processing tools and/or optical-efficient experimental implementations. The use of a 3D engineered PSF, the Double-Helix (DH-PSF), is applied as one solution for three-dimensional, super-resolution fluorescence microscopy. The DH-PSF is a tailored PSF which was engineered to have enhanced information transfer for the task of localizing point sources in three dimensions. Both an information- and optical-efficient implementation of the DH-PSF microscope are demonstrated here for the first time. This microscope is applied to image single-molecules and micro-tubules located within a biological sample. A joint imaging/axial-ranging modality is demonstrated for application to quantifying sources of extended transverse and axial extent. The proposed implementation has improved optical-efficiency relative to prior designs due to the use of serialized cycling through select engineered PSFs. This system is demonstrated for passive-ranging, extended Depth-of-Field imaging and digital refocusing of random objects under broadband illumination. Although the serialized engineered PSF solution is an improvement over prior designs for the joint imaging/passive-ranging modality, it requires the use of multiple PSFs---a potentially significant constraint. Therefore an alternative design is proposed, the Single-Helix PSF, where only one engineered PSF is necessary and the chromatic behavior of objects under broadband illumination provides the necessary information transfer. The matched estimation algorithms are introduced along with an optically-efficient experimental system to image and passively estimate the distance to a test object. An engineered PSF solution is proposed for improving the sensitivity of optical wave-front sensing using a Shack-Hartmann Wave-front Sensor (SHWFS). The performance limits of the classical SHWFS design are evaluated and the engineered PSF system design is demonstrated to enhance performance. This system is fabricated and the mechanism for additional information transfer is identified.

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...

Microscopic Image of Martian Surface Material on a Silicone Substrate

NASA Technical Reports Server (NTRS)

2008-01-01

[figure removed for brevity, see original site] Click on image for larger version of Figure 1

This image taken by the Optical Microscope on NASA's Phoenix Mars Lander shows soil sprinkled from the lander's Robot Arm scoop onto a silicone substrate. The substrate was then rotated in front of the microscope. This is the first sample collected and delivered for instrumental analysis onboard a planetary lander since NASA's Viking Mars missions of the 1970s. It is also the highest resolution image yet seen of Martian soil.

The image is dominated by fine particles close to the resolution of the microscope. These particles have formed clumps, which may be a smaller scale version of what has been observed by Phoenix during digging of the surface material.

The microscope took this image during Phoenix's Sol 17 (June 11), or the 17th Martian day after landing. The scale bar is 1 millimeter (0.04 inch).

Zooming in on the Martian Soil

In figure 1, three zoomed-in portions are shown with an image of Martian soil particles taken by the Optical Microscope on NASA's Phoenix Mars Lander.

The left zoom box shows a composite particle. The top of the particle has a green tinge, possibly indicating olivine. The bottom of the particle has been reimaged at a different focus position in black and white (middle zoom box), showing that this is a clump of finer particles.

The right zoom box shows a rounded, glassy particle, similar to those which have also been seen in an earlier sample of airfall dust collected on a surface exposed during landing.

The shadows at the bottom of image are of the beams of the Atomic Force Microscope.

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.

Intraoperative assessment of laryngeal pathologies with optical coherence tomography integrated into a surgical microscope.

PubMed

Englhard, Anna S; Betz, Tom; Volgger, Veronika; Lankenau, Eva; Ledderose, Georg J; Stepp, Herbert; Homann, Christian; Betz, Christian S

2017-07-01

Endoscopic examination followed by tissue biopsy is the gold standard in the evaluation of lesions of the upper aerodigestive tract. However, it can be difficult to distinguish between healthy mucosa, dysplasia, and invasive carcinoma. Optical coherence tomography (OCT) is a non-invasive technique which acquires high-resolution, cross-sectional images of tissue in vivo. Integrated into a surgical microscope, it allows the intraoperative evaluation of lesions simultaneously with microscopic visualization. In a prospective case series, we evaluated the use of OCT integrated into a surgical microscope during microlaryngoscopy to help differentiating various laryngeal pathologies. 33 patients with laryngeal pathologies were examined with an OCT- microscope (OPMedT iOCT-camera, HS Hi-R 1000G-microscope, Haag-Streit Surgical GmbH, Wedel, Germany) during microlaryngoscopy. The suspected intraoperative diagnoses were compared to the histopathological reports of subsequent tissue biopsies. Hands-free non-contact OCT revealed high-resolution images of the larynx with a varying penetration depth of up to 1.2 mm and an average of 0.6 mm. Picture quality was variable. OCT showed disorders of horizontal tissue layering in dysplasias with a disruption of the basement membrane in carcinomas. When comparing the suspected diagnosis during OCT-supported microlaryngoscopy with histology, 79% of the laryngeal lesions could be correctly identified. Premalignant lesions were difficult to diagnose and falsely classified as carcinoma. OCT integrated into a surgical microscope seems to be a promising adjunct tool to discriminate pathologies of the upper aerodigestive tract intraoperatively. However, picture quality and penetration depth were variable. Although premalignant lesions were difficult to diagnose, the system proved overall helpful for the intraoperative discrimination of benign and malignant tumors. Further studies will be necessary to define its value in the future. Lasers Surg. Med. 49:490-497, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Biological applications of an LCoS-based programmable array microscope (PAM)

NASA Astrophysics Data System (ADS)

Hagen, Guy M.; Caarls, Wouter; Thomas, Martin; Hill, Andrew; Lidke, Keith A.; Rieger, Bernd; Fritsch, Cornelia; van Geest, Bert; Jovin, Thomas M.; Arndt-Jovin, Donna J.

2007-02-01

We report on a new generation, commercial prototype of a programmable array optical sectioning fluorescence microscope (PAM) for rapid, light efficient 3D imaging of living specimens. The stand-alone module, including light source(s) and detector(s), features an innovative optical design and a ferroelectric liquid-crystal-on-silicon (LCoS) spatial light modulator (SLM) instead of the DMD used in the original PAM design. The LCoS PAM (developed in collaboration with Cairn Research, Ltd.) can be attached to a port of a(ny) unmodified fluorescence microscope. The prototype system currently operated at the Max Planck Institute incorporates a 6-position high-intensity LED illuminator, modulated laser and lamp light sources, and an Andor iXon emCCD camera. The module is mounted on an Olympus IX71 inverted microscope with 60-150X objectives with a Prior Scientific x,y, and z high resolution scanning stages. Further enhancements recently include: (i) point- and line-wise spectral resolution and (ii) lifetime imaging (FLIM) in the frequency domain. Multiphoton operation and other nonlinear techniques should be feasible. The capabilities of the PAM are illustrated by several examples demonstrating single molecule as well as lifetime imaging in live cells, and the unique capability to perform photoconversion with arbitrary patterns and high spatial resolution. Using quantum dot coupled ligands we show real-time binding and subsequent trafficking of individual ligand-growth factor receptor complexes on and in live cells with a temporal resolution and sensitivity exceeding those of conventional CLSM systems. The combined use of a blue laser and parallel LED or visible laser sources permits photoactivation and rapid kinetic analysis of cellular processes probed by photoswitchable visible fluorescent proteins such as DRONPA.

x-y-recording in transmission electron microscopy. A versatile and inexpensive interface to personal computers with application to stereology.

PubMed

Rickmann, M; Siklós, L; Joó, F; Wolff, J R

1990-09-01

An interface for IBM XT/AT-compatible computers is described which has been designed to read the actual specimen stage position of electron microscopes. The complete system consists of (i) optical incremental encoders attached to the x- and y-stage drivers of the microscope, (ii) two keypads for operator input, (iii) an interface card fitted to the bus of the personal computer, (iv) a standard configuration IBM XT (or compatible) personal computer optionally equipped with a (v) HP Graphic Language controllable colour plotter. The small size of the encoders and their connection to the stage drivers by simple ribbed belts allows an easy adaptation of the system to most electron microscopes. Operation of the interface card itself is supported by any high-level language available for personal computers. By the modular concept of these languages, the system can be customized to various applications, and no computer expertise is needed for actual operation. The present configuration offers an inexpensive attachment, which covers a wide range of applications from a simple notebook to high-resolution (200-nm) mapping of tissue. Since section coordinates can be processed in real-time, stereological estimations can be derived directly "on microscope". This is exemplified by an application in which particle numbers were determined by the disector method.

Imaging pancreatobiliary ductal system with optical coherence tomography: A review

PubMed Central

Mahmud, Mohammad S; May, Gray R; Kamal, Mohammad M; Khwaja, Ahmed S; Sun, Carry; Vitkin, Alex; Yang, Victor XD

2013-01-01

An accurate, noninvasive and cost-effective method of in situ tissue evaluation during endoscopy would be highly advantageous for the detection of dysplasia or early cancer and for identifying different disease stages. Optical coherence tomography (OCT) is a noninvasive, high-resolution (1-10 μm) emerging optical imaging method with potential for identifying microscopic subsurface features in the pancreatic and biliary ductal system. Tissue microstructure of pancreaticobiliary ductal system has been successfully imaged by inserting an OCT probe through a standard endoscope operative channel. High-resolution OCT images and the technique’s endoscopic compatibility have allowed for the microstructural diagnostic of the pancreatobiliary diseases. In this review, we discussed currently available pancreaticobiliary ductal imaging systems to assess the pancreatobiliary tissue microstructure and to evaluate varieties of pancreaticobiliary disorders and diseases. Results show that OCT can improve the quality of images of pancreatobiliary system during endoscopic retrograde cholangiopancheatography procedure, which may be important in distinguishing between the neoplastic and non-neoplastic lesions. PMID:24255746

New solutions to realize complex optical systems by a combination of diffractive and refractive optical components

NASA Astrophysics Data System (ADS)

Brunner, Robert; Steiner, Reinhard; Dobschal, Hans-Juergen; Martin, Dietrich; Burkhardt, Matthias; Helgert, Michael

2003-11-01

Diffractive optical elements (DOEs) have a great potential in the complete or partial substitution of refractive or reflective optical elements in imaging systems. The greater design flexibility compared to an all-refractive/reflective solution allows a more convenient realization of the optical systems and additionally opens up new possibilities for optimizing the performance or compactness. To demonstrate the opportunities of the hybrid optical concept we discuss different imaging systems for various applications. We present the lens design of a hybrid microscope objective which is especially applicable for wafer inspection technologies. Meeting the requirements for such a system used in the deep-UV regime (248 nm) is very challenging. The short wavelength limits the material selection and demands cement free optical groups. The additional requirement of an autofocus system, working at a wavelength in the near infrared region, is fulfilled by the special combination of two selected and adjusted DOEs. Furthermore, we discuss the opportunities of the hybrid concept c of a slit lamp used for ophthalmologic examinations. The DOEs are the basic elements of this hybrid concept. We demonstrate that holographic lithography is an appropriate technology to realize a wide variety of elements with different profile geometries. We address in particular the additional possibilities of an UV-laser system as an exposure tool. Additionally to the high spatial frequencies, the 266 nm exposure wavelength allows the use of novel photo resists with advantageous development behavior.

Near-Field Scanning Optical Microscopy and Raman Microscopy.

NASA Astrophysics Data System (ADS)

Harootunian, Alec Tate

1987-09-01

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

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.

Microscopic Comparison of Airfall Dust to Martian Soil

NASA Technical Reports Server (NTRS)

2008-01-01

This pair of images taken by the Optical Microscope on NASA's Phoenix Mars Lander offers a side-by-side comparison of an airfall dust sample collected on a substrate exposed during landing (left) and a soil sample scooped up from the surface of the ground beside the lander. In both cases the sample is collected on a silicone substrate, which provides a sticky surface holding sample particles for observation by the microscope.

Similar fine particles at the resolution limit of the microscope are seen in both samples, indicating that the soil has formed from settling of dust.

The microscope took the image on the left during Phoenix's Sol 9 (June 3, 2008), or the ninth Martian day after landing. It took the image on the right during Sol 17 (June 11, 2008).

The scale bar is 1 millimeter (0.04 inch).

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.

Development of a SEM-based low-energy in-line electron holography microscope for individual particle imaging.

PubMed

Adaniya, Hidehito; Cheung, Martin; Cassidy, Cathal; Yamashita, Masao; Shintake, Tsumoru

2018-05-01

A new SEM-based in-line electron holography microscope has been under development. The microscope utilizes conventional SEM and BF-STEM functionality to allow for rapid searching of the specimen of interest, seamless interchange between SEM, BF-STEM and holographic imaging modes, and makes use of coherent low-energy in-line electron holography to obtain low-dose, high-contrast images of light element materials. We report here an overview of the instrumentation and first experimental results on gold nano-particles and carbon nano-fibers for system performance tests. Reconstructed images obtained from the holographic imaging mode of the new microscope show substantial image contrast and resolution compared to those acquired by SEM and BF-STEM modes, demonstrating the feasibility of high-contrast imaging via low-energy in-line electron holography. The prospect of utilizing the new microscope to image purified biological specimens at the individual particle level is discussed and electron optical issues and challenges to further improve resolution and contrast are considered. Copyright © 2018 Elsevier B.V. All rights reserved.

Schematic Animation of Phoenix's Microscope Station

NASA Technical Reports Server (NTRS)

2008-01-01

[figure removed for brevity, see original site] Click on image for animation

This animation shows the workings of the microscope station of the Microscopy, Electrochemistry and Conductivity Analyzer (MECA) instrument suite of NASA's Phoenix Mars Lander.

Samples are delivered to the horizontal portion of the sample wheel (yellow) that pokes outside an opening in the box enclosure. The wheel rotates to present the sample to the microscopes. The Optical Microscope (red) can see particles a little smaller than one-tenth the diameter of a human hair. The Atomic Force Microscope (pink) can see particles forty time smaller. The samples are on a variety of substrate surfaces, the small circles on the beveled edge of the sample wheel. For scale, the diameter of the wheel is about 14 centimeters (5.5 inches). Each substrate is a circle 3 millimeters (0.1 inch) in diameter.

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.

Membrane Mirrors With Bimorph Shape Actuators

NASA Technical Reports Server (NTRS)

Yang, Eui-Hyeok

2003-01-01

Deformable mirrors of a proposed type would be equipped with relatively-large-stroke microscopic piezoelectric actuators that would be used to maintain their reflective surfaces in precise shapes. These mirrors would be members of the class of MEMS-DM (for microelectromechanical system deformable mirror) devices, which offer potential for a precise optical control in adaptive-optics applications in such diverse fields as astronomy and vision science. The proposed mirror would be fabricated, in part, by use of a membrane-transfer technique. The actuator design would contain bimorph-type piezoelectric actuators.

Optical Tweezers for Sample Fixing in Micro-Diffraction Experiments

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

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

2007-01-19

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

Note: Tormenta: An open source Python-powered control software for camera based optical microscopy.

PubMed

Barabas, Federico M; Masullo, Luciano A; Stefani, Fernando D

2016-12-01

Until recently, PC control and synchronization of scientific instruments was only possible through closed-source expensive frameworks like National Instruments' LabVIEW. Nowadays, efficient cost-free alternatives are available in the context of a continuously growing community of open-source software developers. Here, we report on Tormenta, a modular open-source software for the control of camera-based optical microscopes. Tormenta is built on Python, works on multiple operating systems, and includes some key features for fluorescence nanoscopy based on single molecule localization.

Note: Tormenta: An open source Python-powered control software for camera based optical microscopy

NASA Astrophysics Data System (ADS)

Barabas, Federico M.; Masullo, Luciano A.; Stefani, Fernando D.

2016-12-01

Until recently, PC control and synchronization of scientific instruments was only possible through closed-source expensive frameworks like National Instruments' LabVIEW. Nowadays, efficient cost-free alternatives are available in the context of a continuously growing community of open-source software developers. Here, we report on Tormenta, a modular open-source software for the control of camera-based optical microscopes. Tormenta is built on Python, works on multiple operating systems, and includes some key features for fluorescence nanoscopy based on single molecule localization.

Photon theory hypothesis about photon tunneling microscope's subwavelength resolution

NASA Astrophysics Data System (ADS)

Zhu, Yanbin; Ma, Junfu

1995-09-01

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

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)

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.