Infrared absorption nano-spectroscopy using sample photoexpansion induced by tunable quantum cascade lasers.

PubMed

Lu, Feng; Belkin, Mikhail A

2011-10-10

We report a simple technique that allows obtaining mid-infrared absorption spectra with nanoscale spatial resolution under low-power illumination from tunable quantum cascade lasers. Light absorption is detected by measuring associated sample thermal expansion with an atomic force microscope. To detect minute thermal expansion we tune the repetition frequency of laser pulses in resonance with the mechanical frequency of the atomic force microscope cantilever. Spatial resolution of better than 50 nm is experimentally demonstrated.

Determination of scattering structures from spatial coherence measurements.

PubMed

Zarubin, A M

1996-03-01

A new method of structure determination and microscopic imaging with short-wavelength radiations (charged particles, X-rays, neutrons), based on measurements of the modulus and the phase of the degree of spatial coherence of the scattered radiation, is developed. The underlying principle of the method--transfer of structural information about the scattering potential via spatial coherence of the secondary (scattering) source of radiation formed by this potential--is expressed by the generalization of the van Cittert-Zernike theorem to wave and particle scattering [A.M. Zarubin, Opt. Commun. 100 (1993) 491; Opt. Commun. 102 (1993) 543]. Shearing interferometric techniques are proposed for implementing the above measurements; the limits of spatial resolution attainable by reconstruction of the absolute square of a 3D scattering potential and its 2D projections from the measurements are analyzed. It is shown theoretically that 3D imaging with atomic resolution can be realized in a "synthetic aperture" electron or ion microscope and that a 3D resolution of about 6 nm can be obtained with a "synthetic aperture" X-ray microscope. A proof-of-principle optical experiment is presented.

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.

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.

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

PubMed

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

2018-05-01

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

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

PubMed

Jin, P; Li, X

2015-12-01

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

A 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.

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

NASA Astrophysics Data System (ADS)

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

2011-09-01

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

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.

Interference Confocal Microscope Integrated with Spatial Phase Shifter.

PubMed

Wang, Weibo; Gu, Kang; You, Xiaoyu; Tan, Jiubin; Liu, Jian

2016-08-24

We present an interference confocal microscope (ICM) with a new single-body four-step simultaneous phase-shifter device designed to obtain high immunity to vibration. The proposed ICM combines the respective advantages of simultaneous phase shifting interferometry and bipolar differential confocal microscopy to obtain high axis resolution, large dynamic range, and reduce the sensitivity to vibration and reflectance disturbance seamlessly. A compact single body spatial phase shifter is added to capture four phase-shifted interference signals simultaneously without time delay and construct a stable and space-saving simplified interference confocal microscope system. The test result can be obtained by combining the interference phase response and the bipolar property of differential confocal microscopy without phase unwrapping. Experiments prove that the proposed microscope is capable of providing stable measurements with 1 nm of axial depth resolution for either low- or high-numerical aperture objective lenses.

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.

High spatial resolution detection of low-energy electrons using an event-counting method, application to point projection microscopy

NASA Astrophysics Data System (ADS)

Salançon, Evelyne; Degiovanni, Alain; Lapena, Laurent; Morin, Roger

2018-04-01

An event-counting method using a two-microchannel plate stack in a low-energy electron point projection microscope is implemented. 15 μm detector spatial resolution, i.e., the distance between first-neighbor microchannels, is demonstrated. This leads to a 7 times better microscope resolution. Compared to previous work with neutrons [Tremsin et al., Nucl. Instrum. Methods Phys. Res., Sect. A 592, 374 (2008)], the large number of detection events achieved with electrons shows that the local response of the detector is mainly governed by the angle between the hexagonal structures of the two microchannel plates. Using this method in point projection microscopy offers the prospect of working with a greater source-object distance (350 nm instead of 50 nm), advancing toward atomic resolution.

Nonlinear vibrational microscopy

DOEpatents

Holtom, Gary R.; Xie, Xiaoliang Sunney; Zumbusch, Andreas

2000-01-01

The present invention is a method and apparatus for microscopic vibrational imaging using coherent Anti-Stokes Raman Scattering or Sum Frequency Generation. Microscopic imaging with a vibrational spectroscopic contrast is achieved by generating signals in a nonlinear optical process and spatially resolved detection of the signals. The spatial resolution is attained by minimizing the spot size of the optical interrogation beams on the sample. Minimizing the spot size relies upon a. directing at least two substantially co-axial laser beams (interrogation beams) through a microscope objective providing a focal spot on the sample; b. collecting a signal beam together with a residual beam from the at least two co-axial laser beams after passing through the sample; c. removing the residual beam; and d. detecting the signal beam thereby creating said pixel. The method has significantly higher spatial resolution then IR microscopy and higher sensitivity than spontaneous Raman microscopy with much lower average excitation powers. CARS and SFG microscopy does not rely on the presence of fluorophores, but retains the resolution and three-dimensional sectioning capability of confocal and two-photon fluorescence microscopy. Complementary to these techniques, CARS and SFG microscopy provides a contrast mechanism based on vibrational spectroscopy. This vibrational contrast mechanism, combined with an unprecedented high sensitivity at a tolerable laser power level, provides a new approach for microscopic investigations of chemical and biological samples.

Development of scanning x-ray fluorescence microscope with spatial resolution of 30 nm using Kirkpatrick-Baez mirror optics

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

Matsuyama, S.; Mimura, H.; Yumoto, H.

We developed a high-spatial-resolution scanning x-ray fluorescence microscope (SXFM) using Kirkpatrick-Baez mirrors. As a result of two-dimensional focusing tests at BL29XUL of SPring-8, the full width at half maximum of the focused beam was achieved to be 50x30 nm{sup 2} (VxH) under the best focusing conditions. The measured beam profiles were in good agreement with simulated results. Moreover, beam size was controllable within the wide range of 30-1400 nm by changing the virtual source size, although photon flux and size were in a trade-off relationship. To demonstrate SXFM performance, a fine test chart fabricated using focused ion beam system wasmore » observed to determine the best spatial resolution. The element distribution inside a logo mark of SPring-8 in the test chart, which has a minimum linewidth of approximately 50-60 nm, was visualized with a spatial resolution better than 30 nm using the smallest focused x-ray beam.« less

Binary pseudo-random patterned structures for modulation transfer function calibration and resolution characterization of a full-field transmission soft x-ray microscope

DOE PAGES

Yashchuk, V. V.; Fischer, P. J.; Chan, E. R.; ...

2015-12-09

We present a modulation transfer function (MTF) calibration method based on binary pseudo-random (BPR) one-dimensional sequences and two-dimensional arrays as an effective method for spectral characterization in the spatial frequency domain of a broad variety of metrology instrumentation, including interferometric microscopes, scatterometers, phase shifting Fizeau interferometers, scanning and transmission electron microscopes, and at this time, x-ray microscopes. The inherent power spectral density of BPR gratings and arrays, which has a deterministic white-noise-like character, allows a direct determination of the MTF with a uniform sensitivity over the entire spatial frequency range and field of view of an instrument. We demonstrate themore » MTF calibration and resolution characterization over the full field of a transmission soft x-ray microscope using a BPR multilayer (ML) test sample with 2.8 nm fundamental layer thickness. We show that beyond providing a direct measurement of the microscope's MTF, tests with the BPRML sample can be used to fine tune the instrument's focal distance. Finally, our results confirm the universality of the method that makes it applicable to a large variety of metrology instrumentation with spatial wavelength bandwidths from a few nanometers to hundreds of millimeters.« less

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.

Design and commissioning of an aberration-corrected ultrafast spin-polarized low energy electron microscope with multiple electron sources.

PubMed

Wan, Weishi; Yu, Lei; Zhu, Lin; Yang, Xiaodong; Wei, Zheng; Liu, Jefferson Zhe; Feng, Jun; Kunze, Kai; Schaff, Oliver; Tromp, Ruud; Tang, Wen-Xin

2017-03-01

We describe the design and commissioning of a novel aberration-corrected low energy electron microscope (AC-LEEM). A third magnetic prism array (MPA) is added to the standard AC-LEEM with two prism arrays, allowing the incorporation of an ultrafast spin-polarized electron source alongside the standard cold field emission electron source, without degrading spatial resolution. The high degree of symmetries of the AC-LEEM are utilized while we design the electron optics of the ultrafast spin-polarized electron source, so as to minimize the deleterious effect of time broadening, while maintaining full control of electron spin. A spatial resolution of 2nm and temporal resolution of 10ps (ps) are expected in the future time resolved aberration-corrected spin-polarized LEEM (TR-AC-SPLEEM). The commissioning of the three-prism AC-LEEM has been successfully finished with the cold field emission source, with a spatial resolution below 2nm. Copyright © 2017 Elsevier B.V. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

2017-09-01

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

A STED-FLIM microscope applied to imaging the natural killer cell immune synapse

NASA Astrophysics Data System (ADS)

Lenz, M. O.; Brown, A. C. N.; Auksorius, E.; Davis, D. M.; Dunsby, C.; Neil, M. A. A.; French, P. M. W.

2011-03-01

We present a stimulated emission depletion (STED) fluorescence lifetime imaging (FLIM) microscope, excited by a microstructured optical fibre supercontinuum source that is pumped by a femtosecond Ti:Sapphire-laser, which is also used for depletion. Implemented using a piezo-scanning stage on a laser scanning confocal fluorescence microscope system with FLIM realised using time correlated single photon counting (TCSPC), this provides convenient switching between confocal and STED-FLIM with spatial resolution down to below 60 nm. We will present our design considerations to make a robust instrument for biological applications including a comparison between fixed phase plate and spatial light modulator (SLM) approaches to shape the STED beam and the correlation of STED and confocal FLIM microscopy. Following our previous application of FLIM-FRET to study intercellular signalling at the immunological synapse (IS), we are employing STED microscopy to characterize the spatial distribution of cellular molecules with subdiffraction resolution at the IS. In particular, we are imaging cytoskeletal structure at the Natural Killer cell activated immune synapse. We will also present our progress towards multilabel STED microscopy to determine how relative spatial molecular organization, previously undetectable by conventional microscopy techniques, is important for NK cell cytotoxic function. Keywords: STED, Stimulated Emission Depletion Microscopy, Natural Killer (NK) cell, Fluorescence lifetime imaging, FLIM, Super resolution microscopy.

Imaging nanoscale spatial modulation of a relativistic electron beam with a MeV ultrafast electron microscope

NASA Astrophysics Data System (ADS)

Lu, Chao; Jiang, Tao; Liu, Shengguang; Wang, Rui; Zhao, Lingrong; Zhu, Pengfei; Liu, Yaqi; Xu, Jun; Yu, Dapeng; Wan, Weishi; Zhu, Yimei; Xiang, Dao; Zhang, Jie

2018-03-01

An accelerator-based MeV ultrafast electron microscope (MUEM) has been proposed as a promising tool to the study structural dynamics at the nanometer spatial scale and the picosecond temporal scale. Here, we report experimental tests of a prototype MUEM where high quality images with nanoscale fine structures were recorded with a pulsed ˜3 MeV picosecond electron beam. The temporal and spatial resolutions of the MUEM operating in the single-shot mode are about 4 ps (FWHM) and 100 nm (FWHM), corresponding to a temporal-spatial resolution of 4 × 10-19 s m, about 2 orders of magnitude higher than that achieved with state-of-the-art single-shot keV UEM. Using this instrument, we offer the demonstration of visualizing the nanoscale periodic spatial modulation of an electron beam, which may be converted into longitudinal density modulation through emittance exchange to enable production of high-power coherent radiation at short wavelengths. Our results mark a great step towards single-shot nanometer-resolution MUEMs and compact intense x-ray sources that may have widespread applications in many areas of science.

Ultrafast scanning probe microscopy

DOEpatents

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

1995-05-16

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

Ultrafast scanning probe microscopy

DOEpatents

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

1995-01-01

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

Midinfrared absorption measured at a lambda/400 resolution with an atomic force microscope.

PubMed

Houel, Julien; Homeyer, Estelle; Sauvage, Sébastien; Boucaud, Philippe; Dazzi, Alexandre; Prazeres, Rui; Ortéga, Jean-Michel

2009-06-22

Midinfrared absorption can be locally measured using a detection combining an atomic force microscope and a pulsed excitation. This is illustrated for the midinfrared bulk GaAs phonon absorption and for the midinfrared absorption of thin SiO(2) microdisks. We show that the signal given by the cantilever oscillation amplitude of the atomic force microscope follows the spectral dependence of the bulk material absorption. The absorption spatial resolution achieved with microdisks is around 50 nanometer for an optical excitation around 22 micrometer wavelength.

Note: Tandem Kirkpatrick-Baez microscope with sixteen channels for high-resolution laser-plasma diagnostics

NASA Astrophysics Data System (ADS)

Yi, Shengzhen; Zhang, Zhe; Huang, Qiushi; Zhang, Zhong; Wang, Zhanshan; Wei, Lai; Liu, Dongxiao; Cao, Leifeng; Gu, Yuqiu

2018-03-01

Multi-channel Kirkpatrick-Baez (KB) microscopes, which have better resolution and collection efficiency than pinhole cameras, have been widely used in laser inertial confinement fusion to diagnose time evolution of the target implosion. In this study, a tandem multi-channel KB microscope was developed to have sixteen imaging channels with the precise control of spatial resolution and image intervals. This precise control was created using a coarse assembly of mirror pairs with high-accuracy optical prisms, followed by precise adjustment in real-time x-ray imaging experiments. The multilayers coated on the KB mirrors were designed to have substantially the same reflectivity to obtain a uniform brightness of different images for laser-plasma temperature analysis. The study provides a practicable method to achieve the optimum performance of the microscope for future high-resolution applications in inertial confinement fusion experiments.

Visible-to-visible four-photon ultrahigh resolution microscopic imaging with 730-nm diode laser excited nanocrystals.

PubMed

Wang, Baoju; Zhan, Qiuqiang; Zhao, Yuxiang; Wu, Ruitao; Liu, Jing; He, Sailing

2016-01-25

Further development of multiphoton microscopic imaging is confronted with a number of limitations, including high-cost, high complexity and relatively low spatial resolution due to the long excitation wavelength. To overcome these problems, for the first time, we propose visible-to-visible four-photon ultrahigh resolution microscopic imaging by using a common cost-effective 730-nm laser diode to excite the prepared Nd(3+)-sensitized upconversion nanoparticles (Nd(3+)-UCNPs). An ordinary multiphoton scanning microscope system was built using a visible CW diode laser and the lateral imaging resolution as high as 161-nm was achieved via the four-photon upconversion process. The demonstrated large saturation excitation power for Nd(3+)-UCNPs would be more practical and facilitate the four-photon imaging in the application. A sample with fine structure was imaged to demonstrate the advantages of visible-to-visible four-photon ultrahigh resolution microscopic imaging with 730-nm diode laser excited nanocrystals. Combining the uniqueness of UCNPs, the proposed visible-to-visible four-photon imaging would be highly promising and attractive in the field of multiphoton imaging.

Field-Portable Pixel Super-Resolution Colour Microscope

PubMed Central

Greenbaum, Alon; Akbari, Najva; Feizi, Alborz; Luo, Wei; Ozcan, Aydogan

2013-01-01

Based on partially-coherent digital in-line holography, we report a field-portable microscope that can render lensfree colour images over a wide field-of-view of e.g., >20 mm2. This computational holographic microscope weighs less than 145 grams with dimensions smaller than 17×6×5 cm, making it especially suitable for field settings and point-of-care use. In this lensfree imaging design, we merged a colorization algorithm with a source shifting based multi-height pixel super-resolution technique to mitigate ‘rainbow’ like colour artefacts that are typical in holographic imaging. This image processing scheme is based on transforming the colour components of an RGB image into YUV colour space, which separates colour information from brightness component of an image. The resolution of our super-resolution colour microscope was characterized using a USAF test chart to confirm sub-micron spatial resolution, even for reconstructions that employ multi-height phase recovery to handle dense and connected objects. To further demonstrate the performance of this colour microscope Papanicolaou (Pap) smears were also successfully imaged. This field-portable and wide-field computational colour microscope could be useful for tele-medicine applications in resource poor settings. PMID:24086742

Field-portable pixel super-resolution colour microscope.

PubMed

Greenbaum, Alon; Akbari, Najva; Feizi, Alborz; Luo, Wei; Ozcan, Aydogan

2013-01-01

Based on partially-coherent digital in-line holography, we report a field-portable microscope that can render lensfree colour images over a wide field-of-view of e.g., >20 mm(2). This computational holographic microscope weighs less than 145 grams with dimensions smaller than 17×6×5 cm, making it especially suitable for field settings and point-of-care use. In this lensfree imaging design, we merged a colorization algorithm with a source shifting based multi-height pixel super-resolution technique to mitigate 'rainbow' like colour artefacts that are typical in holographic imaging. This image processing scheme is based on transforming the colour components of an RGB image into YUV colour space, which separates colour information from brightness component of an image. The resolution of our super-resolution colour microscope was characterized using a USAF test chart to confirm sub-micron spatial resolution, even for reconstructions that employ multi-height phase recovery to handle dense and connected objects. To further demonstrate the performance of this colour microscope Papanicolaou (Pap) smears were also successfully imaged. This field-portable and wide-field computational colour microscope could be useful for tele-medicine applications in resource poor settings.

High-resolution scanning precession electron diffraction: Alignment and spatial resolution.

PubMed

Barnard, Jonathan S; Johnstone, Duncan N; Midgley, Paul A

2017-03-01

Methods are presented for aligning the pivot point of a precessing electron probe in the scanning transmission electron microscope (STEM) and for assessing the spatial resolution in scanning precession electron diffraction (SPED) experiments. The alignment procedure is performed entirely in diffraction mode, minimising probe wander within the bright-field (BF) convergent beam electron diffraction (CBED) disk and is used to obtain high spatial resolution SPED maps. Through analysis of the power spectra of virtual bright-field images extracted from the SPED data, the precession-induced blur was measured as a function of precession angle. At low precession angles, SPED spatial resolution was limited by electronic noise in the scan coils; whereas at high precession angles SPED spatial resolution was limited by tilt-induced two-fold astigmatism caused by the positive spherical aberration of the probe-forming lens. Copyright © 2016 Elsevier B.V. All rights reserved.

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

Rosch, R.; Boutin, J. Y.; Le Breton, J. P.

This article describes x-ray imaging with grazing-incidence microscopes, developed for the experimental program carried out on the Ligne d'Integration Laser (LIL) facility [J. P. Le Breton et al., Inertial Fusion Sciences and Applications 2001 (Elsevier, Paris, 2002), pp. 856-862] (24 kJ, UV--0.35 nm). The design includes a large target-to-microscope (400-700 mm) distance required by the x-ray ablation issues anticipated on the Laser MegaJoule facility [P. A. Holstein et al., Laser Part. Beams 17, 403 (1999)] (1.8 MJ) which is under construction. Two eight-image Kirkpatrick-Baez microscopes [P. Kirkpatrick and A. V. Baez J. Opt. Soc. Am. 38, 766 (1948)] with differentmore » spectral wavelength ranges and with a 400 mm source-to-mirror distance image the target on a custom-built framing camera (time resolution of {approx}80 ps). The soft x-ray version microscope is sensitive below 1 keV and its spatial resolution is better than 30 {mu}m over a 2-mm-diam region. The hard x-ray version microscope has a 10 {mu}m resolution over an 800-{mu}m-diam region and is sensitive in the 1-5 keV energy range. Two other x-ray microscopes based on an association of toroidal/spherical surfaces (T/S microscopes) produce an image on a streak camera with a spatial resolution better than 30 {mu}m over a 3 mm field of view in the direction of the camera slit. Both microscopes have been designed to have, respectively, a maximum sensitivity in the 0.1-1 and 1-5 keV energy range. We present the original design of these four microscopes and their test on a dc x-ray tube in the laboratory. The diagnostics were successfully used on LIL first experiments early in 2005. Results of soft x-ray imaging of a radiative jet during conical shaped laser interaction are shown.« less

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.

Imaging nanoscale spatial modulation of a relativistic electron beam with a MeV ultrafast electron microscope

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

Lu, Chao; Jiang, Tao; Liu, Shengguang

Here, an accelerator-based MeV ultrafast electron microscope (MUEM) has been proposed as a promising tool to the study structural dynamics at the nanometer spatial scale and the picosecond temporal scale. Here, we report experimental tests of a prototype MUEM where high quality images with nanoscale fine structures were recorded with a pulsed ~3 MeV picosecond electron beam. The temporal and spatial resolutions of the MUEM operating in the single-shot mode are about 4 ps (FWHM) and 100 nm (FWHM), corresponding to a temporal-spatial resolution of 4 × 10 –19 sm, about 2 orders of magnitude higher than that achieved withmore » state-of-the-art single-shot keV UEM. Using this instrument, we offer the demonstration of visualizing the nanoscale periodic spatial modulation of an electron beam, which may be converted into longitudinal density modulation through emittance exchange to enable production of high-power coherent radiation at short wavelengths. Our results mark a great step towards single-shot nanometer-resolution MUEMs and compact intense x-ray sources that may have widespread applications in many areas of science.« less

Imaging nanoscale spatial modulation of a relativistic electron beam with a MeV ultrafast electron microscope

DOE PAGES

Lu, Chao; Jiang, Tao; Liu, Shengguang; ...

2018-03-12

Here, an accelerator-based MeV ultrafast electron microscope (MUEM) has been proposed as a promising tool to the study structural dynamics at the nanometer spatial scale and the picosecond temporal scale. Here, we report experimental tests of a prototype MUEM where high quality images with nanoscale fine structures were recorded with a pulsed ~3 MeV picosecond electron beam. The temporal and spatial resolutions of the MUEM operating in the single-shot mode are about 4 ps (FWHM) and 100 nm (FWHM), corresponding to a temporal-spatial resolution of 4 × 10 –19 sm, about 2 orders of magnitude higher than that achieved withmore » state-of-the-art single-shot keV UEM. Using this instrument, we offer the demonstration of visualizing the nanoscale periodic spatial modulation of an electron beam, which may be converted into longitudinal density modulation through emittance exchange to enable production of high-power coherent radiation at short wavelengths. Our results mark a great step towards single-shot nanometer-resolution MUEMs and compact intense x-ray sources that may have widespread applications in many areas of science.« less

Scanning capacitance microscope as a tool for the characterization of integrated circuits

NASA Astrophysics Data System (ADS)

Born, A.; Wiesendanger, R.

With the decreasing size of integrated circuits (ICs), there is an increasing demand for the measurement of doping profiles with high spatial resolution. The scanning capacitance microscope (SCM) offers the possibility of measuring 2D dopant profiles with spatial resolution of less than 20 nm. A great problem of the SCM technique is the influence of previous measurements on subsequent ones. We have observed hysteresis in the SCM images and measured low-frequency C-V curves with high-frequency equipment. A theoretical model was developed to understand this phenomenon. We are now undertaking the first steps using the SCM as a standard device for the characterization of ICs.

Note: Electron energy spectroscopy mapping of surface with scanning tunneling microscope.

PubMed

Li, Meng; Xu, Chunkai; Zhang, Panke; Li, Zhean; Chen, Xiangjun

2016-08-01

We report a novel scanning probe electron energy spectrometer (SPEES) which combines a double toroidal analyzer with a scanning tunneling microscope to achieve both topography imaging and electron energy spectroscopy mapping of surface in situ. The spatial resolution of spectroscopy mapping is determined to be better than 0.7 ± 0.2 μm at a tip sample distance of 7 μm. Meanwhile, the size of the field emission electron beam spot on the surface is also measured, and is about 3.6 ± 0.8 μm in diameter. This unambiguously demonstrates that the spatial resolution of SPEES technique can be much better than the size of the incident electron beam.

Interior tomography in microscopic CT with image reconstruction constrained by full field of view scan at low spatial resolution

NASA Astrophysics Data System (ADS)

Luo, Shouhua; Shen, Tao; Sun, Yi; Li, Jing; Li, Guang; Tang, Xiangyang

2018-04-01

In high resolution (microscopic) CT applications, the scan field of view should cover the entire specimen or sample to allow complete data acquisition and image reconstruction. However, truncation may occur in projection data and results in artifacts in reconstructed images. In this study, we propose a low resolution image constrained reconstruction algorithm (LRICR) for interior tomography in microscopic CT at high resolution. In general, the multi-resolution acquisition based methods can be employed to solve the data truncation problem if the project data acquired at low resolution are utilized to fill up the truncated projection data acquired at high resolution. However, most existing methods place quite strict restrictions on the data acquisition geometry, which greatly limits their utility in practice. In the proposed LRICR algorithm, full and partial data acquisition (scan) at low and high resolutions, respectively, are carried out. Using the image reconstructed from sparse projection data acquired at low resolution as the prior, a microscopic image at high resolution is reconstructed from the truncated projection data acquired at high resolution. Two synthesized digital phantoms, a raw bamboo culm and a specimen of mouse femur, were utilized to evaluate and verify performance of the proposed LRICR algorithm. Compared with the conventional TV minimization based algorithm and the multi-resolution scout-reconstruction algorithm, the proposed LRICR algorithm shows significant improvement in reduction of the artifacts caused by data truncation, providing a practical solution for high quality and reliable interior tomography in microscopic CT applications. The proposed LRICR algorithm outperforms the multi-resolution scout-reconstruction method and the TV minimization based reconstruction for interior tomography in microscopic CT.

Technique to quantitatively measure magnetic properties of thin structures at <10 NM spatial resolution

DOEpatents

Bajt, Sasa

2003-07-08

A highly sensitive and high resolution magnetic microscope images magnetic properties quantitatively. Imaging is done with a modified transmission electron microscope that allows imaging of the sample in a zero magnetic field. Two images from closely spaced planes, one in focus and one slightly out of focus, are sufficient to calculate the absolute values of the phase change imparted to the electrons, and hence obtain the magnetization vector field distribution.

Automated high resolution full-field spatial coherence tomography for quantitative phase imaging of human red blood cells

NASA Astrophysics Data System (ADS)

Singla, Neeru; Dubey, Kavita; Srivastava, Vishal; Ahmad, Azeem; Mehta, D. S.

2018-02-01

We developed an automated high-resolution full-field spatial coherence tomography (FF-SCT) microscope for quantitative phase imaging that is based on the spatial, rather than the temporal, coherence gating. The Red and Green color laser light was used for finding the quantitative phase images of unstained human red blood cells (RBCs). This study uses morphological parameters of unstained RBCs phase images to distinguish between normal and infected cells. We recorded the single interferogram by a FF-SCT microscope for red and green color wavelength and average the two phase images to further reduced the noise artifacts. In order to characterize anemia infected from normal cells different morphological features were extracted and these features were used to train machine learning ensemble model to classify RBCs with high accuracy.

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

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

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

1992-03-02

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

Design of a normal incidence multilayer imaging X-ray microscope

NASA Astrophysics Data System (ADS)

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

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

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.

Nonlinear hybridization of the fundamental eigenmodes of microscopic ferromagnetic ellipses.

PubMed

Demidov, V E; Buchmeier, M; Rott, K; Krzysteczko, P; Münchenberger, J; Reiss, G; Demokritov, S O

2010-05-28

We have studied experimentally with high spatial resolution the nonlinear eigenmodes of microscopic Permalloy elliptical elements. We show that the nonlinearity affects the frequencies of the edge and the center modes in an essentially different way. This leads to repulsion of corresponding resonances and to nonlinear mode hybridization resulting in qualitative modifications of the spatial characteristics of the modes. We find that the nonlinear counterparts of the edge and the center modes simultaneously exhibit features specific for both their linear analogues.

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

Note: Electron energy spectroscopy mapping of surface with scanning tunneling microscope

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

Li, Meng; Xu, Chunkai, E-mail: xuck@ustc.edu.cn, E-mail: xjun@ustc.edu.cn; Zhang, Panke

We report a novel scanning probe electron energy spectrometer (SPEES) which combines a double toroidal analyzer with a scanning tunneling microscope to achieve both topography imaging and electron energy spectroscopy mapping of surface in situ. The spatial resolution of spectroscopy mapping is determined to be better than 0.7 ± 0.2 μm at a tip sample distance of 7 μm. Meanwhile, the size of the field emission electron beam spot on the surface is also measured, and is about 3.6 ± 0.8 μm in diameter. This unambiguously demonstrates that the spatial resolution of SPEES technique can be much better than themore » size of the incident electron beam.« less

Elemental and topographical imaging of microscopic variations in deposition on NSTX-U and DIII-D samples

NASA Astrophysics Data System (ADS)

Skinner, C. H.; Kaita, R.; Koel, B. E.; Chrobak, C. P.; Wampler, W. R.

2017-10-01

Tokamak plasma facing components (PFCs) have surface roughness that can cause microscopic spatial variations in erosion and deposition and hence influence material migration. Previous RBS measurements showed indirect evidence for this but the spatial (0.5mm) resolution was insufficient for direct imaging. We will present elemental images at sub-micron resolution of deposition on NSTX-U and DiMES samples that show strong microscopic variations and correlate this with 3D topographical maps of surface irregularities. The elemental imaging is performed with a Scanning Auger Microprobe (SAM) that measures element-specific Auger electrons excited by an SEM electron beam. 3D topographical maps of the samples are performed with a Leica DCM 3D confocal light microscope and compared to the elemental deposition pattern. The initial results appear consistent with erosion at the downstream edges of the surface pores exposed to the incident ion flux, whereas the deeper regions are shadowed and serve as deposition traps. Support was provided through DOE Contract Numbers DE-AC02-09CH11466, DE-FC02-04ER54698 and DE-NA0003525.

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

PubMed

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

2015-07-13

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

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.

FIMic: design for ultimate 3D-integral microscopy of in-vivo biological samples

PubMed Central

Scrofani, G.; Sola-Pikabea, J.; Llavador, A.; Sanchez-Ortiga, E.; Barreiro, J. C.; Saavedra, G.; Garcia-Sucerquia, J.; Martínez-Corral, M.

2017-01-01

In this work, Fourier integral microscope (FIMic), an ultimate design of 3D-integral microscopy, is presented. By placing a multiplexing microlens array at the aperture stop of the microscope objective of the host microscope, FIMic shows extended depth of field and enhanced lateral resolution in comparison with regular integral microscopy. As FIMic directly produces a set of orthographic views of the 3D-micrometer-sized sample, it is suitable for real-time imaging. Following regular integral-imaging reconstruction algorithms, a 2.75-fold enhanced depth of field and 2-time better spatial resolution in comparison with conventional integral microscopy is reported. Our claims are supported by theoretical analysis and experimental images of a resolution test target, cotton fibers, and in-vivo 3D-imaging of biological specimens. PMID:29359107

Swept Field Laser Confocal Microscopy for Enhanced Spatial and Temporal Resolution in Live-Cell Imaging

PubMed Central

Castellano-Muñoz, Manuel; Peng, Anthony Wei; Salles, Felipe T.; Ricci, Anthony J.

2013-01-01

Confocal fluorescence microscopy is a broadly used imaging technique that enhances the signal-to-noise ratio by removing out of focal plane fluorescence. Confocal microscopes come with a variety of modifications depending on the particular experimental goals. Microscopes, illumination pathways, and light collection were originally focused upon obtaining the highest resolution image possible, typically on fixed tissue. More recently, live-cell confocal imaging has gained importance. Since measured signals are often rapid or transient, thus requiring higher sampling rates, specializations are included to enhance spatial and temporal resolution while maintaining tissue viability. Thus, a balance between image quality, temporal resolution, and tissue viability is needed. A subtype of confocal imaging, termed swept field confocal (SFC) microscopy, can image live cells at high rates while maintaining confocality. SFC systems can use a pinhole array to obtain high spatial resolution, similar to spinning disc systems. In addition, SFC imaging can achieve faster rates by using a slit to sweep the light across the entire image plane, thus requiring a single scan to generate an image. Coupled to a high-speed charge-coupled device camera and a laser illumination source, images can be obtained at greater than 1,000 frames per second while maintaining confocality. PMID:22831554

High Resolution Tissue Imaging Using the Single-probe Mass Spectrometry under Ambient Conditions

NASA Astrophysics Data System (ADS)

Rao, Wei; Pan, Ning; Yang, Zhibo

2015-06-01

Ambient mass spectrometry imaging (MSI) is an emerging field with great potential for the detailed spatial analysis of biological samples with minimal pretreatment. We have developed a miniaturized sampling and ionization device, the Single-probe, which uses in-situ surface micro-extraction to achieve high detection sensitivity and spatial resolution during MSI experiments. The Single-probe was coupled to a Thermo LTQ Orbitrap XL mass spectrometer and was able to create high spatial and high mass resolution MS images at 8 ± 2 and 8.5 μm on flat polycarbonate microscope slides and mouse kidney sections, respectively, which are among the highest resolutions available for ambient MSI techniques. Our proof-of-principle experiments indicate that the Single-probe MSI technique has the potential to obtain ambient MS images with very high spatial resolutions with minimal sample preparation, which opens the possibility for subcellular ambient tissue MSI to be performed in the future.

Cross-Scale Molecular Analysis of Chemical Heterogeneity in Shale Rocks

DOE PAGES

Hao, Zhao; Bechtel, Hans A.; Kneafsey, Timothy; ...

2018-02-07

The organic and mineralogical heterogeneity in shale at micrometer and nanometer spatial scales contributes to the quality of gas reserves, gas flow mechanisms and gas production. Here, we demonstrate two molecular imaging approaches based on infrared spectroscopy to obtain mineral and kerogen information at these mesoscale spatial resolutions in large-sized shale rock samples. The first method is a modified microscopic attenuated total reflectance measurement that utilizes a large germanium hemisphere combined with a focal plane array detector to rapidly capture chemical images of shale rock surfaces spanning hundreds of micrometers with micrometer spatial resolution. The second method, synchrotron infrared nano-spectroscopy,more » utilizes a metallic atomic force microscope tip to obtain chemical images of micrometer dimensions but with nanometer spatial resolution. This chemically "deconvoluted" imaging at the nano-pore scale is then used to build a machine learning model to generate a molecular distribution map across scales with a spatial span of 1000 times, which enables high-throughput geochemical characterization in greater details across the nano-pore and micro-grain scales and allows us to identify co-localization of mineral phases with chemically distinct organics and even with gas phase sorbents. Finally, this characterization is fundamental to understand mineral and organic compositions affecting the behavior of shales.« less

Cross-Scale Molecular Analysis of Chemical Heterogeneity in Shale Rocks

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

Hao, Zhao; Bechtel, Hans A.; Kneafsey, Timothy

The organic and mineralogical heterogeneity in shale at micrometer and nanometer spatial scales contributes to the quality of gas reserves, gas flow mechanisms and gas production. Here, we demonstrate two molecular imaging approaches based on infrared spectroscopy to obtain mineral and kerogen information at these mesoscale spatial resolutions in large-sized shale rock samples. The first method is a modified microscopic attenuated total reflectance measurement that utilizes a large germanium hemisphere combined with a focal plane array detector to rapidly capture chemical images of shale rock surfaces spanning hundreds of micrometers with micrometer spatial resolution. The second method, synchrotron infrared nano-spectroscopy,more » utilizes a metallic atomic force microscope tip to obtain chemical images of micrometer dimensions but with nanometer spatial resolution. This chemically "deconvoluted" imaging at the nano-pore scale is then used to build a machine learning model to generate a molecular distribution map across scales with a spatial span of 1000 times, which enables high-throughput geochemical characterization in greater details across the nano-pore and micro-grain scales and allows us to identify co-localization of mineral phases with chemically distinct organics and even with gas phase sorbents. Finally, this characterization is fundamental to understand mineral and organic compositions affecting the behavior of shales.« less

Coherent x-ray diffraction imaging with nanofocused illumination.

PubMed

Schroer, C G; Boye, P; Feldkamp, J M; Patommel, J; Schropp, A; Schwab, A; Stephan, S; Burghammer, M; Schöder, S; Riekel, C

2008-08-29

Coherent x-ray diffraction imaging is an x-ray microscopy technique with the potential of reaching spatial resolutions well beyond the diffraction limits of x-ray microscopes based on optics. However, the available coherent dose at modern x-ray sources is limited, setting practical bounds on the spatial resolution of the technique. By focusing the available coherent flux onto the sample, the spatial resolution can be improved for radiation-hard specimens. A small gold particle (size <100 nm) was illuminated with a hard x-ray nanobeam (E=15.25 keV, beam dimensions approximately 100 x 100 nm2) and is reconstructed from its coherent diffraction pattern. A resolution of about 5 nm is achieved in 600 s exposure time.

Scanning photoelectron microscope for nanoscale three-dimensional spatial-resolved electron spectroscopy for chemical analysis.

PubMed

Horiba, K; Nakamura, Y; Nagamura, N; Toyoda, S; Kumigashira, H; Oshima, M; Amemiya, K; Senba, Y; Ohashi, H

2011-11-01

In order to achieve nondestructive observation of the three-dimensional spatially resolved electronic structure of solids, we have developed a scanning photoelectron microscope system with the capability of depth profiling in electron spectroscopy for chemical analysis (ESCA). We call this system 3D nano-ESCA. For focusing the x-ray, a Fresnel zone plate with a diameter of 200 μm and an outermost zone width of 35 nm is used. In order to obtain the angular dependence of the photoelectron spectra for the depth-profile analysis without rotating the sample, we adopted a modified VG Scienta R3000 analyzer with an acceptance angle of 60° as a high-resolution angle-resolved electron spectrometer. The system has been installed at the University-of-Tokyo Materials Science Outstation beamline, BL07LSU, at SPring-8. From the results of the line-scan profiles of the poly-Si/high-k gate patterns, we achieved a total spatial resolution better than 70 nm. The capability of our system for pinpoint depth-profile analysis and high-resolution chemical state analysis is demonstrated. © 2011 American Institute of Physics

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

NASA Astrophysics Data System (ADS)

Iyer, Vijay; Saggau, Peter

2003-10-01

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

Light-sheet enhanced resolution of light field microscopy for rapid imaging of large volumes

NASA Astrophysics Data System (ADS)

Madrid Wolff, Jorge; Castro, Diego; Arbeláez, Pablo; Forero-Shelton, Manu

2018-02-01

Whole-brain imaging is challenging because it demands microscopes with high temporal and spatial resolution, which are often at odds, especially in the context of large fields of view. We have designed and built a light-sheet microscope with digital micromirror illumination and light-field detection. On the one hand, light sheets provide high resolution optical sectioning on live samples without compromising their viability. On the other hand, light field imaging makes it possible to reconstruct full volumes of relatively large fields of view from a single camera exposure; however, its enhanced temporal resolution comes at the expense of spatial resolution, limiting its applicability. We present an approach to increase the resolution of light field images using DMD-based light sheet illumination. To that end, we develop a method to produce synthetic resolution targets for light field microscopy and a procedure to correct the depth at which planes are refocused with rendering software. We measured the axial resolution as a function of depth and show a three-fold potential improvement with structured illumination, albeit by sacrificing some temporal resolution, also three-fold. This results in an imaging system that may be adjusted to specific needs without having to reassemble and realign it. This approach could be used to image relatively large samples at high rates.

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

NASA Technical Reports Server (NTRS)

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

1999-01-01

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

A stand-alone compact EUV microscope based on gas-puff target source.

PubMed

Torrisi, Alfio; Wachulak, Przemyslaw; Węgrzyński, Łukasz; Fok, Tomasz; Bartnik, Andrzej; Parkman, Tomáš; Vondrová, Šárka; Turňová, Jana; Jankiewicz, Bartłomiej J; Bartosewicz, Bartosz; Fiedorowicz, Henryk

2017-02-01

We report on a very compact desk-top transmission extreme ultraviolet (EUV) microscope based on a laser-plasma source with a double stream gas-puff target, capable of acquiring magnified images of objects with a spatial (half-pitch) resolution of sub-50 nm. A multilayer ellipsoidal condenser is used to focus and spectrally narrow the radiation from the plasma, producing a quasi-monochromatic EUV radiation (λ = 13.8 nm) illuminating the object, whereas a Fresnel zone plate objective forms the image. Design details, development, characterization and optimization of the EUV source and the microscope are described and discussed. Test object and other samples were imaged to demonstrate superior resolution compared to visible light microscopy. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

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.

Optofluidic microscope with 3D spatial resolution.

PubMed

Vig, Asger Laurburg; Marie, Rodolphe; Jensen, Eric; Kristensen, Anders

2010-03-01

This paper reports on-chip based optical detection with three-dimensional spatial resolution by integration of an optofluidic microscope (OFM) in a microfluidic pinched flow fractionation (PFF) separation device. This setup also enables on-chip particle image velocimetry (PIV). The position in the plane perpendicular to the flow direction and the velocity along the flow direction of separated fluorescent labeled polystyrene microspheres with diameters of 1 microm , 2.1 microm , 3 microm and 4 microm is determined by the OFM. These results are bench marked against those obtained with a PFF device using conventional fluorescence microscope readout. The size separated microspheres are detected by OFM with an accuracy of

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.

Mapping the layer count of few-layer hexagonal boron nitride at high lateral spatial resolutions

NASA Astrophysics Data System (ADS)

Mohsin, Ali; Cross, Nicholas G.; Liu, Lei; Watanabe, Kenji; Taniguchi, Takashi; Duscher, Gerd; Gu, Gong

2018-01-01

Layer count control and uniformity of two dimensional (2D) layered materials are critical to the investigation of their properties and to their electronic device applications, but methods to map 2D material layer count at nanometer-level lateral spatial resolutions have been lacking. Here, we demonstrate a method based on two complementary techniques widely available in transmission electron microscopes (TEMs) to map the layer count of multilayer hexagonal boron nitride (h-BN) films. The mass-thickness contrast in high-angle annular dark-field (HAADF) imaging in the scanning transmission electron microscope (STEM) mode allows for thickness determination in atomically clean regions with high spatial resolution (sub-nanometer), but is limited by surface contamination. To complement, another technique based on the boron K ionization edge in the electron energy loss spectroscopy spectrum (EELS) of h-BN is developed to quantify the layer count so that surface contamination does not cause an overestimate, albeit at a lower spatial resolution (nanometers). The two techniques agree remarkably well in atomically clean regions with discrepancies within  ±1 layer. For the first time, the layer count uniformity on the scale of nanometers is quantified for a 2D material. The methodology is applicable to layer count mapping of other 2D layered materials, paving the way toward the synthesis of multilayer 2D materials with homogeneous layer count.

High temperature superconductor dc SQUID micro-susceptometer for room temperature objects

NASA Astrophysics Data System (ADS)

Faley, M. I.; Pratt, K.; Reineman, R.; Schurig, D.; Gott, S.; Atwood, C. G.; Sarwinski, R. E.; Paulson, D. N.; Starr, T. N.; Fagaly, R. L.

2004-05-01

We have developed a scanning magnetic microscope (SMM) with 25 µm resolution in spatial position for the magnetic features of room temperature objects. The microscope consists of a high-temperature superconductor (HTS) dc SQUID sensor, suspended in vacuum with a self-adjusting standoff, close spaced liquid nitrogen Dewar, X-Y scanning stage and a computer control system. The HTS SQUIDs were optimized for better spatial and field resolutions for operation at liquid nitrogen temperature. Measured inside a magnetic shield, the 10 pT Hz-1/2 typical noise of the SQUIDs is white down to frequencies of about 10 Hz, increasing up to about 20 pT Hz-1/2 at 1 Hz. The microscope is mounted on actively damped platforms, which negate vibrations from the environment as well as damping internal stepper motor noises. A high-resolution video telescope and a 1 µm precision z-axis positioning system allow a close positioning of the sample under the sensor. The ability of the sensors to operate in unshielded environmental conditions with magnetic fields up to about 15 G allowed us to perform 2D mapping of the local ac and dc susceptibility of the objects.

Fluorescence microscopy.

PubMed

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

2014-10-01

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

Ultra-large field-of-view two-photon microscopy.

PubMed

Tsai, Philbert S; Mateo, Celine; Field, Jeffrey J; Schaffer, Chris B; Anderson, Matthew E; Kleinfeld, David

2015-06-01

We present a two-photon microscope that images the full extent of murine cortex with an objective-limited spatial resolution across an 8 mm by 10 mm field. The lateral resolution is approximately 1 µm and the maximum scan speed is 5 mm/ms. The scan pathway employs large diameter compound lenses to minimize aberrations and performs near theoretical limits. We demonstrate the special utility of the microscope by recording resting-state vasomotion across both hemispheres of the murine brain through a transcranial window and by imaging histological sections without the need to stitch.

Fluorescence Microscopy

PubMed Central

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

2016-01-01

Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. PMID:25275114

Application of laser scanning speckle-microscopy for high-resolution express diagnostics of chlamydial infection

NASA Astrophysics Data System (ADS)

Ulyanov, Sergey; Larionova, Olga; Ulianova, Onega; Zaitsev, Sergey; Saltykov, Yury; Polyanina, Tatiana; Lyapina, Anna; Filonova, Nadezhda; Subbotina, Irina; Kalduzova, Irina; Utz, Sergey; Moiseeva, Yulia; Feodorova, Valentina

2018-04-01

Method of speckle-microscopy has been adapted to the problem of detection of Chlamydia trachomatis microbial cells in clinical samples. Prototype of laser scanning speckle-microscope has been designed. Spatial resolution and output characteristics of this microscope have been analyzed for the case of scanning of C. trachomatis bacteria inclusions - Elementary Bodies (EBs) inside the human cells, fixed on the glass. It has been demonstrated, that presence of C. trachomatis microbial cells in the sample can be easily detected using speckle microscopy.

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 (

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

NASA Astrophysics Data System (ADS)

Upputuri, Paul Kumar; Pramanik, Manojit

2017-12-01

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

Studies of mechanisms of decay and recovery in organic dye-doped polymers using spatially resolved white light interferometry

NASA Astrophysics Data System (ADS)

Anderson, Benjamin; Bernhardt, Elizabeth; Kuzyk, Mark

2012-10-01

Several organic dyes have been shown to self heal when doped in a polymer matrix. Most measurements to date use optical absorbance, amplified spontaneous emission, or digital imaging as a probe. Each method determines a subset of the relevant parameters. We have constructed a white light interferometric microscope, which measures the absorption spectrum and change in refractive index during decay and recovery simultaneously at multiple points in the material. We report on preliminary measurements and results concerning the microscopes spatial resolution.

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).

Spatially resolved D-T(2) correlation NMR of porous media.

PubMed

Zhang, Yan; Blümich, Bernhard

2014-05-01

Within the past decade, 2D Laplace nuclear magnetic resonance (NMR) has been developed to analyze pore geometry and diffusion of fluids in porous media on the micrometer scale. Many objects like rocks and concrete are heterogeneous on the macroscopic scale, and an integral analysis of microscopic properties provides volume-averaged information. Magnetic resonance imaging (MRI) resolves this spatial average on the contrast scale set by the particular MRI technique. Desirable contrast parameters for studies of fluid transport in porous media derive from the pore-size distribution and the pore connectivity. These microscopic parameters are accessed by 1D and 2D Laplace NMR techniques. It is therefore desirable to combine MRI and 2D Laplace NMR to image functional information on fluid transport in porous media. Because 2D Laplace resolved MRI demands excessive measuring time, this study investigates the possibility to restrict the 2D Laplace analysis to the sum signals from low-resolution pixels, which correspond to pixels of similar amplitude in high-resolution images. In this exploratory study spatially resolved D-T2 correlation maps from glass beads and mortar are analyzed. Regions of similar contrast are first identified in high-resolution images to locate corresponding pixels in low-resolution images generated with D-T2 resolved MRI for subsequent pixel summation to improve the signal-to-noise ratio of contrast-specific D-T2 maps. This method is expected to contribute valuable information on correlated sample heterogeneity from the macroscopic and the microscopic scales in various types of porous materials including building materials and rock. Copyright © 2014 Elsevier Inc. All rights reserved.

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.

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

PubMed

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

2012-04-09

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

Near-field microscopy with a microfabricated solid immersion lens

NASA Astrophysics Data System (ADS)

Fletcher, Daniel Alden

2001-07-01

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

Imaging single atoms using secondary electrons with an aberration-corrected electron microscope.

PubMed

Zhu, Y; Inada, H; Nakamura, K; Wall, J

2009-10-01

Aberration correction has embarked on a new frontier in electron microscopy by overcoming the limitations of conventional round lenses, providing sub-angstrom-sized probes. However, improvement of spatial resolution using aberration correction so far has been limited to the use of transmitted electrons both in scanning and stationary mode, with an improvement of 20-40% (refs 3-8). In contrast, advances in the spatial resolution of scanning electron microscopes (SEMs), which are by far the most widely used instrument for surface imaging at the micrometre-nanometre scale, have been stagnant, despite several recent efforts. Here, we report a new SEM, with aberration correction, able to image single atoms by detecting electrons emerging from its surface as a result of interaction with the small probe. The spatial resolution achieved represents a fourfold improvement over the best-reported resolution in any SEM (refs 10-12). Furthermore, we can simultaneously probe the sample through its entire thickness with transmitted electrons. This ability is significant because it permits the selective visualization of bulk atoms and surface ones, beyond a traditional two-dimensional projection in transmission electron microscopy. It has the potential to revolutionize the field of microscopy and imaging, thereby opening the door to a wide range of applications, especially when combined with simultaneous nanoprobe spectroscopy.

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.

Copper Decoration of Carbon Nanotubes and High Resolution Electron Microscopy

NASA Astrophysics Data System (ADS)

Probst, Camille

A new process of decorating carbon nanotubes with copper was developed for the fabrication of nanocomposite aluminum-nanotubes. The process consists of three stages: oxidation, activation and electroless copper plating on the nanotubes. The oxidation step was required to create chemical function on the nanotubes, essential for the activation step. Then, catalytic nanoparticles of tin-palladium were deposited on the tubes. Finally, during the electroless copper plating, copper particles with a size between 20 and 60 nm were uniformly deposited on the nanotubes surface. The reproducibility of the process was shown by using another type of carbon nanotube. The fabrication of nanocomposites aluminum-nanotubes was tested by aluminum vacuum infiltration. Although the infiltration of carbon nanotubes did not produce the expected results, an interesting electron microscopy sample was discovered during the process development: the activated carbon nanotubes. Secondly, scanning transmitted electron microscopy (STEM) imaging in SEM was analysed. The images were obtained with a new detector on the field emission scanning electron microscope (Hitachi S-4700). Various parameters were analysed with the use of two different samples: the activated carbon nanotubes (previously obtained) and gold-palladium nanodeposits. Influences of working distance, accelerating voltage or sample used on the spatial resolution of images obtained with SMART (Scanning Microscope Assessment and Resolution Testing) were analysed. An optimum working distance for the best spatial resolution related to the sample analysed was found for the imaging in STEM mode. Finally, relation between probe size and spatial resolution of backscattered electrons (BSE) images was studied. An image synthesis method was developed to generate the BSE images from backscattered electrons coefficients obtained with CASINO software. Spatial resolution of images was determined using SMART. The analysis shown that using a probe size smaller than the size of the observed object (sample features) does not improve the spatial resolution. In addition, the effects of the accelerating voltage, the current intensity and the sample geometry and composition were analysed.

Spatially-controlled illumination with rescan confocal microscopy enhances image quality, resolution and reduces photodamage

NASA Astrophysics Data System (ADS)

Krishnaswami, Venkataraman; De Luca, Giulia M. R.; Breedijk, Ronald M. P.; Van Noorden, Cornelis J. F.; Manders, Erik M. M.; Hoebe, Ron A.

2017-02-01

Fluorescence microscopy is an important tool in biomedical imaging. An inherent trade-off lies between image quality and photodamage. Recently, we have introduced rescan confocal microscopy (RCM) that improves the lateral resolution of a confocal microscope down to 170 nm. Previously, we have demonstrated that with controlled-light exposure microscopy, spatial control of illumination reduces photodamage without compromising image quality. Here, we show that the combination of these two techniques leads to high resolution imaging with reduced photodamage without compromising image quality. Implementation of spatially-controlled illumination was carried out in RCM using a line scanning-based approach. Illumination is spatially-controlled for every line during imaging with the help of a prediction algorithm that estimates the spatial profile of the fluorescent specimen. The estimation is based on the information available from previously acquired line images. As a proof-of-principle, we show images of N1E-115 neuroblastoma cells, obtained by this new setup with reduced illumination dose, improved resolution and without compromising image quality.

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.

Pan-neuronal calcium imaging with cellular resolution in freely swimming zebrafish.

PubMed

Kim, Dal Hyung; Kim, Jungsoo; Marques, João C; Grama, Abhinav; Hildebrand, David G C; Gu, Wenchao; Li, Jennifer M; Robson, Drew N

2017-11-01

Calcium imaging with cellular resolution typically requires an animal to be tethered under a microscope, which substantially restricts the range of behaviors that can be studied. To expand the behavioral repertoire amenable to imaging, we have developed a tracking microscope that enables whole-brain calcium imaging with cellular resolution in freely swimming larval zebrafish. This microscope uses infrared imaging to track a target animal in a behavior arena. On the basis of the predicted trajectory of the animal, we applied optimal control theory to a motorized stage system to cancel brain motion in three dimensions. We combined this motion-cancellation system with differential illumination focal filtering, a variant of HiLo microscopy, which enabled us to image the brain of a freely swimming larval zebrafish for more than an hour. This work expands the repertoire of natural behaviors that can be studied with cellular-resolution calcium imaging to potentially include spatial navigation, social behavior, feeding and reward.

Enhancing image contrast of carbon nanotubes on cellular background using helium ion microscope by varying helium ion fluence.

PubMed

Dykas, M M; Poddar, K; Yoong, S L; Viswanathan, V; Mathew, S; Patra, A; Saha, S; Pastorin, G; Venkatesan, T

2018-01-01

Carbon nanotubes (CNTs) have become an important nano entity for biomedical applications. Conventional methods of their imaging, often cannot be applied in biological samples due to an inadequate spatial resolution or poor contrast between the CNTs and the biological sample. Here we report a unique and effective detection method, which uses differences in conductivities of carbon nanotubes and HeLa cells. The technique involves the use of a helium ion microscope to image the sample with the surface charging artefacts created by the He + and neutralised by electron flood gun. This enables us to obtain a few nanometre resolution images of CNTs in HeLa Cells with high contrast, which was achieved by tailoring the He + fluence. Charging artefacts can be efficiently removed for conductive CNTs by a low amount of electrons, the fluence of which is not adequate to discharge the cell surface, resulting in high image contrast. Thus, this technique enables rapid detection of any conducting nano structures on insulating cellular background even in large fields of view and fine spatial resolution. The technique demonstrated has wider applications for researchers seeking enhanced contrast and high-resolution imaging of any conducting entity in a biological matrix - a commonly encountered issue of importance in drug delivery, tissue engineering and toxicological studies. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Nanoscale Spatial Organization of Prokaryotic Cells Studied by Super-Resolution Optical Microscopy

NASA Astrophysics Data System (ADS)

McEvoy, Andrea Lynn

All cells spatially organize their interiors, and this arrangement is necessary for cell viability. Until recently, it was believed that only eukaryotic cells spatially segregate their components. However, it is becoming increasingly clear that bacteria also assemble their proteins into complex patterns. In eukaryotic cells, spatial organization arises from membrane bound organelles as well as motor transport proteins which can move cargos within the cell. To date, there are no known motor transport proteins in bacteria and most microbes lack membrane bound organelles, so it remains a mystery how bacterial spatial organization emerges. In hind-sight it is not surprising that bacteria also exhibit complex spatial organization considering much of what we have learned about the basic processes that take place in all cells, such as transcription and translation was first discovered in prokaryotic cells. Perhaps the fundamental principles that govern spatial organization in prokaryotic cells may be applicable in eukaryotic cells as well. In addition, bacteria are attractive model organism for spatial organization studies because they are genetically tractable, grow quickly and much biochemical and structural data is known about them. A powerful tool for observing spatial organization in cells is the fluorescence microscope. By specifically tagging a protein of interest with a fluorescent probe, it is possible to examine how proteins organize and dynamically assemble inside cells. A significant disadvantage of this technology is its spatial resolution (approximately 250 nm laterally and 500 nm axially). This limitation on resolution causes closely spaced proteins to look blurred making it difficult to observe the fine structure within the complexes. This resolution limit is especially problematic within small cells such as bacteria. With the recent invention of new optical microscopies, we now can surpass the existing limits of fluorescence imaging. In some cases, we can now see individual proteins inside of large complexes or observe structures with ten times the resolution of conventional imaging. These techniques are known as super-resolution microscopes. In this dissertation, I use super-resolution microscopes to understand how a model microbe, Escherichia coli, assembles complex protein structures. I focus on two spatially organized systems, the chemotaxis network and the cell division machinery. These assembly mechanisms could be general mechanisms for protein assembly in all organisms. I also characterize new fluorescent probes for use in multiple super-resolution imaging modalities and discuss the practicalities of using different super-resolution microscopes. The chemotaxis network in E. coli is the best understood signal transduction network in biology. Chemotaxis receptors cluster into complexes of thousands of proteins located at the cell poles and are used to move bacteria towards favorable stimuli in the environment. In these dense clusters, the receptors can bind each other and communicate to filter out noise and amplify weak signals. It is surprising that chemotaxis receptors are spatially segregated and the mechanism for polar localization of these complexes remains unclear. Using data from PALM images, we develop a model to understand how bacteria organize their receptors into large clusters. The model, stochastic cluster nucleation, is surprising in that is generates micron-scale periodic patterns without the need for accessory proteins to provide scaffolding or active transport. This model may be a general mechanism that cells utilize to organize small and large complexes of proteins. During cell division, E. coli must elongate, replicate its DNA and position its components properly prior to binary fission. Prior to septum formation, a ubiquitous protein called FtsZ, assembles into a ring at mid-cell (Z-ring) which constricts during cell division and recruits the remaining proteins necessary for cytokinesis. Though many details have been revealed about FtsZ, the detailed in vivo structure of the Z-ring is not well understood, and many questions remain about how ring constriction occurs. Using multiple super-resolution imaging modalities, in combination with conventional time-lapse fluorescence imaging, we show that the Z-ring does not form a long uniform filament around the circumference of the bacterium. We detail how this structure changes during division and how removal of proteins that help to position FtsZ affects the Z-ring as it proceeds through cytokinesis. Ultimately we present a simple model for Z-ring constriction during division.

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.

A sensitive EUV Schwarzschild microscope for plasma studies with sub-micrometer resolution

DOE PAGES

Zastrau, U.; Rodel, C.; Nakatsutsumi, M.; ...

2018-02-05

We present an extreme ultraviolet (EUV) microscope using a Schwarzschild objective which is optimized for single-shot sub-micrometer imaging of laser-plasma targets. The microscope has been designed and constructed for imaging the scattering from an EUV-heated solid-density hydrogen jet. Here, imaging of a cryogenic hydrogen target was demonstrated using single pulses of the free-electron laser in Hamburg (FLASH) free-electron laser at a wavelength of 13.5 nm. In a single exposure, we observe a hydrogen jet with ice fragments with a spatial resolution in the sub-micrometer range. In situ EUV imaging is expected to enable novel experimental capabilities for warm dense mattermore » studies of micrometer-sized samples in laser-plasma experiments.« less

A sensitive EUV Schwarzschild microscope for plasma studies with sub-micrometer resolution

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

Zastrau, U.; Rodel, C.; Nakatsutsumi, M.

We present an extreme ultraviolet (EUV) microscope using a Schwarzschild objective which is optimized for single-shot sub-micrometer imaging of laser-plasma targets. The microscope has been designed and constructed for imaging the scattering from an EUV-heated solid-density hydrogen jet. Here, imaging of a cryogenic hydrogen target was demonstrated using single pulses of the free-electron laser in Hamburg (FLASH) free-electron laser at a wavelength of 13.5 nm. In a single exposure, we observe a hydrogen jet with ice fragments with a spatial resolution in the sub-micrometer range. In situ EUV imaging is expected to enable novel experimental capabilities for warm dense mattermore » studies of micrometer-sized samples in laser-plasma experiments.« less

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.

Magnetoacoustic microscopic imaging of conductive objects and nanoparticles distribution

NASA Astrophysics Data System (ADS)

Liu, Siyu; Zhang, Ruochong; Luo, Yunqi; Zheng, Yuanjin

2017-09-01

Magnetoacoustic tomography has been demonstrated as a powerful and low-cost multi-wave imaging modality. However, due to limited spatial resolution and detection efficiency of magnetoacoustic signal, full potential of the magnetoacoustic imaging remains to be tapped. Here we report a high-resolution magnetoacoustic microscopy method, where magnetic stimulation is provided by a compact solenoid resonance coil connected with a matching network, and acoustic reception is realized by using a high-frequency focused ultrasound transducer. Scanning the magnetoacoustic microscopy system perpendicularly to the acoustic axis of the focused transducer would generate a two-dimensional microscopic image with acoustically determined lateral resolution. It is analyzed theoretically and demonstrated experimentally that magnetoacoustic generation in this microscopic system depends on the conductivity profile of conductive objects and localized distribution of superparamagnetic iron magnetic nanoparticles, based on two different but related implementations. The lateral resolution is characterized. Directional nature of magnetoacoustic vibration and imaging sensitivity for mapping magnetic nanoparticles are also discussed. The proposed microscopy system offers a high-resolution method that could potentially map intrinsic conductivity distribution in biological tissue and extraneous magnetic nanoparticles.

Ultra-large field-of-view two-photon microscopy

PubMed Central

Tsai, Philbert S.; Mateo, Celine; Field, Jeffrey J.; Schaffer, Chris B.; Anderson, Matthew E.; Kleinfeld, David

2015-01-01

We present a two-photon microscope that images the full extent of murine cortex with an objective-limited spatial resolution across an 8 mm by 10 mm field. The lateral resolution is approximately 1 µm and the maximum scan speed is 5 mm/ms. The scan pathway employs large diameter compound lenses to minimize aberrations and performs near theoretical limits. We demonstrate the special utility of the microscope by recording resting-state vasomotion across both hemispheres of the murine brain through a transcranial window and by imaging histological sections without the need to stitch. PMID:26072755

Spectral confocal reflection microscopy using a white light source

NASA Astrophysics Data System (ADS)

Booth, M.; Juškaitis, R.; Wilson, T.

2008-08-01

We present a reflection confocal microscope incorporating a white light supercontinuum source and spectral detection. The microscope provides images resolved spatially in three-dimensions, in addition to spectral resolution covering the wavelength range 450-650nm. Images and reflection spectra of artificial and natural specimens are presented, showing features that are not normally revealed in conventional microscopes or confocal microscopes using discrete line lasers. The specimens include thin film structures on semiconductor chips, iridescent structures in Papilio blumei butterfly scales, nacre from abalone shells and opal gemstones. Quantitative size and refractive index measurements of transparent beads are derived from spectral interference bands.

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.

Improved resolution in practical light microscopy by means of a glass-fiber 2 π-tilting device

NASA Astrophysics Data System (ADS)

Bradl, Joachim; Rinke, Bernd; Schneider, Bernhard; Hausmann, Michael; Cremer, Christoph G.

1996-01-01

The spatial resolution of a conventional light microscope or a confocal laser scanning microscope can be determined by calculating the point spread function for the objective used. Normally, ideal conditions are assumed for these calculations. Such conditions, however, are often not fulfilled in biological applications especially in those cases where biochemical requirements (e.g. buffer conditions) influence the specimen preparation on the microscope slide (i.e. 'practical' light microscopy). It has been shown that the problem of a reduced z- resolution in 3D-microscopy (optical sectioning) can be overcome by a capillary in a 2(pi) - tilting device that allows object rotation into an optimal perspective. The application of the glass capillary instead of a standard slide has an additional influence on the imaging properties of the microscope. Therefore, another 2(pi) -tilting device was developed, using a glass fiber for object fixation and rotation. Such a fiber could be covered by standard cover glasses. To estimate the resolution of this setup, point spread functions were measured under different conditions using fluorescent microspheres of subwavelength dimensions. Results obtained from standard slide setups were compared to the glass fiber setup. These results showed that in practice rotation leads to an overall 3D-resolution improvement.

Focal depth measurement of scanning helium ion microscope

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

Guo, Hongxuan, E-mail: Guo.hongxuan@nims.go.jp; Itoh, Hiroshi; Wang, Chunmei

2014-07-14

When facing the challenges of critical dimension measurement of complicated nanostructures, such as of the three dimension integrated circuit, characterization of the focal depth of microscopes is important. In this Letter, we developed a method for characterizing the focal depth of a scanning helium ion microscope (HIM) by using an atomic force microscope tip characterizer (ATC). The ATC was tilted in a sample chamber at an angle to the scanning plan. Secondary electron images (SEIs) were obtained at different positions of the ATC. The edge resolution of the SEIs shows the nominal diameters of the helium ion beam at differentmore » focal levels. With this method, the nominal shapes of the helium ion beams were obtained with different apertures. Our results show that a small aperture is necessary to get a high spatial resolution and high depth of field images with HIM. This work provides a method for characterizing and improving the performance of HIM.« less

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

Focal depth measurement of scanning helium ion microscope

NASA Astrophysics Data System (ADS)

Guo, Hongxuan; Itoh, Hiroshi; Wang, Chunmei; Zhang, Han; Fujita, Daisuke

2014-07-01

When facing the challenges of critical dimension measurement of complicated nanostructures, such as of the three dimension integrated circuit, characterization of the focal depth of microscopes is important. In this Letter, we developed a method for characterizing the focal depth of a scanning helium ion microscope (HIM) by using an atomic force microscope tip characterizer (ATC). The ATC was tilted in a sample chamber at an angle to the scanning plan. Secondary electron images (SEIs) were obtained at different positions of the ATC. The edge resolution of the SEIs shows the nominal diameters of the helium ion beam at different focal levels. With this method, the nominal shapes of the helium ion beams were obtained with different apertures. Our results show that a small aperture is necessary to get a high spatial resolution and high depth of field images with HIM. This work provides a method for characterizing and improving the performance of HIM.

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

Contribution of the channel electron multiplier to the race of vacuum tubes towards picosecond resolution time

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

Pietri, G.

1977-02-01

The ability to tightly pack millions of microscopic secondary emitting channels into a two-dimensional, very thin, array known as a microchannel plate (MCP) provides excellent electrical charge or current amplification associated with an extremely short response time as well as very good spatial resolution. The ultimate performances in spatial and temporal resolutions achieved by MCP-based vacuum devices are discussed and illustrated by the description of a large range of experimental prototypes (photomultipliers, oscilloscope tubes, streak camera tubes, etc.) designed and produced at LEP, then tested in cooperation with Nuclear Research and Plasma Physics Centers in Europe and USA.

Development and demonstration of a water-window soft x-ray microscope using a Z-pinching capillary discharge source

NASA Astrophysics Data System (ADS)

Nawaz, M. F.; Jancarek, Alexandr; Nevrkla, Michal; Duda, Martin Jakub; Pina, Ladislav

2017-05-01

The development and demonstration of a soft X-ray (SXR) microscope, based on a Z-pinching capillary discharge source has been realized. The Z-pinching plasma acts as a source of SXR radiation. A ceramic capacitor bank is pulsed charged up to 80 kV, and discharged through a pre- ionized nitrogen filled ceramic capillary. The discharge current has an amplitude of 25 kA. Working within the water-window spectral region (λ = 2.88 nm), corresponding to the 1s2-1s2p quantum transition of helium-like nitrogen (N5+), the microscope has a potential in exploiting the natural contrast existing between the K-absorption edges of carbon and oxygen as the main constituents of biological materials, and hence imaging them with high spatial resolution. The SXR microscope uses the grazing incidence ellipsoidal condenser mirror for the illumination, and the Fresnel zone plate optics for the imaging of samples onto a BI-CCD camera. The half- pitch spatial resolution of 100 nm [1] was achieved, as demonstrated by the knife-edge test. In order to enhance the photon-flux at the sample plane, a new scheme for focusing the radiation, from multiple capillary sources has been investigated. Details about the source, and the construction of the microscope are presented and discussed.

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

Yashchuk, V. V.; Fischer, P. J.; Chan, E. R.

We present a modulation transfer function (MTF) calibration method based on binary pseudo-random (BPR) one-dimensional sequences and two-dimensional arrays as an effective method for spectral characterization in the spatial frequency domain of a broad variety of metrology instrumentation, including interferometric microscopes, scatterometers, phase shifting Fizeau interferometers, scanning and transmission electron microscopes, and at this time, x-ray microscopes. The inherent power spectral density of BPR gratings and arrays, which has a deterministic white-noise-like character, allows a direct determination of the MTF with a uniform sensitivity over the entire spatial frequency range and field of view of an instrument. We demonstrate themore » MTF calibration and resolution characterization over the full field of a transmission soft x-ray microscope using a BPR multilayer (ML) test sample with 2.8 nm fundamental layer thickness. We show that beyond providing a direct measurement of the microscope's MTF, tests with the BPRML sample can be used to fine tune the instrument's focal distance. Finally, our results confirm the universality of the method that makes it applicable to a large variety of metrology instrumentation with spatial wavelength bandwidths from a few nanometers to hundreds of millimeters.« less

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.

Estimation of reactogenicity of preparations produced on the basis of photoinactivated live vaccines against brucellosis and tularaemia on the organismic level.2. Using the method of speckle-microscopy with high spatial resolution

NASA Astrophysics Data System (ADS)

Ulianova, O. V.; Uianov, S. S.; Li, Pengcheng; Luo, Qingming

2011-04-01

The method of speckle microscopy was adapted to estimate the reactogenicity of the prototypes of vaccine preparations against extremely dangerous infections. The theory is proposed to describe the mechanism of formation of the output signal from the super-high spatial resolution speckle microscope. The experimental studies show that bacterial suspensions, irradiated in different regimes of inactivation, do not exert negative influence on the blood microcirculations in laboratory animals.

Scanning near-field optical microscopy.

PubMed

Vobornik, Dusan; Vobornik, Slavenka

2008-02-01

An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today's science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM) is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution.

Electron microscopy of whole cells in liquid with nanometer resolution

PubMed Central

de Jonge, N.; Peckys, D. B.; Kremers, G. J.; Piston, D. W.

2009-01-01

Single gold-tagged epidermal growth factor (EGF) molecules bound to cellular EGF receptors of fixed fibroblast cells were imaged in liquid with a scanning transmission electron microscope (STEM). The cells were placed in buffer solution in a microfluidic device with electron transparent windows inside the vacuum of the electron microscope. A spatial resolution of 4 nm and a pixel dwell time of 20 μs were obtained. The liquid layer was sufficiently thick to contain the cells with a thickness of 7 ± 1 μm. The experimental findings are consistent with a theoretical calculation. Liquid STEM is a unique approach for imaging single molecules in whole cells with significantly improved resolution and imaging speed over existing methods. PMID:19164524

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.

Enhanced EDX images by fusion of multimodal SEM images using pansharpening techniques.

PubMed

Franchi, G; Angulo, J; Moreaud, M; Sorbier, L

2018-01-01

The goal of this paper is to explore the potential interest of image fusion in the context of multimodal scanning electron microscope (SEM) imaging. In particular, we aim at merging the backscattered electron images that usually have a high spatial resolution but do not provide enough discriminative information to physically classify the nature of the sample, with energy-dispersive X-ray spectroscopy (EDX) images that have discriminative information but a lower spatial resolution. The produced images are named enhanced EDX. To achieve this goal, we have compared the results obtained with classical pansharpening techniques for image fusion with an original approach tailored for multimodal SEM fusion of information. Quantitative assessment is obtained by means of two SEM images and a simulated dataset produced by a software based on PENELOPE. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Tip-enhanced Raman mapping with top-illumination AFM.

PubMed

Chan, K L Andrew; Kazarian, Sergei G

2011-04-29

Tip-enhanced Raman mapping is a powerful, emerging technique that offers rich chemical information and high spatial resolution. Currently, most of the successes in tip-enhanced Raman scattering (TERS) measurements are based on the inverted configuration where tips and laser are approaching the sample from opposite sides. This results in the limitation of measurement for transparent samples only. Several approaches have been developed to obtain tip-enhanced Raman mapping in reflection mode, many of which involve certain customisations of the system. We have demonstrated in this work that it is also possible to obtain TERS nano-images using an upright microscope (top-illumination) with a gold-coated Si atomic force microscope (AFM) cantilever without significant modification to the existing integrated AFM/Raman system. A TERS image of a single-walled carbon nanotube has been achieved with a spatial resolution of ∼ 20-50 nm, demonstrating the potential of this technique for studying non-transparent nanoscale materials.

Micromirror structured illumination microscope for high-speed in vivo drosophila brain imaging.

PubMed

Masson, A; Pedrazzani, M; Benrezzak, S; Tchenio, P; Preat, T; Nutarelli, D

2014-01-27

Genetic tools and especially genetically encoded fluorescent reporters have given a special place to optical microscopy in drosophila neurobiology research. In order to monitor neural networks activity, high speed and sensitive techniques, with high spatial resolution are required. Structured illumination microscopies are wide-field approaches with optical sectioning ability. Despite the large progress made with the introduction of the HiLo principle, they did not meet the criteria of speed and/or spatial resolution for drosophila brain imaging. We report on a new implementation that took advantage of micromirror matrix technology to structure the illumination. Thus, we showed that the developed instrument exhibits a spatial resolution close to that of confocal microscopy but it can record physiological responses with a speed improved by more than an order a magnitude.

Resolution improvement by nonconfocal theta microscopy.

PubMed

Lindek, S; Stelzer, E H

1999-11-01

We present a novel scanning fluorescence microscopy technique, nonconfocal theta microscopy (NCTM), that provides almost isotropic resolution. In NCTM, multiphoton absorption from two orthogonal illumination directions is used to induce fluorescence emission. Therefore the point-spread function of the microscope is described by the product of illumination point-spread functions with reduced spatial overlap, which provides the resolution improvement and the more isotropic observation volume. We discuss the technical details of this new method.

Spectro-microscopy of living plant cells.

PubMed

Harter, Klaus; Meixner, Alfred J; Schleifenbaum, Frank

2012-01-01

Spectro-microscopy, a combination of fluorescence microscopy with spatially resolved spectroscopic techniques, provides new and exciting tools for functional cell biology in living organisms. This review focuses on recent developments in spectro-microscopic applications for the investigation of living plant cells in their native tissue context. The application of spectro-microscopic methods led to the recent discovery of a fast signal response pathway for the brassinosteroide receptor BRI1 in the plasma membrane of living plant cells. Moreover, the competence of different plant cell types to respond to environmental or endogenous stimuli was determined in vivo by correlation analysis of different optical and spectroscopic readouts such as fluorescence lifetime (FLT). Furthermore, a new spectro-microscopic technique, fluorescence intensity decay shape analysis microscopy (FIDSAM), has been developed. FIDSAM is capable of imaging low-expressed fluorophore-tagged proteins at high spatial resolution and precludes the misinterpretation of autofluorescence artifacts. In addition, FIDSAM provides a very effective and sensitive tool on the basis of Förster resonance energy transfer (FRET) for the qualitative and quantitative determination of protein-protein interaction. Finally, we report on the quantitative analysis of the photosystem I and II (PSI/PSII) ratio in the chloroplasts of living Arabidopsis plants at room temperature, using high-resolution, spatially resolved fluorescence spectroscopy. With this technique, it was not only possible to measure PSI/PSII ratios, but also to demonstrate the differential competence of wild-type and carbohydrate-deficient plants to adapt the PSI/PSII ratio to different light conditions. In summary, the information content of standard microscopic images is extended by several dimensions by the use of spectro-microscopic approaches. Therefore, novel cell physiological and molecular topics can be addressed and valuable insights into molecular and subcellular processes can be obtained in living plants.

Quantitative measurement of piezoelectric coefficient of thin film using a scanning evanescent microwave microscope.

PubMed

Zhao, Zhenli; Luo, Zhenlin; Liu, Chihui; Wu, Wenbin; Gao, Chen; Lu, Yalin

2008-06-01

This article describes a new approach to quantitatively measure the piezoelectric coefficients of thin films at the microscopic level using a scanning evanescent microwave microscope. This technique can resolve 10 pm deformation caused by the piezoelectric effect and has the advantages of high scanning speed, large scanning area, submicron spatial resolution, and a simultaneous accessibility to many other related properties. Results from the test measurements on the longitudinal piezoelectric coefficient of PZT thin film agree well with those from other techniques listed in literatures.

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

PubMed

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

1997-07-01

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

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

PubMed Central

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

1997-01-01

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

Ultrafast 2D IR microscopy

PubMed Central

Baiz, Carlos R.; Schach, Denise; Tokmakoff, Andrei

2014-01-01

We describe a microscope for measuring two-dimensional infrared (2D IR) spectra of heterogeneous samples with μm-scale spatial resolution, sub-picosecond time resolution, and the molecular structure information of 2D IR, enabling the measurement of vibrational dynamics through correlations in frequency, time, and space. The setup is based on a fully collinear “one beam” geometry in which all pulses propagate along the same optics. Polarization, chopping, and phase cycling are used to isolate the 2D IR signals of interest. In addition, we demonstrate the use of vibrational lifetime as a contrast agent for imaging microscopic variations in molecular environments. PMID:25089490

SEM-microphotogrammetry, a new take on an old method for generating high-resolution 3D models from SEM images.

PubMed

Ball, A D; Job, P A; Walker, A E L

2017-08-01

The method we present here uses a scanning electron microscope programmed via macros to automatically capture dozens of images at suitable angles to generate accurate, detailed three-dimensional (3D) surface models with micron-scale resolution. We demonstrate that it is possible to use these Scanning Electron Microscope (SEM) images in conjunction with commercially available software originally developed for photogrammetry reconstructions from Digital Single Lens Reflex (DSLR) cameras and to reconstruct 3D models of the specimen. These 3D models can then be exported as polygon meshes and eventually 3D printed. This technique offers the potential to obtain data suitable to reconstruct very tiny features (e.g. diatoms, butterfly scales and mineral fabrics) at nanometre resolution. Ultimately, we foresee this as being a useful tool for better understanding spatial relationships at very high resolution. However, our motivation is also to use it to produce 3D models to be used in public outreach events and exhibitions, especially for the blind or partially sighted. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Optical high-resolution analysis of rotational movement: testing circular spatial filter velocimetry (CSFV) with rotating biological cells

NASA Astrophysics Data System (ADS)

Schaeper, M.; Schmidt, R.; Kostbade, R.; Damaschke, N.; Gimsa, J.

2016-07-01

Circular spatial filtering velocimetry (CSFV) was tested during the microscopic registration of the individual rotations of baker’s yeast cells. Their frequency-dependent rotation (electrorotation; ER) was induced in rotating electric fields, which were generated in a glass chip chamber with four electrodes (600 μm tip-to-tip distance). The electrodes were driven with sinusoidal quadrature signals of 5 or 8 V PP with frequencies up to 3 MHz. The observed cell rotation was of the order of 1-100 s per revolution. At each measuring frequency, the independent rotations of up to 20 cells were simultaneously recorded with a high-speed camera. CSFV was software-implemented using circular spatial filters with harmonic gratings. ER was proportional to the phase shift between the values of the spatial filtering signal of consecutive frames. ER spectra obtained by CSFV from the rotation velocities at different ER-field frequencies agreed well with manual measurements and theoretical spectra. Oscillations in the rotation velocity of a single cell in the elliptically polarized field near an electrode, which were resolved by CSFV, could not be visually discerned. ER step responses after field-on were recorded at 2500 frames per second. Analysis proved the high temporal resolution of CSFV and revealed a largely linear torque-friction relation during the acceleration phase of ER. Future applications of CSFV will allow for the simple and cheap automated high-resolution analysis of rotational movements where mechanical detection has too low a resolution or is not possible, e.g. in polluted environments or for gas and fluid vortices, microscopic objects, etc.

Soft x-ray imaging with incoherent sources

NASA Astrophysics Data System (ADS)

Wachulak, P.; Torrisi, A.; Ayele, M.; Bartnik, A.; Czwartos, J.; Wegrzyński, Ł.; Fok, T.; Parkman, T.; Vondrová, Š.; Turnová, J.; Odstrcil, M.; Fiedorowicz, H.

2017-05-01

In this work we present experimental, compact desk-top SXR microscope, the EUV microscope which is at this stage a technology demonstrator, and finally, the SXR contact microscope. The systems are based on laser-plasma EUV and SXR sources, employing a double stream gas puff target. The EUV and SXR full field microscopes, operating at 13.8 nm and 2.88 nm wavelengths, respectively, are capable of imaging nanostructures with a sub-50 nm spatial resolution with relatively short (seconds) exposure times. The SXR contact microscope operates in the "water-window" spectral range, to produce an imprint of the internal structure of the sample in a thin layer of SXR light sensitive photoresist. Applications of such desk-top EUV and SXR microscopes for studies of variety of different samples - test objects for resolution assessment and other objects such as carbon membranes, DNA plasmid samples, organic and inorganic thin layers, diatoms, algae and carcinoma cells, are also presented. Details about the sources, the microscopes as well as the imaging results for various objects will be presented and discussed. The development of such compact imaging systems may be important to the new research related to biological, material science and nanotechnology applications.

High-resolution spatiotemporal strain mapping reveals non-uniform deformation in micropatterned elastomers

NASA Astrophysics Data System (ADS)

Aksoy, B.; Rehman, A.; Bayraktar, H.; Alaca, B. E.

2017-04-01

Micropatterns are generated on a vast selection of polymeric substrates for various applications ranging from stretchable electronics to cellular mechanobiological systems. When these patterned substrates are exposed to external loading, strain field is primarily affected by the presence of microfabricated structures and similarly by fabrication-related defects. The capturing of such nonhomogeneous strain fields is of utmost importance in cases where study of the mechanical behavior with a high spatial resolution is necessary. Image-based non-contact strain measurement techniques are favorable and have recently been extended to scanning tunneling microscope and scanning electron microscope images for the characterization of mechanical properties of metallic materials, e.g. steel and aluminum, at the microscale. A similar real-time analysis of strain heterogeneity in elastomers is yet to be achieved during the entire loading sequence. The available measurement methods for polymeric materials mostly depend on cross-head displacement or precalibrated strain values. Thus, they suffer either from the lack of any real-time analysis, spatiotemporal distribution or high resolution in addition to a combination of these factors. In this work, these challenges are addressed by integrating a tensile stretcher with an inverted optical microscope and developing a subpixel particle tracking algorithm. As a proof of concept, the patterns with a critical dimension of 200 µm are generated on polydimethylsiloxane substrates and strain distribution in the vicinity of the patterns is captured with a high spatiotemporal resolution. In the field of strain measurement, there is always a tradeoff between minimum measurable strain value and spatial resolution. Current noncontact techniques on elastomers can deliver a strain resolution of 0.001% over a minimum length of 5 cm. More importantly, inhomogeneities within this quite large region cannot be captured. The proposed technique can overcome this challenge and provides a displacement measurement resolution of 116 nm and a strain resolution of 0.04% over a gage length of 300 µm. Similarly, the ability to capture inhomogeneities is demonstrated by mapping strain around a thru-hole. The robustness of the technique is also evaluated, where no appreciable change in strain measurement is observed despite the significant variations imposed on the measurement mesh. The proposed approach introduces critical improvements for the determination of displacement and strain gradients in elastomers regarding the real-time nature of strain mapping with a microscale spatial resolution.

A Mach-Zender digital holographic microscope with sub-micrometer resolution for imaging and tracking of marine micro-organisms

NASA Astrophysics Data System (ADS)

Kühn, Jonas; Niraula, Bimochan; Liewer, Kurt; Kent Wallace, J.; Serabyn, Eugene; Graff, Emilio; Lindensmith, Christian; Nadeau, Jay L.

2014-12-01

Digital holographic microscopy is an ideal tool for investigation of microbial motility. However, most designs do not exhibit sufficient spatial resolution for imaging bacteria. In this study we present an off-axis Mach-Zehnder design of a holographic microscope with spatial resolution of better than 800 nm and the ability to resolve bacterial samples at varying densities over a 380 μm × 380 μm × 600 μm three-dimensional field of view. Larger organisms, such as protozoa, can be resolved in detail, including cilia and flagella. The instrument design and performance are presented, including images and tracks of bacterial and protozoal mixed samples and pure cultures of six selected species. Organisms as small as 1 μm (bacterial spores) and as large as 60 μm (Paramecium bursaria) may be resolved and tracked without changes in the instrument configuration. Finally, we present a dilution series investigating the maximum cell density that can be imaged, a type of analysis that has not been presented in previous holographic microscopy studies.

MPGD for breast cancer prevention: a high resolution and low dose radiation medical imaging

NASA Astrophysics Data System (ADS)

Gutierrez, R. M.; Cerquera, E. A.; Mañana, G.

2012-07-01

Early detection of small calcifications in mammograms is considered the best preventive tool of breast cancer. However, existing digital mammography with relatively low radiation skin exposure has limited accessibility and insufficient spatial resolution for small calcification detection. Micro Pattern Gaseous Detectors (MPGD) and associated technologies, increasingly provide new information useful to generate images of microscopic structures and make more accessible cutting edge technology for medical imaging and many other applications. In this work we foresee and develop an application for the new information provided by a MPGD camera in the form of highly controlled images with high dynamical resolution. We present a new Super Detail Image (S-DI) that efficiently profits of this new information provided by the MPGD camera to obtain very high spatial resolution images. Therefore, the method presented in this work shows that the MPGD camera with SD-I, can produce mammograms with the necessary spatial resolution to detect microcalcifications. It would substantially increase efficiency and accessibility of screening mammography to highly improve breast cancer prevention.

Progress on PEEM3 -- An Aberration Corrected X-Ray Photoemission Electron Microscope at the ALS

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

MacDowell, A. A.; Feng, J.; DeMello, A.

2007-01-19

A new ultrahigh-resolution photoemission electron microscope called PEEM3 is being developed and built at the Advanced Light Source (ALS). An electron mirror combined with a much-simplified magnetic dipole separator is to be used to provide simultaneous correction of spherical and chromatic aberrations. It is installed on an elliptically polarized undulator (EPU) beamline, and will be operated with very high spatial resolution and high flux to study the composition, structure, electric and magnetic properties of complex materials. The instrument has been designed and is described. The instrumental hardware is being deployed in 2 phases. The first phase is the deployment ofmore » a standard PEEM type microscope consisting of the standard linear array of electrostatic electron lenses. The second phase will be the installation of the aberration corrected upgrade to improve resolution and throughput. This paper describes progress as the instrument enters the commissioning part of the first phase.« less

Progress on PEEM3 - An Aberration Corrected X-Ray PhotoemissionElectron Microscope at the ALS

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

MacDowell, Alastair A.; Feng, J.; DeMello, A.

2006-05-20

A new ultrahigh-resolution photoemission electron microscope called PEEM3 is being developed and built at the Advanced Light Source (ALS). An electron mirror combined with a much-simplified magnetic dipole separator is to be used to provide simultaneous correction of spherical and chromatic aberrations. It is installed on an elliptically polarized undulator (EPU) beamline, and will be operated with very high spatial resolution and high flux to study the composition, structure, electric and magnetic properties of complex materials. The instrument has been designed and is described. The instrumental hardware is being deployed in 2 phases. The first phase is the deployment ofmore » a standard PEEM type microscope consisting of the standard linear array of electrostatic electron lenses. The second phase will be the installation of the aberration corrected upgrade to improve resolution and throughput. This paper describes progress as the instrument enters the commissioning part of the first phase.« less

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

NASA Astrophysics Data System (ADS)

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

2009-11-01

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

SCANNING NEAR-FIELD OPTICAL MICROSCOPY

PubMed Central

Vobornik, Dušan; Vobornik, Slavenka

2008-01-01

An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today’s science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM) is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution. PMID:18318675

Wide field video-rate two-photon imaging by using spinning disk beam scanner

NASA Astrophysics Data System (ADS)

Maeda, Yasuhiro; Kurokawa, Kazuo; Ito, Yoko; Wada, Satoshi; Nakano, Akihiko

2018-02-01

The microscope technology with wider view field, deeper penetration depth, higher spatial resolution and higher imaging speed are required to investigate the intercellular dynamics or interactions of molecules and organs in cells or a tissue in more detail. The two-photon microscope with a near infrared (NIR) femtosecond laser is one of the technique to improve the penetration depth and spatial resolution. However, the video-rate or high-speed imaging with wide view field is difficult to perform with the conventional two-photon microscope. Because point-to-point scanning method is used in conventional one, so it's difficult to achieve video-rate imaging. In this study, we developed a two-photon microscope with spinning disk beam scanner and femtosecond NIR fiber laser with around 10 W average power for the microscope system to achieve above requirements. The laser is consisted of an oscillator based on mode-locked Yb fiber laser, a two-stage pre-amplifier, a main amplifier based on a Yb-doped photonic crystal fiber (PCF), and a pulse compressor with a pair of gratings. The laser generates a beam with maximally 10 W average power, 300 fs pulse width and 72 MHz repetition rate. And the beam incident to a spinning beam scanner (Yokogawa Electric) optimized for two-photon imaging. By using this system, we achieved to obtain the 3D images with over 1mm-penetration depth and video-rate image with 350 x 350 um view field from the root of Arabidopsis thaliana.

Micro-Mirrors for Nanoscale Three-Dimensional Microscopy

PubMed Central

Seale, Kevin; Janetopoulos, Chris; Wikswo, John

2013-01-01

A research-grade optical microscope is capable of resolving fine structures in two-dimensional images. However, three-dimensional resolution, or the ability of the microscope to distinguish between objects lying above or below the focal plane from in-focus objects, is not nearly as good as in-plane resolution. In this issue of ACS Nano, McMahon et al. report the use of mirrored pyramidal wells with a conventional microscope for rapid, 3D localization and tracking of nanoparticles. Mirrors have been used in microscopy before, but recent work with MPWs is unique because it enables the rapid determination of the x-, y-, and z-position of freely diffusing nanoparticles and cellular nanostructures with unprecedented speed and spatial accuracy. As inexpensive tools for 3D visualization, mirrored pyramidal wells may prove to be invaluable aids in nanotechnology and engineering of nanomaterials. PMID:19309167

Estimation of reactogenicity of preparations produced on the basis of photoinactivated live vaccines against brucellosis and tularaemia on the organismic level. 2. Using the method of speckle-microscopy with high spatial resolution

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

Ulianova, O V; Uianov, S S; Li Pengcheng

2011-04-30

The method of speckle microscopy was adapted to estimate the reactogenicity of the prototypes of vaccine preparations against extremely dangerous infections. The theory is proposed to describe the mechanism of formation of the output signal from the super-high spatial resolution speckle microscope. The experimental studies show that bacterial suspensions, irradiated in different regimes of inactivation, do not exert negative influence on the blood microcirculations in laboratory animals. (optical technologies in biophysics and medicine)

Novel microfabrication stage allowing for one-photon and multi-photon light assisted molecular immobilization and for multi-photon microscope

NASA Astrophysics Data System (ADS)

Gonçalves, Odete; Snider, Scott; Zadoyan, Ruben; Nguyen, Quoc-Thang; Vorum, Henrik; Petersen, Steffen B.; Neves-Petersen, Maria Teresa

2017-02-01

Light Assisted Molecular Immobilization (LAMI) results in spatially oriented and localized covalent coupling of biomolecules onto thiol reactive surfaces. LAMI is possible due to the conserved spatial proximity between aromatic residues and disulfide bridges in proteins. When aromatic residues are excited with UV light (275-295nm), disulphide bridges are disrupted and the formed thiol groups covalently bind to surfaces. Immobilization hereby reported is achieved in a microfabrication stage coupled to a fs-laser, through one- or multi-photon excitation. The fundamental 840nm output is tripled to 280nm and focused onto the sample, leading to one-photon excitation and molecular immobilization. The sample rests on a xyz-stage with micrometer step resolution and is illuminated according to a pattern uploaded to the software controlling the stage and the shutter. Molecules are immobilized according to such pattern, with micrometer spatial resolution. Spatial masks inserted in the light path lead to light diffraction patterns used to immobilize biomolecules with submicrometer spatial resolution. Light diffraction patterns are imaged by an inbuilt microscope. Two-photon microscopy and imaging of the fluorescent microbeads is shown. Immobilization of proteins, e.g. C-reactive protein, and of an engineered molecular beacon has been successfully achieved. The beacon was coupled to a peptide containing a disulfide bridge neighboring a tryptophan residue, being this way possible to immobilize the beacon on a surface using one-photon LAMI. This technology is being implemented in the creation of point-of-care biosensors aiming at the detection of cancer and cardiovascular disease markers.

Microspectroscopic imaging of solution plasma: How do its physical properties and chemical species evolve in atmospheric-pressure water vapor bubbles?

NASA Astrophysics Data System (ADS)

Yui, Hiroharu; Banno, Motohiro

2018-01-01

In this article, we review the development of scientific instruments for obtaining information on the evolution of physical properties and chemical species of solution plasma (SP). When a pulsed high voltage is applied between electrodes immersed in an aqueous solution, SP is formed in water vapor bubbles transiently generated in the solution under atmospheric pressure. To clarify how SP emerges in water vapor bubbles and is sustained in solutions, an instrument with micrometer spatial resolution and nanosecond temporal resolution is required. To meet these requirements, a microscopic system with a custom-made optical discharge cell was newly developed, where the working distance between the SP and the microscopic objective lens was minimized. A hollow electrode equipped in the discharge cell also enabled us to control the chemical composition in water vapor bubbles. To study the spatial and temporal evolutions of chemical species in micrometer and nano- to microsecond regions, a streak camera with a spectrometer and a CCD detector with a time-gated electronic device were combined with the microscope system. The developed instrument is expected to contribute to providing a new means of developing new schemes for chemical reactions and material syntheses.

Microscopy illumination engineering using a low-cost liquid crystal display.

PubMed

Guo, Kaikai; Bian, Zichao; Dong, Siyuan; Nanda, Pariksheet; Wang, Ying Min; Zheng, Guoan

2015-02-01

Illumination engineering is critical for obtaining high-resolution, high-quality images in microscope settings. In a typical microscope, the condenser lens provides sample illumination that is uniform and free from glare. The associated condenser diaphragm can be manually adjusted to obtain the optimal illumination numerical aperture. In this paper, we report a programmable condenser lens for active illumination control. In our prototype setup, we used a $15 liquid crystal display as a transparent spatial light modulator and placed it at the back focal plane of the condenser lens. By setting different binary patterns on the display, we can actively control the illumination and the spatial coherence of the microscope platform. We demonstrated the use of such a simple scheme for multimodal imaging, including bright-field microscopy, darkfield microscopy, phase-contrast microscopy, polarization microscopy, 3D tomographic imaging, and super-resolution Fourier ptychographic imaging. The reported illumination engineering scheme is cost-effective and compatible with most existing platforms. It enables a turnkey solution with high flexibility for researchers in various communities. From the engineering point-of-view, the reported illumination scheme may also provide new insights for the development of multimodal microscopy and Fourier ptychographic imaging.

Dynamic full-field infrared imaging with multiple synchrotron beams

PubMed Central

Stavitski, Eli; Smith, Randy J.; Bourassa, Megan W.; Acerbo, Alvin S.; Carr, G. L.; Miller, Lisa M.

2013-01-01

Microspectroscopic imaging in the infrared (IR) spectral region allows for the examination of spatially resolved chemical composition on the microscale. More than a decade ago, it was demonstrated that diffraction limited spatial resolution can be achieved when an apertured, single pixel IR microscope is coupled to the high brightness of a synchrotron light source. Nowadays, many IR microscopes are equipped with multi-pixel Focal Plane Array (FPA) detectors, which dramatically improve data acquisition times for imaging large areas. Recently, progress been made toward efficiently coupling synchrotron IR beamlines to multi-pixel detectors, but they utilize expensive and highly customized optical schemes. Here we demonstrate the development and application of a simple optical configuration that can be implemented on most existing synchrotron IR beamlines in order to achieve full-field IR imaging with diffraction-limited spatial resolution. Specifically, the synchrotron radiation fan is extracted from the bending magnet and split into four beams that are combined on the sample, allowing it to fill a large section of the FPA. With this optical configuration, we are able to oversample an image by more than a factor of two, even at the shortest wavelengths, making image restoration through deconvolution algorithms possible. High chemical sensitivity, rapid acquisition times, and superior signal-to-noise characteristics of the instrument are demonstrated. The unique characteristics of this setup enabled the real time study of heterogeneous chemical dynamics with diffraction-limited spatial resolution for the first time. PMID:23458231

Real-time terahertz near-field microscope.

PubMed

Blanchard, F; Doi, A; Tanaka, T; Hirori, H; Tanaka, H; Kadoya, Y; Tanaka, K

2011-04-25

We report a terahertz near-field microscope with a high dynamic range that can capture images of a 370 x 740 μm2 area at 35 frames per second. We achieve high spatial resolution (14 μm corresponding to λ/30 for a center frequency at 0.7 THz) on a large area by combining two novel techniques: terahertz generation by tilted-pulse-front excitation and electro-optic balanced imaging detection using a thin crystal. To demonstrate the microscope capability, we reveal the field enhancement at the gap position of a dipole antenna after the irradiation of a terahertz pulse.

Localization-based super-resolution imaging of cellular structures.

PubMed

Kanchanawong, Pakorn; Waterman, Clare M

2013-01-01

Fluorescence microscopy allows direct visualization of fluorescently tagged proteins within cells. However, the spatial resolution of conventional fluorescence microscopes is limited by diffraction to ~250 nm, prompting the development of super-resolution microscopy which offers resolution approaching the scale of single proteins, i.e., ~20 nm. Here, we describe protocols for single molecule localization-based super-resolution imaging, using focal adhesion proteins as an example and employing either photoswitchable fluorophores or photoactivatable fluorescent proteins. These protocols should also be easily adaptable to imaging a broad array of macromolecular assemblies in cells whose components can be fluorescently tagged and assemble into high density structures.

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

Yashchuk, V. V., E-mail: VVYashchuk@lbl.gov; Chan, E. R.; Lacey, I.

We present a modulation transfer function (MTF) calibration method based on binary pseudo-random (BPR) one-dimensional sequences and two-dimensional arrays as an effective method for spectral characterization in the spatial frequency domain of a broad variety of metrology instrumentation, including interferometric microscopes, scatterometers, phase shifting Fizeau interferometers, scanning and transmission electron microscopes, and at this time, x-ray microscopes. The inherent power spectral density of BPR gratings and arrays, which has a deterministic white-noise-like character, allows a direct determination of the MTF with a uniform sensitivity over the entire spatial frequency range and field of view of an instrument. We demonstrate themore » MTF calibration and resolution characterization over the full field of a transmission soft x-ray microscope using a BPR multilayer (ML) test sample with 2.8 nm fundamental layer thickness. We show that beyond providing a direct measurement of the microscope’s MTF, tests with the BPRML sample can be used to fine tune the instrument’s focal distance. Our results confirm the universality of the method that makes it applicable to a large variety of metrology instrumentation with spatial wavelength bandwidths from a few nanometers to hundreds of millimeters.« less

Large image microscope array for the compilation of multimodality whole organ image databases.

PubMed

Namati, Eman; De Ryk, Jessica; Thiesse, Jacqueline; Towfic, Zaid; Hoffman, Eric; Mclennan, Geoffrey

2007-11-01

Three-dimensional, structural and functional digital image databases have many applications in education, research, and clinical medicine. However, to date, apart from cryosectioning, there have been no reliable means to obtain whole-organ, spatially conserving histology. Our aim was to generate a system capable of acquiring high-resolution images, featuring microscopic detail that could still be spatially correlated to the whole organ. To fulfill these objectives required the construction of a system physically capable of creating very fine whole-organ sections and collecting high-magnification and resolution digital images. We therefore designed a large image microscope array (LIMA) to serially section and image entire unembedded organs while maintaining the structural integrity of the tissue. The LIMA consists of several integrated components: a novel large-blade vibrating microtome, a 1.3 megapixel peltier cooled charge-coupled device camera, a high-magnification microscope, and a three axis gantry above the microtome. A custom control program was developed to automate the entire sectioning and automated raster-scan imaging sequence. The system is capable of sectioning unembedded soft tissue down to a thickness of 40 microm at specimen dimensions of 200 x 300 mm to a total depth of 350 mm. The LIMA system has been tested on fixed lung from sheep and mice, resulting in large high-quality image data sets, with minimal distinguishable disturbance in the delicate alveolar structures. Copyright 2007 Wiley-Liss, Inc.

Numerical restoration of surface vortices in Nb films measured by a scanning SQUID microscope

NASA Astrophysics Data System (ADS)

Ito, Atsuki; Thanh Huy, Ho; Dang, Vu The; Miyoshi, Hiroki; Hayashi, Masahiko; Ishida, Takekazu

2017-07-01

In the present work, we investigated a vortex profile appeared on a pure Nb film (500 nm in thickness, 10 mm x 10 mm) by using a scanning SQUID microscope. We found that the local magnetic distribution thus observed is broadened compared to a true vortex profile in the superconducting film. We therefore applied the numerical method to improve a spatial resolution of the scanning SQUID microscope. The method is based on the inverse Biot-Savart law and the Fourier transformation to recover a real-space image. We found that the numerical analyses give a smaller vortex than the raw vortex profile observed by the scanning microscope.

High resolution projection X-ray microscope equipped with fluorescent X-ray analyzer and its applications

NASA Astrophysics Data System (ADS)

Minami, K.; Saito, Y.; Kai, H.; Shirota, K.; Yada, K.

2009-09-01

We have newly developed an open type fine-focus X-ray tube "TX-510" to realize a spatial resolution of 50nm and to radiate low energy characteristic X-rays for giving high absorption contrast to images of microscopic organisms. The "TX-510" employs a ZrO/W(100) Schottky emitter and an "In-Lens Field Emission Gun". The key points of the improvements are (1) reduced spherical aberration coefficient of magnetic objective lens, (2) easy and accurate focusing, (3) newly designed astigmatism compensator, (4) segmented thin film target for interchanging the target materials by electron beam shift and (5) fluorescent X-ray analysis system.

Portable microscopy platform for the clinical and environmental monitoring

NASA Astrophysics Data System (ADS)

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

2016-04-01

Light microscopy can not only address various diagnosis needs such as aquatic parasites and bacteria such as E. coli in water, but also provide a method for the screening of red tide. Traditional microscope based on the smartphone created by adding lens couldn't keep the tradeoff between field-of-view(FOV) and the resolution. In this paper, we demonstrate a non-contact, light and cost-effective microscope platform, that can image highly dense samples with a spatial resolution of ~0.8um over a field-of-view(FOV) of >1mm2. After captured the direct images, we performed the pixel super-resolution algorithm to improve the image resolution and overcome the hardware interference. The system would be a good point-of-care diagnostic solution in resource limited settings. We validated the performance of the system by imaging resolution test targets, the squamous cell cancer(SqCC) and green algae that necessary to detect the squamous carcinoma and red tide

Compact three-dimensional super-resolution system based on fluorescence emission difference microscopy

NASA Astrophysics Data System (ADS)

Zhu, Dazhao; Chen, Youhua; Fang, Yue; Hussain, Anwar; Kuang, Cuifang; Zhou, Xiaoxu; Xu, Yingke; Liu, Xu

2017-12-01

A compact microscope system for three-dimensional (3-D) super-resolution imaging is presented. The super-resolution capability of the system is based on a size-reduced effective 3-D point spread function generated through the fluorescence emission difference (FED) method. The appropriate polarization direction distribution and manipulation allows the panel active area of the spatial light modulator to be fully utilized. This allows simultaneous modulation of the incident light by two kinds of phase masks to be performed with a single spatial light modulator in order to generate a 3-D negative spot. The system is more compact than standard 3-D FED systems while maintaining all the advantages of 3-D FED microscopy. The experimental results demonstrated the improvement in 3-D resolution by nearly 1.7 times and 1.6 times compared to the classic confocal resolution in the lateral and axial directions, respectively.

Diffraction-limited IR Microspectroscopy with IRENI

Treesearch

J. Sedlmair; B. Illman; M. Unger; C. Hirschmugl

2012-01-01

In a unique way, IRENI (Infrared environmental Imaging), operated at the Synchrotron Radiation Center in Madison, combines IR spectroscopy and IR imaging, revealing the chemical morphology of a sample. Most storage ring based IR confocal microscopes have to overcome a trade-off between spatial resolution versus...

Unraveling submicron-scale mechanical heterogeneity by three-dimensional X-ray microdiffraction

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

Li, Runguang; Xie, Qingge; Wang, Yan-Dong

Shear banding is a ubiquitous phenomenon of severe plastic deformation, and damage accumulation in shear bands often results in the catastrophic failure of a material. Despite extensive studies, the microscopic mechanisms of strain localization and deformation damage in shear bands remain elusive due to their spatial-temporal complexities embedded in bulk materials. Here we conducted synchrotron-based X-ray microdiffraction (μXRD) experiments to map out the 3D lattice strain field with a submicron resolution around fatigue shear bands in a stainless steel. Both in situ and postmortem μXRD results revealed large lattice strain gradients at intersections of the primary and secondary shear bands.more » Such strain gradients resulted in severe mechanical heterogeneities across the fatigue shear bands, leading to reduced fatigue limits in the high-cycle regime. The ability to spatially quantify the localized strain gradients with submicron resolution through μXRD opens opportunities for understanding the microscopic mechanisms of damage and failure in bulk materials.« less

Unraveling submicron-scale mechanical heterogeneity by three-dimensional X-ray microdiffraction

DOE PAGES

Li, Runguang; Xie, Qingge; Wang, Yan-Dong; ...

2017-12-28

Shear banding is a ubiquitous phenomenon of severe plastic deformation, and damage accumulation in shear bands often results in the catastrophic failure of a material. Despite extensive studies, the microscopic mechanisms of strain localization and deformation damage in shear bands remain elusive due to their spatial-temporal complexities embedded in bulk materials. Here we conducted synchrotron-based X-ray microdiffraction (μXRD) experiments to map out the 3D lattice strain field with a submicron resolution around fatigue shear bands in a stainless steel. Both in situ and postmortem μXRD results revealed large lattice strain gradients at intersections of the primary and secondary shear bands.more » Such strain gradients resulted in severe mechanical heterogeneities across the fatigue shear bands, leading to reduced fatigue limits in the high-cycle regime. The ability to spatially quantify the localized strain gradients with submicron resolution through μXRD opens opportunities for understanding the microscopic mechanisms of damage and failure in bulk materials.« less

Mechanism of Prism-Coupled Scanning Tunneling Microscope Light Emission

NASA Astrophysics Data System (ADS)

Iida, Wataru; Ahamed, Jamal U.; Katano, Satoshi; Uehara, Yoichi

2011-09-01

We have investigated the mechanism of scanning tunneling microscope light emission (STM-LE) in a prism-coupled configuration using finite difference time domain analysis. In this configuration, the sample is a metallic thin film evaporated on the bottom surface of a hemispherical glass prism. STM light emitted into the prism (prism-side emission) through the metallic film is measured. Since both localized surface plasmons (LSP) and surface plasmon polaritons (SPP) contribute to prism-side emission, this emission is stronger than that in conventional STM-LE measured from the sample surface side, which is radiated by LSP alone. We show that the spatial resolution of prism-side emission is determined not by the propagation length of SPP, but by the lateral size of LSP, similarly to conventional (i.e., tip side) STM-LE. Thus, we conclude that, by using the prism-coupled configuration, the signal level of STM-LE improves without the loss of spatial resolution attained in tip side emission.

Hard X-ray Microscopy with sub 30 nm Spatial Resolution

NASA Astrophysics Data System (ADS)

Tang, Mau-Tsu; Song, Yen-Fang; Yin, Gung-Chian; Chen, Fu-Rong; Chen, Jian-Hua; Chen, Yi-Ming; Liang, Keng S.; Duewer, F.; Yun, Wenbing

2007-01-01

A transmission X-ray microscope (TXM) has been installed at the BL01B beamline at National Synchrotron Radiation Research Center in Taiwan. This state-of-the-art TXM operational in a range 8-11 keV provides 2D images and 3D tomography with spatial resolution 60 nm, and with the Zernike-phase contrast mode for imaging light materials such as biological specimens. A spatial resolution of the TXM better than 30 nm, apparently the best result in hard X-ray microscopy, has been achieved by employing the third diffraction order of the objective zone plate. The TXM has been applied in diverse research fields, including analysis of failure mechanisms in microelectronic devices, tomographic structures of naturally grown photonic specimens, and the internal structure of fault zone gouges from an earthquake core. Here we discuss the scope and prospects of the project, and the progress of the TXM in NSRRC.

Isotope analysis in the transmission electron microscope.

PubMed

Susi, Toma; Hofer, Christoph; Argentero, Giacomo; Leuthner, Gregor T; Pennycook, Timothy J; Mangler, Clemens; Meyer, Jannik C; Kotakoski, Jani

2016-10-10

The Ångström-sized probe of the scanning transmission electron microscope can visualize and collect spectra from single atoms. This can unambiguously resolve the chemical structure of materials, but not their isotopic composition. Here we differentiate between two isotopes of the same element by quantifying how likely the energetic imaging electrons are to eject atoms. First, we measure the displacement probability in graphene grown from either 12 C or 13 C and describe the process using a quantum mechanical model of lattice vibrations coupled with density functional theory simulations. We then test our spatial resolution in a mixed sample by ejecting individual atoms from nanoscale areas spanning an interface region that is far from atomically sharp, mapping the isotope concentration with a precision better than 20%. Although we use a scanning instrument, our method may be applicable to any atomic resolution transmission electron microscope and to other low-dimensional materials.

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

PubMed Central

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

2007-01-01

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

Long-distance super-resolution imaging assisted by enhanced spatial Fourier transform.

PubMed

Tang, Heng-He; Liu, Pu-Kun

2015-09-07

A new gradient-index (GRIN) lens that can realize enhanced spatial Fourier transform (FT) over optically long distances is demonstrated. By using an anisotropic GRIN metamaterial with hyperbolic dispersion, evanescent wave in free space can be transformed into propagating wave in the metamaterial and then focused outside due to negative-refraction. Both the results based on the ray tracing and the finite element simulation show that the spatial frequency bandwidth of the spatial FT can be extended to 2.7k(0) (k(0) is the wave vector in free space). Furthermore, assisted by the enhanced spatial FT, a new long-distance (in the optical far-field region) super-resolution imaging scheme is also proposed and the super resolved capability of λ/5 (λ is the wavelength in free space) is verified. The work may provide technical support for designing new-type high-speed microscopes with long working distances.

Concurrent in situ ion irradiation transmission electron microscope

DOE PAGES

Hattar, K.; Bufford, D. C.; Buller, D. L.

2014-08-29

An in situ ion irradiation transmission electron microscope has been developed and is operational at Sandia National Laboratories. This facility permits high spatial resolution, real time observation of electron transparent samples under ion irradiation, implantation, mechanical loading, corrosive environments, and combinations thereof. This includes the simultaneous implantation of low-energy gas ions (0.8–30 keV) during high-energy heavy ion irradiation (0.8–48 MeV). In addition, initial results in polycrystalline gold foils are provided to demonstrate the range of capabilities.

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

PubMed

Sun, Zhe; Zheng, Desheng; Baldelli, Steven

2017-02-21

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

Distinction of nuclear spin states with the scanning tunneling microscope.

PubMed

Natterer, Fabian Donat; Patthey, François; Brune, Harald

2013-10-25

We demonstrate rotational excitation spectroscopy with the scanning tunneling microscope for physisorbed H(2) and its isotopes HD and D(2). The observed excitation energies are very close to the gas phase values and show the expected scaling with the moment of inertia. Since these energies are characteristic for the molecular nuclear spin states we are able to identify the para and ortho species of hydrogen and deuterium, respectively. We thereby demonstrate nuclear spin sensitivity with unprecedented spatial resolution.

Spectral Interferometry with Electron Microscopes

PubMed Central

Talebi, Nahid

2016-01-01

Interference patterns are not only a defining characteristic of waves, but also have several applications; characterization of coherent processes and holography. Spatial holography with electron waves, has paved the way towards space-resolved characterization of magnetic domains and electrostatic potentials with angstrom spatial resolution. Another impetus in electron microscopy has been introduced by ultrafast electron microscopy which uses pulses of sub-picosecond durations for probing a laser induced excitation of the sample. However, attosecond temporal resolution has not yet been reported, merely due to the statistical distribution of arrival times of electrons at the sample, with respect to the laser time reference. This is however, the very time resolution which will be needed for performing time-frequency analysis. These difficulties are addressed here by proposing a new methodology to improve the synchronization between electron and optical excitations through introducing an efficient electron-driven photon source. We use focused transition radiation of the electron as a pump for the sample. Due to the nature of transition radiation, the process is coherent. This technique allows us to perform spectral interferometry with electron microscopes, with applications in retrieving the phase of electron-induced polarizations and reconstructing dynamics of the induced vector potential. PMID:27649932

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

PubMed

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

2017-01-11

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

Photon event distribution sampling: an image formation technique for scanning microscopes that permits tracking of sub-diffraction particles with high spatial and temporal resolutions.

PubMed

Larkin, J D; Publicover, N G; Sutko, J L

2011-01-01

In photon event distribution sampling, an image formation technique for scanning microscopes, the maximum likelihood position of origin of each detected photon is acquired as a data set rather than binning photons in pixels. Subsequently, an intensity-related probability density function describing the uncertainty associated with the photon position measurement is applied to each position and individual photon intensity distributions are summed to form an image. Compared to pixel-based images, photon event distribution sampling images exhibit increased signal-to-noise and comparable spatial resolution. Photon event distribution sampling is superior to pixel-based image formation in recognizing the presence of structured (non-random) photon distributions at low photon counts and permits use of non-raster scanning patterns. A photon event distribution sampling based method for localizing single particles derived from a multi-variate normal distribution is more precise than statistical (Gaussian) fitting to pixel-based images. Using the multi-variate normal distribution method, non-raster scanning and a typical confocal microscope, localizations with 8 nm precision were achieved at 10 ms sampling rates with acquisition of ~200 photons per frame. Single nanometre precision was obtained with a greater number of photons per frame. In summary, photon event distribution sampling provides an efficient way to form images when low numbers of photons are involved and permits particle tracking with confocal point-scanning microscopes with nanometre precision deep within specimens. © 2010 The Authors Journal of Microscopy © 2010 The Royal Microscopical Society.

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

NASA Astrophysics Data System (ADS)

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

2018-07-01

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

Tuning donut profile for spatial resolution in stimulated emission depletion microscopy.

PubMed

Neupane, Bhanu; Chen, Fang; Sun, Wei; Chiu, Daniel T; Wang, Gufeng

2013-04-01

In stimulated emission depletion (STED)-based or up-conversion depletion-based super-resolution optical microscopy, the donut-shaped depletion beam profile is of critical importance to its resolution. In this study, we investigate the transformation of the donut-shaped depletion beam focused by a high numerical aperture (NA) microscope objective, and model STED point spread function (PSF) as a function of donut beam profile. We show experimentally that the intensity profile of the dark kernel of the donut can be approximated as a parabolic function, whose slope is determined by the donut beam size before the objective back aperture, or the effective NA. Based on this, we derive the mathematical expression for continuous wave (CW) STED PSF as a function of focal plane donut and excitation beam profiles, as well as dye properties. We find that the effective NA and the residual intensity at the center are critical factors for STED imaging quality and the resolution. The effective NA is critical for STED resolution in that it not only determines the donut shape but also the area the depletion laser power is dispersed. An improperly expanded depletion beam will have negligible improvement in resolution. The polarization of the depletion beam also plays an important role as it affects the residual intensity in the center of the donut. Finally, we construct a CW STED microscope operating at 488 nm excitation and 592 nm depletion with a resolution of 70 nm. Our study provides detailed insight to the property of donut beam, and parameters that are important for the optimal performance of STED microscopes. This paper will provide a useful guide for the construction and future development of STED microscopes.

Super-resolution imaging of ciliary microdomains in isolated olfactory sensory neurons using a custom two-color stimulated emission depletion microscope

NASA Astrophysics Data System (ADS)

Meyer, Stephanie A.; Ozbay, Baris N.; Potcoava, Mariana; Salcedo, Ernesto; Restrepo, Diego; Gibson, Emily A.

2016-06-01

We performed stimulated emission depletion (STED) imaging of isolated olfactory sensory neurons (OSNs) using a custom-built microscope. The STED microscope uses a single pulsed laser to excite two separate fluorophores, Atto 590 and Atto 647N. A gated timing circuit combined with temporal interleaving of the different color excitation/STED laser pulses filters the two channel detection and greatly minimizes crosstalk. We quantified the instrument resolution to be ˜81 and ˜44 nm, for the Atto 590 and Atto 647N channels. The spatial separation between the two channels was measured to be under 10 nm, well below the resolution limit. The custom-STED microscope is incorporated onto a commercial research microscope allowing brightfield, differential interference contrast, and epifluorescence imaging on the same field of view. We performed immunolabeling of OSNs in mice to image localization of ciliary membrane proteins involved in olfactory transduction. We imaged Ca2+-permeable cyclic nucleotide gated (CNG) channel (Atto 594) and adenylyl cyclase type III (ACIII) (Atto 647N) in distinct cilia. STED imaging resolved well-separated subdiffraction limited clusters for each protein. We quantified the size of each cluster to have a mean value of 88±48 nm and 124±43 nm, for CNG and ACIII, respectively. STED imaging showed separated clusters that were not resolvable in confocal images.

Performance evaluation of a quasi-microscope for planetary landers

NASA Technical Reports Server (NTRS)

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

1977-01-01

Spatial resolutions achieved with cameras on lunar and planetary landers have been limited to about 1 mm, whereas microscopes of the type proposed for such landers could have obtained resolutions of about 1 um but were never accepted because of their complexity and weight. The quasi-microscope evaluated in this paper could provide intermediate resolutions of about 10 um with relatively simple optics that would augment a camera, such as the Viking lander camera, without imposing special design requirements on the camera of limiting its field of view of the terrain. Images of natural particulate samples taken in black and white and in color show that grain size, shape, and texture are made visible for unconsolidated materials in a 50- to 500-um size range. Such information may provide broad outlines of planetary surface mineralogy and allow inferences to be made of grain origin and evolution. The mineralogical descriptions of single grains would be aided by the reflectance spectra that could, for example, be estimated from the six-channel multispectral data of the Viking lander camera.

Adaptive pixel-super-resolved lensfree in-line digital holography for wide-field on-chip microscopy.

PubMed

Zhang, Jialin; Sun, Jiasong; Chen, Qian; Li, Jiaji; Zuo, Chao

2017-09-18

High-resolution wide field-of-view (FOV) microscopic imaging plays an essential role in various fields of biomedicine, engineering, and physical sciences. As an alternative to conventional lens-based scanning techniques, lensfree holography provides a new way to effectively bypass the intrinsical trade-off between the spatial resolution and FOV of conventional microscopes. Unfortunately, due to the limited sensor pixel-size, unpredictable disturbance during image acquisition, and sub-optimum solution to the phase retrieval problem, typical lensfree microscopes only produce compromised imaging quality in terms of lateral resolution and signal-to-noise ratio (SNR). Here, we propose an adaptive pixel-super-resolved lensfree imaging (APLI) method which can solve, or at least partially alleviate these limitations. Our approach addresses the pixel aliasing problem by Z-scanning only, without resorting to subpixel shifting or beam-angle manipulation. Automatic positional error correction algorithm and adaptive relaxation strategy are introduced to enhance the robustness and SNR of reconstruction significantly. Based on APLI, we perform full-FOV reconstruction of a USAF resolution target (~29.85 mm 2 ) and achieve half-pitch lateral resolution of 770 nm, surpassing 2.17 times of the theoretical Nyquist-Shannon sampling resolution limit imposed by the sensor pixel-size (1.67µm). Full-FOV imaging result of a typical dicot root is also provided to demonstrate its promising potential applications in biologic imaging.

Resolution enhancement of pump-probe microscope with an inverse-annular filter

NASA Astrophysics Data System (ADS)

Kobayashi, Takayoshi; Kawasumi, Koshi; Miyazaki, Jun; Nakata, Kazuaki

2018-04-01

Optical pump-probe microscopy can provide images by detecting changes in probe light intensity induced by stimulated emission, photoinduced absorbance change, or photothermal-induced refractive index change in either transmission or reflection mode. Photothermal microscopy, which is one type of optical pump-probe microscopy, has intrinsically super resolution capability due to the bilinear dependence of signal intensity of pump and probe. We introduce new techniques for further resolution enhancement and fast imaging in photothermal microscope. First, we introduce a new pupil filter, an inverse-annular pupil filter in a pump-probe photothermal microscope, which provides resolution enhancement in three dimensions. The resolutions are proved to be improved in lateral and axial directions by imaging experiment using 20-nm gold nanoparticles. The improvement in X (perpendicular to the common pump and probe polarization direction), Y (parallel to the polarization direction), and Z (axial direction) are by 15 ± 6, 8 ± 8, and 21 ± 2% from the resolution without a pupil filter. The resolution enhancement is even better than the calculation using vector field, which predicts the corresponding enhancement of 11, 8, and 6%. The discussion is made to explain the unexpected results. We also demonstrate the photothermal imaging of thick biological samples (cells from rabbit intestine and kidney) stained with hematoxylin and eosin dye with the inverse-annular filter. Second, a fast, high-sensitivity photothermal microscope is developed by implementing a spatially segmented balanced detection scheme into a laser scanning microscope using a Galvano mirror. We confirm a 4.9 times improvement in signal-to-noise ratio in the spatially segmented balanced detection compared with that of conventional detection. The system demonstrates simultaneous bi-modal photothermal and confocal fluorescence imaging of transgenic mouse brain tissue with a pixel dwell time of 20 µs. The fluorescence image visualizes neurons expressing yellow fluorescence proteins, while the photothermal signal detected endogenous chromophores in the mouse brain, allowing 3D visualization of the distribution of various features such as blood cells and fine structures most probably due to lipids. This imaging modality was constructed using compact and cost-effective laser diodes, and will thus be widely useful in the life and medical sciences. Third, we have made further resolution improvement of high-sensitivity laser scanning photothermal microscopy by applying non-linear detection. By this, the new method has super resolution with 61 and 42% enhancement from the diffraction limit values of the probe and pump wavelengths, respectively, by a second-order non-linear scheme and a high-frame rate in a laser scanning microscope. The maximum resolution is determined to be 160 nm in the second-order non-linear detection mode and 270 nm in the linear detection mode by the PT signal of GNPs. The pixel rate and frame rate for 300 × 300 pixel image are 50 µs and 4.5 s, respectively. The pixel and frame rate are shorter than the rates, those are 1 ms and 100 s, using the piezo-driven stage system.

Table-top soft x-ray microscope using laser-induced plasma from a pulsed gas jet.

PubMed

Müller, Matthias; Mey, Tobias; Niemeyer, Jürgen; Mann, Klaus

2014-09-22

An extremely compact soft x-ray microscope operating in the "water window" region at the wavelength λ = 2.88 nm is presented, making use of a long-term stable and nearly debris-free laser-induced plasma from a pulsed nitrogen gas jet target. The well characterized soft x-ray radiation is focused by an ellipsoidal grazing incidence condenser mirror. Imaging of a sample onto a CCD camera is achieved with a Fresnel zone plate using magnifications up to 500x. The spatial resolution of the recorded microscopic images is about 100 nm as demonstrated for a Siemens star test pattern.

CHAMP (Camera, Handlens, and Microscope Probe)

NASA Technical Reports Server (NTRS)

Mungas, Greg S.; Boynton, John E.; Balzer, Mark A.; Beegle, Luther; Sobel, Harold R.; Fisher, Ted; Klein, Dan; Deans, Matthew; Lee, Pascal; Sepulveda, Cesar A.

2005-01-01

CHAMP (Camera, Handlens And Microscope Probe)is a novel field microscope capable of color imaging with continuously variable spatial resolution from infinity imaging down to diffraction-limited microscopy (3 micron/pixel). As a robotic arm-mounted imager, CHAMP supports stereo imaging with variable baselines, can continuously image targets at an increasing magnification during an arm approach, can provide precision rangefinding estimates to targets, and can accommodate microscopic imaging of rough surfaces through a image filtering process called z-stacking. CHAMP was originally developed through the Mars Instrument Development Program (MIDP) in support of robotic field investigations, but may also find application in new areas such as robotic in-orbit servicing and maintenance operations associated with spacecraft and human operations. We overview CHAMP'S instrument performance and basic design considerations below.

Attenuated total reflection-Fourier transform infrared imaging of large areas using inverted prism crystals and combining imaging and mapping.

PubMed

Chan, K L Andrew; Kazarian, Sergei G

2008-10-01

Attenuated total reflection-Fourier transform infrared (ATR-FT-IR) imaging is a very useful tool for capturing chemical images of various materials due to the simple sample preparation and the ability to measure wet samples or samples in an aqueous environment. However, the size of the array detector used for image acquisition is often limited and there is usually a trade off between spatial resolution and the field of view (FOV). The combination of mapping and imaging can be used to acquire images with a larger FOV without sacrificing spatial resolution. Previous attempts have demonstrated this using an infrared microscope and a Germanium hemispherical ATR crystal to achieve images of up to 2.5 mm x 2.5 mm but with varying spatial resolution and depth of penetration across the imaged area. In this paper, we demonstrate a combination of mapping and imaging with a different approach using an external optics housing for large ATR accessories and inverted ATR prisms to achieve ATR-FT-IR images with a large FOV and reasonable spatial resolution. The results have shown that a FOV of 10 mm x 14 mm can be obtained with a spatial resolution of approximately 40-60 microm when using an accessory that gives no magnification. A FOV of 1.3 mm x 1.3 mm can be obtained with spatial resolution of approximately 15-20 microm when using a diamond ATR imaging accessory with 4x magnification. No significant change in image quality such as spatial resolution or depth of penetration has been observed across the whole FOV with this method and the measurement time was approximately 15 minutes for an image consisting of 16 image tiles.

Ultrafast, large-field multiphoton microscopy based on an acousto-optic deflector and a spatial light modulator.

PubMed

Shao, Yonghong; Qin, Wan; Liu, Honghai; Qu, Junle; Peng, Xiang; Niu, Hanben; Gao, Bruce Z

2012-07-01

We present an ultrafast, large-field multiphoton excitation fluorescence microscope with high lateral and axial resolutions based on a two-dimensional (2-D) acousto-optical deflector (AOD) scanner and spatial light modulator (SLM). When a phase-only SLM is used to shape the near-infrared light from a mode-locked titanium:sapphire laser into a multifocus array including the 0-order beam, a 136 μm × 136 μm field of view is achieved with a 60× objective using a 2-D AOD scanner without any mechanical scan element. The two-photon fluorescence image of a neuronal network that was obtained using this system demonstrates that our microscopy permits observation of dynamic biological events in a large field with high-temporal and -spatial resolution.

Development of confocal laser microscope system for examination of microscopic characteristics of radiophotoluminescence glass dosemeters.

PubMed

Maki, Daisuke; Ishii, Tetsuya; Sato, Fuminobu; Kato, Yushi; Yamamoto, Takayoshi; Iida, Toshiyuki

2011-03-01

A confocal laser microscope system was developed for the measurement of radiophotoluminescence (RPL) photons emitted from a minute alpha-ray-irradiated area in an RPL glass dosemeter. The system was composed mainly of an inverted-type microscope, an ultraviolet laser, an XY movable stage and photon-counting circuits. The photon-counting circuits were effective in the reduction of the background noise level in the measurement of RPL photons. The performance of this microscope system was examined by the observation of standard RPL glass samples irradiated using (241)Am alpha rays. The spatial resolution of this system was ∼ 3 μm, and with regard to the sensitivity of this system, a hit of more than four to five alpha rays in unit area produced enough amount of RPL photons to construct the image.

Color digital lensless holographic microscopy: laser versus LED illumination.

PubMed

Garcia-Sucerquia, Jorge

2016-08-20

A comparison of the performance of color digital lensless holographic microscopy (CDLHM) as utilized for illumination of RGB lasers or a super-bright white-light LED with a set of spectral filters is presented. As the use of lasers in CDLHM conceals the possibility of having a compact, lightweight, portable, and low cost microscope, and additionally the limited available laser radiation wavelengths limit a real multispectral imaging microscope, here we present the use of super-bright white-light LED and spectral filters for illuminating the sample. The performance of RGB laser-CDLHM and LED-CDLHM is evaluated on imaging a section of the head of a Drosophila melanogaster fly. This comparison shows that there is trade-off between the spatial resolution of the microscope and the light sources utilized, which can be understood with regard to the coherence properties of the illuminating light. Despite the smaller spatial coherence features of LED-CDLHM in comparison with laser-CDLHM, the former shows promise as a portable RGB digital lensless holographic microscope that could be extended to other wavelengths by the use of different spectral filters.

High-resolution room-temperature sample scanning superconducting quantum interference device microscope configurable for geological and biomagnetic applications

NASA Astrophysics Data System (ADS)

Fong, L. E.; Holzer, J. R.; McBride, K. K.; Lima, E. A.; Baudenbacher, F.; Radparvar, M.

2005-05-01

We have developed a scanning superconducting quantum interference device (SQUID) microscope system with interchangeable sensor configurations for imaging magnetic fields of room-temperature (RT) samples with submillimeter resolution. The low-critical-temperature (Tc) niobium-based monolithic SQUID sensors are mounted on the tip of a sapphire and thermally anchored to the helium reservoir. A 25μm sapphire window separates the vacuum space from the RT sample. A positioning mechanism allows us to adjust the sample-to-sensor spacing from the top of the Dewar. We achieved a sensor-to-sample spacing of 100μm, which could be maintained for periods of up to four weeks. Different SQUID sensor designs are necessary to achieve the best combination of spatial resolution and field sensitivity for a given source configuration. For imaging thin sections of geological samples, we used a custom-designed monolithic low-Tc niobium bare SQUID sensor, with an effective diameter of 80μm, and achieved a field sensitivity of 1.5pT/Hz1/2 and a magnetic moment sensitivity of 5.4×10-18Am2/Hz1/2 at a sensor-to-sample spacing of 100μm in the white noise region for frequencies above 100Hz. Imaging action currents in cardiac tissue requires a higher field sensitivity, which can only be achieved by compromising spatial resolution. We developed a monolithic low-Tc niobium multiloop SQUID sensor, with sensor sizes ranging from 250μm to 1mm, and achieved sensitivities of 480-180fT /Hz1/2 in the white noise region for frequencies above 100Hz, respectively. For all sensor configurations, the spatial resolution was comparable to the effective diameter and limited by the sensor-to-sample spacing. Spatial registration allowed us to compare high-resolution images of magnetic fields associated with action currents and optical recordings of transmembrane potentials to study the bidomain nature of cardiac tissue or to match petrography to magnetic field maps in thin sections of geological samples.

Plasmonic Imaging of Electrochemical Reactions of Single Nanoparticles.

PubMed

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

2016-11-15

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

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.

Improving Secondary Ion Mass Spectrometry Image Quality with Image Fusion

NASA Astrophysics Data System (ADS)

Tarolli, Jay G.; Jackson, Lauren M.; Winograd, Nicholas

2014-12-01

The spatial resolution of chemical images acquired with cluster secondary ion mass spectrometry (SIMS) is limited not only by the size of the probe utilized to create the images but also by detection sensitivity. As the probe size is reduced to below 1 μm, for example, a low signal in each pixel limits lateral resolution because of counting statistics considerations. Although it can be useful to implement numerical methods to mitigate this problem, here we investigate the use of image fusion to combine information from scanning electron microscope (SEM) data with chemically resolved SIMS images. The advantage of this approach is that the higher intensity and, hence, spatial resolution of the electron images can help to improve the quality of the SIMS images without sacrificing chemical specificity. Using a pan-sharpening algorithm, the method is illustrated using synthetic data, experimental data acquired from a metallic grid sample, and experimental data acquired from a lawn of algae cells. The results show that up to an order of magnitude increase in spatial resolution is possible to achieve. A cross-correlation metric is utilized for evaluating the reliability of the procedure.

X-ray Tomography and Chemical Imaging within Butterfly Wing Scales

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

Chen Jianhua; Lee Yaochang; Tang, M.-T.

2007-01-19

The rainbow like color of butterfly wings is associated with the internal and surface structures of the wing scales. While the photonic structure of the scales is believed to diffract specific lights at different angle, there is no adequate probe directly answering the 3-D structures with sufficient spatial resolution. The NSRRC nano-transmission x-ray microscope (nTXM) with tens nanometers spatial resolution is able to image biological specimens without artifacts usually introduced in sophisticated sample staining processes. With the intrinsic deep penetration of x-rays, the nTXM is capable of nondestructively investigating the internal structures of fragile and soft samples. In this study,more » we imaged the structure of butterfly wing scales in 3-D view with 60 nm spatial resolution. In addition, synchrotron-radiation-based Fourier transform Infrared (FT-IR) microspectroscopy was employed to analyze the chemical components with spatial information of the butterfly wing scales. Based on the infrared spectral images, we suggest that the major components of scale structure were rich in protein and polysaccharide.« less

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

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

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

2016-01-28

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

Ballistic-Electron-Emission Microscope

NASA Technical Reports Server (NTRS)

Kaiser, William J.; Bell, L. Douglas

1990-01-01

Ballistic-electron-emission microscope (BEEM) employs scanning tunneling-microscopy (STM) methods for nondestructive, direct electrical investigation of buried interfaces, such as interface between semiconductor and thin metal film. In BEEM, there are at least three electrodes: emitting tip, biasing electrode, and collecting electrode, receiving current crossing interface under investigation. Signal-processing device amplifies electrode signals and converts them into form usable by computer. Produces spatial images of surface by scanning tip; in addition, provides high-resolution images of buried interface under investigation. Spectroscopic information extracted by measuring collecting-electrode current as function of one of interelectrode voltages.

Indentation-Enabled In Situ Mechanical Characterization of Micro/Nanopillars in Electron Microscopes

NASA Astrophysics Data System (ADS)

Guo, Qiang; Fu, Xidan; Guo, Xiaolei; Liu, Zhiying; Shi, Yan; Zhang, Di

2018-04-01

Indentation-enabled micro/nanomechanical characterization of small-scale specimens provides powerful new tools for probing materials properties that were once unattainable by conventional experimental methods. Recent advancement in instrumentation further allows mechanical testing to be carried out in situ in electron microscopes, with high spatial and temporal resolution. This review discusses the recent development of nanoindentation-enabled in situ mechanical testing in electron microscopes, with an emphasis on the study of micro/nanopillars. Focus is given to novel applications beyond simple compressive and tensile testing that have been developed in the past few years, and limitations and possible future research directions in this field are proposed and discussed.

Mesoscopic-microscopic spatial stochastic simulation with automatic system partitioning.

PubMed

Hellander, Stefan; Hellander, Andreas; Petzold, Linda

2017-12-21

The reaction-diffusion master equation (RDME) is a model that allows for efficient on-lattice simulation of spatially resolved stochastic chemical kinetics. Compared to off-lattice hard-sphere simulations with Brownian dynamics or Green's function reaction dynamics, the RDME can be orders of magnitude faster if the lattice spacing can be chosen coarse enough. However, strongly diffusion-controlled reactions mandate a very fine mesh resolution for acceptable accuracy. It is common that reactions in the same model differ in their degree of diffusion control and therefore require different degrees of mesh resolution. This renders mesoscopic simulation inefficient for systems with multiscale properties. Mesoscopic-microscopic hybrid methods address this problem by resolving the most challenging reactions with a microscale, off-lattice simulation. However, all methods to date require manual partitioning of a system, effectively limiting their usefulness as "black-box" simulation codes. In this paper, we propose a hybrid simulation algorithm with automatic system partitioning based on indirect a priori error estimates. We demonstrate the accuracy and efficiency of the method on models of diffusion-controlled networks in 3D.

In vivo correlation mapping microscopy

NASA Astrophysics Data System (ADS)

McGrath, James; Alexandrov, Sergey; Owens, Peter; Subhash, Hrebesh; Leahy, Martin

2016-04-01

To facilitate regular assessment of the microcirculation in vivo, noninvasive imaging techniques such as nailfold capillaroscopy are required in clinics. Recently, a correlation mapping technique has been applied to optical coherence tomography (OCT), which extends the capabilities of OCT to microcirculation morphology imaging. This technique, known as correlation mapping optical coherence tomography, has been shown to extract parameters, such as capillary density and vessel diameter, and key clinical markers associated with early changes in microvascular diseases. However, OCT has limited spatial resolution in both the transverse and depth directions. Here, we extend this correlation mapping technique to other microscopy modalities, including confocal microscopy, and take advantage of the higher spatial resolution offered by these modalities. The technique is achieved as a processing step on microscopy images and does not require any modification to the microscope hardware. Results are presented which show that this correlation mapping microscopy technique can extend the capabilities of conventional microscopy to enable mapping of vascular networks in vivo with high spatial resolution in both the transverse and depth directions.

Photoluminescence Imaging and LBIC Characterization of Defects in mc-Si Solar Cells

NASA Astrophysics Data System (ADS)

Sánchez, L. A.; Moretón, A.; Guada, M.; Rodríguez-Conde, S.; Martínez, O.; González, M. A.; Jiménez, J.

2018-05-01

Today's photovoltaic market is dominated by multicrystalline silicon (mc-Si) based solar cells with around 70% of worldwide production. In order to improve the quality of the Si material, a proper characterization of the electrical activity in mc-Si solar cells is essential. A full-wafer characterization technique such as photoluminescence imaging (PLi) provides a fast inspection of the wafer defects, though at the expense of the spatial resolution. On the other hand, a study of the defects at a microscopic scale can be achieved through the light-beam induced current technique. The combination of these macroscopic and microscopic resolution techniques allows a detailed study of the electrical activity of defects in mc-Si solar cells. In this work, upgraded metallurgical-grade Si solar cells are studied using these two techniques.

Real-Time Nanoscopy by Using Blinking Enhanced Quantum Dots

PubMed Central

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

2010-01-01

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

Report on the Installation and Preparedness of a Protochips Fusion in-situ Heating Holder for TEM

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

Edmondson, Philip D.

2017-03-01

This brief report documents the procurement and installation of a Protochips Fusion (formerly Aduro) high-temperature, high stability transmission electron microscopy (TEM) specimen holder that allows for the high spatial resolution characterization of material specimens at high temperature in situ of an electron microscope. This specimen holder was specifically procured for use with The FEI Talos F200X Scanning/Transmission Electron Microscope (STEM) in Oak Ridge National Laboratory’s (ORNL’s) Low Activation Materials Development and Analysis (LAMDA) Laboratory. The Protochips Fusion holder will enable high-resolution structural and chemical analysis of irradiated materials at high temperature, becoming a unique capability worldwide, and would encourage high-qualitymore » in situ experiments to be conducted on irradiated materials.« less

Recording High Resolution 3D Lagrangian Motions In Marine Dinoflagellates using Digital Holographic Microscopic Cinematography

NASA Astrophysics Data System (ADS)

Sheng, J.; Malkiel, E.; Katz, J.; Place, A. R.; Belas, R.

2006-11-01

Detailed data on swimming behavior and locomotion for dense population of dinoflagellates constitutes a key component to understanding cell migration, cell-cell interactions and predator-prey dynamics, all of which affect algae bloom dynamics. Due to the multi-dimensional nature of flagellated cell motions, spatial-temporal Lagrangian measurements of multiple cells in high concentration are very limited. Here we present detailed data on 3D Lagrangian motions for three marine dinoflagellates: Oxyrrhis marina, Karlodinium veneficum, and Pfiesteria piscicida, using digital holographic microscopic cinematography. The measurements are performed in a 5x5x25mm cuvette with cell densities varying from 50,000 ˜ 90,000 cells/ml. Approximately 200-500 cells are tracked simultaneously for 12s at 60fps in a sample volume of 1x1x5 mm at a spatial resolution of 0.4x0.4x2 μm. We fully resolve the longitudinal flagella (˜200nm) along with the Lagrangian trajectory of each organism. Species dependent swimming behavior are identified and categorized quantitatively by velocities, radii of curvature, and rotations of pitch. Statistics on locomotion, temporal & spatial scales, and diffusion rate show substantial differences between species. The scaling between turning radius and cell dimension can be explained by a distributed stokeslet model for a self-propelled body.

Low-temperature-compatible tunneling-current-assisted scanning microwave microscope utilizing a rigid coaxial resonator.

PubMed

Takahashi, Hideyuki; Imai, Yoshinori; Maeda, Atsutaka

2016-06-01

We present a design for a tunneling-current-assisted scanning near-field microwave microscope. For stable operation at cryogenic temperatures, making a small and rigid microwave probe is important. Our coaxial resonator probe has a length of approximately 30 mm and can fit inside the 2-in. bore of a superconducting magnet. The probe design includes an insulating joint, which separates DC and microwave signals without degrading the quality factor. By applying the SMM to the imaging of an electrically inhomogeneous superconductor, we obtain the spatial distribution of the microwave response with a spatial resolution of approximately 200 nm. Furthermore, we present an analysis of our SMM probe based on a simple lumped-element circuit model along with the near-field microwave measurements of silicon wafers having different conductivities.

Low-temperature-compatible tunneling-current-assisted scanning microwave microscope utilizing a rigid coaxial resonator

NASA Astrophysics Data System (ADS)

Takahashi, Hideyuki; Imai, Yoshinori; Maeda, Atsutaka

2016-06-01

We present a design for a tunneling-current-assisted scanning near-field microwave microscope. For stable operation at cryogenic temperatures, making a small and rigid microwave probe is important. Our coaxial resonator probe has a length of approximately 30 mm and can fit inside the 2-in. bore of a superconducting magnet. The probe design includes an insulating joint, which separates DC and microwave signals without degrading the quality factor. By applying the SMM to the imaging of an electrically inhomogeneous superconductor, we obtain the spatial distribution of the microwave response with a spatial resolution of approximately 200 nm. Furthermore, we present an analysis of our SMM probe based on a simple lumped-element circuit model along with the near-field microwave measurements of silicon wafers having different conductivities.

Light propagation and interaction observed with electrons.

PubMed

Word, Robert C; Fitzgerald, J P S; Könenkamp, R

2016-01-01

We discuss possibilities for a microscopic optical characterization of thin films and surfaces based on photoemission electron microscopy. We show that propagating light with wavelengths across the visible range can readily be visualized, and linear and non-linear materials properties can be evaluated non-invasively with nanometer spatial resolution. While femtosecond temporal resolution can be achieved in pump-probe-type experiments, the interferometric approach presented here has typical image frame times of ~200 fs. Copyright © 2015 Elsevier B.V. All rights reserved.

Dynamic microscale temperature gradient in a gold nanorod solution measured by diffraction-limited nanothermometry

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

Li, Chengmingyue; Gan, Xiaosong; Li, Xiangping

2015-09-21

We quantify the dynamic microscale temperature gradient in a gold nanorod solution using quantum-dot-based microscopic fluorescence nanothermometry. By incorporating CdSe quantum dots into the solution as a nanothermometer, precise temperature mapping with diffraction-limited spatial resolution and sub-degree temperature resolution is achieved. The acquired data on heat generation and dissipation show an excellent agreement with theoretical simulations. This work reveals an effective approach for noninvasive temperature regulation with localized nanoheaters in microfluidic environment.

Multifocal Fluorescence Microscope for Fast Optical Recordings of Neuronal Action Potentials

PubMed Central

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

2015-01-01

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

Visualizing chemical functionality in plant cell walls

DOE PAGES

Zeng, Yining; Himmel, Michael E.; Ding, Shi-You

2017-11-30

Understanding plant cell wall cross-linking chemistry and polymeric architecture is key to the efficient utilization of biomass in all prospects from rational genetic modification to downstream chemical and biological conversion to produce fuels and value chemicals. In fact, the bulk properties of cell wall recalcitrance are collectively determined by its chemical features over a wide range of length scales from tissue, cellular to polymeric architectures. Microscopic visualization of cell walls from the nanometer to the micrometer scale offers an in situ approach to study their chemical functionality considering its spatial and chemical complexity, particularly the capabilities of characterizing biomass non-destructivelymore » and in real-time during conversion processes. Microscopic characterization has revealed heterogeneity in the distribution of chemical features, which would otherwise be hidden in bulk analysis. Key microscopic features include cell wall type, wall layering, and wall composition - especially cellulose and lignin distributions. Microscopic tools, such as atomic force microscopy, stimulated Raman scattering microscopy, and fluorescence microscopy, have been applied to investigations of cell wall structure and chemistry from the native wall to wall treated by thermal chemical pretreatment and enzymatic hydrolysis. While advancing our current understanding of plant cell wall recalcitrance and deconstruction, microscopic tools with improved spatial resolution will steadily enhance our fundamental understanding of cell wall function.« less

Visualizing chemical functionality in plant cell walls

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

Zeng, Yining; Himmel, Michael E.; Ding, Shi-You

Understanding plant cell wall cross-linking chemistry and polymeric architecture is key to the efficient utilization of biomass in all prospects from rational genetic modification to downstream chemical and biological conversion to produce fuels and value chemicals. In fact, the bulk properties of cell wall recalcitrance are collectively determined by its chemical features over a wide range of length scales from tissue, cellular to polymeric architectures. Microscopic visualization of cell walls from the nanometer to the micrometer scale offers an in situ approach to study their chemical functionality considering its spatial and chemical complexity, particularly the capabilities of characterizing biomass non-destructivelymore » and in real-time during conversion processes. Microscopic characterization has revealed heterogeneity in the distribution of chemical features, which would otherwise be hidden in bulk analysis. Key microscopic features include cell wall type, wall layering, and wall composition - especially cellulose and lignin distributions. Microscopic tools, such as atomic force microscopy, stimulated Raman scattering microscopy, and fluorescence microscopy, have been applied to investigations of cell wall structure and chemistry from the native wall to wall treated by thermal chemical pretreatment and enzymatic hydrolysis. While advancing our current understanding of plant cell wall recalcitrance and deconstruction, microscopic tools with improved spatial resolution will steadily enhance our fundamental understanding of cell wall function.« less

Visualizing chemical functionality in plant cell walls.

PubMed

Zeng, Yining; Himmel, Michael E; Ding, Shi-You

2017-01-01

Understanding plant cell wall cross-linking chemistry and polymeric architecture is key to the efficient utilization of biomass in all prospects from rational genetic modification to downstream chemical and biological conversion to produce fuels and value chemicals. In fact, the bulk properties of cell wall recalcitrance are collectively determined by its chemical features over a wide range of length scales from tissue, cellular to polymeric architectures. Microscopic visualization of cell walls from the nanometer to the micrometer scale offers an in situ approach to study their chemical functionality considering its spatial and chemical complexity, particularly the capabilities of characterizing biomass non-destructively and in real-time during conversion processes. Microscopic characterization has revealed heterogeneity in the distribution of chemical features, which would otherwise be hidden in bulk analysis. Key microscopic features include cell wall type, wall layering, and wall composition-especially cellulose and lignin distributions. Microscopic tools, such as atomic force microscopy, stimulated Raman scattering microscopy, and fluorescence microscopy, have been applied to investigations of cell wall structure and chemistry from the native wall to wall treated by thermal chemical pretreatment and enzymatic hydrolysis. While advancing our current understanding of plant cell wall recalcitrance and deconstruction, microscopic tools with improved spatial resolution will steadily enhance our fundamental understanding of cell wall function.

Evaluation of multiple-scale 3D characterization for coal physical structure with DCM method and synchrotron X-ray CT.

PubMed

Wang, Haipeng; Yang, Yushuang; Yang, Jianli; Nie, Yihang; Jia, Jing; Wang, Yudan

2015-01-01

Multiscale nondestructive characterization of coal microscopic physical structure can provide important information for coal conversion and coal-bed methane extraction. In this study, the physical structure of a coal sample was investigated by synchrotron-based multiple-energy X-ray CT at three beam energies and two different spatial resolutions. A data-constrained modeling (DCM) approach was used to quantitatively characterize the multiscale compositional distributions at the two resolutions. The volume fractions of each voxel for four different composition groups were obtained at the two resolutions. Between the two resolutions, the difference for DCM computed volume fractions of coal matrix and pores is less than 0.3%, and the difference for mineral composition groups is less than 0.17%. This demonstrates that the DCM approach can account for compositions beyond the X-ray CT imaging resolution with adequate accuracy. By using DCM, it is possible to characterize a relatively large coal sample at a relatively low spatial resolution with minimal loss of the effect due to subpixel fine length scale structures.

Nanoimaging using soft X-ray and EUV laser-plasma sources

NASA Astrophysics Data System (ADS)

Wachulak, Przemyslaw; Torrisi, Alfio; Ayele, Mesfin; Bartnik, Andrzej; Czwartos, Joanna; Węgrzyński, Łukasz; Fok, Tomasz; Fiedorowicz, Henryk

2018-01-01

In this work we present three experimental, compact desk-top imaging systems: SXR and EUV full field microscopes and the SXR contact microscope. The systems are based on laser-plasma EUV and SXR sources based on a double stream gas puff target. The EUV and SXR full field microscopes, operating at 13.8 nm and 2.88 nm wavelengths are capable of imaging nanostructures with a sub-50 nm spatial resolution and short (seconds) exposure times. The SXR contact microscope operates in the "water-window" spectral range and produces an imprint of the internal structure of the imaged sample in a thin layer of SXR sensitive photoresist. Applications of such desk-top EUV and SXR microscopes, mostly for biological samples (CT26 fibroblast cells and Keratinocytes) are also presented. Details about the sources, the microscopes as well as the imaging results for various objects will be presented and discussed. The development of such compact imaging systems may be important to the new research related to biological, material science and nanotechnology applications.

Three dimensional live-cell STED microscopy at increased depth using a water immersion objective

NASA Astrophysics Data System (ADS)

Heine, Jörn; Wurm, Christian A.; Keller-Findeisen, Jan; Schönle, Andreas; Harke, Benjamin; Reuss, Matthias; Winter, Franziska R.; Donnert, Gerald

2018-05-01

Modern fluorescence superresolution microscopes are capable of imaging living cells on the nanometer scale. One of those techniques is stimulated emission depletion (STED) which increases the microscope's resolution many times in the lateral and the axial directions. To achieve these high resolutions not only close to the coverslip but also at greater depths, the choice of objective becomes crucial. Oil immersion objectives have frequently been used for STED imaging since their high numerical aperture (NA) leads to high spatial resolutions. But during live-cell imaging, especially at great penetration depths, these objectives have a distinct disadvantage. The refractive index mismatch between the immersion oil and the usually aqueous embedding media of living specimens results in unwanted spherical aberrations. These aberrations distort the point spread functions (PSFs). Notably, during z- and 3D-STED imaging, the resolution increase along the optical axis is majorly hampered if at all possible. To overcome this limitation, we here use a water immersion objective in combination with a spatial light modulator for z-STED measurements of living samples at great depths. This compact design allows for switching between objectives without having to adapt the STED beam path and enables on the fly alterations of the STED PSF to correct for aberrations. Furthermore, we derive the influence of the NA on the axial STED resolution theoretically and experimentally. We show under live-cell imaging conditions that a water immersion objective leads to far superior results than an oil immersion objective at penetration depths of 5-180 μm.

Micrometer-scale magnetic imaging of geological samples using a quantum diamond microscope

NASA Astrophysics Data System (ADS)

Glenn, D. R.; Fu, R. R.; Kehayias, P.; Le Sage, D.; Lima, E. A.; Weiss, B. P.; Walsworth, R. L.

2017-08-01

Remanent magnetization in geological samples may record the past intensity and direction of planetary magnetic fields. Traditionally, this magnetization is analyzed through measurements of the net magnetic moment of bulk millimeter to centimeter sized samples. However, geological samples are often mineralogically and texturally heterogeneous at submillimeter scales, with only a fraction of the ferromagnetic grains carrying the remanent magnetization of interest. Therefore, characterizing this magnetization in such cases requires a technique capable of imaging magnetic fields at fine spatial scales and with high sensitivity. To address this challenge, we developed a new instrument, based on nitrogen-vacancy centers in diamond, which enables direct imaging of magnetic fields due to both remanent and induced magnetization, as well as optical imaging, of room-temperature geological samples with spatial resolution approaching the optical diffraction limit. We describe the operating principles of this device, which we call the quantum diamond microscope (QDM), and report its optimized image-area-normalized magnetic field sensitivity (20 µTṡµm/Hz1/2), spatial resolution (5 µm), and field of view (4 mm), as well as trade-offs between these parameters. We also perform an absolute magnetic field calibration for the device in different modes of operation, including three-axis (vector) and single-axis (projective) magnetic field imaging. Finally, we use the QDM to obtain magnetic images of several terrestrial and meteoritic rock samples, demonstrating its ability to resolve spatially distinct populations of ferromagnetic carriers.

Design principles and applications of a cooled CCD camera for electron microscopy.

PubMed

Faruqi, A R

1998-01-01

Cooled CCD cameras offer a number of advantages in recording electron microscope images with CCDs rather than film which include: immediate availability of the image in a digital format suitable for further computer processing, high dynamic range, excellent linearity and a high detective quantum efficiency for recording electrons. In one important respect however, film has superior properties: the spatial resolution of CCD detectors tested so far (in terms of point spread function or modulation transfer function) are inferior to film and a great deal of our effort has been spent in designing detectors with improved spatial resolution. Various instrumental contributions to spatial resolution have been analysed and in this paper we discuss the contribution of the phosphor-fibre optics system in this measurement. We have evaluated the performance of a number of detector components and parameters, e.g. different phosphors (and a scintillator), optical coupling with lens or fibre optics with various demagnification factors, to improve the detector performance. The camera described in this paper, which is based on this analysis, uses a tapered fibre optics coupling between the phosphor and the CCD and is installed on a Philips CM12 electron microscope equipped to perform cryo-microscopy. The main use of the camera so far has been in recording electron diffraction patterns from two dimensional crystals of bacteriorhodopsin--from wild type and from different trapped states during the photocycle. As one example of the type of data obtained with the CCD camera a two dimensional Fourier projection map from the trapped O-state is also included. With faster computers, it will soon be possible to undertake this type of work on an on-line basis. Also, with improvements in detector size and resolution, CCD detectors, already ideal for diffraction, will be able to compete with film in the recording of high resolution images.

Dynamic x-ray imaging of laser-driven nanoplasmas

NASA Astrophysics Data System (ADS)

Fennel, Thomas

2016-05-01

A major promise of current x-ray science at free electron lasers is the realization of unprecedented imaging capabilities for resolving the structure and ultrafast dynamics of matter with nanometer spatial and femtosecond temporal resolution or even below via single-shot x-ray diffraction. Laser-driven atomic clusters and nanoparticles provide an ideal platform for developing and demonstrating the required technology to extract the ultrafast transient spatiotemporal dynamics from the diffraction images. In this talk, the perspectives and challenges of dynamic x-ray imaging will be discussed using complete self-consistent microscopic electromagnetic simulations of IR pump x-ray probe imaging for the example of clusters. The results of the microscopic particle-in-cell simulations (MicPIC) enable the simulation-assisted reconstruction of corresponding experimental data. This capability is demonstrated by converting recently measured LCLS data into a ultrahigh resolution movie of laser-induced plasma expansion. Finally, routes towards reaching attosecond time resolution in the visualization of complex dynamical processes in matter by x-ray diffraction will be discussed.

MRI of articular cartilage at microscopic resolution

PubMed Central

Xia, Y.

2013-01-01

This review briefly summarises some of the definitive studies of articular cartilage by microscopic MRI (µMRI) that were conducted with the highest spatial resolutions. The article has four major sections. The first section introduces the cartilage tissue, MRI and µMRI, and the concept of image contrast in MRI. The second section describes the characteristic profiles of three relaxation times (T1, T2 and T1ρ) and self-diffusion in healthy articular cartilage. The third section discusses several factors that can influence the visualisation of articular cartilage and the detection of cartilage lesion by MRI and µMRI. These factors include image resolution, image analysis strategies, visualisation of the total tissue, topographical variations of the tissue properties, surface fibril ambiguity, deformation of the articular cartilage, and cartilage lesion. The final section justifies the values of multidisciplinary imaging that correlates MRI with other technical modalities, such as optical imaging. Rather than an exhaustive review to capture all activities in the literature, the studies cited in this review are merely illustrative. PMID:23610697

Commissioning of the PRIOR proton microscope

DOE PAGES

Varentsov, D.; Antonov, O.; Bakhmutova, A.; ...

2016-02-18

Recently, a new high energy proton microscopy facility PRIOR (Proton Microscope for FAIR Facility for Anti-proton and Ion Research) has been designed, constructed, and successfully commissioned at GSI Helmholtzzentrum für Schwerionenforschung (Darmstadt, Germany). As a result of the experiments with 3.5–4.5 GeV proton beams delivered by the heavy ion synchrotron SIS-18 of GSI, 30 μm spatial and 10 ns temporal resolutions of the proton microscope have been demonstrated. A new pulsed power setup for studying properties of matter under extremes has been developed for the dynamic commissioning of the PRIOR facility. This study describes the PRIOR setup as well asmore » the results of the first static and dynamic protonradiography experiments performed at GSI.« less

Commissioning of the PRIOR proton microscope

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

Varentsov, D.; Antonov, O.; Bakhmutova, A.

Recently, a new high energy proton microscopy facility PRIOR (Proton Microscope for FAIR Facility for Anti-proton and Ion Research) has been designed, constructed, and successfully commissioned at GSI Helmholtzzentrum für Schwerionenforschung (Darmstadt, Germany). As a result of the experiments with 3.5–4.5 GeV proton beams delivered by the heavy ion synchrotron SIS-18 of GSI, 30 μm spatial and 10 ns temporal resolutions of the proton microscope have been demonstrated. A new pulsed power setup for studying properties of matter under extremes has been developed for the dynamic commissioning of the PRIOR facility. This study describes the PRIOR setup as well asmore » the results of the first static and dynamic protonradiography experiments performed at GSI.« less

Camera array based light field microscopy

PubMed Central

Lin, Xing; Wu, Jiamin; Zheng, Guoan; Dai, Qionghai

2015-01-01

This paper proposes a novel approach for high-resolution light field microscopy imaging by using a camera array. In this approach, we apply a two-stage relay system for expanding the aperture plane of the microscope into the size of an imaging lens array, and utilize a sensor array for acquiring different sub-apertures images formed by corresponding imaging lenses. By combining the rectified and synchronized images from 5 × 5 viewpoints with our prototype system, we successfully recovered color light field videos for various fast-moving microscopic specimens with a spatial resolution of 0.79 megapixels at 30 frames per second, corresponding to an unprecedented data throughput of 562.5 MB/s for light field microscopy. We also demonstrated the use of the reported platform for different applications, including post-capture refocusing, phase reconstruction, 3D imaging, and optical metrology. PMID:26417490

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

High spatial resolution soft-x-ray microscopy

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

Meyer-Ilse, W.; Medecki, H.; Brown, J.T.

1997-04-01

A new soft x-ray microscope (XM-1) with high spatial resolution has been constructed by the Center for X-ray Optics. It uses bending magnet radiation from beamline 6.1 at the Advanced Light Source, and is used in a variety of projects and applications in the life and physical sciences. Most of these projects are ongoing. The instrument uses zone plate lenses and achieves a resolution of 43 nm, measured over 10% to 90% intensity with a knife edge test sample. X-ray microscopy permits the imaging of relatively thick samples, up to 10 {mu}m thick, in water. XM-1 has an easy tomore » use interface, that utilizes visible light microscopy to precisely position and focus the specimen. The authors describe applications of this device in the biological sciences, as well as in studying industrial applications including structured polymer samples.« less

Optical digital microscopy for cyto- and hematological studies in vitro

NASA Astrophysics Data System (ADS)

Ganilova, Yu. A.; Dolmashkin, A. A.; Doubrovski, V. A.; Yanina, I. Yu.; Tuchin, V. V.

2013-08-01

The dependence of the spatial resolution and field of view of an optical microscope equipped with a CCD camera on the objective magnification has been experimentally investigated. Measurement of these characteristics has shown that a spatial resolution of 20-25 px/μm at a field of view of about 110 μm is quite realistic; this resolution is acceptable for a detailed study of the processes occurring in cell. It is proposed to expand the dynamic range of digital camera by measuring and approximating its light characteristics with subsequent plotting of the corresponding calibration curve. The biological objects of study were human adipose tissue cells, as well as erythrocytes and their immune complexes in human blood; both objects have been investigated in vitro. Application of optical digital microscopy for solving specific problems of cytology and hematology can be useful in both biomedical studies in experiments with objects of nonbiological origin.

Development of first ever scanning probe microscopy capabilities for plutonium

NASA Astrophysics Data System (ADS)

Beaux, Miles F.; Cordoba, Miguel Santiago; Zocco, Adam T.; Vodnik, Douglas R.; Ramos, Michael; Richmond, Scott; Moore, David P.; Venhaus, Thomas J.; Joyce, Stephen A.; Usov, Igor O.

2017-04-01

Scanning probe microscopy capabilities have been developed for plutonium and its derivative compounds. Specifically, a scanning tunneling microscope and an atomic force microscope housed in an ultra-high vacuum system and an inert atmosphere glove box, respectively, were prepared for the introduction of small non-dispersible δ-Pu coupons. Experimental details, procedures, and preliminary imaging of δ-Pu coupons are presented to demonstrate the functionality of these new capabilities. These first of a kind capabilities for plutonium represent a significant step forward in the ability to characterize and understand plutonium surfaces with high spatial resolution.

Development of first ever scanning probe microscopy capabilities for plutonium

DOE PAGES

Beaux, Miles F.; Cordoba, Miguel Santiago; Zocco, Adam T.; ...

2017-04-01

Scanning probe microscopy capabilities have been developed for plutonium and its derivative compounds. Specifically, a scanning tunneling microscope and an atomic force microscope housed in an ultra-high vacuum system and an inert atmosphere glove box, respectively, were prepared for the introduction of small non-dispersible δ-Pu coupons. Experimental details, procedures, and preliminary imaging of δ-Pu coupons are presented to demonstrate the functionality of these new capabilities. In conclusion, these first of a kind capabilities for plutonium represent a significant step forward in the ability to characterize and understand plutonium surfaces with high spatial resolution.

Evanescent-Wave Filtering in Images Using Remote Terahertz Structured Illumination

NASA Astrophysics Data System (ADS)

Flammini, M.; Pontecorvo, E.; Giliberti, V.; Rizza, C.; Ciattoni, A.; Ortolani, M.; DelRe, E.

2017-11-01

Imaging with structured illumination allows for the retrieval of subwavelength features of an object by conversion of evanescent waves into propagating waves. In conditions in which the object plane and the structured-illumination plane do not coincide, this conversion process is subject to progressive filtering of the components with high spatial frequency when the distance between the two planes increases, until the diffraction-limited lateral resolution is restored when the distance exceeds the extension of evanescent waves. We study the progressive filtering of evanescent waves by developing a remote super-resolution terahertz imaging system operating at a wavelength λ =1.00 mm , based on a freestanding knife edge and a reflective confocal terahertz microscope. In the images recorded with increasing knife-edge-to-object-plane distance, we observe the transition from a super-resolution of λ /17 ≃60 μ m to the diffraction-limited lateral resolution of Δ x ≃λ expected for our confocal microscope. The extreme nonparaxial conditions are analyzed in detail, exploiting the fact that, in the terahertz frequency range, the knife edge can be positioned at a variable subwavelength distance from the object plane. Electromagnetic simulations of radiation scattering by the knife edge reproduce the experimental super-resolution achieved.

Pixel-super-resolved lensfree holography using adaptive relaxation factor and positional error correction

NASA Astrophysics Data System (ADS)

Zhang, Jialin; Chen, Qian; Sun, Jiasong; Li, Jiaji; Zuo, Chao

2018-01-01

Lensfree holography provides a new way to effectively bypass the intrinsical trade-off between the spatial resolution and field-of-view (FOV) of conventional lens-based microscopes. Unfortunately, due to the limited sensor pixel-size, unpredictable disturbance during image acquisition, and sub-optimum solution to the phase retrieval problem, typical lensfree microscopes only produce compromised imaging quality in terms of lateral resolution and signal-to-noise ratio (SNR). In this paper, we propose an adaptive pixel-super-resolved lensfree imaging (APLI) method to address the pixel aliasing problem by Z-scanning only, without resorting to subpixel shifting or beam-angle manipulation. Furthermore, an automatic positional error correction algorithm and adaptive relaxation strategy are introduced to enhance the robustness and SNR of reconstruction significantly. Based on APLI, we perform full-FOV reconstruction of a USAF resolution target across a wide imaging area of {29.85 mm2 and achieve half-pitch lateral resolution of 770 nm, surpassing 2.17 times of the theoretical Nyquist-Shannon sampling resolution limit imposed by the sensor pixel-size (1.67 μm). Full-FOV imaging result of a typical dicot root is also provided to demonstrate its promising potential applications in biologic imaging.

Nanoscale visualization of redox activity at lithium-ion battery cathodes.

PubMed

Takahashi, Yasufumi; Kumatani, Akichika; Munakata, Hirokazu; Inomata, Hirotaka; Ito, Komachi; Ino, Kosuke; Shiku, Hitoshi; Unwin, Patrick R; Korchev, Yuri E; Kanamura, Kiyoshi; Matsue, Tomokazu

2014-11-17

Intercalation and deintercalation of lithium ions at electrode surfaces are central to the operation of lithium-ion batteries. Yet, on the most important composite cathode surfaces, this is a rather complex process involving spatially heterogeneous reactions that have proved difficult to resolve with existing techniques. Here we report a scanning electrochemical cell microscope based approach to define a mobile electrochemical cell that is used to quantitatively visualize electrochemical phenomena at the battery cathode material LiFePO4, with resolution of ~100 nm. The technique measures electrode topography and different electrochemical properties simultaneously, and the information can be combined with complementary microscopic techniques to reveal new perspectives on structure and activity. These electrodes exhibit highly spatially heterogeneous electrochemistry at the nanoscale, both within secondary particles and at individual primary nanoparticles, which is highly dependent on the local structure and composition.

Extracellular oxygen concentration mapping with a confocal multiphoton laser scanning microscope and TCSPC card

NASA Astrophysics Data System (ADS)

Hosny, Neveen A.; Lee, David A.; Knight, Martin M.

2010-02-01

Extracellular oxygen concentrations influence cell metabolism and tissue function. Fluorescence Lifetime Imaging Microscopy (FLIM) offers a non-invasive method for quantifying local oxygen concentrations. However, existing methods show limited spatial resolution and/or require custom made systems. This study describes a new optimised approach for quantitative extracellular oxygen detection, providing an off-the-shelf system with high spatial resolution and an improved lifetime determination over previous techniques, while avoiding systematic photon pile-up. Fluorescence lifetime detection of an oxygen sensitive fluorescent dye, tris(2,2'-bipyridyl)ruthenium(II) chloride hexahydrate [Ru(bipy)3]2+, was measured using a Becker&Hickl time-correlated single photon counting (TCSPC) card with excitation provided by a multi-photon laser. This technique was able to identify a subpopulation of isolated chondrocyte cells, seeded in three-dimensional agarose gel, displaying a significant spatial oxygen gradient. Thus this technique provides a powerful tool for quantifying spatial oxygen gradients within three-dimensional cellular models.

Scanning magnetic tunnel junction microscope for high-resolution imaging of remanent magnetization fields

NASA Astrophysics Data System (ADS)

Lima, E. A.; Bruno, A. C.; Carvalho, H. R.; Weiss, B. P.

2014-10-01

Scanning magnetic microscopy is a new methodology for mapping magnetic fields with high spatial resolution and field sensitivity. An important goal has been to develop high-performance instruments that do not require cryogenic technology due to its high cost, complexity, and limitation on sensor-to-sample distance. Here we report the development of a low-cost scanning magnetic microscope based on commercial room-temperature magnetic tunnel junction (MTJ) sensors that typically achieves spatial resolution better than 7 µm. By comparing different bias and detection schemes, optimal performance was obtained when biasing the MTJ sensor with a modulated current at 1.0 kHz in a Wheatstone bridge configuration while using a lock-in amplifier in conjunction with a low-noise custom-made preamplifier. A precision horizontal (x-y) scanning stage comprising two coupled nanopositioners controls the position of the sample and a linear actuator adjusts the sensor-to-sample distance. We obtained magnetic field sensitivities better than 150 nT/Hz1/2 between 0.1 and 10 Hz, which is a critical frequency range for scanning magnetic microscopy. This corresponds to a magnetic moment sensitivity of 10-14 A m2, a factor of 100 better than achievable with typical commercial superconducting moment magnetometers. It also represents an improvement in sensitivity by a factor between 10 and 30 compared to similar scanning MTJ microscopes based on conventional bias-detection schemes. To demonstrate the capabilities of the instrument, two polished thin sections of representative geological samples were scanned along with a synthetic sample containing magnetic microparticles. The instrument is usable for a diversity of applications that require mapping of samples at room temperature to preserve magnetic properties or viability, including paleomagnetism and rock magnetism, nondestructive evaluation of materials, and biological assays.

Theory of the spatial resolution of (scanning) transmission electron microscopy in liquid water or ice layers.

PubMed

de Jonge, Niels

2018-04-01

The sample dependent spatial resolution was calculated for transmission electron microscopy (TEM) and scanning TEM (STEM) of objects (e.g., nanoparticles, proteins) embedded in a layer of liquid water or amorphous ice. The theoretical model includes elastic- and inelastic scattering, beam broadening, and chromatic aberration. Different contrast mechanisms were evaluated as function of the electron dose, the detection angle, and the sample configuration. It was found that the spatial resolution scales with the electron dose to the -1/4th power. Gold- and carbon nanoparticles were examined in the middle of water layers ranging from 0.01--10 µm thickness representing relevant classes of experiments in both materials science and biology. The optimal microscope settings differ between experimental configurations. STEM performs the best for gold nanoparticles for all layer thicknesses, while carbon is best imaged with phase-contrast TEM for thin layers but bright field STEM is preferred for thicker layers. The resolution was also calculated for a water layer enclosed between thin membranes. The influence of chromatic aberration correction for TEM was examined as well. The theory is broadly applicable to other types of materials and sample configurations. Copyright © 2018 Elsevier B.V. All rights reserved.

Improving Secondary Ion Mass Spectrometry Image Quality with Image Fusion

PubMed Central

Tarolli, Jay G.; Jackson, Lauren M.; Winograd, Nicholas

2014-01-01

The spatial resolution of chemical images acquired with cluster secondary ion mass spectrometry (SIMS) is limited not only by the size of the probe utilized to create the images, but also by detection sensitivity. As the probe size is reduced to below 1 µm, for example, a low signal in each pixel limits lateral resolution due to counting statistics considerations. Although it can be useful to implement numerical methods to mitigate this problem, here we investigate the use of image fusion to combine information from scanning electron microscope (SEM) data with chemically resolved SIMS images. The advantage of this approach is that the higher intensity and, hence, spatial resolution of the electron images can help to improve the quality of the SIMS images without sacrificing chemical specificity. Using a pan-sharpening algorithm, the method is illustrated using synthetic data, experimental data acquired from a metallic grid sample, and experimental data acquired from a lawn of algae cells. The results show that up to an order of magnitude increase in spatial resolution is possible to achieve. A cross-correlation metric is utilized for evaluating the reliability of the procedure. PMID:24912432

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.

High-throughput isotropic mapping of whole mouse brain using multi-view light-sheet microscopy

NASA Astrophysics Data System (ADS)

Nie, Jun; Li, Yusha; Zhao, Fang; Ping, Junyu; Liu, Sa; Yu, Tingting; Zhu, Dan; Fei, Peng

2018-02-01

Light-sheet fluorescence microscopy (LSFM) uses an additional laser-sheet to illuminate selective planes of the sample, thereby enabling three-dimensional imaging at high spatial-temporal resolution. These advantages make LSFM a promising tool for high-quality brain visualization. However, even by the use of LSFM, the spatial resolution remains insufficient to resolve the neural structures across a mesoscale whole mouse brain in three dimensions. At the same time, the thick-tissue scattering prevents a clear observation from the deep of brain. Here we use multi-view LSFM strategy to solve this challenge, surpassing the resolution limit of standard light-sheet microscope under a large field-of-view (FOV). As demonstrated by the imaging of optically-cleared mouse brain labelled with thy1-GFP, we achieve a brain-wide, isotropic cellular resolution of 3μm. Besides the resolution enhancement, multi-view braining imaging can also recover complete signals from deep tissue scattering and attenuation. The identification of long distance neural projections across encephalic regions can be identified and annotated as a result.

Magnetic microscopic imaging with an optically pumped magnetometer and flux guides

DOE PAGES

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

2017-01-23

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

CHAMP - Camera, Handlens, and Microscope Probe

NASA Technical Reports Server (NTRS)

Mungas, G. S.; Beegle, L. W.; Boynton, J.; Sepulveda, C. A.; Balzer, M. A.; Sobel, H. R.; Fisher, T. A.; Deans, M.; Lee, P.

2005-01-01

CHAMP (Camera, Handlens And Microscope Probe) is a novel field microscope capable of color imaging with continuously variable spatial resolution from infinity imaging down to diffraction-limited microscopy (3 micron/pixel). As an arm-mounted imager, CHAMP supports stereo-imaging with variable baselines, can continuously image targets at an increasing magnification during an arm approach, can provide precision range-finding estimates to targets, and can accommodate microscopic imaging of rough surfaces through a image filtering process called z-stacking. Currently designed with a filter wheel with 4 different filters, so that color and black and white images can be obtained over the entire Field-of-View, future designs will increase the number of filter positions to include 8 different filters. Finally, CHAMP incorporates controlled white and UV illumination so that images can be obtained regardless of sun position, and any potential fluorescent species can be identified so the most astrobiologically interesting samples can be identified.

External and internal gelation of pectin solutions: microscopic dynamics versus macroscopic rheology

NASA Astrophysics Data System (ADS)

Secchi, E.; Munarin, F.; Alaimo, M. D.; Bosisio, S.; Buzzaccaro, S.; Ciccarella, G.; Vergaro, V.; Petrini, P.; Piazza, R.

2014-11-01

Pectin is a natural biopolymer that forms, in the presence of divalent cations, ionic-bound gels typifying a large class of biological gels stabilized by non-covalent cross-links. We investigate and compare the kinetics of formation and aging of pectin gels obtained either through external gelation via perfusion of free Ca2+ ions, or by internal gelation due to the supply of the same ions from the dissolution of CaCO3 nanoparticles. The microscopic dynamics obtained with photon correlation imaging, a novel optical technique that allows obtaining the microscopic dynamics of the sample while retaining the spatial resolution of imaging techniques, is contrasted with macroscopic rheological measurements at constant strain. Pectin gelation is found to display peculiar two-stage kinetics, highlighted by non-monotonic growth in time of both microscopic correlations and gel mechanical strength. These results are compared to those found for alginate, another biopolymer extensively used in food formulation.

Field-portable lensfree tomographic microscope.

PubMed

Isikman, Serhan O; Bishara, Waheb; Sikora, Uzair; Yaglidere, Oguzhan; Yeah, John; Ozcan, Aydogan

2011-07-07

We present a field-portable lensfree tomographic microscope, which can achieve sectional imaging of a large volume (∼20 mm(3)) on a chip with an axial resolution of <7 μm. In this compact tomographic imaging platform (weighing only ∼110 grams), 24 light-emitting diodes (LEDs) that are each butt-coupled to a fibre-optic waveguide are controlled through a cost-effective micro-processor to sequentially illuminate the sample from different angles to record lensfree holograms of the sample that is placed on the top of a digital sensor array. In order to generate pixel super-resolved (SR) lensfree holograms and hence digitally improve the achievable lateral resolution, multiple sub-pixel shifted holograms are recorded at each illumination angle by electromagnetically actuating the fibre-optic waveguides using compact coils and magnets. These SR projection holograms obtained over an angular range of ±50° are rapidly reconstructed to yield projection images of the sample, which can then be back-projected to compute tomograms of the objects on the sensor-chip. The performance of this compact and light-weight lensfree tomographic microscope is validated by imaging micro-beads of different dimensions as well as a Hymenolepis nana egg, which is an infectious parasitic flatworm. Achieving a decent three-dimensional spatial resolution, this field-portable on-chip optical tomographic microscope might provide a useful toolset for telemedicine and high-throughput imaging applications in resource-poor settings. This journal is © The Royal Society of Chemistry 2011

Spatially multiplexed interferometric microscopy with partially coherent illumination

NASA Astrophysics Data System (ADS)

Picazo-Bueno, José Ángel; Zalevsky, Zeev; García, Javier; Ferreira, Carlos; Micó, Vicente

2016-10-01

We have recently reported on a simple, low cost, and highly stable way to convert a standard microscope into a holographic one [Opt. Express 22, 14929 (2014)]. The method, named spatially multiplexed interferometric microscopy (SMIM), proposes an off-axis holographic architecture implemented onto a regular (nonholographic) microscope with minimum modifications: the use of coherent illumination and a properly placed and selected one-dimensional diffraction grating. In this contribution, we report on the implementation of partially (temporally reduced) coherent illumination in SMIM as a way to improve quantitative phase imaging. The use of low coherence sources forces the application of phase shifting algorithm instead of off-axis holographic recording to recover the sample's phase information but improves phase reconstruction due to coherence noise reduction. In addition, a less restrictive field of view limitation (1/2) is implemented in comparison with our previously reported scheme (1/3). The proposed modification is experimentally validated in a regular Olympus BX-60 upright microscope considering a wide range of samples (resolution test, microbeads, swine sperm cells, red blood cells, and prostate cancer cells).

Calibrated thermal microscopy of the tool-chip interface in machining

NASA Astrophysics Data System (ADS)

Yoon, Howard W.; Davies, Matthew A.; Burns, Timothy J.; Kennedy, M. D.

2000-03-01

A critical parameter in predicting tool wear during machining and in accurate computer simulations of machining is the spatially-resolved temperature at the tool-chip interface. We describe the development and the calibration of a nearly diffraction-limited thermal-imaging microscope to measure the spatially-resolved temperatures during the machining of an AISI 1045 steel with a tungsten-carbide tool bit. The microscope has a target area of 0.5 mm X 0.5 mm square region with a < 5 micrometers spatial resolution and is based on a commercial InSb 128 X 128 focal plane array with an all reflective microscope objective. The minimum frame image acquisition time is < 1 ms. The microscope is calibrated using a standard blackbody source from the radiance temperature calibration laboratory at the National Institute of Standards and Technology, and the emissivity of the machined material is deduced from the infrared reflectivity measurements. The steady-state thermal images from the machining of 1045 steel are compared to previous determinations of tool temperatures from micro-hardness measurements and are found to be in agreement with those studies. The measured average chip temperatures are also in agreement with the temperature rise estimated from energy balance considerations. From these calculations and the agreement between the experimental and the calculated determinations of the emissivity of the 1045 steel, the standard uncertainty of the temperature measurements is estimated to be about 45 degree(s)C at 900 degree(s)C.

Spherical aberration correction in a scanning transmission electron microscope using a sculpted thin film.

PubMed

Shiloh, Roy; Remez, Roei; Lu, Peng-Han; Jin, Lei; Lereah, Yossi; Tavabi, Amir H; Dunin-Borkowski, Rafal E; Arie, Ady

2018-06-01

Nearly eighty years ago, Scherzer showed that rotationally symmetric, charge-free, static electron lenses are limited by an unavoidable, positive spherical aberration. Following a long struggle, a major breakthrough in the spatial resolution of electron microscopes was reached two decades ago by abandoning the first of these conditions, with the successful development of multipole aberration correctors. Here, we use a refractive silicon nitride thin film to tackle the second of Scherzer's constraints and demonstrate an alternative method for correcting spherical aberration in a scanning transmission electron microscope. We reveal features in Si and Cu samples that cannot be resolved in an uncorrected microscope. Our thin film corrector can be implemented as an immediate low cost upgrade to existing electron microscopes without re-engineering of the electron column or complicated operation protocols and can be extended to the correction of additional aberrations. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Real-space measurement of potential distribution in PECVD ONO electrets by Kelvin probe force microscopy.

PubMed

Emmerich, F; Thielemann, C

2016-05-20

Multilayers of silicon oxide/silicon nitride/silicon oxide (ONO) are known for their good electret properties due to deep energy traps near the material interfaces, facilitating charge storage. However, measurement of the space charge distribution in such multilayers is a challenge for conventional methods if layer thickness dimensions shrink below 1 μm. In this paper, we propose an atomic force microscope based method to determine charge distributions in ONO layers with spatial resolution below 100 nm. By applying Kelvin probe force microscopy (KPFM) on freshly cleaved, corona-charged multilayers, the surface potential is measured directly along the z-axis and across the interfaces. This new method gives insights into charge distribution and charge movement in inorganic electrets with a high spatial resolution.

Active chiral control of GHz acoustic whispering-gallery modes

NASA Astrophysics Data System (ADS)

Mezil, Sylvain; Fujita, Kentaro; Otsuka, Paul H.; Tomoda, Motonobu; Clark, Matt; Wright, Oliver B.; Matsuda, Osamu

2017-10-01

We selectively generate chiral surface-acoustic whispering-gallery modes in the gigahertz range on a microscopic disk by means of an ultrafast time-domain technique incorporating a spatial light modulator. Active chiral control is achieved by making use of an optical pump spatial profile in the form of a semicircular arc, positioned on the sample to break the symmetry of clockwise- and counterclockwise-propagating modes. Spatiotemporal Fourier transforms of the interferometrically monitored two-dimensional acoustic fields measured to micron resolution allow individual chiral modes and their azimuthal mode order, both positive and negative, to be distinguished. In particular, for modes with 15-fold rotational symmetry, we demonstrate ultrafast chiral control of surface acoustic waves in a micro-acoustic system with picosecond temporal resolution. Applications include nondestructive testing and surface acoustic wave devices.

High-Rate Capable Floating Strip Micromegas

NASA Astrophysics Data System (ADS)

Bortfeldt, Jonathan; Bender, Michael; Biebel, Otmar; Danger, Helge; Flierl, Bernhard; Hertenberger, Ralf; Lösel, Philipp; Moll, Samuel; Parodi, Katia; Rinaldi, Ilaria; Ruschke, Alexander; Zibell, André

2016-04-01

We report on the optimization of discharge insensitive floating strip Micromegas (MICRO-MEsh GASeous) detectors, fit for use in high-energy muon spectrometers. The suitability of these detectors for particle tracking is shown in high-background environments and at very high particle fluxes up to 60 MHz/cm2. Measurement and simulation of the microscopic discharge behavior have demonstrated the excellent discharge tolerance. A floating strip Micromegas with an active area of 48 cm × 50 cm with 1920 copper anode strips exhibits in 120 GeV pion beams a spatial resolution of 50 μm at detection efficiencies above 95%. Pulse height, spatial resolution and detection efficiency are homogeneous over the detector. Reconstruction of particle track inclination in a single detector plane is discussed, optimum angular resolutions below 5° are observed. Systematic deviations of this μTPC-method are fully understood. The reconstruction capabilities for minimum ionizing muons are investigated in a 6.4 cm × 6.4 cm floating strip Micromegas under intense background irradiation of the whole active area with 20 MeV protons at a rate of 550 kHz. The spatial resolution for muons is not distorted by space charge effects. A 6.4 cm × 6.4 cm floating strip Micromegas doublet with low material budget is investigated in highly ionizing proton and carbon ion beams at particle rates between 2 MHz and 2 GHz. Stable operation up to the highest rates is observed, spatial resolution, detection efficiencies, the multi-hit and high-rate capability are discussed.

Performance of bent-crystal x-ray microscopes for high energy density physics research

DOE PAGES

Schollmeier, Marius S.; Geissel, Matthias; Shores, Jonathon E.; ...

2015-05-29

We present calculations for the field of view (FOV), image fluence, image monochromaticity, spectral acceptance, and image aberrations for spherical crystal microscopes, which are used as self-emission imaging or backlighter systems at large-scale high energy density physics facilities. Our analytic results are benchmarked with ray-tracing calculations as well as with experimental measurements from the 6.151 keV backlighter system at Sandia National Laboratories. Furthermore, the analytic expressions can be used for x-ray source positions anywhere between the Rowland circle and object plane. We discovered that this enables quick optimization of the performance of proposed but untested, bent-crystal microscope systems to findmore » the best compromise between FOV, image fluence, and spatial resolution for a particular application.« less

Video-mosaicking of in vivo reflectance confocal microscopy images for noninvasive examination of skin lesion (Conference Presentation)

NASA Astrophysics Data System (ADS)

Kose, Kivanc; Gou, Mengran; Yelamos, Oriol; Cordova, Miguel A.; Rossi, Anthony; Nehal, Kishwer S.; Camps, Octavia I.; Dy, Jennifer G.; Brooks, Dana H.; Rajadhyaksha, Milind

2017-02-01

In this report we describe a computer vision based pipeline to convert in-vivo reflectance confocal microscopy (RCM) videos collected with a handheld system into large field of view (FOV) mosaics. For many applications such as imaging of hard to access lesions, intraoperative assessment of MOHS margins, or delineation of lesion margins beyond clinical borders, raster scan based mosaicing techniques have clinically significant limitations. In such cases, clinicians often capture RCM videos by freely moving a handheld microscope over the area of interest, but the resulting videos lose large-scale spatial relationships. Videomosaicking is a standard computational imaging technique to register, and stitch together consecutive frames of videos into large FOV high resolution mosaics. However, mosaicing RCM videos collected in-vivo has unique challenges: (i) tissue may deform or warp due to physical contact with the microscope objective lens, (ii) discontinuities or "jumps" between consecutive images and motion blur artifacts may occur, due to manual operation of the microscope, and (iii) optical sectioning and resolution may vary between consecutive images due to scattering and aberrations induced by changes in imaging depth and tissue morphology. We addressed these challenges by adapting or developing new algorithmic methods for videomosaicking, specifically by modeling non-rigid deformations, followed by automatically detecting discontinuities (cut locations) and, finally, applying a data-driven image stitching approach that fully preserves resolution and tissue morphologic detail without imposing arbitrary pre-defined boundaries. We will present example mosaics obtained by clinical imaging of both melanoma and non-melanoma skin cancers. The ability to combine freehand mosaicing for handheld microscopes with preserved cellular resolution will have high impact application in diverse clinical settings, including low-resource healthcare systems.

High-energy proton imaging for biomedical applications

DOE PAGES

Prall, Matthias; Durante, Marco; Berger, Thomas; ...

2016-06-10

The charged particle community is looking for techniques exploiting proton interactions instead of X-ray absorption for creating images of human tissue. Due to multiple Coulomb scattering inside the measured object it has shown to be highly non-trivial to achieve sufficient spatial resolution. We present imaging of biological tissue with a proton microscope. This device relies on magnetic optics, distinguishing it from most published proton imaging methods. For these methods reducing the data acquisition time to a clinically acceptable level has turned out to be challenging. In a proton microscope, data acquisition and processing are much simpler. This device even allowsmore » imaging in real time. The primary medical application will be image guidance in proton radiosurgery. Proton images demonstrating the potential for this application are presented. As a result, tomographic reconstructions are included to raise awareness of the possibility of high-resolution proton tomography using magneto-optics.« less

High-energy proton imaging for biomedical applications

NASA Astrophysics Data System (ADS)

Prall, M.; Durante, M.; Berger, T.; Przybyla, B.; Graeff, C.; Lang, P. M.; Latessa, C.; Shestov, L.; Simoniello, P.; Danly, C.; Mariam, F.; Merrill, F.; Nedrow, P.; Wilde, C.; Varentsov, D.

2016-06-01

The charged particle community is looking for techniques exploiting proton interactions instead of X-ray absorption for creating images of human tissue. Due to multiple Coulomb scattering inside the measured object it has shown to be highly non-trivial to achieve sufficient spatial resolution. We present imaging of biological tissue with a proton microscope. This device relies on magnetic optics, distinguishing it from most published proton imaging methods. For these methods reducing the data acquisition time to a clinically acceptable level has turned out to be challenging. In a proton microscope, data acquisition and processing are much simpler. This device even allows imaging in real time. The primary medical application will be image guidance in proton radiosurgery. Proton images demonstrating the potential for this application are presented. Tomographic reconstructions are included to raise awareness of the possibility of high-resolution proton tomography using magneto-optics.

Analytical scanning evanescent microwave microscope and control stage

DOEpatents

Xiang, Xiao-Dong; Gao, Chen; Duewer, Fred; Yang, Hai Tao; Lu, Yalin

2013-01-22

A scanning evanescent microwave microscope (SEMM) that uses near-field evanescent electromagnetic waves to probe sample properties is disclosed. The SEMM is capable of high resolution imaging and quantitative measurements of the electrical properties of the sample. The SEMM has the ability to map dielectric constant, loss tangent, conductivity, electrical impedance, and other electrical parameters of materials. Such properties are then used to provide distance control over a wide range, from to microns to nanometers, over dielectric and conductive samples for a scanned evanescent microwave probe, which enable quantitative non-contact and submicron spatial resolution topographic and electrical impedance profiling of dielectric, nonlinear dielectric and conductive materials. The invention also allows quantitative estimation of microwave impedance using signals obtained by the scanned evanescent microwave probe and quasistatic approximation modeling. The SEMM can be used to measure electrical properties of both dielectric and electrically conducting materials.

Analytical scanning evanescent microwave microscope and control stage

DOEpatents

Xiang, Xiao-Dong; Gao, Chen; Duewer, Fred; Yang, Hai Tao; Lu, Yalin

2009-06-23

A scanning evanescent microwave microscope (SEMM) that uses near-field evanescent electromagnetic waves to probe sample properties is disclosed. The SEMM is capable of high resolution imaging and quantitative measurements of the electrical properties of the sample. The SEMM has the ability to map dielectric constant, loss tangent, conductivity, electrical impedance, and other electrical parameters of materials. Such properties are then used to provide distance control over a wide range, from to microns to nanometers, over dielectric and conductive samples for a scanned evanescent microwave probe, which enable quantitative non-contact and submicron spatial resolution topographic and electrical impedance profiling of dielectric, nonlinear dielectric and conductive materials. The invention also allows quantitative estimation of microwave impedance using signals obtained by the scanned evanescent microwave probe and quasistatic approximation modeling. The SEMM can be used to measure electrical properties of both dielectric and electrically conducting materials.

High-energy proton imaging for biomedical applications

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

Prall, Matthias; Durante, Marco; Berger, Thomas

The charged particle community is looking for techniques exploiting proton interactions instead of X-ray absorption for creating images of human tissue. Due to multiple Coulomb scattering inside the measured object it has shown to be highly non-trivial to achieve sufficient spatial resolution. We present imaging of biological tissue with a proton microscope. This device relies on magnetic optics, distinguishing it from most published proton imaging methods. For these methods reducing the data acquisition time to a clinically acceptable level has turned out to be challenging. In a proton microscope, data acquisition and processing are much simpler. This device even allowsmore » imaging in real time. The primary medical application will be image guidance in proton radiosurgery. Proton images demonstrating the potential for this application are presented. As a result, tomographic reconstructions are included to raise awareness of the possibility of high-resolution proton tomography using magneto-optics.« less

Influence of imaging resolution on color fidelity in digital archiving.

PubMed

Zhang, Pengchang; Toque, Jay Arre; Ide-Ektessabi, Ari

2015-11-01

Color fidelity is of paramount importance in digital archiving. In this paper, the relationship between color fidelity and imaging resolution was explored by calculating the color difference of an IT8.7/2 color chart with a CIELAB color difference formula for scanning and simulation images. Microscopic spatial sampling was used in selecting the image pixels for the calculations to highlight the loss of color information. A ratio, called the relative imaging definition (RID), was defined to express the correlation between image resolution and color fidelity. The results show that in order for color differences to remain unrecognizable, the imaging resolution should be at least 10 times higher than the physical dimension of the smallest feature in the object being studied.

Porcine pulmonary angiotensin I-converting enzyme--biochemical characterization and spatial arrangement of the N- and C-domains by three-dimensional electron microscopic reconstruction.

PubMed

Chen, Hui-Ling; Lünsdorf, Heinrich; Hecht, Hans-Jürgen; Tsai, Hsin

2010-08-01

The somatic angiotensin I-converting enzyme (sACE; peptidyl-dipeptidase A; EC 3.4.15.1) was isolated from pig lung and purified to homogeneity. The purified enzyme has a molecular mass of about 180 kDa. Upon proteolytic cleavage, two approximately 90 kDa fragments were obtained and identified by amino-terminal sequence analysis as the N- and C-domains of sACE. Both purified domains were shown to be catalytically active. A 2.3 nm resolution model of sACE was obtained by three-dimensional electron microscopic reconstruction of negatively stained sACE particles, based on atomic X-ray data fitting. Our model shows for the first time the relative orientation of the sACE catalytically active domains and their spatial distance. (c) 2010 Elsevier Ltd. All rights reserved.

Comparison of grain to grain orientation and stiffness mapping by spatially resolved acoustic spectroscopy and EBSD.

PubMed

Mark, A F; Li, W; Sharples, S; Withers, P J

2017-07-01

Our aim was to establish the capability of spatially resolved acoustic spectroscopy (SRAS) to map grain orientations and the anisotropy in stiffness at the sub-mm to micron scale by comparing the method with electron backscatter diffraction (EBSD) undertaken within a scanning electron microscope. In the former the grain orientations are deduced by measuring the spatial variation in elastic modulus; conversely, in EBSD the elastic anisotropy is deduced from direct measurements of the crystal orientations. The two test-cases comprise mapping the fusion zones for large TIG and MMA welds in thick power plant austenitic and ferritic steels, respectively; these are technologically important because, among other things, elastic anisotropy can cause ultrasonic weld inspection methods to become inaccurate because it causes bending in the paths of sound waves. The spatial resolution of SRAS is not as good as that for EBSD (∼100 μm vs. ∼a few nm), nor is the angular resolution (∼1.5° vs. ∼0.5°). However the method can be applied to much larger areas (currently on the order of 300 mm square), is much faster (∼5 times), is cheaper and easier to perform, and it could be undertaken on the manufacturing floor. Given these advantages, particularly to industrial users, and the on-going improvements to the method, SRAS has the potential to become a standard method for orientation mapping, particularly in cases where the elastic anisotropy is important over macroscopic/component length scales. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Orientation and phase mapping in the transmission electron microscope using precession-assisted diffraction spot recognition: state-of-the-art results.

PubMed

Viladot, D; Véron, M; Gemmi, M; Peiró, F; Portillo, J; Estradé, S; Mendoza, J; Llorca-Isern, N; Nicolopoulos, S

2013-10-01

A recently developed technique based on the transmission electron microscope, which makes use of electron beam precession together with spot diffraction pattern recognition now offers the possibility to acquire reliable orientation/phase maps with a spatial resolution down to 2 nm on a field emission gun transmission electron microscope. The technique may be described as precession-assisted crystal orientation mapping in the transmission electron microscope, precession-assisted crystal orientation mapping technique-transmission electron microscope, also known by its product name, ASTAR, and consists in scanning the precessed electron beam in nanoprobe mode over the specimen area, thus producing a collection of precession electron diffraction spot patterns, to be thereafter indexed automatically through template matching. We present a review on several application examples relative to the characterization of microstructure/microtexture of nanocrystalline metals, ceramics, nanoparticles, minerals and organics. The strengths and limitations of the technique are also discussed using several application examples. ©2013 The Authors. Journal of Microscopy published by John Wiley & Sons Ltd on behalf of Royal Microscopical Society.

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

Koenenkamp, Rolf

We report on the design, assembly, operation and application of an aberration-corrected photoemission electron microscope. The instrument used novel hyperbolic mirror-correctors with two and three electrodes that allowed simultaneous correction of spherical and chromatic aberrations. A spatial resolution of 5.4nm was obtained with this instrument in 2009, and 4.7nm in subsequent years. New imaging methodology was introduced involving interferometric imaging of light diffraction. This methodology was applied in nano-photonics and in the characterization of surface-plasmon polaritons. Photonic crystals and waveguides, optical antennas and new plasmonic devices such as routers, localizers and filters were designed and demonstrated using the new capabilitiesmore » offered by the microscope.« less

Refractive index profiles of Ge-doped optical fibers with nanometer spatial resolution using atomic force microscopy.

PubMed

Pace, P; Huntington, Shane; Lyytikäinen, K; Roberts, A; Love, J

2004-04-05

We show a quantitative connection between Refractive Index Profiles (RIP) and measurements made by an Atomic Force Microscope (AFM). Germanium doped fibers were chemically etched in hydrofluoric acid solution (HF) and the wet etching characteristics of germanium were studied using an AFM. The AFM profiles were compared to both a concentration profile of the preform determined using a Scanning Electron Microscope (SEM) and a RIP of the fiber measured using a commercial profiling instrument, and were found to be in excellent agreement. It is now possible to calculate the RIP of a germanium doped fiber directly from an AFM profile.

Scanning tip microwave near field microscope

DOEpatents

Xiang, X.D.; Schultz, P.G.; Wei, T.

1998-10-13

A microwave near field microscope has a novel microwave probe structure wherein the probing field of evanescent radiation is emitted from a sharpened metal tip instead of an aperture or gap. This sharpened tip, which is electrically and mechanically connected to a central electrode, extends through and beyond an aperture in an end wall of a microwave resonating device such as a microwave cavity resonator or a microwave stripline resonator. Since the field intensity at the tip increases as the tip sharpens, the total energy which is radiated from the tip and absorbed by the sample increases as the tip sharpens. The result is improved spatial resolution without sacrificing sensitivity. 17 figs.

Scanning tip microwave near field microscope

DOEpatents

Xiang, Xiao-Dong; Schultz, Peter G.; Wei, Tao

1998-01-01

A microwave near field microscope has a novel microwave probe structure wherein the probing field of evanescent radiation is emitted from a sharpened metal tip instead of an aperture or gap. This sharpened tip, which is electrically and mechanically connected to a central electrode, extends through and beyond an aperture in an endwall of a microwave resonating device such as a microwave cavity resonator or a microwave stripline resonator. Since the field intensity at the tip increases as the tip sharpens, the total energy which is radiated from the tip and absorbed by the sample increases as the tip sharpens. The result is improved spatial resolution without sacrificing sensitivity.

Zone plate lenses for X-ray microscopy

NASA Astrophysics Data System (ADS)

Vladimirsky, Y.; Kern, D. P.; Chang, T. H. P.; Attwood, D. T.; Iskander, N.; Rothman, S.; McQuaide, K.; Kirz, J.; Ade, H.; McNulty, I.; Rarback, H.; Shu, D.

1988-04-01

Fresnel zone plate lenses with feature sizes as small as 50 nm have been constructed and used in the Stony Brook/NSLS scanning X-ray microscope with 3.1 nm radiation from Brookhaven's X-17 mini-undulator. The zone plates were fabricated at IBM using electron beam writing techniques, moiré pattern techniques to monitor ellipticity, and a double development/double plating technique to provide additional thickness in the central region. A spatial resolution down to 75 nm was measured in the microscope. Using these zone plates, biological images were obtained of unaltered subcellular components. The images highlight protein concentration in unsectioned, unfixed, and unstained enzymatic granules in an aqueous environment.

Implementing an Accurate and Rapid Sparse Sampling Approach for Low-Dose Atomic Resolution STEM Imaging

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

Kovarik, Libor; Stevens, Andrew J.; Liyu, Andrey V.

Aberration correction for scanning transmission electron microscopes (STEM) has dramatically increased spatial image resolution for beam-stable materials, but it is the sample stability rather than the microscope that often limits the practical resolution of STEM images. To extract physical information from images of beam sensitive materials it is becoming clear that there is a critical dose/dose-rate below which the images can be interpreted as representative of the pristine material, while above it the observation is dominated by beam effects. Here we describe an experimental approach for sparse sampling in the STEM and in-painting image reconstruction in order to reduce themore » electron dose/dose-rate to the sample during imaging. By characterizing the induction limited rise-time and hysteresis in scan coils, we show that sparse line-hopping approach to scan randomization can be implemented that optimizes both the speed of the scan and the amount of the sample that needs to be illuminated by the beam. The dose and acquisition time for the sparse sampling is shown to be effectively decreased by factor of 5x relative to conventional acquisition, permitting imaging of beam sensitive materials to be obtained without changing the microscope operating parameters. As a result, the use of sparse line-hopping scan to acquire STEM images is demonstrated with atomic resolution aberration corrected Z-contrast images of CaCO 3, a material that is traditionally difficult to image by TEM/STEM because of dose issues.« less

Implementing an Accurate and Rapid Sparse Sampling Approach for Low-Dose Atomic Resolution STEM Imaging

DOE PAGES

Kovarik, Libor; Stevens, Andrew J.; Liyu, Andrey V.; ...

2016-10-17

Aberration correction for scanning transmission electron microscopes (STEM) has dramatically increased spatial image resolution for beam-stable materials, but it is the sample stability rather than the microscope that often limits the practical resolution of STEM images. To extract physical information from images of beam sensitive materials it is becoming clear that there is a critical dose/dose-rate below which the images can be interpreted as representative of the pristine material, while above it the observation is dominated by beam effects. Here we describe an experimental approach for sparse sampling in the STEM and in-painting image reconstruction in order to reduce themore » electron dose/dose-rate to the sample during imaging. By characterizing the induction limited rise-time and hysteresis in scan coils, we show that sparse line-hopping approach to scan randomization can be implemented that optimizes both the speed of the scan and the amount of the sample that needs to be illuminated by the beam. The dose and acquisition time for the sparse sampling is shown to be effectively decreased by factor of 5x relative to conventional acquisition, permitting imaging of beam sensitive materials to be obtained without changing the microscope operating parameters. The use of sparse line-hopping scan to acquire STEM images is demonstrated with atomic resolution aberration corrected Z-contrast images of CaCO3, a material that is traditionally difficult to image by TEM/STEM because of dose issues.« less

Performance Evaluation of 18F Radioluminescence Microscopy Using Computational Simulation

PubMed Central

Wang, Qian; Sengupta, Debanti; Kim, Tae Jin; Pratx, Guillem

2017-01-01

Purpose Radioluminescence microscopy can visualize the distribution of beta-emitting radiotracers in live single cells with high resolution. Here, we perform a computational simulation of 18F positron imaging using this modality to better understand how radioluminescence signals are formed and to assist in optimizing the experimental setup and image processing. Methods First, the transport of charged particles through the cell and scintillator and the resulting scintillation is modeled using the GEANT4 Monte-Carlo simulation. Then, the propagation of the scintillation light through the microscope is modeled by a convolution with a depth-dependent point-spread function, which models the microscope response. Finally, the physical measurement of the scintillation light using an electron-multiplying charge-coupled device (EMCCD) camera is modeled using a stochastic numerical photosensor model, which accounts for various sources of noise. The simulated output of the EMCCD camera is further processed using our ORBIT image reconstruction methodology to evaluate the endpoint images. Results The EMCCD camera model was validated against experimentally acquired images and the simulated noise, as measured by the standard deviation of a blank image, was found to be accurate within 2% of the actual detection. Furthermore, point-source simulations found that a reconstructed spatial resolution of 18.5 μm can be achieved near the scintillator. As the source is moved away from the scintillator, spatial resolution degrades at a rate of 3.5 μm per μm distance. These results agree well with the experimentally measured spatial resolution of 30–40 μm (live cells). The simulation also shows that the system sensitivity is 26.5%, which is also consistent with our previous experiments. Finally, an image of a simulated sparse set of single cells is visually similar to the measured cell image. Conclusions Our simulation methodology agrees with experimental measurements taken with radioluminescence microscopy. This in silico approach can be used to guide further instrumentation developments and to provide a framework for improving image reconstruction. PMID:28273348

Enhanced resolution imaging of ultrathin ZnO layers on Ag(111) by multiple hydrogen molecules in a scanning tunneling microscope junction

NASA Astrophysics Data System (ADS)

Liu, Shuyi; Shiotari, Akitoshi; Baugh, Delroy; Wolf, Martin; Kumagai, Takashi

2018-05-01

Molecular hydrogen in a scanning tunneling microscope (STM) junction has been found to enhance the lateral spatial resolution of the STM imaging, referred to as scanning tunneling hydrogen microscopy (STHM). Here we report atomic resolution imaging of 2- and 3-monolayer (ML) thick ZnO layers epitaxially grown on Ag(111) using STHM. The enhanced resolution can be obtained at a relatively large tip to surface distance and resolves a more defective structure exhibiting dislocation defects for 3-ML-thick ZnO than for 2 ML. In order to elucidate the enhanced imaging mechanism, the electric and mechanical properties of the hydrogen molecular junction (HMJ) are investigated by a combination of STM and atomic force microscopy. It is found that the HMJ shows multiple kinklike features in the tip to surface distance dependence of the conductance and frequency shift curves, which are absent in a hydrogen-free junction. Based on a simple modeling, we propose that the junction contains several hydrogen molecules and sequential squeezing of the molecules out of the junction results in the kinklike features in the conductance and frequency shift curves. The model also qualitatively reproduces the enhanced resolution image of the ZnO films.

Studying dynamic processes in liquids by TEM/STEM/DTEM

NASA Astrophysics Data System (ADS)

Abellan, Patricia; Evans, James; Woehl, Taylor; Jungjohann, Katherine; Parent, Lucas; Arslan, Ilke; Ristenpart, William; Browning, Nigel; Mater. Sci. Group Team; Microsc. Group Team; Catal. Sci. Group Collaboration; Ristenpart Res. Group Collaboration

2013-03-01

In order to study dynamic phenomena such as corrosion or catalysis, extreme environmental conditions must be reproduced around the specimen - these include high-temperatures, high-pressures, specific oxidizing/reducing atmospheres or a liquid environment. The use of environmental stages specifically designed to fit in any transmission electron microscope (TEM) allows us to apply the distinct capabilities of each instrument to study dynamic processes. Localized gas/fluid conditions are created around the sample and separated from the high vacuum inside the microscope using hermetically sealed windowed-cells. Advanced capabilities of these techniques include spatial resolutions of ~1 Angstrom or better in aberration corrected instruments or temporal resolutions in the microsecond-nanosecond range in a dynamic TEM (DTEM). Here, unique qualities of the DTEM that benefit the in-situ experiments with gas/fluid environmental cells will be discussed. We also present our results with a liquid stage allowing atomic resolution imaging of nanomaterials in a colloidal suspension, core EEL spectra acquisition, continuous flow, controlled growth of nanocrystals and systematic calibration of the effect of the electron dose on silver nuclei formation.

Open-source image reconstruction of super-resolution structured illumination microscopy data in ImageJ

PubMed Central

Müller, Marcel; Mönkemöller, Viola; Hennig, Simon; Hübner, Wolfgang; Huser, Thomas

2016-01-01

Super-resolved structured illumination microscopy (SR-SIM) is an important tool for fluorescence microscopy. SR-SIM microscopes perform multiple image acquisitions with varying illumination patterns, and reconstruct them to a super-resolved image. In its most frequent, linear implementation, SR-SIM doubles the spatial resolution. The reconstruction is performed numerically on the acquired wide-field image data, and thus relies on a software implementation of specific SR-SIM image reconstruction algorithms. We present fairSIM, an easy-to-use plugin that provides SR-SIM reconstructions for a wide range of SR-SIM platforms directly within ImageJ. For research groups developing their own implementations of super-resolution structured illumination microscopy, fairSIM takes away the hurdle of generating yet another implementation of the reconstruction algorithm. For users of commercial microscopes, it offers an additional, in-depth analysis option for their data independent of specific operating systems. As a modular, open-source solution, fairSIM can easily be adapted, automated and extended as the field of SR-SIM progresses. PMID:26996201

Developments in Scanning Hall Probe Microscopy

NASA Astrophysics Data System (ADS)

Chouinard, Taras; Chu, Ricky; David, Nigel; Broun, David

2009-05-01

Low temperature scanning Hall probe microscopy is a sensitive means of imaging magnetic structures with high spatial resolution and magnetic flux sensitivity approaching that of a Superconducting Quantum Interference Device. We have developed a scanning Hall probe microscope with novel features, including highly reliable coarse positioning, in situ optimization of sensor-sample alignment and capacitive transducers for linear, long range positioning measurement. This has been motivated by the need to reposition accurately above fabricated nanostructures such as small superconducting rings. Details of the design and performance will be presented as well as recent progress towards time-resolved measurements with sub nanosecond resolution.

Finding the patterns in complex specimens by improving the acquisition and analysis of x-ray spectromicroscopy data

NASA Astrophysics Data System (ADS)

Lerotic, Mirna

Soft x-ray spectromicroscopy provides spectral data on the chemical speciation of light elements at sub-100 nanometer spatial resolution. The high resolution imaging places a strong demand on the microscope stability and on the reproducibility of the scanned image field, and the volume of data necessitates the need for improved data analysis methods. This dissertation concerns two developments in extending the capability of soft x-ray transmission microscopes to carry out studies of chemical speciation at high spatial resolution. One development involves an improvement in x-ray microscope instrumentation: a new Stony Brook scanning transmission x-ray microscope which incorporates laser interferometer feedback in scanning stage positions. The interferometer is used to control the position between the sample and focusing optics, and thus improve the stability of the system. A second development concerns new analysis methods for the study of chemical speciation of complex specimens, such as those in biological and environmental science studies. When all chemical species in a specimen are known and separately characterized, existing approaches can be used to measure the concentration of each component at each pixel. In other cases (such as often occur in biology or environmental science), where the specimen may be too complicated or provide at least some unknown spectral signatures, other approaches must be used. We describe here an approach that uses principal component analysis (similar to factor analysis) to orthogonalize and noise-filter spectromicroscopy data. We then use cluster analysis (a form of unsupervised pattern matching) to classify pixels according to spectral similarity, to extract representative, cluster-averaged spectra with good signal-to-noise ratio, and to obtain gradations of concentration of these representative spectra at each pixel. The method is illustrated with a simulated data set of organic compounds, and a mixture of lutetium in hematite used to understand colloidal transport properties of radionuclides. Also, we describe here an extension of that work employing an angle distance measure; this measure provides better classification based on spectral signatures alone in specimens with significant thickness variations. The method is illustrated using simulated data, and also to examine sporulation in the bacterium Clostridium sp.

Functional imaging of hippocampal place cells at cellular resolution during virtual navigation

PubMed Central

Dombeck, Daniel A.; Harvey, Christopher D.; Tian, Lin; Looger, Loren L.; Tank, David W.

2010-01-01

Spatial navigation is a widely employed behavior in rodent studies of neuronal circuits underlying cognition, learning and memory. In vivo microscopy combined with genetically-encoded indicators provides important new tools to study neuronal circuits, but has been technically difficult to apply during navigation. We describe methods to image the activity of hippocampal CA1 neurons with sub-cellular resolution in behaving mice. Neurons expressing the genetically encoded calcium indicator GCaMP3 were imaged through a chronic hippocampal window. Head-fixed mice performed spatial behaviors within a setup combining a virtual reality system and a custom built two-photon microscope. Populations of place cells were optically identified, and the correlation between the location of their place fields in the virtual environment and their anatomical location in the local circuit was measured. The combination of virtual reality and high-resolution functional imaging should allow for a new generation of studies to probe neuronal circuit dynamics during behavior. PMID:20890294

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

PubMed

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

2017-12-01

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

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

NASA Astrophysics Data System (ADS)

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

2017-12-01

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

Computational imaging of defects in commercial substrates for electronic and photonic devices

NASA Astrophysics Data System (ADS)

Fukuzawa, Masayuki; Kashiwagi, Ryo; Yamada, Masayoshi

2012-03-01

Computational defect imaging has been performed in commercial substrates for electronic and photonic devices by combining the transmission profile acquired with an imaging type of linear polariscope and the computational algorithm to extract a small amount of birefringence. The computational images of phase retardation δ exhibited spatial inhomogeneity of defect-induced birefringence in GaP, LiNbO3, and SiC substrates, which were not detected by conventional 'visual inspection' based on simple optical refraction or transmission because of poor sensitivity. The typical imaging time was less than 30 seconds for 3-inch diameter substrate with the spatial resolution of 200 μm, while that by scanning polariscope was 2 hours to get the same spatial resolution. Since our proposed technique have been achieved high sensitivity, short imaging time, and wide coverage of substrate materials, which are practical advantages over the laboratory-scale apparatus such as X-ray topography and electron microscope, it is useful for nondestructive inspection of various commercial substrates in production of electronic and photonic devices.

Nonlinear Focal Modulation Microscopy.

PubMed

Zhao, Guangyuan; Zheng, Cheng; Kuang, Cuifang; Zhou, Renjie; Kabir, Mohammad M; Toussaint, Kimani C; Wang, Wensheng; Xu, Liang; Li, Haifeng; Xiu, Peng; Liu, Xu

2018-05-11

We demonstrate nonlinear focal modulation microscopy (NFOMM) to achieve superresolution imaging. Traditional approaches to superresolution that utilize point scanning often rely on spatially reducing the size of the emission pattern by directly narrowing (e.g., through minimizing the detection pinhole in Airyscan, Zeiss) or indirectly peeling its outer profiles [e.g., through depleting the outer emission region in stimulated emission depletion (STED) microscopy]. We show that an alternative conceptualization that focuses on maximizing the optical system's frequency shifting ability offers advantages in further improving resolution while reducing system complexity. In NFOMM, a spatial light modulator and a suitably intense laser illumination are used to implement nonlinear focal-field modulation to achieve a transverse spatial resolution of ∼60  nm (∼λ/10). We show that NFOMM is comparable with STED microscopy and suitable for fundamental biology studies, as evidenced in imaging nuclear pore complexes, tubulin and vimentin in Vero cells. Since NFOMM is readily implemented as an add-on module to a laser-scanning microscope, we anticipate wide utility of this new imaging technique.

Nonlinear Focal Modulation Microscopy

NASA Astrophysics Data System (ADS)

Zhao, Guangyuan; Zheng, Cheng; Kuang, Cuifang; Zhou, Renjie; Kabir, Mohammad M.; Toussaint, Kimani C.; Wang, Wensheng; Xu, Liang; Li, Haifeng; Xiu, Peng; Liu, Xu

2018-05-01

We demonstrate nonlinear focal modulation microscopy (NFOMM) to achieve superresolution imaging. Traditional approaches to superresolution that utilize point scanning often rely on spatially reducing the size of the emission pattern by directly narrowing (e.g., through minimizing the detection pinhole in Airyscan, Zeiss) or indirectly peeling its outer profiles [e.g., through depleting the outer emission region in stimulated emission depletion (STED) microscopy]. We show that an alternative conceptualization that focuses on maximizing the optical system's frequency shifting ability offers advantages in further improving resolution while reducing system complexity. In NFOMM, a spatial light modulator and a suitably intense laser illumination are used to implement nonlinear focal-field modulation to achieve a transverse spatial resolution of ˜60 nm (˜λ /10 ). We show that NFOMM is comparable with STED microscopy and suitable for fundamental biology studies, as evidenced in imaging nuclear pore complexes, tubulin and vimentin in Vero cells. Since NFOMM is readily implemented as an add-on module to a laser-scanning microscope, we anticipate wide utility of this new imaging technique.

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

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

Baumbach, S., E-mail: baumbach@rheinahrcampus.de; Wilhein, T.; Kanngießer, B.

2015-08-15

This article introduces a laboratory setup of a transmission full-field x-ray microscope at 8 keV photon energy. The microscope operates in bright and dark field imaging mode with a maximum field of view of 50 μm. Since the illumination geometry determines whether the sample is illuminated homogeneously and moreover, if different imaging methods can be applied, the condenser optic is one of the most significant parts. With a new type of x-ray condenser, a polycapillary optic, we realized bright field imaging and for the first time dark field imaging at 8 keV photon energy in a laboratory setup. A detectormore » limited spatial resolution of 210 nm is measured on x-ray images of Siemens star test patterns.« less

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

PubMed

Baumbach, S; Kanngießer, B; Malzer, W; Stiel, H; Wilhein, T

2015-08-01

This article introduces a laboratory setup of a transmission full-field x-ray microscope at 8 keV photon energy. The microscope operates in bright and dark field imaging mode with a maximum field of view of 50 μm. Since the illumination geometry determines whether the sample is illuminated homogeneously and moreover, if different imaging methods can be applied, the condenser optic is one of the most significant parts. With a new type of x-ray condenser, a polycapillary optic, we realized bright field imaging and for the first time dark field imaging at 8 keV photon energy in a laboratory setup. A detector limited spatial resolution of 210 nm is measured on x-ray images of Siemens star test patterns.

Imaging of current density distributions with a Nb weak-link scanning nano-SQUID microscope

PubMed Central

Shibata, Yusuke; Nomura, Shintaro; Kashiwaya, Hiromi; Kashiwaya, Satoshi; Ishiguro, Ryosuke; Takayanagi, Hideaki

2015-01-01

Superconducting quantum interference devices (SQUIDs) are accepted as one of the highest magnetic field sensitive probes. There are increasing demands to image local magnetic fields to explore spin properties and current density distributions in a two-dimensional layer of semiconductors or superconductors. Nano-SQUIDs have recently attracting much interest for high spatial resolution measurements in nanometer-scale samples. Whereas weak-link Dayem Josephson junction nano-SQUIDs are suitable to miniaturization, hysteresis in current-voltage (I-V) characteristics that is often observed in Dayem Josephson junction is not desirable for a scanning microscope. Here we report on our development of a weak-link nano-SQUIDs scanning microscope with small hysteresis in I-V curve and on reconstructions of two-dimensional current density vector in two-dimensional electron gas from measured magnetic field. PMID:26459874

Imaging of current density distributions with a Nb weak-link scanning nano-SQUID microscope

NASA Astrophysics Data System (ADS)

Shibata, Yusuke; Nomura, Shintaro; Kashiwaya, Hiromi; Kashiwaya, Satoshi; Ishiguro, Ryosuke; Takayanagi, Hideaki

2015-10-01

Superconducting quantum interference devices (SQUIDs) are accepted as one of the highest magnetic field sensitive probes. There are increasing demands to image local magnetic fields to explore spin properties and current density distributions in a two-dimensional layer of semiconductors or superconductors. Nano-SQUIDs have recently attracting much interest for high spatial resolution measurements in nanometer-scale samples. Whereas weak-link Dayem Josephson junction nano-SQUIDs are suitable to miniaturization, hysteresis in current-voltage (I-V) characteristics that is often observed in Dayem Josephson junction is not desirable for a scanning microscope. Here we report on our development of a weak-link nano-SQUIDs scanning microscope with small hysteresis in I-V curve and on reconstructions of two-dimensional current density vector in two-dimensional electron gas from measured magnetic field.

Imaging ion and molecular transport at subcellular resolution by secondary ion mass spectrometry

NASA Astrophysics Data System (ADS)

Chandra, Subhash; Morrison, George H.

1995-05-01

The transport of K+, Na+, and Ca2+ were imaged in individual cells with a Cameca IMS-3f ion microscope. Strict cryogenic frozen freeze-dry sample preparations were employed. Ion redistribution artifacts in conventional chemical preparations are discussed. Cryogenically prepared freeze-fractured freeze-dried cultured cells allowed the three-dimensional ion microscopic imaging of elements. As smaller structures in calcium images can be resolved with the 0.5 [mu]m spatial resolution, correlative techniques are needed to confirm their identity. The potentials of reflected light microscopy, scanning electron microscopy and laser scanning confocal microscopy are discussed for microfeature recognition in freeze-fractured freeze-dried cells. The feasibility of using frozen freeze-dried cells for imaging molecular transport at subcellular resolution was tested. Ion microscopy successfully imaged the transport of the isotopically tagged (13C, 15N) amino acid, -arginine. The labeled amino acid was imaged at mass 28 with a Cs+ primary ion beam as the 28(13C15N)- species. After a 4 h exposure of LLC-PK1 kidney cells to 4 mM labeled arginine, the amino acid was localized throughout the cell with a preferential incorporation into the nucleus and nucleolus. An example is also shown of the ion microscopic imaging of sodium borocaptate, an experimental therapeutic drug for brain tumors, in cryogenically prepared frozen freeze-dried Swiss 3T3 cells.

Field-portable lensfree tomographic microscope†

PubMed Central

Isikman, Serhan O.; Bishara, Waheb; Sikora, Uzair; Yaglidere, Oguzhan; Yeah, John; Ozcan, Aydogan

2011-01-01

We present a field-portable lensfree tomographic microscope, which can achieve sectional imaging of a large volume (~20 mm3) on a chip with an axial resolution of <7 μm. In this compact tomographic imaging platform (weighing only ~110 grams), 24 light-emitting diodes (LEDs) that are each butt-coupled to a fibre-optic waveguide are controlled through a cost-effective micro-processor to sequentially illuminate the sample from different angles to record lensfree holograms of the sample that is placed on the top of a digital sensor array. In order to generate pixel super-resolved (SR) lensfree holograms and hence digitally improve the achievable lateral resolution, multiple sub-pixel shifted holograms are recorded at each illumination angle by electromagnetically actuating the fibre-optic waveguides using compact coils and magnets. These SR projection holograms obtained over an angular range of ~50° are rapidly reconstructed to yield projection images of the sample, which can then be back-projected to compute tomograms of the objects on the sensor-chip. The performance of this compact and light-weight lensfree tomographic microscope is validated by imaging micro-beads of different dimensions as well as a Hymenolepis nana egg, which is an infectious parasitic flatworm. Achieving a decent three-dimensional spatial resolution, this field-portable on-chip optical tomographic microscope might provide a useful toolset for telemedicine and high-throughput imaging applications in resource-poor settings. PMID:21573311

Low-loss electron energy loss spectroscopy: An atomic-resolution complement to optical spectroscopies and application to graphene

DOE PAGES

Kapetanakis, Myron; Zhou, Wu; Oxley, Mark P.; ...

2015-09-25

Photon-based spectroscopies have played a central role in exploring the electronic properties of crystalline solids and thin films. They are a powerful tool for probing the electronic properties of nanostructures, but they are limited by lack of spatial resolution. On the other hand, electron-based spectroscopies, e.g., electron energy loss spectroscopy (EELS), are now capable of subangstrom spatial resolution. Core-loss EELS, a spatially resolved analog of x-ray absorption, has been used extensively in the study of inhomogeneous complex systems. In this paper, we demonstrate that low-loss EELS in an aberration-corrected scanning transmission electron microscope, which probes low-energy excitations, combined with amore » theoretical framework for simulating and analyzing the spectra, is a powerful tool to probe low-energy electron excitations with atomic-scale resolution. The theoretical component of the method combines density functional theory–based calculations of the excitations with dynamical scattering theory for the electron beam. We apply the method to monolayer graphene in order to demonstrate that atomic-scale contrast is inherent in low-loss EELS even in a perfectly periodic structure. The method is a complement to optical spectroscopy as it probes transitions entailing momentum transfer. The theoretical analysis identifies the spatial and orbital origins of excitations, holding the promise of ultimately becoming a powerful probe of the structure and electronic properties of individual point and extended defects in both crystals and inhomogeneous complex nanostructures. The method can be extended to probe magnetic and vibrational properties with atomic resolution.« less

High spatial resolution and high brightness ion beam probe for in-situ elemental and isotopic analysis

NASA Astrophysics Data System (ADS)

Long, Tao; Clement, Stephen W. J.; Bao, Zemin; Wang, Peizhi; Tian, Di; Liu, Dunyi

2018-03-01

A high spatial resolution and high brightness ion beam from a cold cathode duoplasmatron source and primary ion optics are presented and applied to in-situ analysis of micro-scale geological material with complex structural and chemical features. The magnetic field in the source as well as the influence of relative permeability of magnetic materials on source performance was simulated using COMSOL to confirm the magnetic field strength of the source. Based on SIMION simulation, a high brightness and high spatial resolution negative ion optical system has been developed to achieve Critical (Gaussian) illumination mode. The ion source and primary column are installed on a new Time-of-Flight secondary ion mass spectrometer for analysis of geological samples. The diameter of the ion beam was measured by the knife-edge method and a scanning electron microscope (SEM). Results show that an O2- beam of ca. 5 μm diameter with a beam intensity of ∼5 nA and an O- beam of ca. 5 μm diameter with a beam intensity of ∼50 nA were obtained, respectively. This design will open new possibilities for in-situ elemental and isotopic analysis in geological studies.

Near-field transport imaging applied to photovoltaic materials

DOE PAGES

Xiao, Chuanxiao; Jiang, Chun -Sheng; Moseley, John; ...

2017-05-26

We developed and applied a new analytical technique - near-field transport imaging (NF-TI or simply TI) - to photovoltaic materials. Charge-carrier transport is an important factor in solar cell performance, and TI is an innovative approach that integrates a scanning electron microscope with a near-field scanning optical microscope, providing the possibility to study luminescence associated with recombination and transport with high spatial resolution. In this paper, we describe in detail the technical barriers we had to overcome to develop the technique for routine application and the data-fitting procedure used to calculate minority-carrier diffusion length values. The diffusion length measured bymore » TI agrees well with the results calculated by time-resolved photoluminescence on well-controlled gallium arsenide (GaAs) thin-film samples. We report for the first time on measurements on thin-film cadmium telluride using this technique, including the determination of effective carrier diffusion length, as well as the first near-field imaging of the effect of a single localized defect on carrier transport and recombination in a GaAs heterostructure. Furthermore, by changing the scanning setup, we were able to demonstrate near-field cathodoluminescence (CL), and correlated the results with standard CL measurements. In conclusion, the TI technique shows great potential for mapping transport properties in solar cell materials with high spatial resolution.« less

Synchrotron-based X-ray microscopic studies for bioeffects of nanomaterials.

PubMed

Zhu, Ying; Cai, Xiaoqing; Li, Jiang; Zhong, Zengtao; Huang, Qing; Fan, Chunhai

2014-04-01

There have been increasing interests in studying biological effects of nanomaterials, which are nevertheless faced up with many challenges due to the nanoscale dimensions and unique chemical properties of nanomaterials. Synchrotron-based X-ray microscopy, an advanced imaging technology with high spatial resolution and excellent elemental specificity, provides a new platform for studying interactions between nanomaterials and living systems. In this article, we review the recent progress of X-ray microscopic studies on bioeffects of nanomaterials in several living systems including cells, model organisms, animals and plants. We aim to provide an overview of the state of the art, and the advantages of using synchrotron-based X-ray microscopy for characterizing in vitro and in vivo behaviors and biodistribution of nanomaterials. We also expect that the use of a combination of new synchrotron techniques should offer unprecedented opportunities for better understanding complex interactions at the nano-biological interface and accounting for unique bioeffects of nanomaterials. Synchrotron-based X-ray microscopy is a non-destructive imaging technique that enables high resolution spatial mapping of metals with elemental level detection methods. This review summarizes the current use and perspectives of this novel technique in studying the biology and tissue interactions of nanomaterials. Copyright © 2014 Elsevier Inc. All rights reserved.

Ultrawidefield microscope for high-speed fluorescence imaging and targeted optogenetic stimulation.

PubMed

Werley, Christopher A; Chien, Miao-Ping; Cohen, Adam E

2017-12-01

The rapid increase in the number and quality of fluorescent reporters and optogenetic actuators has yielded a powerful set of tools for recording and controlling cellular state and function. To achieve the full benefit of these tools requires improved optical systems with high light collection efficiency, high spatial and temporal resolution, and patterned optical stimulation, in a wide field of view (FOV). Here we describe our 'Firefly' microscope, which achieves these goals in a Ø6 mm FOV. The Firefly optical system is optimized for simultaneous photostimulation and fluorescence imaging in cultured cells. All but one of the optical elements are commercially available, yet the microscope achieves 10-fold higher light collection efficiency at its design magnification than the comparable commercially available microscope using the same objective. The Firefly microscope enables all-optical electrophysiology ('Optopatch') in cultured neurons with a throughput and information content unmatched by other neuronal phenotyping systems. This capability opens possibilities in disease modeling and phenotypic drug screening. We also demonstrate applications of the system to voltage and calcium recordings in human induced pluripotent stem cell derived cardiomyocytes.

Ultrawidefield microscope for high-speed fluorescence imaging and targeted optogenetic stimulation

PubMed Central

Werley, Christopher A.; Chien, Miao-Ping; Cohen, Adam E.

2017-01-01

The rapid increase in the number and quality of fluorescent reporters and optogenetic actuators has yielded a powerful set of tools for recording and controlling cellular state and function. To achieve the full benefit of these tools requires improved optical systems with high light collection efficiency, high spatial and temporal resolution, and patterned optical stimulation, in a wide field of view (FOV). Here we describe our ‘Firefly’ microscope, which achieves these goals in a Ø6 mm FOV. The Firefly optical system is optimized for simultaneous photostimulation and fluorescence imaging in cultured cells. All but one of the optical elements are commercially available, yet the microscope achieves 10-fold higher light collection efficiency at its design magnification than the comparable commercially available microscope using the same objective. The Firefly microscope enables all-optical electrophysiology (‘Optopatch’) in cultured neurons with a throughput and information content unmatched by other neuronal phenotyping systems. This capability opens possibilities in disease modeling and phenotypic drug screening. We also demonstrate applications of the system to voltage and calcium recordings in human induced pluripotent stem cell derived cardiomyocytes. PMID:29296505

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.

Immunogold labeling reveals subcellular localisation of silica nanoparticles in a human blood-brain barrier model

NASA Astrophysics Data System (ADS)

Ye, Dong; Anguissola, Sergio; O'Neill, Tiina; Dawson, Kenneth A.

2015-05-01

Subcellular location of nanoparticles has been widely investigated with fluorescence microscopy, via fluorescently labeled antibodies to visualise target antigens in cells. However, fluorescence microscopy, such as confocal or live cell imaging, has generally limited 3D spatial resolution. Conventional electron microscopy can be useful in bridging resolution gap, but still not ideal in resolving subcellular organelle identities. Using the pre-embedding immunogold electron microscopic imaging, we performed accurate examination of the intracellular trafficking and gathered further evidence of transport mechanisms of silica nanoparticles across a human in vitro blood-brain barrier model. Our approach can effectively immunolocalise a variety of intracellular compartments and provide new insights into the uptake and subcellular transport of nanoparticles.Subcellular location of nanoparticles has been widely investigated with fluorescence microscopy, via fluorescently labeled antibodies to visualise target antigens in cells. However, fluorescence microscopy, such as confocal or live cell imaging, has generally limited 3D spatial resolution. Conventional electron microscopy can be useful in bridging resolution gap, but still not ideal in resolving subcellular organelle identities. Using the pre-embedding immunogold electron microscopic imaging, we performed accurate examination of the intracellular trafficking and gathered further evidence of transport mechanisms of silica nanoparticles across a human in vitro blood-brain barrier model. Our approach can effectively immunolocalise a variety of intracellular compartments and provide new insights into the uptake and subcellular transport of nanoparticles. Electronic supplementary information (ESI) available: Nanoparticle characterisation data, preservation of cellular structures, staining controls, optimisation of size amplification via the silver enhancement, and more imaging results from anti-clathrin and anti-caveolin 1 immunolabeling. See DOI: 10.1039/c5nr01539a

Multipurpose Hyperspectral Imaging System

NASA Technical Reports Server (NTRS)

Mao, Chengye; Smith, David; Lanoue, Mark A.; Poole, Gavin H.; Heitschmidt, Jerry; Martinez, Luis; Windham, William A.; Lawrence, Kurt C.; Park, Bosoon

2005-01-01

A hyperspectral imaging system of high spectral and spatial resolution that incorporates several innovative features has been developed to incorporate a focal plane scanner (U.S. Patent 6,166,373). This feature enables the system to be used for both airborne/spaceborne and laboratory hyperspectral imaging with or without relative movement of the imaging system, and it can be used to scan a target of any size as long as the target can be imaged at the focal plane; for example, automated inspection of food items and identification of single-celled organisms. The spectral resolution of this system is greater than that of prior terrestrial multispectral imaging systems. Moreover, unlike prior high-spectral resolution airborne and spaceborne hyperspectral imaging systems, this system does not rely on relative movement of the target and the imaging system to sweep an imaging line across a scene. This compact system (see figure) consists of a front objective mounted at a translation stage with a motorized actuator, and a line-slit imaging spectrograph mounted within a rotary assembly with a rear adaptor to a charged-coupled-device (CCD) camera. Push-broom scanning is carried out by the motorized actuator which can be controlled either manually by an operator or automatically by a computer to drive the line-slit across an image at a focal plane of the front objective. To reduce the cost, the system has been designed to integrate as many as possible off-the-shelf components including the CCD camera and spectrograph. The system has achieved high spectral and spatial resolutions by using a high-quality CCD camera, spectrograph, and front objective lens. Fixtures for attachment of the system to a microscope (U.S. Patent 6,495,818 B1) make it possible to acquire multispectral images of single cells and other microscopic objects.

Nanoscale magnetic imaging using picosecond thermal gradients

NASA Astrophysics Data System (ADS)

Fuchs, Gregory

Research and development in spintronics is challenged by the lack of table-top magnetic imaging technologies that posses the simultaneous temporal resolution and spatial resolution to characterize magnetization dynamics in emerging spintronic devices. In addition, many of the most exciting magnetic material systems for spintronics are difficult to image with any method. To address this challenge, we developed a spatiotemporal magnetic microscope based on picosecond heat pulses that stroboscopically transduces an in-plane magnetization into a voltage signal. When the magnetic device contains a magnetic metal like FeCoB or NiFe, we use the time-resolved anomalous Nernst effect. When it contains a magnetic insulator/normal metal bilayer like yttrium iron garnet/platinum, we use the combination of the time-resolved longitudinal spin Seebeck effect and the inverse spin Hall effect. We demonstrate that these imaging modalities have time resolutions in the range of 10-100 ps and sensitivities in the range of 0.1 - 0.3° /√{Hz} , which enables not only static magnetic imaging, but also phase-sensitive ferromagnetic resonance imaging. One application of this technology is for magnetic torque vector imaging, which we apply to a spin Hall device. We find an unexpected variation in the spin torque vector that suggests conventional, all-electrical FMR measurements of spin torque vectors can produce a systematic error as large as 30% when quantifying the spin Hall efficiency. Finally, I will describe how time-resolved magnetic imaging can greatly exceed the spatial resolution of optical diffraction. We demonstrate scanning a sharp gold tip to create near-field thermal transfer from a picosecond laser pulse to a magnetic sample as the basis of a nanoscale spatiotemporal microscope. We gratefully acknowledge support from the AFOSR (FA9550-14-1-0243) and the NSF through the Cornell Center for Materials Research (DMR-1120296).

RGB digital lensless holographic microscopy

NASA Astrophysics Data System (ADS)

Garcia-Sucerquia, Jorge

2013-11-01

The recent introduction of color digital lensless holographic microscopy (CDLHM) has shown the possibility of imaging microscopic specimens at full color without the need of lenses. Owing to the simplicity, robustness, and compactness of the digital lensless holographic microscopes (DLHM), they have been presented as the ideal candidates to being developed into portable holographic microscopes. However, in the case of CDLHM the utilization of three independent lasers hinders the portability option for this microscope. In this contribution an alternative to reduce the complexity of CDLHM aimed to recover the portability of this microscopy technology is presented. A super-bright white-light light-emitting diode (LED) is spectrally and spatially filtered to produce the needed illumination by CDLHM to work. CDLHM with LED illumination is used to image at full color a section of the head of a drosophila melanogaster fly (fruit fly). The LED-CDLHM method shows the capability of imaging objects of 2μm size in comparison with the micrometer resolution reported for LASER-CDLHM.

Laboratory-size three-dimensional x-ray microscope with Wolter type I mirror optics and an electron-impact water window x-ray source

NASA Astrophysics Data System (ADS)

Ohsuka, Shinji; Ohba, Akira; Onoda, Shinobu; Nakamoto, Katsuhiro; Nakano, Tomoyasu; Miyoshi, Motosuke; Soda, Keita; Hamakubo, Takao

2014-09-01

We constructed a laboratory-size three-dimensional water window x-ray microscope that combines wide-field transmission x-ray microscopy with tomographic reconstruction techniques, and observed bio-medical samples to evaluate its applicability to life science research fields. It consists of a condenser and an objective grazing incidence Wolter type I mirror, an electron-impact type oxygen Kα x-ray source, and a back-illuminated CCD for x-ray imaging. A spatial resolution limit of around 1.0 line pairs per micrometer was obtained for two-dimensional transmission images, and 1-μm scale three-dimensional fine structures were resolved.

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

Schollmeier, Marius S.; Geissel, Matthias; Shores, Jonathon E.

We present calculations for the field of view (FOV), image fluence, image monochromaticity, spectral acceptance, and image aberrations for spherical crystal microscopes, which are used as self-emission imaging or backlighter systems at large-scale high energy density physics facilities. Our analytic results are benchmarked with ray-tracing calculations as well as with experimental measurements from the 6.151 keV backlighter system at Sandia National Laboratories. Furthermore, the analytic expressions can be used for x-ray source positions anywhere between the Rowland circle and object plane. We discovered that this enables quick optimization of the performance of proposed but untested, bent-crystal microscope systems to findmore » the best compromise between FOV, image fluence, and spatial resolution for a particular application.« less

Laboratory-size three-dimensional x-ray microscope with Wolter type I mirror optics and an electron-impact water window x-ray source.

PubMed

Ohsuka, Shinji; Ohba, Akira; Onoda, Shinobu; Nakamoto, Katsuhiro; Nakano, Tomoyasu; Miyoshi, Motosuke; Soda, Keita; Hamakubo, Takao

2014-09-01

We constructed a laboratory-size three-dimensional water window x-ray microscope that combines wide-field transmission x-ray microscopy with tomographic reconstruction techniques, and observed bio-medical samples to evaluate its applicability to life science research fields. It consists of a condenser and an objective grazing incidence Wolter type I mirror, an electron-impact type oxygen Kα x-ray source, and a back-illuminated CCD for x-ray imaging. A spatial resolution limit of around 1.0 line pairs per micrometer was obtained for two-dimensional transmission images, and 1-μm scale three-dimensional fine structures were resolved.

Assessing resolution in live cell structured illumination microscopy

NASA Astrophysics Data System (ADS)

Pospíšil, Jakub; Fliegel, Karel; Klíma, Miloš

2017-12-01

Structured Illumination Microscopy (SIM) is a powerful super-resolution technique, which is able to enhance the resolution of optical microscope beyond the Abbe diffraction limit. In the last decade, numerous SIM methods that achieve the resolution of 100 nm in the lateral dimension have been developed. The SIM setups with new high-speed cameras and illumination pattern generators allow rapid acquisition of the live specimen. Therefore, SIM is widely used for investigation of the live structures in molecular and live cell biology. Quantitative evaluation of resolution enhancement in a real sample is essential to describe the efficiency of super-resolution microscopy technique. However, measuring the resolution of a live cell sample is a challenging task. Based on our experimental findings, the widely used Fourier ring correlation (FRC) method does not seem to be well suited for measuring the resolution of SIM live cell video sequences. Therefore, the resolution assessing methods based on Fourier spectrum analysis are often used. We introduce a measure based on circular average power spectral density (PSDca) estimated from a single SIM image (one video frame). PSDca describes the distribution of the power of a signal with respect to its spatial frequency. Spatial resolution corresponds to the cut-off frequency in Fourier space. In order to estimate the cut-off frequency from a noisy signal, we use a spectral subtraction method for noise suppression. In the future, this resolution assessment approach might prove useful also for single-molecule localization microscopy (SMLM) live cell imaging.

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.

Probing Chemical Properties of Interstitial Micro-fluids in Ice

NASA Astrophysics Data System (ADS)

Cheng, J.; Colussi, A. J.; Hoffmann, M. R.

2007-12-01

Liquid is present as microscopic channels in polycrystalline ice at sub-freezing and even sub-eutectic temperatures. Not only do chemicals tend to concentrate substantially in this microscopic liquid phase, but local physicochemical properties may also differ widely from the bulk counterparts, therefore critically affecting the thermodynamics and kinetics of chemical processes occurring in frozen media such as snow, frost, and frost- flowers. This phenomenon has important implications in atmospheric chemistry such as affecting the composition of the atmospheric boundary layer in snow-covered regions. A method using con-focal laser scanning microscope equipped with a cryostat has been developed to measure physicochemical properties of the microscopic liquid phase in ice that are not readily extrapolated from the bulk data. The experimental setup allows for monitoring the freezing process of an aqueous solution with a sub- second time resolution and a submicron 3D spatial resolution. The physicochemical properties (e.g. viscosity, polarity, and acidity) can, in theory, be deduced from features of the fluorescence spectra of particular fluorescent indicators. For example, the acidity change during the freezing and melting process of electrolyte solutions has been monitored in real time by a pH-dependent dual emission fluorescent probe C-SNARF-1. The effects of temperature, freezing rate, and added electrolytes such as ammonium sulfate, sodium chloride and zwitterions are also examined. The findings complement the theory and previous experimental evidence of freezing hydrolysis.

A portable fluorescence microscopic imaging system for cholecystectomy

NASA Astrophysics Data System (ADS)

Ye, Jian; Yang, Chaoyu; Gan, Qi; Ma, Rong; Zhang, Zeshu; Chang, Shufang; Shao, Pengfei; Zhang, Shiwu; Liu, Chenhai; Xu, Ronald

2016-03-01

In this paper we proposed a portable fluorescence microscopic imaging system to prevent iatrogenic biliary injuries from occurring during cholecystectomy due to misidentification of the cystic structures. The system consisted of a light source module, a CMOS camera, a Raspberry Pi computer and a 5 inch HDMI LCD. Specifically, the light source module was composed of 690 nm and 850 nm LEDs, allowing the CMOS camera to simultaneously acquire both fluorescence and background images. The system was controlled by Raspberry Pi using Python programming with the OpenCV library under Linux. We chose Indocyanine green(ICG) as a fluorescent contrast agent and then tested fluorescence intensities of the ICG aqueous solution at different concentration levels by our fluorescence microscopic system compared with the commercial Xenogen IVIS system. The spatial resolution of the proposed fluorescence microscopic imaging system was measured by a 1951 USAF resolution target and the dynamic response was evaluated quantitatively with an automatic displacement platform. Finally, we verified the technical feasibility of the proposed system in mouse models of bile duct, performing both correct and incorrect gallbladder resection. Our experiments showed that the proposed system can provide clear visualization of the confluence between the cystic duct and common bile duct or common hepatic duct, suggesting that this is a potential method for guiding cholecystectomy. The proposed portable system only cost a total of $300, potentially promoting its use in resource-limited settings.

The National Ignition Facility modular Kirkpatrick-Baez microscope

DOE PAGES

Pickworth, L. A.; Ayers, J.; Bell, P.; ...

2016-08-10

Current two-dimensional X-ray imaging at the National Ignition Facility (NIF) uses time resolved pinhole cameras with ~10-25µm pinholes. This method has limitations in the smallest resolvable features that can be imaged with reasonable photon statistics for inertial confinement fusion (ICF) applications. ICF sources have a broadband self-emission spectrum that causes the pinhole images obtained, through thin foil filters, to contain a similarly broadband spectrum complicating the interpretation of structure in the source. In order to study phenomena on the scale of ~5 µm, such as dopant mix in the ICF capsule, a narrow energy band, higher spatial resolution microscope systemmore » with improved signal/noise has been developed using X-ray optics. Utilizing grazing incidence mirrors in a Kirkpatrick-Baez microscope (KBM) configuration, an X-ray microscope has been designed and fielded on NIF with four imaging channels. The KBM has ~12x magnification, <8 µm resolution and higher throughput in comparison to similar pinhole systems. The first KBM mirrors are coated with a multilayer mirror to allow a ‘narrow band’ energy response at 10.2keV with ΔE~3keV. By adjusting the mirror coating only, the energy response can be matched to future experimental requirements. Here, several mirror packs have been commissioned and are interchangeable in the diagnostic snout.« less

A framed, 16-image Kirkpatrick–Baez x-ray microscope

DOE PAGES

Marshall, F. J.; Bahr, R. E.; Goncharov, V. N.; ...

2017-09-08

A 16-image Kirkpatrick–Baez (KB)–type x-ray microscope consisting of compact KB mirrors has been assembled for the first time with mirrors aligned to allow it to be coupled to a high-speed framing camera. The high-speed framing camera has four independently gated strips whose emission sampling interval is ~30 ps. Images are arranged four to a strip with ~60-ps temporal spacing between frames on a strip. By spacing the timing of the strips, a frame spacing of ~15 ps is achieved. A framed resolution of ~6-um is achieved with this combination in a 400-um region of laser–plasma x-ray emission in the 2-more » to 8-keV energy range. A principal use of the microscope is to measure the evolution of the implosion stagnation region of cryogenic DT target implosions on the University of Rochester’s OMEGA Laser System. The unprecedented time and spatial resolution achieved with this framed, multi-image KB microscope have made it possible to accurately determine the cryogenic implosion core emission size and shape at the peak of stagnation. In conclusion, these core size measurements, taken in combination with those of ion temperature, neutron-production temporal width, and neutron yield allow for inference of core pressures, currently exceeding 50 GBar in OMEGA cryogenic target implosions.« less

A framed, 16-image Kirkpatrick–Baez x-ray microscope

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

Marshall, F. J.; Bahr, R. E.; Goncharov, V. N.

A 16-image Kirkpatrick–Baez (KB)–type x-ray microscope consisting of compact KB mirrors has been assembled for the first time with mirrors aligned to allow it to be coupled to a high-speed framing camera. The high-speed framing camera has four independently gated strips whose emission sampling interval is ~30 ps. Images are arranged four to a strip with ~60-ps temporal spacing between frames on a strip. By spacing the timing of the strips, a frame spacing of ~15 ps is achieved. A framed resolution of ~6-um is achieved with this combination in a 400-um region of laser–plasma x-ray emission in the 2-more » to 8-keV energy range. A principal use of the microscope is to measure the evolution of the implosion stagnation region of cryogenic DT target implosions on the University of Rochester’s OMEGA Laser System. The unprecedented time and spatial resolution achieved with this framed, multi-image KB microscope have made it possible to accurately determine the cryogenic implosion core emission size and shape at the peak of stagnation. In conclusion, these core size measurements, taken in combination with those of ion temperature, neutron-production temporal width, and neutron yield allow for inference of core pressures, currently exceeding 50 GBar in OMEGA cryogenic target implosions.« less

The National Ignition Facility modular Kirkpatrick-Baez microscope

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

Pickworth, L. A.; Ayers, J.; Bell, P.

Current two-dimensional X-ray imaging at the National Ignition Facility (NIF) uses time resolved pinhole cameras with ~10-25µm pinholes. This method has limitations in the smallest resolvable features that can be imaged with reasonable photon statistics for inertial confinement fusion (ICF) applications. ICF sources have a broadband self-emission spectrum that causes the pinhole images obtained, through thin foil filters, to contain a similarly broadband spectrum complicating the interpretation of structure in the source. In order to study phenomena on the scale of ~5 µm, such as dopant mix in the ICF capsule, a narrow energy band, higher spatial resolution microscope systemmore » with improved signal/noise has been developed using X-ray optics. Utilizing grazing incidence mirrors in a Kirkpatrick-Baez microscope (KBM) configuration, an X-ray microscope has been designed and fielded on NIF with four imaging channels. The KBM has ~12x magnification, <8 µm resolution and higher throughput in comparison to similar pinhole systems. The first KBM mirrors are coated with a multilayer mirror to allow a ‘narrow band’ energy response at 10.2keV with ΔE~3keV. By adjusting the mirror coating only, the energy response can be matched to future experimental requirements. Here, several mirror packs have been commissioned and are interchangeable in the diagnostic snout.« less

The National Ignition Facility modular Kirkpatrick-Baez microscope.

PubMed

Pickworth, L A; Ayers, J; Bell, P; Brejnholt, N F; Buscho, J G; Bradley, D; Decker, T; Hau-Riege, S; Kilkenny, J; McCarville, T; Pardini, T; Vogel, J; Walton, C

2016-11-01

Current two-dimensional X-ray imaging at the National Ignition Facility (NIF) uses time resolved pinhole cameras with ∼10-25 μm pinholes. This method has limitations in the smallest resolvable features that can be imaged with reasonable photon statistics for inertial confinement fusion (ICF) applications. ICF sources have a broadband self-emission spectrum that causes the pinhole images obtained, through thin foil filters, to contain a similarly broadband spectrum complicating the interpretation of structure in the source. In order to study phenomena on the scale of ∼5 μm, such as dopant mix in the ICF capsule, a narrow energy band, higher spatial resolution microscope system with improved signal/noise has been developed using X-ray optics. Utilizing grazing incidence mirrors in a Kirkpatrick-Baez microscope (KBM) configuration [P. Kirkpatrick and A. V. Baez, J. Opt. Soc. Am. 38, 766-774 (1948)], an X-ray microscope has been designed and fielded on NIF with four imaging channels. The KBM has ∼12 × magnification, <8 μm resolution, and higher throughput in comparison to similar pinhole systems. The first KBM mirrors are coated with a multilayer mirror to allow a "narrow band" energy response at 10.2 keV with ΔE ∼ 3 keV. By adjusting the mirror coating only, the energy response can be matched to the future experimental requirements. Several mirror packs have been commissioned and are interchangeable in the diagnostic snout.

Portable, battery-operated, fluorescence field microscope for the developing world

NASA Astrophysics Data System (ADS)

Miller, Andrew R.; Davis, Gregory; Pierce, Mark; Oden, Z. Maria; Richards-Kortum, Rebecca

2010-02-01

In many areas of the world, current methods for diagnosis of infectious diseases such as malaria and tuberculosis involve microscopic evaluation of a patient specimen. Advances in fluorescence microscopy can improve diagnostic sensitivity and reduce time and expertise necessary to interpret diagnostic results. However, modern research-grade microscopes are neither available nor appropriate for use in many settings in the developing world. To address this need, we designed, fabricated, and tested a portable, battery-powered, bright field and fluorescence inverted field microscope, optimized for infrastructural constraints of the developing world. We characterized an initial prototype constructed with rapidprototyping techniques, which utilized low-cost, over-the-counter components such as a battery-powered LED flashlight as the light source. The microscope exhibited suitable spatial resolution (0.8 μm) in fluorescence mode to resolve M. tuberculosis bacilli. In bright field mode, malaria parasites were resolvable at 1000x magnification. The initial prototype cost 480 USD and we estimate that the microscope can be manufactured for 230 USD. While future studies are planned to evaluate ease-of-use and reliability, our current system serves as a proof of concept that combined fluorescence and bright field microscopy is possible in a low-cost and portable system.

Micromega/IR: Design and status of a near-infrared spectral microscope for in situ analysis of Mars samples

NASA Astrophysics Data System (ADS)

Leroi, Vaitua; Bibring, Jean-Pierre; Berthe, Michel

2009-07-01

MicrOmega is an ultra miniaturized spectral microscope for in situ analysis of samples. It is composed of 2 microscopes; one with a spatial sampling less or equal to 4 μm, working in 4 colors in the visible range: MicrOmega/VIS, and a NIR hyperspectral microscope working in the spectral range 0.9-4 μm with a spatial sampling of 20 μm per pixel: MicrOmega/IR (described in this paper). MicrOmega/IR illuminates and images samples a few mm in size and acquires the NIR spectrum of each resolved pixel in up to 320 contiguous spectral channels. The goal of this instrument is to analyze in situ the composition of collected samples at almost their grain size scale, in a non-destructive way. With the chosen spectral range and resolution, a wide variety of constituents can be identified: minerals, such as pyroxene and olivine, ferric oxides, hydrated phyllosilicates, sulfates and carbonates and ices and organics. The composition of the various phases within a given sample is a critical record of its formation and evolution. Coupled to the mapping information, it provides unique clues to describe the history of the parent body (planet, satellite and small body). In particular, the capability to identify hydrated grains and to characterize their adjacent phases has a huge potential in the search for possible bio-relics.

A resolution measure for three-dimensional microscopy

PubMed Central

Chao, Jerry; Ram, Sripad; Abraham, Anish V.; Ward, E. Sally; Ober, Raimund J.

2009-01-01

A three-dimensional (3D) resolution measure for the conventional optical microscope is introduced which overcomes the drawbacks of the classical 3D (axial) resolution limit. Formulated within the context of a parameter estimation problem and based on the Cramer-Rao lower bound, this 3D resolution measure indicates the accuracy with which a given distance between two objects in 3D space can be determined from the acquired image. It predicts that, given enough photons from the objects of interest, arbitrarily small distances of separation can be estimated with prespecified accuracy. Using simulated images of point source pairs, we show that the maximum likelihood estimator is capable of attaining the accuracy predicted by the resolution measure. We also demonstrate how different factors, such as extraneous noise sources and the spatial orientation of the imaged object pair, can affect the accuracy with which a given distance of separation can be determined. PMID:20161040

Tip-enhanced near-field optical microscope with side-on and ATR-mode sample excitation for super-resolution Raman imaging of surfaces

NASA Astrophysics Data System (ADS)

Heilman, A. L.; Gordon, M. J.

2016-06-01

A tip-enhanced near-field optical microscope with side-on and attenuated total reflectance (ATR) excitation and collection is described and used to demonstrate sub-diffraction-limited (super-resolution) optical and chemical characterization of surfaces. ATR illumination is combined with an Au optical antenna tip to show that (i) the tip can quantitatively transduce the optical near-field (evanescent waves) above the surface by scattering photons into the far-field, (ii) the ATR geometry enables excitation and characterization of surface plasmon polaritons (SPPs), whose associated optical fields are shown to enhance Raman scattering from a thin layer of copper phthalocyanine (CuPc), and (iii) SPPs can be used to plasmonically excite the tip for super-resolution chemical imaging of patterned CuPc via tip-enhanced Raman spectroscopy (TERS). ATR-illumination TERS is also quantitatively compared with the more conventional side-on illumination scheme. In both cases, spatial resolution was better than 40 nm and tip on/tip off Raman enhancement factors were >6500. Furthermore, ATR illumination was shown to provide similar Raman signal levels at lower "effective" pump powers due to additional optical energy delivered by SPPs to the active region in the tip-surface gap.

Tip-enhanced near-field optical microscope with side-on and ATR-mode sample excitation for super-resolution Raman imaging of surfaces

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

Heilman, A. L.; Gordon, M. J.

A tip-enhanced near-field optical microscope with side-on and attenuated total reflectance (ATR) excitation and collection is described and used to demonstrate sub-diffraction-limited (super-resolution) optical and chemical characterization of surfaces. ATR illumination is combined with an Au optical antenna tip to show that (i) the tip can quantitatively transduce the optical near-field (evanescent waves) above the surface by scattering photons into the far-field, (ii) the ATR geometry enables excitation and characterization of surface plasmon polaritons (SPPs), whose associated optical fields are shown to enhance Raman scattering from a thin layer of copper phthalocyanine (CuPc), and (iii) SPPs can be used tomore » plasmonically excite the tip for super-resolution chemical imaging of patterned CuPc via tip-enhanced Raman spectroscopy (TERS). ATR-illumination TERS is also quantitatively compared with the more conventional side-on illumination scheme. In both cases, spatial resolution was better than 40 nm and tip on/tip off Raman enhancement factors were >6500. Furthermore, ATR illumination was shown to provide similar Raman signal levels at lower “effective” pump powers due to additional optical energy delivered by SPPs to the active region in the tip-surface gap.« less

A high speed multifocal multiphoton fluorescence lifetime imaging microscope for live-cell FRET imaging

PubMed Central

Poland, Simon P.; Krstajić, Nikola; Monypenny, James; Coelho, Simao; Tyndall, David; Walker, Richard J.; Devauges, Viviane; Richardson, Justin; Dutton, Neale; Barber, Paul; Li, David Day-Uei; Suhling, Klaus; Ng, Tony; Henderson, Robert K.; Ameer-Beg, Simon M.

2015-01-01

We demonstrate diffraction limited multiphoton imaging in a massively parallel, fully addressable time-resolved multi-beam multiphoton microscope capable of producing fluorescence lifetime images with sub-50ps temporal resolution. This imaging platform offers a significant improvement in acquisition speed over single-beam laser scanning FLIM by a factor of 64 without compromising in either the temporal or spatial resolutions of the system. We demonstrate FLIM acquisition at 500 ms with live cells expressing green fluorescent protein. The applicability of the technique to imaging protein-protein interactions in live cells is exemplified by observation of time-dependent FRET between the epidermal growth factor receptor (EGFR) and the adapter protein Grb2 following stimulation with the receptor ligand. Furthermore, ligand-dependent association of HER2-HER3 receptor tyrosine kinases was observed on a similar timescale and involved the internalisation and accumulation or receptor heterodimers within endosomes. These data demonstrate the broad applicability of this novel FLIM technique to the spatio-temporal dynamics of protein-protein interaction. PMID:25780724

Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning of thick tissues

NASA Astrophysics Data System (ADS)

Cheng, Li-Chung; Chang, Chia-Yuan; Yen, Wei-Chung; Chen, Shean-Jen

2012-10-01

Conventional multiphoton microscopy employs beam scanning; however, in this study a microscope based on spatiotemporal focusing offering widefield multiphoton excitation has been developed to provide fast optical sectioning images. The microscope integrates a 10 kHz repetition rate ultrafast amplifier featuring strong instantaneous peak power (maximum 400 μJ/pulse at 90 fs pulse width) with a TE-cooled, ultra-sensitive photon detecting, electron multiplying charge-coupled device camera. This configuration can produce multiphoton excited images with an excitation area larger than 200 × 100 μm2 at a frame rate greater than 100 Hz. Brownian motions of fluorescent microbeads as small as 0.5 μm have been instantaneously observed with a lateral spatial resolution of less than 0.5 μm and an axial resolution of approximately 3.5 μm. Moreover, we combine the widefield multiphoton microscopy with structure illuminated technique named HiLo to reject the background scattering noise to get better quality for bioimaging.

Rewritable ghost floating gates by tunnelling triboelectrification for two-dimensional electronics

PubMed Central

Kim, Seongsu; Kim, Tae Yun; Lee, Kang Hyuck; Kim, Tae-Ho; Cimini, Francesco Arturo; Kim, Sung Kyun; Hinchet, Ronan; Kim, Sang-Woo; Falconi, Christian

2017-01-01

Gates can electrostatically control charges inside two-dimensional materials. However, integrating independent gates typically requires depositing and patterning suitable insulators and conductors. Moreover, after manufacturing, gates are unchangeable. Here we introduce tunnelling triboelectrification for localizing electric charges in very close proximity of two-dimensional materials. As representative materials, we use chemical vapour deposition graphene deposited on a SiO2/Si substrate. The triboelectric charges, generated by friction with a Pt-coated atomic force microscope tip and injected through defects, are trapped at the air–SiO2 interface underneath graphene and act as ghost floating gates. Tunnelling triboelectrification uniquely permits to create, modify and destroy p and n regions at will with the spatial resolution of atomic force microscopes. As a proof of concept, we draw rewritable p/n+ and p/p+ junctions with resolutions as small as 200 nm. Our results open the way to time-variant two-dimensional electronics where conductors, p and n regions can be defined on demand. PMID:28649986

Unraveling submicron-scale mechanical heterogeneity by three-dimensional X-ray microdiffraction

PubMed Central

Li, Runguang; Xie, Qingge; Wang, Yan-Dong; Liu, Wenjun; Wang, Mingguang; Wu, Guilin; Li, Xiaowu; Zhang, Minghe; Lu, Zhaoping; Geng, Chang; Zhu, Ting

2018-01-01

Shear banding is a ubiquitous phenomenon of severe plastic deformation, and damage accumulation in shear bands often results in the catastrophic failure of a material. Despite extensive studies, the microscopic mechanisms of strain localization and deformation damage in shear bands remain elusive due to their spatial−temporal complexities embedded in bulk materials. Here we conducted synchrotron-based X-ray microdiffraction (μXRD) experiments to map out the 3D lattice strain field with a submicron resolution around fatigue shear bands in a stainless steel. Both in situ and postmortem μXRD results revealed large lattice strain gradients at intersections of the primary and secondary shear bands. Such strain gradients resulted in severe mechanical heterogeneities across the fatigue shear bands, leading to reduced fatigue limits in the high-cycle regime. The ability to spatially quantify the localized strain gradients with submicron resolution through μXRD opens opportunities for understanding the microscopic mechanisms of damage and failure in bulk materials. PMID:29284751

Rewritable ghost floating gates by tunnelling triboelectrification for two-dimensional electronics

NASA Astrophysics Data System (ADS)

Kim, Seongsu; Kim, Tae Yun; Lee, Kang Hyuck; Kim, Tae-Ho; Cimini, Francesco Arturo; Kim, Sung Kyun; Hinchet, Ronan; Kim, Sang-Woo; Falconi, Christian

2017-06-01

Gates can electrostatically control charges inside two-dimensional materials. However, integrating independent gates typically requires depositing and patterning suitable insulators and conductors. Moreover, after manufacturing, gates are unchangeable. Here we introduce tunnelling triboelectrification for localizing electric charges in very close proximity of two-dimensional materials. As representative materials, we use chemical vapour deposition graphene deposited on a SiO2/Si substrate. The triboelectric charges, generated by friction with a Pt-coated atomic force microscope tip and injected through defects, are trapped at the air-SiO2 interface underneath graphene and act as ghost floating gates. Tunnelling triboelectrification uniquely permits to create, modify and destroy p and n regions at will with the spatial resolution of atomic force microscopes. As a proof of concept, we draw rewritable p/n+ and p/p+ junctions with resolutions as small as 200 nm. Our results open the way to time-variant two-dimensional electronics where conductors, p and n regions can be defined on demand.

Thermometry of Silicon Nanoparticles

NASA Astrophysics Data System (ADS)

Mecklenburg, Matthew; Zutter, Brian; Regan, B. C.

2018-01-01

Current thermometry techniques lack the spatial resolution required to see the temperature gradients in typical, highly scaled modern transistors. As a step toward addressing this problem, we measure the temperature dependence of the volume plasmon energy in silicon nanoparticles from room temperature to 1250 °C , using a chip-style heating sample holder in a scanning transmission electron microscope (STEM) equipped with electron energy loss spectroscopy (EELS). The plasmon energy changes as expected for an electron gas subject to the thermal expansion of silicon. Reversing this reasoning, we find that measurements of the plasmon energy provide an independent measure of the nanoparticle temperature consistent with that of the heater chip's macroscopic, dual-function heater-and-thermometer to within the 5% accuracy of the thermometer's calibration. Thus, silicon has the potential to provide its own high-spatial-resolution thermometric readout signal via measurements of its volume plasmon energy. Furthermore, nanoparticles can, in general, serve as convenient nanothermometers for in situ electron-microscopy experiments.

Microscopic Views of Martian Soils and Evidence for Incipient Diagenesis

NASA Technical Reports Server (NTRS)

Goetz, W.; Madsen, M. B.; Bridges, N.; Clark, B.; Edgett, K. S.; Fisk, M.; Grotzinger, J. P.; Hviid, S. F.; Meslin, P.-Y.; Ming, D. W.;

Three-dimensional localization of nanoscale battery reactions using soft X-ray tomography.

PubMed

Yu, Young-Sang; Farmand, Maryam; Kim, Chunjoong; Liu, Yijin; Grey, Clare P; Strobridge, Fiona C; Tyliszczak, Tolek; Celestre, Rich; Denes, Peter; Joseph, John; Krishnan, Harinarayan; Maia, Filipe R N C; Kilcoyne, A L David; Marchesini, Stefano; Leite, Talita Perciano Costa; Warwick, Tony; Padmore, Howard; Cabana, Jordi; Shapiro, David A

2018-03-02

Battery function is determined by the efficiency and reversibility of the electrochemical phase transformations at solid electrodes. The microscopic tools available to study the chemical states of matter with the required spatial resolution and chemical specificity are intrinsically limited when studying complex architectures by their reliance on two-dimensional projections of thick material. Here, we report the development of soft X-ray ptychographic tomography, which resolves chemical states in three dimensions at 11 nm spatial resolution. We study an ensemble of nano-plates of lithium iron phosphate extracted from a battery electrode at 50% state of charge. Using a set of nanoscale tomograms, we quantify the electrochemical state and resolve phase boundaries throughout the volume of individual nanoparticles. These observations reveal multiple reaction points, intra-particle heterogeneity, and size effects that highlight the importance of multi-dimensional analytical tools in providing novel insight to the design of the next generation of high-performance devices.

Distinguishing Individual DNA Bases in a Network by Non-Resonant Tip-Enhanced Raman Scattering.

PubMed

Zhang, Rui; Zhang, Xianbiao; Wang, Huifang; Zhang, Yao; Jiang, Song; Hu, Chunrui; Zhang, Yang; Luo, Yi; Dong, Zhenchao

2017-05-08

The importance of identifying DNA bases at the single-molecule level is well recognized for many biological applications. Although such identification can be achieved by electrical measurements using special setups, it is still not possible to identify single bases in real space by optical means owing to the diffraction limit. Herein, we demonstrate the outstanding ability of scanning tunneling microscope (STM)-controlled non-resonant tip-enhanced Raman scattering (TERS) to unambiguously distinguish two individual complementary DNA bases (adenine and thymine) with a spatial resolution down to 0.9 nm. The distinct Raman fingerprints identified for the two molecules allow to differentiate in real space individual DNA bases in coupled base pairs. The demonstrated ability of non-resonant Raman scattering with super-high spatial resolution will significantly extend the applicability of TERS, opening up new routes for single-molecule DNA sequencing. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

Damage-free vibrational spectroscopy of biological materials in the electron microscope

PubMed Central

Rez, Peter; Aoki, Toshihiro; March, Katia; Gur, Dvir; Krivanek, Ondrej L.; Dellby, Niklas; Lovejoy, Tracy C.; Wolf, Sharon G.; Cohen, Hagai

2016-01-01

Vibrational spectroscopy in the electron microscope would be transformative in the study of biological samples, provided that radiation damage could be prevented. However, electron beams typically create high-energy excitations that severely accelerate sample degradation. Here this major difficulty is overcome using an ‘aloof' electron beam, positioned tens of nanometres away from the sample: high-energy excitations are suppressed, while vibrational modes of energies <1 eV can be ‘safely' investigated. To demonstrate the potential of aloof spectroscopy, we record electron energy loss spectra from biogenic guanine crystals in their native state, resolving their characteristic C–H, N–H and C=O vibrational signatures with no observable radiation damage. The technique opens up the possibility of non-damaging compositional analyses of organic functional groups, including non-crystalline biological materials, at a spatial resolution of ∼10 nm, simultaneously combined with imaging in the electron microscope. PMID:26961578

Damage-free vibrational spectroscopy of biological materials in the electron microscope.

PubMed

Rez, Peter; Aoki, Toshihiro; March, Katia; Gur, Dvir; Krivanek, Ondrej L; Dellby, Niklas; Lovejoy, Tracy C; Wolf, Sharon G; Cohen, Hagai

2016-03-10

Vibrational spectroscopy in the electron microscope would be transformative in the study of biological samples, provided that radiation damage could be prevented. However, electron beams typically create high-energy excitations that severely accelerate sample degradation. Here this major difficulty is overcome using an 'aloof' electron beam, positioned tens of nanometres away from the sample: high-energy excitations are suppressed, while vibrational modes of energies <1 eV can be 'safely' investigated. To demonstrate the potential of aloof spectroscopy, we record electron energy loss spectra from biogenic guanine crystals in their native state, resolving their characteristic C-H, N-H and C=O vibrational signatures with no observable radiation damage. The technique opens up the possibility of non-damaging compositional analyses of organic functional groups, including non-crystalline biological materials, at a spatial resolution of ∼10 nm, simultaneously combined with imaging in the electron microscope.

Design and properties of a cryogenic dip-stick scanning tunneling microscope with capacitive coarse approach control.

PubMed

Schlegel, R; Hänke, T; Baumann, D; Kaiser, M; Nag, P K; Voigtländer, R; Lindackers, D; Büchner, B; Hess, C

2014-01-01

We present the design, setup, and operation of a new dip-stick scanning tunneling microscope. Its special design allows measurements in the temperature range from 4.7 K up to room temperature, where cryogenic vacuum conditions are maintained during the measurement. The system fits into every (4)He vessel with a bore of 50 mm, e.g., a transport dewar or a magnet bath cryostat. The microscope is equipped with a cleaving mechanism for cleaving single crystals in the whole temperature range and under cryogenic vacuum conditions. For the tip approach, a capacitive automated coarse approach is implemented. We present test measurements on the charge density wave system 2H-NbSe2 and the superconductor LiFeAs which demonstrate scanning tunneling microscopy and spectroscopy data acquisition with high stability, high spatial resolution at variable temperatures and in high magnetic fields.

Damage-free vibrational spectroscopy of biological materials in the electron microscope

DOE PAGES

Rez, Peter; Aoki, Toshihiro; March, Katia; ...

2016-03-10

Vibrational spectroscopy in the electron microscope would be transformative in the study of biological samples, provided that radiation damage could be prevented. However, electron beams typically create high-energy excitations that severely accelerate sample degradation. Here this major difficulty is overcome using an ‘aloof’ electron beam, positioned tens of nanometres away from the sample: high-energy excitations are suppressed, while vibrational modes of energies o1 eV can be ‘safely’ investigated. To demonstrate the potential of aloof spectroscopy, we record electron energy loss spectra from biogenic guanine crystals in their native state, resolving their characteristic C–H, N–H and C=O vibrational signatures with nomore » observable radiation damage. Furthermore, the technique opens up the possibility of non-damaging compositional analyses of organic functional groups, including non-crystalline biological materials, at a spatial resolution of ~10nm, simultaneously combined with imaging in the electron microscope.« less

Damage-free vibrational spectroscopy of biological materials in the electron microscope

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

Rez, Peter; Aoki, Toshihiro; March, Katia

Vibrational spectroscopy in the electron microscope would be transformative in the study of biological samples, provided that radiation damage could be prevented. However, electron beams typically create high-energy excitations that severely accelerate sample degradation. Here this major difficulty is overcome using an ‘aloof’ electron beam, positioned tens of nanometres away from the sample: high-energy excitations are suppressed, while vibrational modes of energies o1 eV can be ‘safely’ investigated. To demonstrate the potential of aloof spectroscopy, we record electron energy loss spectra from biogenic guanine crystals in their native state, resolving their characteristic C–H, N–H and C=O vibrational signatures with nomore » observable radiation damage. Furthermore, the technique opens up the possibility of non-damaging compositional analyses of organic functional groups, including non-crystalline biological materials, at a spatial resolution of ~10nm, simultaneously combined with imaging in the electron microscope.« less

Atomic-Resolution Spectrum Imaging of Semiconductor Nanowires.

PubMed

Zamani, Reza R; Hage, Fredrik S; Lehmann, Sebastian; Ramasse, Quentin M; Dick, Kimberly A

2018-03-14

Over the past decade, III-V heterostructure nanowires have attracted a surge of attention for their application in novel semiconductor devices such as tunneling field-effect transistors (TFETs). The functionality of such devices critically depends on the specific atomic arrangement at the semiconductor heterointerfaces. However, most of the currently available characterization techniques lack sufficient spatial resolution to provide local information on the atomic structure and composition of these interfaces. Atomic-resolution spectrum imaging by means of electron energy-loss spectroscopy (EELS) in the scanning transmission electron microscope (STEM) is a powerful technique with the potential to resolve structure and chemical composition with sub-angstrom spatial resolution and to provide localized information about the physical properties of the material at the atomic scale. Here, we demonstrate the use of atomic-resolution EELS to understand the interface atomic arrangement in three-dimensional heterostructures in semiconductor nanowires. We observed that the radial interfaces of GaSb-InAs heterostructure nanowires are atomically abrupt, while the axial interface in contrast consists of an interfacial region where intermixing of the two compounds occurs over an extended spatial region. The local atomic configuration affects the band alignment at the interface and, hence, the charge transport properties of devices such as GaSb-InAs nanowire TFETs. STEM-EELS thus represents a very promising technique for understanding nanowire physical properties, such as differing electrical behavior across the radial and axial heterointerfaces of GaSb-InAs nanowires for TFET applications.

Lensfree super-resolution holographic microscopy using wetting films on a chip

NASA Astrophysics Data System (ADS)

Mudanyali, Onur; Bishara, Waheb; Ozcan, Aydogan

2011-08-01

We investigate the use of wetting films to significantly improve the imaging performance of lensfree pixel super-resolution on-chip microscopy, achieving < 1 μm spatial resolution over a large imaging area of ~24 mm2. Formation of an ultra-thin wetting film over the specimen effectively creates a micro-lens effect over each object, which significantly improves the signal-to-noise-ratio and therefore the resolution of our lensfree images. We validate the performance of this approach through lensfree on-chip imaging of various objects having fine morphological features (with dimensions of e.g., ≤0.5 μm) such as Escherichia coli (E. coli), human sperm, Giardia lamblia trophozoites, polystyrene micro beads as well as red blood cells. These results are especially important for the development of highly sensitive field-portable microscopic analysis tools for resource limited settings.

Polarization- and wavelength-resolved near-field imaging of complex plasmonic modes in Archimedean nanospirals

DOE PAGES

Hachtel, Jordan A.; Davidson, II, Roderick B.; Kovalik, Elena R.; ...

2018-02-15

Asymmetric nanophotonic structures enable a wide range of opportunities in optical nanotechnology because they support efficient optical nonlinearities mediated by multiple plasmon resonances over a broad spectral range. The Archimedean nanospiral is a canonical example of a chiral plasmonic structure because it supports even-order nonlinearities that are not generally accessible in locally symmetric geometries. However, the complex spiral response makes nanoscale experimental characterization of the plasmonic near-field structure highly desirable. As a result, we employ high-efficiency, high-spatial-resolution cathodoluminescence imaging in a scanning transmission electron microscope to describe the spatial, spectral, and polarization response of plasmon modes in the nanospiral geometry.

Polarization- and wavelength-resolved near-field imaging of complex plasmonic modes in Archimedean nanospirals

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

Hachtel, Jordan A.; Davidson, II, Roderick B.; Kovalik, Elena R.

Asymmetric nanophotonic structures enable a wide range of opportunities in optical nanotechnology because they support efficient optical nonlinearities mediated by multiple plasmon resonances over a broad spectral range. The Archimedean nanospiral is a canonical example of a chiral plasmonic structure because it supports even-order nonlinearities that are not generally accessible in locally symmetric geometries. However, the complex spiral response makes nanoscale experimental characterization of the plasmonic near-field structure highly desirable. As a result, we employ high-efficiency, high-spatial-resolution cathodoluminescence imaging in a scanning transmission electron microscope to describe the spatial, spectral, and polarization response of plasmon modes in the nanospiral geometry.

Quantitative Imaging with a Mobile Phone Microscope

PubMed Central

Skandarajah, Arunan; Reber, Clay D.; Switz, Neil A.; Fletcher, Daniel A.

2014-01-01

Use of optical imaging for medical and scientific applications requires accurate quantification of features such as object size, color, and brightness. High pixel density cameras available on modern mobile phones have made photography simple and convenient for consumer applications; however, the camera hardware and software that enables this simplicity can present a barrier to accurate quantification of image data. This issue is exacerbated by automated settings, proprietary image processing algorithms, rapid phone evolution, and the diversity of manufacturers. If mobile phone cameras are to live up to their potential to increase access to healthcare in low-resource settings, limitations of mobile phone–based imaging must be fully understood and addressed with procedures that minimize their effects on image quantification. Here we focus on microscopic optical imaging using a custom mobile phone microscope that is compatible with phones from multiple manufacturers. We demonstrate that quantitative microscopy with micron-scale spatial resolution can be carried out with multiple phones and that image linearity, distortion, and color can be corrected as needed. Using all versions of the iPhone and a selection of Android phones released between 2007 and 2012, we show that phones with greater than 5 MP are capable of nearly diffraction-limited resolution over a broad range of magnifications, including those relevant for single cell imaging. We find that automatic focus, exposure, and color gain standard on mobile phones can degrade image resolution and reduce accuracy of color capture if uncorrected, and we devise procedures to avoid these barriers to quantitative imaging. By accommodating the differences between mobile phone cameras and the scientific cameras, mobile phone microscopes can be reliably used to increase access to quantitative imaging for a variety of medical and scientific applications. PMID:24824072

Electron beam dynamics in an ultrafast transmission electron microscope with Wehnelt electrode.

PubMed

Bücker, K; Picher, M; Crégut, O; LaGrange, T; Reed, B W; Park, S T; Masiel, D J; Banhart, F

2016-12-01

High temporal resolution transmission electron microscopy techniques have shown significant progress in recent years. Using photoelectron pulses induced by ultrashort laser pulses on the cathode, these methods can probe ultrafast materials processes and have revealed numerous dynamic phenomena at the nanoscale. Most recently, the technique has been implemented in standard thermionic electron microscopes that provide a flexible platform for studying material's dynamics over a wide range of spatial and temporal scales. In this study, the electron pulses in such an ultrafast transmission electron microscope are characterized in detail. The microscope is based on a thermionic gun with a Wehnelt electrode and is operated in a stroboscopic photoelectron mode. It is shown that the Wehnelt bias has a decisive influence on the temporal and energy spread of the picosecond electron pulses. Depending on the shape of the cathode and the cathode-Wehnelt distance, different emission patterns with different pulse parameters are obtained. The energy spread of the pulses is determined by space charge and Boersch effects, given by the number of electrons in a pulse. However, filtering effects due to the chromatic aberrations of the Wehnelt electrode allow the extraction of pulses with narrow energy spreads. The temporal spread is governed by electron trajectories of different length and in different electrostatic potentials. High temporal resolution is obtained by excluding shank emission from the cathode and aberration-induced halos in the emission pattern. By varying the cathode-Wehnelt gap, the Wehnelt bias, and the number of photoelectrons in a pulse, tradeoffs between energy and temporal resolution as well as beam intensity can be made as needed for experiments. Based on the characterization of the electron pulses, the optimal conditions for the operation of ultrafast TEMs with thermionic gun assembly are elaborated. Copyright © 2016 Elsevier B.V. All rights reserved.

Data-driven signal-resolving approaches of infrared spectra to explore the macroscopic and microscopic spatial distribution of organic and inorganic compounds in plant.

PubMed

Chen, Jian-bo; Sun, Su-qin; Zhou, Qun

2015-07-01

The nondestructive and label-free infrared (IR) spectroscopy is a direct tool to characterize the spatial distribution of organic and inorganic compounds in plant. Since plant samples are usually complex mixtures, signal-resolving methods are necessary to find the spectral features of compounds of interest in the signal-overlapped IR spectra. In this research, two approaches using existing data-driven signal-resolving methods are proposed to interpret the IR spectra of plant samples. If the number of spectra is small, "tri-step identification" can enhance the spectral resolution to separate and identify the overlapped bands. First, the envelope bands of the original spectrum are interpreted according to the spectra-structure correlations. Then the spectrum is differentiated to resolve the underlying peaks in each envelope band. Finally, two-dimensional correlation spectroscopy is used to enhance the spectral resolution further. For a large number of spectra, "tri-step decomposition" can resolve the spectra by multivariate methods to obtain the structural and semi-quantitative information about the chemical components. Principal component analysis is used first to explore the existing signal types without any prior knowledge. Then the spectra are decomposed by self-modeling curve resolution methods to estimate the spectra and contents of significant chemical components. At last, targeted methods such as partial least squares target can explore the content profiles of specific components sensitively. As an example, the macroscopic and microscopic distribution of eugenol and calcium oxalate in the bud of clove is studied.

Optical magnetic imaging of living cells

PubMed Central

Le Sage, D.; Arai, K.; Glenn, D. R.; DeVience, S. J.; Pham, L. M.; Rahn-Lee, L.; Lukin, M. D.; Yacoby, A.; Komeili, A.; Walsworth, R. L.

2013-01-01

Magnetic imaging is a powerful tool for probing biological and physical systems. However, existing techniques either have poor spatial resolution compared to optical microscopy and are hence not generally applicable to imaging of sub-cellular structure (e.g., magnetic resonance imaging [MRI]1), or entail operating conditions that preclude application to living biological samples while providing sub-micron resolution (e.g., scanning superconducting quantum interference device [SQUID] microscopy2, electron holography3, and magnetic resonance force microscopy [MRFM]4). Here we demonstrate magnetic imaging of living cells (magnetotactic bacteria) under ambient laboratory conditions and with sub-cellular spatial resolution (400 nm), using an optically-detected magnetic field imaging array consisting of a nanoscale layer of nitrogen-vacancy (NV) colour centres implanted at the surface of a diamond chip. With the bacteria placed on the diamond surface, we optically probe the NV quantum spin states and rapidly reconstruct images of the vector components of the magnetic field created by chains of magnetic nanoparticles (magnetosomes) produced in the bacteria, and spatially correlate these magnetic field maps with optical images acquired in the same apparatus. Wide-field sCMOS acquisition allows parallel optical and magnetic imaging of multiple cells in a population with sub-micron resolution and >100 micron field-of-view. Scanning electron microscope (SEM) images of the bacteria confirm that the correlated optical and magnetic images can be used to locate and characterize the magnetosomes in each bacterium. The results provide a new capability for imaging bio-magnetic structures in living cells under ambient conditions with high spatial resolution, and will enable the mapping of a wide range of magnetic signals within cells and cellular networks5, 6. PMID:23619694

Removal of anti-Stokes emission background in STED microscopy by FPGA-based synchronous detection

NASA Astrophysics Data System (ADS)

Castello, M.; Tortarolo, G.; Coto Hernández, I.; Deguchi, T.; Diaspro, A.; Vicidomini, G.

2017-05-01

In stimulated emission depletion (STED) microscopy, the role of the STED beam is to de-excite, via stimulated emission, the fluorophores that have been previously excited by the excitation beam. This condition, together with specific beam intensity distributions, allows obtaining true sub-diffraction spatial resolution images. However, if the STED beam has a non-negligible probability to excite the fluorophores, a strong fluorescent background signal (anti-Stokes emission) reduces the effective resolution. For STED scanning microscopy, different synchronous detection methods have been proposed to remove this anti-Stokes emission background and recover the resolution. However, every method works only for a specific STED microscopy implementation. Here we present a user-friendly synchronous detection method compatible with any STED scanning microscope. It exploits a data acquisition (DAQ) card based on a field-programmable gate array (FPGA), which is progressively used in STED microscopy. In essence, the FPGA-based DAQ card synchronizes the fluorescent signal registration, the beam deflection, and the excitation beam interruption, providing a fully automatic pixel-by-pixel synchronous detection method. We validate the proposed method in both continuous wave and pulsed STED microscope systems.

Fluorescence advantages with microscopic spatiotemporal control

NASA Astrophysics Data System (ADS)

Goswami, Debabrata; Roy, Debjit; De, Arijit K.

2013-03-01

We present a clever design concept of using femtosecond laser pulses in microscopy by selective excitation or de-excitation of one fluorophore over the other overlapping one. Using either a simple pair of femtosecond pulses with variable delay or using a train of laser pulses at 20-50 Giga-Hertz excitation, we show controlled fluorescence excitation or suppression of one of the fluorophores with respect to the other through wave-packet interference, an effect that prevails even after the fluorophore coherence timescale. Such an approach can be used both under the single-photon excitation as well as in the multi-photon excitation conditions resulting in effective higher spatial resolution. Such high spatial resolution advantage with broadband-pulsed excitation is of immense benefit to multi-photon microscopy and can also be an effective detection scheme for trapped nanoparticles with near-infrared light. Such sub-diffraction limit trapping of nanoparticles is challenging and a two-photon fluorescence diagnostics allows a direct observation of a single nanoparticle in a femtosecond high-repetition rate laser trap, which promises new directions to spectroscopy at the single molecule level in solution. The gigantic peak power of femtosecond laser pulses at high repetition rate, even at low average powers, provide huge instantaneous gradient force that most effectively result in a stable optical trap for spatial control at sub-diffraction limit. Such studies have also enabled us to explore simultaneous control of internal and external degrees of freedom that require coupling of various control parameters to result in spatiotemporal control, which promises to be a versatile tool for the microscopic world.

Two-photon polymerization microfabrication of hydrogels: an advanced 3D printing technology for tissue engineering and drug delivery.

PubMed

Xing, Jin-Feng; Zheng, Mei-Ling; Duan, Xuan-Ming

2015-08-07

3D printing technology has attracted much attention due to its high potential in scientific and industrial applications. As an outstanding 3D printing technology, two-photon polymerization (TPP) microfabrication has been applied in the fields of micro/nanophotonics, micro-electromechanical systems, microfluidics, biomedical implants and microdevices. In particular, TPP microfabrication is very useful in tissue engineering and drug delivery due to its powerful fabrication capability for precise microstructures with high spatial resolution on both the microscopic and the nanometric scale. The design and fabrication of 3D hydrogels widely used in tissue engineering and drug delivery has been an important research area of TPP microfabrication. The resolution is a key parameter for 3D hydrogels to simulate the native 3D environment in which the cells reside and the drug is controlled to release with optimal temporal and spatial distribution in vitro and in vivo. The resolution of 3D hydrogels largely depends on the efficiency of TPP initiators. In this paper, we will review the widely used photoresists, the development of TPP photoinitiators, the strategies for improving the resolution and the microfabrication of 3D hydrogels.

Novel detectors for silicon based microdosimetry, their concepts and applications

NASA Astrophysics Data System (ADS)

Rosenfeld, Anatoly B.

2016-02-01

This paper presents an overview of the development of semiconductor microdosimetry and the most current (state-of-the-art) Silicon on Insulator (SOI) detectors for microdosimetry based mainly on research and development carried out at the Centre for Medical Radiation Physics (CMRP) at the University of Wollongong with collaborators over the last 18 years. In this paper every generation of CMRP SOI microdosimeters, including their fabrication, design, and electrical and charge collection characterisation are presented. A study of SOI microdosimeters in various radiation fields has demonstrated that under appropriate geometrical scaling, the response of SOI detectors with the well-known geometry of microscopically sensitive volumes will record the energy deposition spectra representative of tissue cells of an equivalent shape. This development of SOI detectors for microdosimetry with increased complexity has improved the definition of microscopic sensitive volume (SV), which is modelling the deposition of ionising energy in a biological cell, that are led from planar to 3D SOI detectors with an array of segmented microscopic 3D SVs. The monolithic ΔE-E silicon telescope, which is an alternative to the SOI silicon microdosimeter, is presented, and as an example, applications of SOI detectors and ΔE-E monolithic telescope for microdosimetery in proton therapy field and equivalent neutron dose measurements out of field are also presented. An SOI microdosimeter "bridge" with 3D SVs can derive the relative biological effectiveness (RBE) in 12C ion radiation therapy that matches the tissue equivalent proportional counter (TEPC) quite well, but with outstanding spatial resolution. The use of SOI technology in experimental microdosimetry offers simplicity (no gas system or HV supply), high spatial resolution, low cost, high count rates, and the possibility of integrating the system onto a single device with other types of detectors.

High temporal and spatial resolution studies of bone cells using real-time confocal reflection microscopy.

PubMed

Boyde, A; Vesely, P; Gray, C; Jones, S J

1994-01-01

Chick and rat bone-derived cells were mounted in sealed coverslip-covered chambers; individual osteoclasts (but also osteoblasts) were selected and studied at 37 degrees C using three different types of high-speed scanning confocal microscopes: (1) A Noran Tandem Scanning Microscope (TSM) was used with a low light level, cooled CCD camera for image transfer to a Noran TN8502 frame store-based image analysing computer to make time lapse movie sequences using 0.1 s exposure periods, thus losing some of the advantage of the high frame rate of the TSM. Rapid focus adjustment using computer controlled piezo drivers permitted two or more focus planes to be imaged sequentially: thus (with additional light-source shuttering) the reflection confocal image could be alternated with the phase contrast image at a different focus. Individual cells were followed for up to 5 days, suggesting no significant irradiation problem. (2) Exceptional temporal and spatial resolution is available in video rate laser confocal scanning microscopes (VRCSLMs). We used the Noran Odyssey unitary beam VRCSLM with an argon ion laser at 488 nm and acousto-optic deflection (AOD) on the line axis: this instrument is truly and adjustably confocal in the reflection mode. (3) We also used the Lasertec 1LM11 line scan instrument, with an He-Ne laser at 633 nm, and AOD for the frame scan. We discuss the technical problems and merits of the different approaches. The VRCSLMs documented rapid, real-time oscillatory motion: all the methods used show rapid net movement of organelles within bone cells. The interference reflection mode gives particularly strong contrasts in confocal instruments. Phase contrast and other interference methods used in the microscopy of living cells can be used simultaneously in the TSM.

Highly charged ion based time of flight emission microscope

DOEpatents

Barnes, Alan V.; Schenkel, Thomas; Hamza, Alex V.; Schneider, Dieter H.; Doyle, Barney

2001-01-01

A highly charged ion based time-of-flight emission microscope has been designed, which improves the surface sensitivity of static SIMS measurements because of the higher ionization probability of highly charged ions. Slow, highly charged ions are produced in an electron beam ion trap and are directed to the sample surface. The sputtered secondary ions and electrons pass through a specially designed objective lens to a microchannel plate detector. This new instrument permits high surface sensitivity (10.sup.10 atoms/cm.sup.2), high spatial resolution (100 nm), and chemical structural information due to the high molecular ion yields. The high secondary ion yield permits coincidence counting, which can be used to enhance determination of chemical and topological structure and to correlate specific molecular species.

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

NASA Astrophysics Data System (ADS)

Shin, Sanghoon; Yu, Younghun

2018-04-01

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

Electronic structure of polycrystalline CVD-graphene revealed by Nano-ARPES

NASA Astrophysics Data System (ADS)

Chen, Chaoyu; Avila, José; Asensio, Maria C.

2017-06-01

The ability to explore electronic structure and their role in determining material’s macroscopic behaviour is essential to explain and engineer functions of material and device. Since its debut in 2004, graphene has attracted global research interest due to its unique properties. Chemical vapor deposition (CVD) has emerged as an important method for the massive preparation and production of graphene for various applications. Here by employing angle-resolved photoemission spectroscopy with nanoscale spatial resolution ˜ 100 nm (Nano-ARPES), we describe the approach to measure the electronic structure of polycrystalline graphene on copper foils, demonstrating the power of Nano-ARPES to detect the electronic structure of microscopic single crystalline domains, being fully compatible with conventional ARPES. Similar analysis could be employed to other microscopic materials

Electric field stimulation setup for photoemission electron microscopes.

PubMed

Buzzi, M; Vaz, C A F; Raabe, J; Nolting, F

2015-08-01

Manipulating magnetisation by the application of an electric field in magnetoelectric multiferroics represents a timely issue due to the potential applications in low power electronics and the novel physics involved. Thanks to its element sensitivity and high spatial resolution, X-ray photoemission electron microscopy is a uniquely suited technique for the investigation of magnetoelectric coupling in multiferroic materials. In this work, we present a setup that allows for the application of in situ electric and magnetic fields while the sample is analysed in the microscope. As an example of the performances of the setup, we present measurements on Ni/Pb(Mg(0.66)Nb(0.33))O3-PbTiO3 and La(0.7)Sr(0.3)MnO3/PMN-PT artificial multiferroic nanostructures.

Neutron Microtomography of MgB2 Superconducting Multifilament Wire

NASA Astrophysics Data System (ADS)

Trtik, Pavel; Scheuerlein, Christian; Alknes, Patrick; Meyer, Michael; Schmid, Florian; Lehmann, Eberhard

Neutron imaging of sub-10-micrometres spatial resolution has been recently achieved in 2D mode within the framework of the Neutron Microscope project at the Paul Scherrer Institut. Here we report on the development of the PSI Neutron Microscope instrument and the results of the first microtomographic imaging experiment of multifilament superconducting MgB2 wire. The sample of MgB2 superconducting 37 multifilaments embedded in copper-nickel matrix was investigated -in microtomographic mode- with the scientific interest regarding the distribution of boron within the individual superconducting filaments (about 40 μm in diameter). The resulting tomographic dataset revealed the distribution of boron within the entire 0.8 mm thick multifilamental wire with the isotropic voxel size of 2.6 micrometres.

Multiplexed phase-space imaging for 3D fluorescence microscopy.

PubMed

Liu, Hsiou-Yuan; Zhong, Jingshan; Waller, Laura

2017-06-26

Optical phase-space functions describe spatial and angular information simultaneously; examples of optical phase-space functions include light fields in ray optics and Wigner functions in wave optics. Measurement of phase-space enables digital refocusing, aberration removal and 3D reconstruction. High-resolution capture of 4D phase-space datasets is, however, challenging. Previous scanning approaches are slow, light inefficient and do not achieve diffraction-limited resolution. Here, we propose a multiplexed method that solves these problems. We use a spatial light modulator (SLM) in the pupil plane of a microscope in order to sequentially pattern multiplexed coded apertures while capturing images in real space. Then, we reconstruct the 3D fluorescence distribution of our sample by solving an inverse problem via regularized least squares with a proximal accelerated gradient descent solver. We experimentally reconstruct a 101 Megavoxel 3D volume (1010×510×500µm with NA 0.4), demonstrating improved acquisition time, light throughput and resolution compared to scanning aperture methods. Our flexible patterning scheme further allows sparsity in the sample to be exploited for reduced data capture.

Microscopic resolution broadband dielectric spectroscopy

NASA Astrophysics Data System (ADS)

Mukherjee, S.; Watson, P.; Prance, R. J.

2011-08-01

Results are presented for a non-contact measurement system capable of micron level spatial resolution. It utilises the novel electric potential sensor (EPS) technology, invented at Sussex, to image the electric field above a simple composite dielectric material. EP sensors may be regarded as analogous to a magnetometer and require no adjustments or offsets during either setup or use. The sample consists of a standard glass/epoxy FR4 circuit board, with linear defects machined into the surface by a PCB milling machine. The sample is excited with an a.c. signal over a range of frequencies from 10 kHz to 10 MHz, from the reverse side, by placing it on a conducting sheet connected to the source. The single sensor is raster scanned over the surface at a constant working distance, consistent with the spatial resolution, in order to build up an image of the electric field, with respect to the reference potential. The results demonstrate that both the surface defects and the internal dielectric variations within the composite may be imaged in this way, with good contrast being observed between the glass mat and the epoxy resin.

Examining nanoparticle assemblies using high spatial resolution x-ray microtomography

NASA Astrophysics Data System (ADS)

Jenneson, P. M.; Luggar, R. D.; Morton, E. J.; Gundogdu, O.; Tüzün, U.

2004-09-01

An experimental system has been designed to examine the assembly of nanoparticles in a variety of process engineering applications. These applications include the harvesting from solutions of nanoparticles into green parts, and the subsequent sintering into finished components. The system is based on an x-ray microtomography with a spatial resolution down to 5μm. The theoretical limitations in x-ray imaging are considered to allow experimental optimization. A standard nondestructive evaluation type apparatus with a small focal-spot x-ray tube, high-resolution complementary metal oxide semiconductor flat-panel pixellated detector, and a mechanical rotational stage is used to image the static systems. Dynamic sintering processes are imaged using the same x-ray source and detector but a custom rotational stage which is contained in an environmental chamber where the temperature, atmospheric pressure, and compaction force can be controlled. Three-dimensional tomographic data sets are presented here for samples from the pharmaceutical, nutraceutical, biotechnology, and nanoparticle handling industries and show the microscopic features and defects which can be resolved with the system.

Multimodal second harmonic generation and two photon fluorescence imaging of microdomain calcium contraction coupling in single cardiomyocytes

NASA Astrophysics Data System (ADS)

Chan, James; Awasthi, Samir; Izu, Leighton; Mao, Ziliang; Jian, Zhong; Landas, Trevor; Lerner, Aaron; Shimkunas, Rafael; Woldeyesus, Rahwa; Bossuyt, Julie; Wood, Brittani; Chen, Yi-Je; Matthews, Dennis; Lieu, Deborah; Chiamvimonvat, Nipavan; Lam, Kit; Chen-Izu, Ye

2016-11-01

The objective of this study was to develop a method for simultaneously measuring the calcium and contraction dynamics of single, live cardiomyocytes at high spatial resolutions. Such measurements are important to investigate local calcium release and the mechanical response at the sarcomere level (i.e. the basic unit of contraction), which have important implications in cardiac dysfunction and arrhythmias in conditions such as hypertension, atrial fibrillation, and myocardial infarction. Here, we describe a multimodal second harmonic generation (SHG) and two photon fluorescence (2PF) microscopy technique that is used to simultaneously measure subsarcomere calcium and contraction events at high spatial and temporal resolutions. The method takes advantage of the label-free nature of SHG for imaging the sarcomeres and the high spatial colocalization of the SHG signal and the fluorescence signal excited from calcium indicators. This microscope was used to measure calcium sparks and waves and associated contractions in subcellular microdomains, leading to the generation of subcellular strain. We anticipate this new imaging tool will play an important role in studying mechanical stress-induced heart disease.

Morphological imaging and quantification of axial xylem tissue in Fraxinus excelsior L. through X-ray micro-computed tomography.

PubMed

Koddenberg, Tim; Militz, Holger

2018-05-05

The popularity of X-ray based imaging methods has continued to increase in research domains. In wood research, X-ray micro-computed tomography (XμCT) is useful for structural studies examining the three-dimensional and complex xylem tissue of trees qualitatively and quantitatively. In this study, XμCT made it possible to visualize and quantify the spatial xylem organization of the angiosperm species Fraxinus excelsior L. on the microscopic level. Through image analysis, it was possible to determine morphological characteristics of the cellular axial tissue (vessel elements, fibers, and axial parenchyma cells) three-dimensionally. X-ray imaging at high resolutions provides very distinct visual insight into the xylem structure. Numerical analyses performed through semi-automatic procedures made it possible to quickly quantify cell characteristics (length, diameter, and volume of cells). Use of various spatial resolutions (0.87-5 μm) revealed boundaries users should be aware of. Nevertheless, our findings, both qualitative and quantitative, demonstrate XμCT to be a valuable tool for studying the spatial cell morphology of F. excelsior. Copyright © 2018. Published by Elsevier Ltd.

[Detection of single-walled carbon nanotube bundles by tip-enhanced Raman spectroscopy].

PubMed

Wu, Xiao-Bin; Wang, Jia; Wang, Rui; Xu, Ji-Ying; Tian, Qian; Yu, Jian-Yuan

2009-10-01

Raman spectroscopy is a powerful technique in the characterization of carbon nanotubes (CNTs). However, this spectral method is subject to two obstacles. One is spatial resolution, namely the diffraction limits of light, and the other is its inherent small Raman cross section and weak signal. To resolve these problems, a new approach has been developed, denoted tip-enhanced Raman spectroscopy (TERS). TERS has been demonstrated to be a powerful spectroscopic and microscopic technique to characterize nanomaterial or nanostructures. Excited by a focused laser beam, an enhanced electric field is generated in the vicinity of a metallic tip because of the surface plasmon polariton (SPP) and lightening rod effect. Consequently, Raman signal from the sample area illuminated by the enhanced field nearby the tip is enhanced. At the same time, the topography is obtained in the nanometer scale. The exact corresponding relationship between the localized Raman and the topography makes the Raman identification at the nanometer scale to be feasible. In the present paper, based on an inverted microscope and a metallic AFM tip, a tip-enhanced Raman system was set up. The radius of the Au-coated metallic tip is about 30 nm. The 532 nm laser passes through a high numerical objective (NA0.95) from the bottom to illuminate the tip to excite the enhanced electric field. Corresponding with the AFM image, the tip-enhanced near-field Raman of a 100 nm diameter single-walled carbon nanotube (SWNT) bundles was obtained. The SWNTs were prepared by arc method. Furthermore, the near-field Raman of about 3 SWNTs of the bundles was received with the spatial resolution beyond the diffraction limit. Compared with the far-field Raman, the enhancement factor of the tip-enhanced Raman is more than 230. With the super-diffraction spatial resolution and the tip-enhanced Raman ability, tip-enhanced Raman spectroscopy will play an important role in the nano-material and nano-structure characterization.

Combined Atomic Force Microscope-Based Topographical Imaging and Nanometer Scale Resolved Proximal Probe Thermal Desorption/Electrospray Ionization-Mass Spectrometry

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

Ovchinnikova, Olga S; Nikiforov, Maxim; Bradshaw, James A

2011-01-01

Nanometer scale proximal probe thermal desorption/electrospray ionization mass spectrometry (TD/ESI-MS) was demonstrated for molecular surface sampling of caffeine from a thin film using a 30 nm diameter nano-thermal analysis (nano-TA) probe tip in an atomic force microscope (AFM) coupled via a vapor transfer line and ESI interface to a MS detection platform. Using a probe temperature of 350 C and a spot sampling time of 30 s, conical desorption craters 250 nm in diameter and 100 nm deep were created as shown through subsequent topographical imaging of the surface within the same system. Automated sampling of a 5 x 2more » array of spots, with 2 m spacing between spots, and real time selective detection of the desorbed caffeine using tandem mass spectrometry was also demonstrated. Estimated from the crater volume (~2x106 nm3), only about 10 amol (2 fg) of caffeine was liberated from each thermal desorption crater in the thin film. These results illustrate a relatively simple experimental setup and means to acquire in automated fashion sub-micrometer scale spatial sampling resolution and mass spectral detection of materials amenable to TD. The ability to achieve MS-based chemical imaging with 250 nm scale spatial resolution with this system is anticipated.« less

Chemical release from single-PMMA microparticles monitored by CARS microscopy

NASA Astrophysics Data System (ADS)

Enejder, Annika; Svedberg, Fredrik; Nordstierna, Lars; Nydén, Magnus

2011-03-01

Microparticles loaded with antigens, proteins, DNA, fungicides, and other functional agents emerge as ideal vehicles for vaccine, drug delivery, genetic therapy, surface- and crop protection. The microscopic size of the particles and their collective large specific surface area enables highly active and localized release of the functional substance. In order to develop designs with release profiles optimized for the specific application, it is desirable to map the distribution of the active substance within the particle and how parameters such as size, material and morphology affect release rates at single particle level. Current imaging techniques are limited in resolution, sensitivity, image acquisition time, or sample treatment, excluding dynamic studies of active agents in microparticles. Here, we demonstrate that the combination of CARS and THG microscopy can successfully be used, by mapping the spatial distribution and release rates of the fungicide and food preservative IPBC from different designs of PMMA microparticles at single-particle level. By fitting a radial diffusion model to the experimental data, single particle diffusion coefficients can be determined. We show that release rates are highly dependent on the size and morphology of the particles. Hence, CARS and THG microscopy provides adequate sensitivity and spatial resolution for quantitative studies on how singleparticle properties affect the diffusion of active agents at microscopic level. This will aid the design of innovative microencapsulating systems for controlled release.

Seeing with the nano-eye: accessing structure, function, and dynamics of matter on its natural length and time scales

NASA Astrophysics Data System (ADS)

Raschke, Markus

2015-03-01

To understand and ultimately control the properties of most functional materials, from molecular soft-matter to quantum materials, requires access to the structure, coupling, and dynamics on the elementary time and length scales that define the microscopic interactions in these materials. To gain the desired nanometer spatial resolution with simultaneous spectroscopic specificity we combine scanning probe microscopy with different optical, including coherent, nonlinear, and ultrafast spectroscopies. The underlying near-field interaction mediated by the atomic-force or scanning tunneling microscope tip provides the desired deep-sub wavelength nano-focusing enabling few-nm spatial resolution. I will introduce our generalization of the approach in terms of the near-field impedance matching to a quantum system based on special optical antenna-tip designs. The resulting enhanced and qualitatively new forms of light-matter interaction enable measurements of quantum dynamics in an interacting environment or to image the electromagnetic local density of states of thermal radiation. Other applications include the inter-molecular coupling and dynamics in soft-matter hetero-structures, surface plasmon interferometry as a probe of electronic structure and dynamics in graphene, and quantum phase transitions in correlated electron materials. These examples highlight the general applicability of the new near-field microscopy approach, complementing emergent X-ray and electron imaging tools, aiming towards the ultimate goal of probing matter on its most elementary spatio-temporal level.

Automated detection of analyzable metaphase chromosome cells depicted on scanned digital microscopic images

NASA Astrophysics Data System (ADS)

Qiu, Yuchen; Wang, Xingwei; Chen, Xiaodong; Li, Yuhua; Liu, Hong; Li, Shibo; Zheng, Bin

2010-02-01

Visually searching for analyzable metaphase chromosome cells under microscopes is quite time-consuming and difficult. To improve detection efficiency, consistency, and diagnostic accuracy, an automated microscopic image scanning system was developed and tested to directly acquire digital images with sufficient spatial resolution for clinical diagnosis. A computer-aided detection (CAD) scheme was also developed and integrated into the image scanning system to search for and detect the regions of interest (ROI) that contain analyzable metaphase chromosome cells in the large volume of scanned images acquired from one specimen. Thus, the cytogeneticists only need to observe and interpret the limited number of ROIs. In this study, the high-resolution microscopic image scanning and CAD performance was investigated and evaluated using nine sets of images scanned from either bone marrow (three) or blood (six) specimens for diagnosis of leukemia. The automated CAD-selection results were compared with the visual selection. In the experiment, the cytogeneticists first visually searched for the analyzable metaphase chromosome cells from specimens under microscopes. The specimens were also automated scanned and followed by applying the CAD scheme to detect and save ROIs containing analyzable cells while deleting the others. The automated selected ROIs were then examined by a panel of three cytogeneticists. From the scanned images, CAD selected more analyzable cells than initially visual examinations of the cytogeneticists in both blood and bone marrow specimens. In general, CAD had higher performance in analyzing blood specimens. Even in three bone marrow specimens, CAD selected 50, 22, 9 ROIs, respectively. Except matching with the initially visual selection of 9, 7, and 5 analyzable cells in these three specimens, the cytogeneticists also selected 41, 15 and 4 new analyzable cells, which were missed in initially visual searching. This experiment showed the feasibility of applying this CAD-guided high-resolution microscopic image scanning system to prescreen and select ROIs that may contain analyzable metaphase chromosome cells. The success and the further improvement of this automated scanning system may have great impact on the future clinical practice in genetic laboratories to detect and diagnose diseases.

Multiscale pore structure and its effect on gas transport in organic-rich shale

NASA Astrophysics Data System (ADS)

Wu, Tianhao; Li, Xiang; Zhao, Junliang; Zhang, Dongxiao

2017-07-01

A systematic investigation of multiscale pore structure in organic-rich shale by means of the combination of various imaging techniques is presented, including the state-of-the-art Helium-Ion-Microscope (HIM). The study achieves insight into the major features at each scale and suggests the affordable techniques for specific objectives from the aspects of resolution, dimension, and cost. The pores, which appear to be isolated, are connected by smaller pores resolved by higher-resolution imaging. This observation provides valuable information, from the microscopic perspective of pore structure, for understanding how gas accumulates and transports from where it is generated. A comprehensive workflow is proposed based on the characteristics acquired from the multiscale pore structure analysis to simulate the gas transport process. The simulations are completed with three levels: the microscopic mechanisms should be taken into consideration at level I; the spatial distribution features of organic matter, inorganic matter, and macropores constitute the major issue at level II; and the microfracture orientation and topological structure are dominant factors at level III. The results of apparent permeability from simulations agree well with the values acquired from experiments. By means of the workflow, the impact of various gas transport mechanisms at different scales can be investigated more individually and precisely than conventional experiments.

Label-free imaging of intracellular motility by low-coherent quantitative phase microscope in reflection geometry

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Iwai, Hidenao; Yamashita, Yutaka

2011-11-01

We demonstrate tomographic imaging of intracellular activity of living cells by a low-coherent quantitative phase microscope. The intracellular organelles, such as the nucleus, nucleolus, and mitochondria, are moving around inside living cells, driven by the cellular physiological activity. In order to visualize the intracellular motility in a label-free manner we have developed a reflection-type quantitative phase microscope which employs the phase shifting interferometric technique with a low-coherent light source. The phase shifting interferometry enables us to quantitatively measure the intensity and phase of the optical field, and the low-coherence interferometry makes it possible to selectively probe a specific sectioning plane in the cell volume. The results quantitatively revealed the depth-resolved fluctuations of intracellular surfaces so that the plasma membrane and the membranes of intracellular organelles were independently measured. The transversal and the vertical spatial resolutions were 0.56 μm and 0.93 μm, respectively, and the mechanical sensitivity of the phase measurement was 1.2 nanometers. The mean-squared displacement was applied as a statistical tool to analyze the temporal fluctuation of the intracellular organelles. To the best of our knowledge, our system visualized depth-resolved intracellular organelles motion for the first time in sub-micrometer resolution without contrast agents.

Hyperspectral imaging with laser-scanning sum-frequency generation microscopy

PubMed Central

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

2017-01-01

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

Nanoscale magnetic imaging with a single nitrogen-vacancy center in diamond

NASA Astrophysics Data System (ADS)

Hong, Sungkun

Magnetic imaging has been playing central roles not only in fundamental sciences but also in engineering and industry. Their numerous applications can be found in various areas, ranging from chemical analysis and biomedical imaging to magnetic data storage technology. An outstanding problem is to develop new magnetic imaging techniques with improved spatial resolutions down to nanoscale, while maintaining their magnetic sensitivities. For instance, if detecting individual electron or nuclear spins with nanomter spatial resolution is possible, it would allow for direct imaging of chemical structures of complex molecules, which then could bring termendous impacts on biological sciences. While realization of such nanoscale magnetic imaging still remains challenging, nitrogen-vacancy (NV) defects in diamond have recently considered as promising magnetic field sensors, as their electron spins show exceptionally long coherence even at room temperature. This thesis presents experimental progress in realizing a nanoscale magnetic imaging apparatus with a single nitrogen-vacancy (NV) color center diamond. We first fabricated diamond nanopillar devices hosting single NV centers at their ends, and incorporated them to a custom-built atomic force microscope (AFM). Our devices showed unprecedented combination of magnetic field sensitivity and spatial resolution for scanning NV systems. We then used these devices to magnetically image a single isolated electronic spin with nanometer resolution, for the first time under ambient condition. We also extended our study to improve and generalize the application of the scanning NV magnetometer we developed. We first introduced magnetic field gradients from a strongly magnetized tip, and demonstrated that the spatial resolution can be further improved by spectrally distinguishing identical spins at different locations. In addition, we developed a method to synchronize the periodic motion of an AFM tip and pulsed microwave sequences controlling an NV spin. This scheme enabled employment of 'AC magnetic field sensing scheme' in imaging samples with static and spatially varying magnetizations.

Unsupervised data mining in nanoscale x-ray spectro-microscopic study of NdFeB magnet

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

Duan, Xiaoyue; Yang, Feifei; Antono, Erin

Novel developments in X-ray based spectro-microscopic characterization techniques have increased the rate of acquisition of spatially resolved spectroscopic data by several orders of magnitude over what was possible a few years ago. This accelerated data acquisition, with high spatial resolution at nanoscale and sensitivity to subtle differences in chemistry and atomic structure, provides a unique opportunity to investigate hierarchically complex and structurally heterogeneous systems found in functional devices and materials systems. However, handling and analyzing the large volume data generated poses significant challenges. Here we apply an unsupervised data-mining algorithm known as DBSCAN to study a rare-earth element based permanentmore » magnet material, Nd 2Fe 14B. We are able to reduce a large spectro-microscopic dataset of over 300,000 spectra to 3, preserving much of the underlying information. Scientists can easily and quickly analyze in detail three characteristic spectra. Our approach can rapidly provide a concise representation of a large and complex dataset to materials scientists and chemists. For instance, it shows that the surface of common Nd 2Fe 14B magnet is chemically and structurally very different from the bulk, suggesting a possible surface alteration effect possibly due to the corrosion, which could affect the material’s overall properties.« less

Unsupervised data mining in nanoscale x-ray spectro-microscopic study of NdFeB magnet

DOE PAGES

Duan, Xiaoyue; Yang, Feifei; Antono, Erin; ...

2016-09-29

Novel developments in X-ray based spectro-microscopic characterization techniques have increased the rate of acquisition of spatially resolved spectroscopic data by several orders of magnitude over what was possible a few years ago. This accelerated data acquisition, with high spatial resolution at nanoscale and sensitivity to subtle differences in chemistry and atomic structure, provides a unique opportunity to investigate hierarchically complex and structurally heterogeneous systems found in functional devices and materials systems. However, handling and analyzing the large volume data generated poses significant challenges. Here we apply an unsupervised data-mining algorithm known as DBSCAN to study a rare-earth element based permanentmore » magnet material, Nd 2Fe 14B. We are able to reduce a large spectro-microscopic dataset of over 300,000 spectra to 3, preserving much of the underlying information. Scientists can easily and quickly analyze in detail three characteristic spectra. Our approach can rapidly provide a concise representation of a large and complex dataset to materials scientists and chemists. For instance, it shows that the surface of common Nd 2Fe 14B magnet is chemically and structurally very different from the bulk, suggesting a possible surface alteration effect possibly due to the corrosion, which could affect the material’s overall properties.« less

An interchangeable scanning Hall probe/scanning SQUID microscope

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

Tang, Chiu-Chun; Lin, Hui-Ting; Wu, Sing-Lin

2014-08-15

We have constructed a scanning probe microscope for magnetic imaging, which can function as a scanning Hall probe microscope (SHPM) and as a scanning SQUID microscope (SSM). The scanning scheme, applicable to SHPM and SSM, consists of a mechanical positioning (sub) micron-XY stage and a flexible direct contact to the sample without a feedback control system for the Z-axis. With the interchangeable capability of operating two distinct scanning modes, our microscope can incorporate the advantageous functionalities of the SHPM and SSM with large scan range up to millimeter, high spatial resolution (⩽4 μm), and high field sensitivity in a widemore » range of temperature (4.2 K-300 K) and magnetic field (10{sup −7} T-1 T). To demonstrate the capabilities of the system, we present magnetic images scanned with SHPM and SSM, including a RbFeB magnet and a nickel grid pattern at room temperature, surface magnetic domain structures of a La{sub 2/3}Ca{sub 1/3}MnO{sub 3} thin film at 77 K, and superconducting vortices in a striped niobium film at 4.2 K.« less

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.

Design and performance of an ultra-high vacuum scanning tunneling microscope operating at dilution refrigerator temperatures and high magnetic fields.

PubMed

Misra, S; Zhou, B B; Drozdov, I K; Seo, J; Urban, L; Gyenis, A; Kingsley, S C J; Jones, H; Yazdani, A

2013-10-01

We describe the construction and performance of a scanning tunneling microscope capable of taking maps of the tunneling density of states with sub-atomic spatial resolution at dilution refrigerator temperatures and high (14 T) magnetic fields. The fully ultra-high vacuum system features visual access to a two-sample microscope stage at the end of a bottom-loading dilution refrigerator, which facilitates the transfer of in situ prepared tips and samples. The two-sample stage enables location of the best area of the sample under study and extends the experiment lifetime. The successful thermal anchoring of the microscope, described in detail, is confirmed through a base temperature reading of 20 mK, along with a measured electron temperature of 250 mK. Atomically resolved images, along with complementary vibration measurements, are presented to confirm the effectiveness of the vibration isolation scheme in this instrument. Finally, we demonstrate that the microscope is capable of the same level of performance as typical machines with more modest refrigeration by measuring spectroscopic maps at base temperature both at zero field and in an applied magnetic field.

Imaging the beating heart in the mouse using intravital microscopy techniques

PubMed Central

Vinegoni, Claudio; Aguirre, Aaron D; Lee, Sungon; Weissleder, Ralph

2017-01-01

Real-time microscopic imaging of moving organs at single-cell resolution represents a major challenge in studying complex biology in living systems. Motion of the tissue from the cardiac and respiratory cycles severely limits intravital microscopy by compromising ultimate spatial and temporal imaging resolution. However, significant recent advances have enabled single-cell resolution imaging to be achieved in vivo. In this protocol, we describe experimental procedures for intravital microscopy based on a combination of thoracic surgery, tissue stabilizers and acquisition gating methods, which enable imaging at the single-cell level in the beating heart in the mouse. Setup of the model is typically completed in 1 h, which allows 2 h or more of continuous cardiac imaging. This protocol can be readily adapted for the imaging of other moving organs, and it will therefore broadly facilitate in vivo high-resolution microscopy studies. PMID:26492138

High-resolution x-ray tomography using laboratory sources

NASA Astrophysics Data System (ADS)

Tkachuk, Andrei; Feser, Michael; Cui, Hongtao; Duewer, Fred; Chang, Hauyee; Yun, Wenbing

2006-08-01

X-ray computed tomography (XCT) is a powerful nondestructive 3D imaging technique, which enables the visualization of the three dimensional structure of complex, optically opaque samples. High resolution XCT using Fresnel zone plate lenses has been confined in the past to synchrotron radiation centers due to the need for a bright and intense source of x-rays. This confinement severely limits the availability and accessibility of x-ray microscopes and the wide proliferation of this methodology. We are describing a sub-50nm resolution XCT system operating at 8 keV in absorption and Zernike phase contrast mode based on a commercially available laboratory x-ray source. The system utilizes high-efficiency Fresnel zone plates with an outermost zone width of 35 nm and 700 nm structure height resulting in a current spatial resolution better than 50 nm. In addition to the technical description of the system and specifications, we present application examples in the semiconductor field.

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.

Pulse compressor with aberration correction

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

Mankos, Marian

In this SBIR project, Electron Optica, Inc. (EOI) is developing an electron mirror-based pulse compressor attachment to new and retrofitted dynamic transmission electron microscopes (DTEMs) and ultrafast electron diffraction (UED) cameras for improving the temporal resolution of these instruments from the characteristic range of a few picoseconds to a few nanoseconds and beyond, into the sub-100 femtosecond range. The improvement will enable electron microscopes and diffraction cameras to better resolve the dynamics of reactions in the areas of solid state physics, chemistry, and biology. EOI’s pulse compressor technology utilizes the combination of electron mirror optics and a magnetic beam separatormore » to compress the electron pulse. The design exploits the symmetry inherent in reversing the electron trajectory in the mirror in order to compress the temporally broadened beam. This system also simultaneously corrects the chromatic and spherical aberration of the objective lens for improved spatial resolution. This correction will be found valuable as the source size is reduced with laser-triggered point source emitters. With such emitters, it might be possible to significantly reduce the illuminated area and carry out ultrafast diffraction experiments from small regions of the sample, e.g. from individual grains or nanoparticles. During phase I, EOI drafted a set of candidate pulse compressor architectures and evaluated the trade-offs between temporal resolution and electron bunch size to achieve the optimum design for two particular applications with market potential: increasing the temporal and spatial resolution of UEDs, and increasing the temporal and spatial resolution of DTEMs. Specialized software packages that have been developed by MEBS, Ltd. were used to calculate the electron optical properties of the key pulse compressor components: namely, the magnetic prism, the electron mirror, and the electron lenses. In the final step, these results were folded into a model describing the key electron-optical parameters of the complete pulse compressor. The simulations reveal that the mirror pulse compressor can reduce the temporal spread of UEDs and DTEMs to the sub-100 femtosecond level for practical electron bunch sizes. EOI’s pulse compressors can be designed and built to attach to different types of UEDs and DTEMs, thus making them suitable for enhancing the study of the structure, composition, and bonding states of new materials at ultrafast time scales to advance material science research in the field of nanotechnology as well as biomedical research.« less

Overview of nanoscale NEXAFS performed with soft X-ray microscopes.

PubMed

Guttmann, Peter; Bittencourt, Carla

2015-01-01

Today, in material science nanoscale structures are becoming more and more important. Not only for the further miniaturization of semiconductor devices like carbon nanotube based transistors, but also for newly developed efficient energy storage devices, gas sensors or catalytic systems nanoscale and functionalized materials have to be analysed. Therefore, analytical tools like near-edge X-ray absorption fine structure (NEXAFS) spectroscopy has to be applied on single nanostructures. Scanning transmission X-ray microscopes (STXM) as well as full-field transmission X-ray microscopes (TXM) allow the required spatial resolution to study individual nanostructures. In the soft X-ray energy range only STXM was used so far for NEXAFS studies. Due to its unique setup, the TXM operated by the Helmholtz-Zentrum Berlin (HZB) at the electron storage ring BESSY II is the first one in the soft X-ray range which can be used for NEXAFS spectroscopy studies which will be shown in this review. Here we will give an overview of the different microscopes used for NEXAFS studies and describe their advantages and disadvantages for different samples.

Ionic channels in Langmuir-Blodgett films imaged by a scanning tunneling microscope.

PubMed Central

Kolomytkin, O V; Golubok, A O; Davydov, D N; Timofeev, V A; Vinogradova, S A; Tipisev SYa

1991-01-01

The molecular structure of channels formed by gramicidin A in a lipid membrane was imaged by a scanning tunneling microscope operating in air. The mono- and bimolecular films of lipid with gramicidin A were deposited onto a highly oriented pyrolitic graphite substrate by the Langmuir-Blodgett technique. It has been shown that under high concentration gramicidin A molecules can form in lipid films a quasi-regular, densely packed structure. Single gramicidin A molecules were imaged for the first time as well. The cavity of 0.4 +/- 0.05 nm in halfwidth was found on the scanning tunneling microscopy image of the gramicidin A molecule. The results of direct observation obtained by means of scanning tunneling microscope are in good agreement with the known molecular model of gramicidin A. It was shown that gramicidin A molecules can exist in a lipid monolayer as individual molecules or combined into clusters. The results demonstrate that scanning tunneling microscope can be used for high spatial resolution study of ionic channel structure. Images FIGURE 1 FIGURE 2 FIGURE 4 FIGURE 5 PMID:1712239

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.

High-resolution imaging of magnetic fields using scanning superconducting quantum interference device (SQUID) microscopy

NASA Astrophysics Data System (ADS)

Fong de Los Santos, Luis E.

Development of a scanning superconducting quantum interference device (SQUID) microscope system with interchangeable sensor configurations for imaging magnetic fields of room-temperature (RT) samples with sub-millimeter resolution. The low-critical-temperature (Tc) niobium-based monolithic SQUID sensor is mounted in the tip of a sapphire rod and thermally anchored to the cryostat helium reservoir. A 25 mum sapphire window separates the vacuum space from the RT sample. A positioning mechanism allows adjusting the sample-to-sensor spacing from the top of the Dewar. I have achieved a sensor-to-sample spacing of 100 mum, which could be maintained for periods of up to 4 weeks. Different SQUID sensor configurations are necessary to achieve the best combination of spatial resolution and field sensitivity for a given magnetic source. For imaging thin sections of geological samples, I used a custom-designed monolithic low-Tc niobium bare SQUID sensor, with an effective diameter of 80 mum, and achieved a field sensitivity of 1.5 pT/Hz1/2 and a magnetic moment sensitivity of 5.4 x 10-18 Am2/Hz1/2 at a sensor-to-sample spacing of 100 mum in the white noise region for frequencies above 100 Hz. Imaging action currents in cardiac tissue requires higher field sensitivity, which can only be achieved by compromising spatial resolution. I developed a monolithic low-Tc niobium multiloop SQUID sensor, with sensor sizes ranging from 250 mum to 1 mm, and achieved sensitivities of 480 - 180 fT/Hz1/2 in the white noise region for frequencies above 100 Hz, respectively. For all sensor configurations, the spatial resolution was comparable to the effective diameter and limited by the sensor-to-sample spacing. Spatial registration allowed us to compare high-resolution images of magnetic fields associated with action currents and optical recordings of transmembrane potentials to study the bidomain nature of cardiac tissue or to match petrography to magnetic field maps in thin sections of geological samples.

The 2015 super-resolution microscopy roadmap

NASA Astrophysics Data System (ADS)

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

2015-11-01

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

Aplanatic and quasi-aplanatic diffraction gratings

DOEpatents

Hettrick, M.C.

1987-09-14

A reflection diffraction grating having a series of transverse minute grooves of progressively varying spacing along a concave surface enables use of such gratings for x-ray or longer wavelength imaging of objects. The variable groove spacing establishes aplanatism or substantially uniform magnetification across the optical aperture. The grating may be sued, for example, in x-ray microscopes or telescopes of the imaging type and in x-ray microprobed. Increased spatial resolution and field of view may be realized in x-ray imaging. 5 figs.

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 light scattering microscopy

NASA Astrophysics Data System (ADS)

Dzakpasu, Rhonda

An optical microscope technique, dynamic light scattering microscopy (DLSM) that images dynamically scattered light fluctuation decay rates is introduced. Using physical optics we show theoretically that within the optical resolution of the microscope, relative motions between scattering centers are sufficient to produce significant phase variations resulting in interference intensity fluctuations in the image plane. The time scale for these intensity fluctuations is predicted. The spatial coherence distance defining the average distance between constructive and destructive interference in the image plane is calculated and compared with the pixel size. We experimentally tested DLSM on polystyrene latex nanospheres and living macrophage cells. In order to record these rapid fluctuations, on a slow progressive scan CCD camera, we used a thin laser line of illumination on the sample such that only a single column of pixels in the CCD camera is illuminated. This allowed the use of the rate of the column-by-column readout transfer process as the acquisition rate of the camera. This manipulation increased the data acquisition rate by at least an order of magnitude in comparison to conventional CCD cameras rates defined by frames/s. Analysis of the observed fluctuations provides information regarding the rates of motion of the scattering centers. These rates, acquired from each position on the sample are used to create a spatial map of the fluctuation decay rates. Our experiments show that with this technique, we are able to achieve a good signal-to-noise ratio and can monitor fast intensity fluctuations, on the order of milliseconds. DLSM appears to provide dynamic information about fast motions within cells at a sub-optical resolution scale and provides a new kind of spatial contrast.

Super-Resolution Scanning Laser Microscopy Based on Virtually Structured Detection

PubMed Central

Zhi, Yanan; Wang, Benquan; Yao, Xincheng

2016-01-01

Light microscopy plays a key role in biological studies and medical diagnosis. The spatial resolution of conventional optical microscopes is limited to approximately half the wavelength of the illumination light as a result of the diffraction limit. Several approaches—including confocal microscopy, stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy, and structured illumination microscopy—have been established to achieve super-resolution imaging. However, none of these methods is suitable for the super-resolution ophthalmoscopy of retinal structures because of laser safety issues and inevitable eye movements. We recently experimentally validated virtually structured detection (VSD) as an alternative strategy to extend the diffraction limit. Without the complexity of structured illumination, VSD provides an easy, low-cost, and phase artifact–free strategy to achieve super-resolution in scanning laser microscopy. In this article we summarize the basic principles of the VSD method, review our demonstrated single-point and line-scan super-resolution systems, and discuss both technical challenges and the potential of VSD-based instrumentation for super-resolution ophthalmoscopy of the retina. PMID:27480461

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

NASA Astrophysics Data System (ADS)

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

2018-04-01

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

Microscopic time-resolved imaging of singlet oxygen by delayed fluorescence in living cells.

PubMed

Scholz, Marek; Dědic, Roman; Hála, Jan

2017-11-08

Singlet oxygen is a highly reactive species which is involved in a number of processes, including photodynamic therapy of cancer. Its very weak near-infrared emission makes imaging of singlet oxygen in biological systems a long-term challenge. We address this challenge by introducing Singlet Oxygen Feedback Delayed Fluorescence (SOFDF) as a novel modality for semi-direct microscopic time-resolved wide-field imaging of singlet oxygen in biological systems. SOFDF has been investigated in individual fibroblast cells incubated with a well-known photosensitizer aluminium phthalocyanine tetrasulfonate. The SOFDF emission from the cells is several orders of magnitude stronger and much more readily detectable than the very weak near-infrared phosphorescence of singlet oxygen. Moreover, the analysis of SOFDF kinetics enables us to estimate the lifetimes of the involved excited states. Real-time SOFDF images with micrometer spatial resolution and submicrosecond temporal-resolution have been recorded. Interestingly, a steep decrease in the SOFDF intensity after the photodynamically induced release of a photosensitizer from lysosomes has been demonstrated. This effect could be potentially employed as a valuable diagnostic tool for monitoring and dosimetry in photodynamic therapy.

What is the diffraction limit? From Airy to Abbe using direct numerical integration

NASA Astrophysics Data System (ADS)

Calm, Y. M.; Merlo, J. M.; Burns, M. J.; Kempa, K.; Naughton, M. J.

The resolution of a conventional optical microscope is sometimes taken from Airy's point spread function (PSF), 0 . 61 λ / NA , and sometimes from Abbe, λ / 2 NA , where NA is the numerical aperture, however modern fluorescence and near-field optical microscopies achieve spatial resolution far better than either of these limits. There is a new category of 2D metamaterials called planar optical elements (POEs), which have a microscopic thickness (< λ), macroscopic transverse dimensions (> 100 λ), and are composed of an array of nanostructured light scatterers. POEs are found in a range of micro- and nano-photonic technologies, and will influence the future optical nanoscopy. With this pretext, we shed some light on the 'diffraction limit' by numerically evaluating Kirchhoff's scalar formulae (in their exact form) and identifying the features of highly non-paraxial, 3D PSFs. We show that the Airy and Abbe criteria are connected, and we comment on the design rules for a particular type of POE: the flat lens. This work is supported by the W. M. Keck Foundation.

Two-way communication with neural networks in vivo using focused light

PubMed Central

Wilson, Nathan R.; Schummers, James; Runyan, Caroline A.; Yan, Sherry; Chen, Robert F.; Deng, Yuting; Sur, Mriganka

2014-01-01

Neuronal networks process information in a distributed, spatially heterogeneous fashion that transcends the layout of electrodes. In contrast, directed and steerable light offers the potential to engage specific cells on demand. We present a unified framework for adapting microscopes to use light for simultaneous in vivo stimulation and recording of cells at fine spatiotemporal resolutions. We utilize straightforward optics to lock onto networks in vivo, steer light to activate circuit elements, and simultaneously record from other cells. We then actualize this “free” augmentation on both an “open” two-photon microscope, and a leading commercial one. Following this protocol, setup of the system takes a few days and the result is a non-invasive interface to brain dynamics based on directed light, at a network resolution that was not previously possible and which will further improve with the rapid advance in development of optical reporters and effectors. This protocol is for physiologists who are competent with computers and wish to extend hardware and software to interface more fluidly with neuronal networks. PMID:23702834

Paleomagnetic Analysis Using SQUID Microscopy

NASA Technical Reports Server (NTRS)

Weiss, Benjamin P.; Lima, Eduardo A.; Fong, Luis E.; Baudenbacher, Franz J.

2007-01-01

Superconducting quantum interference device (SQUID) microscopes are a new generation of instruments that map magnetic fields with unprecedented spatial resolution and moment sensitivity. Unlike standard rock magnetometers, SQUID microscopes map magnetic fields rather than measuring magnetic moments such that the sample magnetization pattern must be retrieved from source model fits to the measured field data. In this paper, we presented the first direct comparison between paleomagnetic analyses on natural samples using joint measurements from SQUID microscopy and moment magnetometry. We demonstrated that in combination with apriori geologic and petrographic data, SQUID microscopy can accurately characterize the magnetization of lunar glass spherules and Hawaiian basalt. The bulk moment magnitude and direction of these samples inferred from inversions of SQUID microscopy data match direct measurements on the same samples using moment magnetometry. In addition, these inversions provide unique constraints on the magnetization distribution within the sample. These measurements are among the most sensitive and highest resolution quantitative paleomagnetic studies of natural remanent magnetization to date. We expect that this technique will be able to extend many other standard paleomagnetic techniques to previously inaccessible microscale samples.

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

NASA Astrophysics Data System (ADS)

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

2017-08-01

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

Multi-images deconvolution improves signal-to-noise ratio on gated stimulated emission depletion microscopy

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

Castello, Marco; DIBRIS, University of Genoa, Via Opera Pia 13, Genoa 16145; Diaspro, Alberto

2014-12-08

Time-gated detection, namely, only collecting the fluorescence photons after a time-delay from the excitation events, reduces complexity, cost, and illumination intensity of a stimulated emission depletion (STED) microscope. In the gated continuous-wave- (CW-) STED implementation, the spatial resolution improves with increased time-delay, but the signal-to-noise ratio (SNR) reduces. Thus, in sub-optimal conditions, such as a low photon-budget regime, the SNR reduction can cancel-out the expected gain in resolution. Here, we propose a method which does not discard photons, but instead collects all the photons in different time-gates and recombines them through a multi-image deconvolution. Our results, obtained on simulated andmore » experimental data, show that the SNR of the restored image improves relative to the gated image, thereby improving the effective resolution.« less

Nuclear magnetic resonance imaging at microscopic resolution

NASA Astrophysics Data System (ADS)

Johnson, G. Allan; Thompson, Morrow B.; Gewalt, Sally L.; Hayes, Cecil E.

Resolution limits in NMR imaging are imposed by bandwidth considerations, available magnetic gradients for spatial encoding, and signal to noise. This work reports modification of a clinical NMR imaging device with picture elements of 500 × 500 × 5000 μm to yield picture elements of 50 × 50 × 1000 μm. Resolution has been increased by using smaller gradient coils permitting gradient fields >0.4 mT/cm. Significant improvements in signal to noise are achieved with smaller rf coils, close attention to choice of bandwidth, and signal averaging. These improvements permit visualization of anatomical structures in the rat brain with an effective diameter of 1 cm with the same definition as is seen in human imaging. The techniques and instrumentation should open a number of basic sciences such as embryology, plant sciences, and teratology to the potentials of NMR imaging.

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.

Electron energy loss spectroscopy on semiconductor heterostructures for optoelectronics and photonics applications.

PubMed

Eljarrat, A; López-Conesa, L; Estradé, S; Peiró, F

2016-05-01

In this work, we present characterization methods for the analysis of nanometer-sized devices, based on silicon and III-V nitride semiconductor materials. These methods are devised in order to take advantage of the aberration corrected scanning transmission electron microscope, equipped with a monochromator. This set-up ensures the necessary high spatial and energy resolution for the characterization of the smallest structures. As with these experiments, we aim to obtain chemical and structural information, we use electron energy loss spectroscopy (EELS). The low-loss region of EELS is exploited, which features fundamental electronic properties of semiconductor materials and facilitates a high data throughput. We show how the detailed analysis of these spectra, using theoretical models and computational tools, can enhance the analytical power of EELS. In this sense, initially, results from the model-based fit of the plasmon peak are presented. Moreover, the application of multivariate analysis algorithms to low-loss EELS is explored. Finally, some physical limitations of the technique, such as spatial delocalization, are mentioned. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Biological tissue imaging with a position and time sensitive pixelated detector.

PubMed

Jungmann, Julia H; Smith, Donald F; MacAleese, Luke; Klinkert, Ivo; Visser, Jan; Heeren, Ron M A

2012-10-01

We demonstrate the capabilities of a highly parallel, active pixel detector for large-area, mass spectrometric imaging of biological tissue sections. A bare Timepix assembly (512 × 512 pixels) is combined with chevron microchannel plates on an ion microscope matrix-assisted laser desorption time-of-flight mass spectrometer (MALDI TOF-MS). The detector assembly registers position- and time-resolved images of multiple m/z species in every measurement frame. We prove the applicability of the detection system to biomolecular mass spectrometry imaging on biologically relevant samples by mass-resolved images from Timepix measurements of a peptide-grid benchmark sample and mouse testis tissue slices. Mass-spectral and localization information of analytes at physiologic concentrations are measured in MALDI-TOF-MS imaging experiments. We show a high spatial resolution (pixel size down to 740 × 740 nm(2) on the sample surface) and a spatial resolving power of 6 μm with a microscope mode laser field of view of 100-335 μm. Automated, large-area imaging is demonstrated and the Timepix' potential for fast, large-area image acquisition is highlighted.

Biocytin-Derived MRI Contrast Agent for Longitudinal Brain Connectivity Studies

PubMed Central

2011-01-01

To investigate the connectivity of brain networks noninvasively and dynamically, we have developed a new strategy to functionalize neuronal tracers and designed a biocompatible probe that can be visualized in vivo using magnetic resonance imaging (MRI). Furthermore, the multimodal design used allows combined ex vivo studies with microscopic spatial resolution by conventional histochemical techniques. We present data on the functionalization of biocytin, a well-known neuronal tract tracer, and demonstrate the validity of the approach by showing brain networks of cortical connectivity in live rats under MRI, together with the corresponding microscopic details, such as fibers and neuronal morphology under light microscopy. We further demonstrate that the developed molecule is the first MRI-visible probe to preferentially trace retrograde connections. Our study offers a new platform for the development of multimodal molecular imaging tools of broad interest in neuroscience, that capture in vivo the dynamics of large scale neural networks together with their microscopic characteristics, thereby spanning several organizational levels. PMID:22860157

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

PubMed Central

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

2009-01-01

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

Real-time imaging of nitric oxide production in living cells with 1,3,5,7-tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacence by invert fluorescence microscope.

PubMed

Huang, Ke-Jing; Wang, Hong; Ma, Ming; Zhang, Xian; Zhang, Hua-Shan

2007-02-01

Although the importance of nitric oxide (NO) as a signalling molecule in many biological processes is becoming increasingly evident, many proposed and potential biological functions of NO still remain unclear. Bioimaging is a good technique to visualize observation of nitric oxide in biological samples. In this report, a fluorescent probe, 1,3,5,7-tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacence (TMDCDABODIPY), has been first applied to real-time image NO produced in PC12 cells, Sf9 cells and human vascular endothelial cells at the presence of l-arginine with inverted fluorescence microscope. NO production in the cells is successfully captured and imaged with fine temporal and spatial resolution. The results prove that the probe combined with inverted fluorescence microscope can be developed into a sensitive and selective method for further study of NO release from cells.

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.

Unsupervised segmentation of low-contrast multichannel images: discrimination of tissue components in microscopic images of unstained specimens

NASA Astrophysics Data System (ADS)

Kopriva, Ivica; Popović Hadžija, Marijana; Hadžija, Mirko; Aralica, Gorana

2015-06-01

Low-contrast images, such as color microscopic images of unstained histological specimens, are composed of objects with highly correlated spectral profiles. Such images are very hard to segment. Here, we present a method that nonlinearly maps low-contrast color image into an image with an increased number of non-physical channels and a decreased correlation between spectral profiles. The method is a proof-of-concept validated on the unsupervised segmentation of color images of unstained specimens, in which case the tissue components appear colorless when viewed under the light microscope. Specimens of human hepatocellular carcinoma, human liver with metastasis from colon and gastric cancer and mouse fatty liver were used for validation. The average correlation between the spectral profiles of the tissue components was greater than 0.9985, and the worst case correlation was greater than 0.9997. The proposed method can potentially be applied to the segmentation of low-contrast multichannel images with high spatial resolution that arise in other imaging modalities.

A versatile indirect detector design for hard X-ray microimaging

NASA Astrophysics Data System (ADS)

Douissard, P.-A.; Cecilia, A.; Rochet, X.; Chapel, X.; Martin, T.; van de Kamp, T.; Helfen, L.; Baumbach, T.; Luquot, L.; Xiao, X.; Meinhardt, J.; Rack, A.

2012-09-01

Indirect X-ray detectors are of outstanding importance for high resolution imaging, especially at synchrotron light sources: while consisting mostly of components which are widely commercially available, they allow for a broad range of applications in terms of the X-ray energy employed, radiation dose to the detector, data acquisition rate and spatial resolving power. Frequently, an indirect detector consists of a thin-film single crystal scintillator and a high-resolution visible light microscope as well as a camera. In this article, a novel modular-based indirect design is introduced, which offers several advantages: it can be adapted for different cameras, i.e. different sensor sizes, and can be trimmed to work either with (quasi-)monochromatic illumination and the correspondingly lower absorbed dose or with intense white beam irradiation. In addition, it allows for a motorized quick exchange between different magnifications / spatial resolutions. Developed within the European project SCINTAX, it is now commercially available. The characteristics of the detector in its different configurations (i.e. for low dose or for high dose irradiation) as measured within the SCINTAX project will be outlined. Together with selected applications from materials research, non-destructive evaluation and life sciences they underline the potential of this design to make high resolution X-ray imaging widely available.

Three-dimensional nanoscale characterisation of materials by atom probe tomography

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

Devaraj, Arun; Perea, Daniel E.; Liu, Jia

The development of three-dimensional (3D), characterization techniques with high spatial and mass resolution is crucial for understanding and developing advanced materials for many engineering applications as well as for understanding natural materials. In recent decades, atom probe tomography (APT) which combines a point projection microscope and time-of-flight mass spectrometer has evolved to be an excellent characterization technique capable of providing 3D nanoscale characterization of materials with sub-nanometer scale spatial resolution, with equal sensitivity for all elements. This review discusses the current state as of beginning of the year 2016 of APT instrumentation, new developments in sample preparation methods, experimental proceduresmore » for different material classes, reconstruction of APT results, the current status of correlative microscopy, and application of APT for microstructural characterization in established scientific areas like structural materials as well as new applications in semiconducting nanowires, semiconductor devices, battery materials, catalyst materials, geological materials and biological materials. Finally, a brief perspective is given regarding the future of APT.« less

Three-dimensional localization of nanoscale battery reactions using soft X-ray tomography

DOE PAGES

Yu, Young-Sang; Farmand, Maryam; Kim, Chunjoong; ...

2018-03-02

Battery function is determined by the efficiency and reversibility of the electrochemical phase transformations at solid electrodes. The microscopic tools available to study the chemical states of matter with the required spatial resolution and chemical specificity are intrinsically limited when studying complex architectures by their reliance on two-dimensional projections of thick material. Here in this paper, we report the development of soft X-ray ptychographic tomography, which resolves chemical states in three dimensions at 11 nm spatial resolution. We study an ensemble of nano-plates of lithium iron phosphate extracted from a battery electrode at 50% state of charge. Using a setmore » of nanoscale tomograms, we quantify the electrochemical state and resolve phase boundaries throughout the volume of individual nanoparticles. These observations reveal multiple reaction points, intra-particle heterogeneity, and size effects that highlight the importance of multi-dimensional analytical tools in providing novel insight to the design of the next generation of high-performance devices.« less

Infrared and Raman Microscopy in Cell Biology

PubMed Central

Matthäus, Christian; Bird, Benjamin; Miljković, Miloš; Chernenko, Tatyana; Romeo, Melissa; Diem, Max

2009-01-01

This chapter presents novel microscopic methods to monitor cell biological processes of live or fixed cells without the use of any dye, stains, or other contrast agent. These methods are based on spectral techniques that detect inherent spectroscopic properties of biochemical constituents of cells, or parts thereof. Two different modalities have been developed for this task. One of them is infrared micro-spectroscopy, in which an average snapshot of a cell’s biochemical composition is collected at a spatial resolution of typically 25 mm. This technique, which is extremely sensitive and can collect such a snapshot in fractions of a second, is particularly suited for studying gross biochemical changes. The other technique, Raman microscopy (also known as Raman micro-spectroscopy), is ideally suited to study variations of cellular composition on the scale of subcellular organelles, since its spatial resolution is as good as that of fluorescence microscopy. Both techniques exhibit the fingerprint sensitivity of vibrational spectroscopy toward biochemical composition, and can be used to follow a variety of cellular processes. PMID:19118679

Diatom Valve Three-Dimensional Representation: A New Imaging Method Based on Combined Microscopies

PubMed Central

Ferrara, Maria Antonietta; De Tommasi, Edoardo; Coppola, Giuseppe; De Stefano, Luca; Rea, Ilaria; Dardano, Principia

2016-01-01

The frustule of diatoms, unicellular microalgae, shows very interesting photonic features, generally related to its complicated and quasi-periodic micro- and nano-structure. In order to simulate light propagation inside and through this natural structure, it is important to develop three-dimensional (3D) models for synthetic replica with high spatial resolution. In this paper, we present a new method that generates images of microscopic diatoms with high definition, by merging scanning electron microscopy and digital holography microscopy or atomic force microscopy data. Starting from two digital images, both acquired separately with standard characterization procedures, a high spatial resolution (Δz = λ/20, Δx = Δy ≅ 100 nm, at least) 3D model of the object has been generated. Then, the two sets of data have been processed by matrix formalism, using an original mathematical algorithm implemented on a commercially available software. The developed methodology could be also of broad interest in the design and fabrication of micro-opto-electro-mechanical systems. PMID:27690008

Micro Cantilever Movement Detection with an Amorphous Silicon Array of Position Sensitive Detectors

PubMed Central

Contreras, Javier; Costa, Daniel; Pereira, Sonia; Fortunato, Elvira; Martins, Rodrigo; Wierzbicki, Rafal; Heerlein, Holger; Ferreira, Isabel

2010-01-01

The movement of a micro cantilever was detected via a self constructed portable data acquisition prototype system which integrates a linear array of 32 1D amorphous silicon position sensitive detectors (PSD). The system was mounted on a microscope using a metal structure platform and the movement of the 30 μm wide by 400 μm long cantilever was tracked by analyzing the signals acquired by the 32 sensor array electronic readout system and the relevant data algorithm. The obtained results show a linear behavior of the photocurrent relating X and Y movement, with a non-linearity of about 3%, a spatial resolution of less than 2 μm along the lateral dimension of the sensor as well as of less than 3 μm along the perpendicular dimension of the sensor, when detecting just the micro-cantilever, and a spatial resolution of less than 1 μm when detecting the holding structure. PMID:22163648

Three-dimensional localization of nanoscale battery reactions using soft X-ray tomography

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

Yu, Young-Sang; Farmand, Maryam; Kim, Chunjoong

Battery function is determined by the efficiency and reversibility of the electrochemical phase transformations at solid electrodes. The microscopic tools available to study the chemical states of matter with the required spatial resolution and chemical specificity are intrinsically limited when studying complex architectures by their reliance on two-dimensional projections of thick material. Here in this paper, we report the development of soft X-ray ptychographic tomography, which resolves chemical states in three dimensions at 11 nm spatial resolution. We study an ensemble of nano-plates of lithium iron phosphate extracted from a battery electrode at 50% state of charge. Using a setmore » of nanoscale tomograms, we quantify the electrochemical state and resolve phase boundaries throughout the volume of individual nanoparticles. These observations reveal multiple reaction points, intra-particle heterogeneity, and size effects that highlight the importance of multi-dimensional analytical tools in providing novel insight to the design of the next generation of high-performance devices.« less

High-resolution optical control of spatiotemporal neuronal activity patterns in zebrafish using a digital micromirror device.

PubMed

Zhu, Peixin; Fajardo, Otto; Shum, Jennifer; Zhang Schärer, Yan-Ping; Friedrich, Rainer W

2012-06-28

Optogenetic approaches allow the manipulation of neuronal activity patterns in space and time by light, particularly in small animals such as zebrafish. However, most techniques cannot control neuronal activity independently at different locations. Here we describe equipment and provide a protocol for single-photon patterned optical stimulation of neurons using a digital micromirror device (DMD). This method can create arbitrary spatiotemporal light patterns with spatial and temporal resolutions in the micrometer and submillisecond range, respectively. Different options to integrate a DMD into a multiphoton microscope are presented and compared. We also describe an ex vivo preparation of the adult zebrafish head that greatly facilitates optogenetic and other experiments. After assembly, the initial alignment takes about one day and the zebrafish preparation takes <30 min. The method has previously been used to activate channelrhodopsin-2 and manipulate oscillatory synchrony among spatially distributed neurons in the zebrafish olfactory bulb. It can be adapted easily to a wide range of other species, optogenetic probes and scientific applications.

Elemental mapping and microimaging by x-ray capillary optics.

PubMed

Hampai, D; Dabagov, S B; Cappuccio, G; Longoni, A; Frizzi, T; Cibin, G; Guglielmotti, V; Sala, M

2008-12-01

Recently, many experiments have highlighted the advantage of using polycapillary optics for x-ray fluorescence studies. We have developed a special confocal scheme for micro x-ray fluorescence measurements that enables us to obtain not only elemental mapping of the sample but also simultaneously its own x-ray imaging. We have designed the prototype of a compact x-ray spectrometer characterized by a spatial resolution of less than 100 microm for fluorescence and less than 10 microm for imaging. A couple of polycapillary lenses in a confocal configuration together with a silicon drift detector allow elemental studies of extended samples (approximately 3 mm) to be performed, while a CCD camera makes it possible to record an image of the same samples with 6 microm spatial resolution, which is limited only by the pixel size of the camera. By inserting a compound refractive lens between the sample and the CCD camera, we hope to develop an x-ray microscope for more enlarged images of the samples under test.

The Extended Range X-Ray Telescope center director's discretionary fund report

NASA Technical Reports Server (NTRS)

Hoover, R. B.; Cumings, N. P.; Hildner, E.; Moore, R. L.; Tandberg-Hanssen, E. A.

1985-01-01

An Extended Range X-Ray Telescope (ERXRT) of high sensitivity and spatial resolution capable of functioning over a broad region of the X-ray/XUV portion of the spectrum has been designed and analyzed. This system has been configured around the glancing-incidence Wolter Type I X-ray mirror system which was flown on the Skylab Apollo Telescope Mount as ATM Experiment S-056. Enhanced sensitivity over a vastly broader spectral range can be realized by the utilization of a thinned, back-illuminated, buried-channel Charge Coupled Device (CCD) as the X-ray/XUV detector rather than photographic film. However, to maintain the high spatial resolution inherent in the X-ray optics when a CCD of 30 micron pixel size is used, it is necessary to increase the telescope plate scale. This can be accomplished by use of a glancing-incidence X-ray microscope to enlarge and re-focus the primary image onto the focal surface of the CCD.

Maximizing the Biochemical Resolving Power of Fluorescence Microscopy

PubMed Central

Esposito, Alessandro; Popleteeva, Marina; Venkitaraman, Ashok R.

2013-01-01

Most recent advances in fluorescence microscopy have focused on achieving spatial resolutions below the diffraction limit. However, the inherent capability of fluorescence microscopy to non-invasively resolve different biochemical or physical environments in biological samples has not yet been formally described, because an adequate and general theoretical framework is lacking. Here, we develop a mathematical characterization of the biochemical resolution in fluorescence detection with Fisher information analysis. To improve the precision and the resolution of quantitative imaging methods, we demonstrate strategies for the optimization of fluorescence lifetime, fluorescence anisotropy and hyperspectral detection, as well as different multi-dimensional techniques. We describe optimized imaging protocols, provide optimization algorithms and describe precision and resolving power in biochemical imaging thanks to the analysis of the general properties of Fisher information in fluorescence detection. These strategies enable the optimal use of the information content available within the limited photon-budget typically available in fluorescence microscopy. This theoretical foundation leads to a generalized strategy for the optimization of multi-dimensional optical detection, and demonstrates how the parallel detection of all properties of fluorescence can maximize the biochemical resolving power of fluorescence microscopy, an approach we term Hyper Dimensional Imaging Microscopy (HDIM). Our work provides a theoretical framework for the description of the biochemical resolution in fluorescence microscopy, irrespective of spatial resolution, and for the development of a new class of microscopes that exploit multi-parametric detection systems. PMID:24204821

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.

Grating-based tomography applications in biomedical engineering

NASA Astrophysics Data System (ADS)

Schulz, Georg; Thalmann, Peter; Khimchenko, Anna; Müller, Bert

2017-10-01

For the investigation of soft tissues or tissues consisting of soft and hard tissues on the microscopic level, hard X-ray phase tomography has become one of the most suitable imaging techniques. Besides other phase contrast methods grating interferometry has the advantage of higher sensitivity than inline methods and the quantitative results. One disadvantage of the conventional double-grating setup (XDGI) compared to inline methods is the limitation of the spatial resolution. This limitation can be overcome by removing the analyser grating resulting in a single-grating setup (XSGI). In order to verify the performance of XSGI concerning contrast and spatial resolution, a quantitative comparison of XSGI and XDGI tomograms of a human nerve was performed. Both techniques provide sufficient contrast to allow for the distinction of tissue types. The spatial resolution of the two-fold binned XSGI data set is improved by a factor of two in comparison to XDGI which underlies its performance in tomography of soft tissues. Another application for grating-based X-ray phase tomography is the simultaneous visualization of soft and hard tissues of a plaque-containing coronary artery. The simultaneous visualization of both tissues is important for the segmentation of the lumen. The segmented data can be used for flow simulations in order to obtain information about the three-dimensional wall shear stress distribution needed for the optimization of mechano-sensitive nanocontainers used for drug delivery.

High-resolution x-ray imaging using a structured scintillator.

PubMed

Hormozan, Yashar; Sychugov, Ilya; Linnros, Jan

2016-02-01

In this study, the authors introduce a new generation of finely structured scintillators with a very high spatial resolution (a few micrometers) compared to conventional scintillators, yet maintaining a thick absorbing layer for improved detectivity. Their concept is based on a 2D array of high aspect ratio pores which are fabricated by ICP etching, with spacings (pitches) of a few micrometers, on silicon and oxidation of the pore walls. The pores were subsequently filled by melting of powdered CsI(Tl), as the scintillating agent. In order to couple the secondary emitted photons of the back of the scintillator array to a CCD device, having a larger pixel size than the pore pitch, an open optical microscope with adjustable magnification was designed and implemented. By imaging a sharp edge, the authors were able to calculate the modulation transfer function (MTF) of this finely structured scintillator. The x-ray images of individually resolved pores suggest that they have been almost uniformly filled, and the MTF measurements show the feasibility of a few microns spatial resolution imaging, as set by the scintillator pore size. Compared to existing techniques utilizing CsI needles as a structured scintillator, their results imply an almost sevenfold improvement in resolution. Finally, high resolution images, taken by their detector, are presented. The presented work successfully shows the functionality of their detector concept for high resolution imaging and further fabrication developments are most likely to result in higher quantum efficiencies.

High-resolution x-ray imaging using a structured scintillator

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

Hormozan, Yashar, E-mail: hormozan@kth.se; Sychugov, Ilya; Linnros, Jan

2016-02-15

Purpose: In this study, the authors introduce a new generation of finely structured scintillators with a very high spatial resolution (a few micrometers) compared to conventional scintillators, yet maintaining a thick absorbing layer for improved detectivity. Methods: Their concept is based on a 2D array of high aspect ratio pores which are fabricated by ICP etching, with spacings (pitches) of a few micrometers, on silicon and oxidation of the pore walls. The pores were subsequently filled by melting of powdered CsI(Tl), as the scintillating agent. In order to couple the secondary emitted photons of the back of the scintillator arraymore » to a CCD device, having a larger pixel size than the pore pitch, an open optical microscope with adjustable magnification was designed and implemented. By imaging a sharp edge, the authors were able to calculate the modulation transfer function (MTF) of this finely structured scintillator. Results: The x-ray images of individually resolved pores suggest that they have been almost uniformly filled, and the MTF measurements show the feasibility of a few microns spatial resolution imaging, as set by the scintillator pore size. Compared to existing techniques utilizing CsI needles as a structured scintillator, their results imply an almost sevenfold improvement in resolution. Finally, high resolution images, taken by their detector, are presented. Conclusions: The presented work successfully shows the functionality of their detector concept for high resolution imaging and further fabrication developments are most likely to result in higher quantum efficiencies.« less

Learning surface molecular structures via machine vision

NASA Astrophysics Data System (ADS)

Ziatdinov, Maxim; Maksov, Artem; Kalinin, Sergei V.

2017-08-01

Recent advances in high resolution scanning transmission electron and scanning probe microscopies have allowed researchers to perform measurements of materials structural parameters and functional properties in real space with a picometre precision. In many technologically relevant atomic and/or molecular systems, however, the information of interest is distributed spatially in a non-uniform manner and may have a complex multi-dimensional nature. One of the critical issues, therefore, lies in being able to accurately identify (`read out') all the individual building blocks in different atomic/molecular architectures, as well as more complex patterns that these blocks may form, on a scale of hundreds and thousands of individual atomic/molecular units. Here we employ machine vision to read and recognize complex molecular assemblies on surfaces. Specifically, we combine Markov random field model and convolutional neural networks to classify structural and rotational states of all individual building blocks in molecular assembly on the metallic surface visualized in high-resolution scanning tunneling microscopy measurements. We show how the obtained full decoding of the system allows us to directly construct a pair density function—a centerpiece in analysis of disorder-property relationship paradigm—as well as to analyze spatial correlations between multiple order parameters at the nanoscale, and elucidate reaction pathway involving molecular conformation changes. The method represents a significant shift in our way of analyzing atomic and/or molecular resolved microscopic images and can be applied to variety of other microscopic measurements of structural, electronic, and magnetic orders in different condensed matter systems.

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.

Spatial Variation in Mobility-Lifetime Product in Bulk TlBr and CZT

NASA Astrophysics Data System (ADS)

Phillips, David; Haegel, Nancy; Blaine, Kevin; Kim, Hadong; Ciampi, Guido; Cirignano, Len

2012-02-01

The energy resolution of a semiconductor radiation detector depends on the charge transport properties of the semiconductor, and the mobility-lifetime (μτ) product is a key figure of merit for charge transport. In this work, we investigate the effects of two impurities, Na and Cu, on the μτ product in bulk thallium bromide (TlBr) using cathodoluminescence (CL) and transport imaging. Transport imaging uses a scanning electron microscope to generate a line of charge carriers on the surface of a bulk sample, and the intensity and spatial distribution of the recombination luminescence are recorded. A Green's function approach is used to model the generation, diffusion, and recombination of charge carriers under steady-state conditions. The luminescence distribution is fit to the model to extract the ambipolar diffusion length and the μτ product, providing a high-resolution correlation between the luminescence variations due to dopants/defects and the quantitative transport behavior. The μτ product has been mapped across a 40 μm segment of TlBr at a resolution of 2 μm. Additionally, this approach has been used to locally map variations in ambipolar diffusion length and μτ product due to extended defects in cadmium zinc telluride (CZT).

Imaging multi-scale dynamics in vivo with spiral volumetric optoacoustic tomography

NASA Astrophysics Data System (ADS)

Deán-Ben, X. Luís.; Fehm, Thomas F.; Ford, Steven J.; Gottschalk, Sven; Razansky, Daniel

2017-03-01

Imaging dynamics in living organisms is essential for the understanding of biological complexity. While multiple imaging modalities are often required to cover both microscopic and macroscopic spatial scales, dynamic phenomena may also extend over different temporal scales, necessitating the use of different imaging technologies based on the trade-off between temporal resolution and effective field of view. Optoacoustic (photoacoustic) imaging has been shown to offer the exclusive capability to link multiple spatial scales ranging from organelles to entire organs of small animals. Yet, efficient visualization of multi-scale dynamics remained difficult with state-of-the-art systems due to inefficient trade-offs between image acquisition and effective field of view. Herein, we introduce a spiral volumetric optoacoustic tomography (SVOT) technique that provides spectrally-enriched high-resolution optical absorption contrast across multiple spatio-temporal scales. We demonstrate that SVOT can be used to monitor various in vivo dynamics, from video-rate volumetric visualization of cardiac-associated motion in whole organs to high-resolution imaging of pharmacokinetics in larger regions. The multi-scale dynamic imaging capability thus emerges as a powerful and unique feature of the optoacoustic technology that adds to the multiple advantages of this technology for structural, functional and molecular imaging.

High Resolution Chemical Study of ALH84001

NASA Technical Reports Server (NTRS)

Conrad, Pamela G.; Douglas, Susanne; Kuhlman, Kimberly R.

2001-01-01

We have studied the chemistry of a sample of the SNC meteorite ALH84001 using an environmental scanning electron microscope (ESEM) with an energy dispersive chemical analytical detector and a focused ion beam secondary ion mass spectrometer (FIB-SIMS). Here we present the chemical data, both spectra and images, from two techniques that do not require sample preparation with a conductive coating, thus eliminating the possibility of preparation-induced textural artifacts. The FIB-SIMS instrument includes a column optimized for SEM with a quadrupole type mass spectrometer. Its spatial and spectral resolution are 20 nm and 0.4 AMU, respectively. The spatial resolution of the ESEM for chemical analysis is about 100 nm. Limits of detection for both instruments are mass dependent. Both the ESEM and the FIB-SIMS instrument revealed contrasting surficial features; crumbled, weathered appearance of the matrix in some regions as well as a rather ubiquitous presence of euhedral halite crystals, often associated with cracks or holes in the surface of the rock. Other halogen elements present in the vicinity of the NaCl crystals include K and Br. In this report, elemental inventories are shown as mass spectra and as X-ray maps.

High-resolution studies of the Majorana atomic chain platform

NASA Astrophysics Data System (ADS)

Feldman, Benjamin E.; Randeria, Mallika T.; Li, Jian; Jeon, Sangjun; Xie, Yonglong; Wang, Zhijun; Drozdov, Ilya K.; Andrei Bernevig, B.; Yazdani, Ali

2017-03-01

Ordered assemblies of magnetic atoms on the surface of conventional superconductors can be used to engineer topological superconducting phases and realize Majorana fermion quasiparticles (MQPs) in a condensed matter setting. Recent experiments have shown that chains of Fe atoms on Pb generically have the required electronic characteristics to form a one-dimensional topological superconductor and have revealed spatially resolved signatures of localized MQPs at the ends of such chains. Here we report higher-resolution measurements of the same atomic chain system performed using a dilution refrigerator scanning tunnelling microscope (STM). With significantly better energy resolution than previous studies, we show that the zero-bias peak (ZBP) in Fe chains has no detectable splitting from hybridization with other states. The measurements also reveal that the ZBP exhibits a distinctive `double eye’ spatial pattern on nanometre length scales. Theoretically we show that this is a general consequence of STM measurements of MQPs with substantial spectral weight in the superconducting substrate, a conclusion further supported by measurements of Pb overlayers deposited on top of the Fe chains. Finally, we report experiments performed with superconducting tips in search of the particle-hole symmetric MQP signature expected in such measurements.

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.

Direct observation of the actin filament by tip-scan atomic force microscopy

PubMed Central

Narita, Akihiro; Usukura, Eiji; Yagi, Akira; Tateyama, Kiyohiko; Akizuki, Shogo; Kikumoto, Mahito; Matsumoto, Tomoharu; Maéda, Yuichiro; Ito, Shuichi; Usukura, Jiro

2016-01-01

Actin filaments, the actin–myosin complex and the actin–tropomyosin complex were observed by a tip-scan atomic force microscope (AFM), which was recently developed by Olympus as the AFM part of a correlative microscope. This newly developed AFM uses cantilevers of similar size as stage-scan AFMs to improve substantially the spatial and temporal resolution. Such an approach has previously never been possible by a tip-scan system, in which a cantilever moves in the x, y and z directions. We evaluated the performance of this developed tip-scan AFM by observing the molecular structure of actin filaments and the actin–tropomyosin complex. In the image of the actin filament, the molecular interval of the actin subunits (∼5.5 nm) was clearly observed as stripes. From the shape of the stripes, the polarity of the actin filament was directly determined and the results were consistent with the polarity determined by myosin binding. In the image of the actin–tropomyosin complex, each tropomyosin molecule (∼2 nm in diameter) on the actin filament was directly observed without averaging images of different molecules. Each tropomyosin molecule on the actin filament has never been directly observed by AFM or electron microscopy. Thus, our developed tip-scan AFM offers significant potential in observing purified proteins and cellular structures at nanometer resolution. Current results represent an important step in the development of a new correlative microscope to observe nm-order structures at an acceptable frame rate (∼10 s/frame) by AFM at the position indicated by the fluorescent dye observed under a light microscope. PMID:27242058

Relative merits and limiting factors for x-ray and electron microscopy of thick, hydrated organic materials

DOE PAGES

Du, Ming; Jacobsen, Chris

2017-10-07

Electron and x-ray microscopes allow one to image the entire, unlabeled structure of hydrated materials at a resolution well beyond what visible light microscopes can achieve. However, both approaches involve ionizing radiation, so that radiation damage must be considered as one of the limits to imaging. Drawing upon earlier work, we describe here a unified approach to estimating the image contrast (and thus the required exposure and corresponding radiation dose) in both x-ray and electron microscopy. This approach accounts for factors such as plural and inelastic scattering, and (in electron microscopy) the use of energy filters to obtain so-called "zeromore » loss" images. As expected, it shows that electron microscopy offers lower dose for specimens thinner than about 1 mu m (such as for studies of macromolecules, viruses, bacteria and archaebacteria, and thin sectioned material), while x-ray microscopy offers superior characteristics for imaging thicker specimen such as whole eukaryotic cells, thick-sectioned tissues, and organs. The required radiation dose scales strongly as a function of the desired spatial resolution, allowing one to understand the limits of live and frozen hydrated specimen imaging. Lastly, we consider the factors limiting x-ray microscopy of thicker materials, suggesting that specimens as thick as a whole mouse brain can be imaged with x-ray microscopes without significant image degradation should appropriate image reconstruction methods be identified.« less

Relative merits and limiting factors for x-ray and electron microscopy of thick, hydrated organic materials

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

Du, Ming; Jacobsen, Chris

Electron and x-ray microscopes allow one to image the entire, unlabeled structure of hydrated materials at a resolution well beyond what visible light microscopes can achieve. However, both approaches involve ionizing radiation, so that radiation damage must be considered as one of the limits to imaging. Drawing upon earlier work, we describe here a unified approach to estimating the image contrast (and thus the required exposure and corresponding radiation dose) in both x-ray and electron microscopy. This approach accounts for factors such as plural and inelastic scattering, and (in electron microscopy) the use of energy filters to obtain so-called "zeromore » loss" images. As expected, it shows that electron microscopy offers lower dose for specimens thinner than about 1 mu m (such as for studies of macromolecules, viruses, bacteria and archaebacteria, and thin sectioned material), while x-ray microscopy offers superior characteristics for imaging thicker specimen such as whole eukaryotic cells, thick-sectioned tissues, and organs. The required radiation dose scales strongly as a function of the desired spatial resolution, allowing one to understand the limits of live and frozen hydrated specimen imaging. Lastly, we consider the factors limiting x-ray microscopy of thicker materials, suggesting that specimens as thick as a whole mouse brain can be imaged with x-ray microscopes without significant image degradation should appropriate image reconstruction methods be identified.« less

MicrOmega: a VIS/NIR hyperspectral microscope for in situ analysis in space

NASA Astrophysics Data System (ADS)

Leroi, V.; Bibring, J. P.; Berthé, M.

2008-07-01

MicrOmega is an ultra miniaturized spectral microscope for in situ analysis of samples. It is composed of 2 microscopes: one with a spatial sampling of 5 μm, working in 4 color in the visible range and one NIR hyperspectral microscope in the spectral range 0.9-4 μm with a spatial sampling of 20 μm per pixel (described in this paper). MicrOmega/NIR illuminates and images samples a few mm in size and acquires the NIR spectrum of each resolved pixel in up to 600 contiguous spectral channels. The goal of this instrument is to analyse in situ the composition of collected samples at almost their grain size scale, in a non destructive way. It should be among the first set of instruments who will analyse the sample and enable other complementary analyses to be performed on it. With the spectral range and resolution chosen, a wide variety of constituents can be identified: minerals, such as pyroxene and olivine, ferric oxides, hydrated phyllosilicates, sulfates and carbonates; ices and organics. The composition of the various phases within a given sample is a critical record of its formation and evolution. Coupled to the mapping information, it provides unique clues to describe the history of the parent body. In particular, the capability to identify hydrated grains and to characterize their adjacent phases has a huge potential in the search for potential bio-relics. We will present the major instrumental principles and specifications of MicrOmega/NIR, and its expected performances in particular for the ESA/ExoMars Mission.

Differential polarization laser scanning microscopy: biological applications

NASA Astrophysics Data System (ADS)

Steinbach, G.; Besson, F.; Pomozi, I.; Garab, G.

2005-09-01

With the aid of a differential polarization (DP) apparatus, developed in our laboratory and attached to our laser scanning confocal microscope, we can measure the magnitude and spatial distribution of 8 different DP quantities: linear and circular dichroism (LD&CD), linear and circular anisotropy of the emission (R and CPL, confocal), fluorescence detected dichroisms (FDLD&FDCD, confocal), linear birefringence (LB), and the degree of polarization of fluorescence emission (P, confocal). The attachment uses high frequency modulation and subsequent demodulation, via lock-in amplifier, of the detected intensity values, and records and displays pixel-by-pixel the measured DP quantity. These microscopic DP data carry important physical information on the molecular architecture of anisotropically organized samples. Microscopic DP measurements are thought to be of particular importance in biology. In most biological samples anisotropy is difficult to determine with conventional, macroscopic DP measurements and microscopic variations are of special significance. In this paper, we describe the method of LB imaging. Using magnetically oriented isolated chloroplasts trapped in polyacrylamide gel, we demonstrate that LB can be determined with high sensitivity and good spatial resolution. Granal thylakoid membranes in edge-aligned orientation exhibited strong LB, with large variations in its sign and magnitude. In face-aligned position LB was considerably weaker, and tended to vanish when averaged for the whole image. The strong local variations are attributed to the inherent heterogeneity of the membranes, i.e. to their internal differentiation into multilamellar, stacked membranes (grana), and single thylakoids (stroma membranes). Further details and applications of our DP-LSM will be published elsewhere.

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

Rosch, R.; Trosseille, C.; Caillaud, T.

The Laser Megajoule (LMJ) facility located at CEA/CESTA started to operate in the early 2014 with two quadruplets (20 kJ at 351 nm) focused on target for the first experimental campaign. We present here the first set of gated x-ray imaging (GXI) diagnostics implemented on LMJ since mid-2014. This set consists of two imaging diagnostics with spatial, temporal, and broadband spectral resolution. These diagnostics will give basic measurements, during the entire life of the facility, such as position, structure, and balance of beams, but they will also be used to characterize gas filled target implosion symmetry and timing, to studymore » x-ray radiography and hydrodynamic instabilities. The design requires a vulnerability approach, because components will operate in a harsh environment induced by neutron fluxes, gamma rays, debris, and shrapnel. Grazing incidence x-ray microscopes are fielded as far as possible away from the target to minimize potential damage and signal noise due to these sources. These imaging diagnostics incorporate microscopes with large source-to-optic distance and large size gated microchannel plate detectors. Microscopes include optics with grazing incidence mirrors, pinholes, and refractive lenses. Spatial, temporal, and spectral performances have been measured on x-ray tubes and UV lasers at CEA-DIF and at Physikalisch-Technische Bundesanstalt BESSY II synchrotron prior to be set on LMJ. GXI-1 and GXI-2 designs, metrology, and first experiments on LMJ are presented here.« less

CAMECA IMS 1300-HR3: The New Generation Ion Microprobe

NASA Astrophysics Data System (ADS)

Peres, P.; Choi, S. Y.; Renaud, L.; Saliot, P.; Larson, D. J.

2016-12-01

The success of secondary ion mass spectrometry (SIMS) in Geo- and Cosmo-chemistry relies on its performance in terms of: 1) very high sensitivity (mandatory for high precision measurements or to achieve low detection limits); 2) a broad mass range of elemental and isotopic species, from low mass (H) to high mass (U and above); 3) in-situ analysis of any solid flat polished surface; and 4) high spatial resolution from tens of microns down to sub-micron scale. The IMS 1300-HR3 (High Reproducibility, High spatial Resolution, High mass Resolution) is the latest generation of CAMECA's large geometry magnetic sector SIMS (or ion microprobe), successor to the internationally recognized IMS 1280-HR. The 1300-HR3delivers unmatched analytical performance for a wide range of applications (stable isotopes, geochronology, trace elements, nuclear safeguards and environmental studies…) due to: • High brightness RF-plasma oxygen ion source with enhanced beam density and current stability, dramatically improving spatial resolution, data reproducibility, and throughput • Automated sample loading system with motorized sample height (Z) adjustment, significantly increasing analysis precision, ease-of-use, and productivity • UV-light microscope for enhanced optical image resolution, together with dedicated software for easy sample navigation (developed by University of Wisconsin, USA) • Low noise 1012Ω resistor Faraday cup preamplifier boards for measuring low signal intensities In addition, improvements in electronics and software have been integrated into the new instrument. In order to meet a growing demand from geochronologists, CAMECA also introduces the KLEORA, which is a fully optimized ion microprobe for advanced mineral dating derived from the IMS 1300-HR3. Instrumental developments as well as data obtained for stable isotope and U-Pb dating applications will be presented in detail.

Multiscale and multi-modality visualization of angiogenesis in a human breast cancer model

PubMed Central

Cebulla, Jana; Kim, Eugene; Rhie, Kevin; Zhang, Jiangyang

2017-01-01

Angiogenesis in breast cancer helps fulfill the metabolic demands of the progressing tumor and plays a critical role in tumor metastasis. Therefore, various imaging modalities have been used to characterize tumor angiogenesis. While micro-CT (μCT) is a powerful tool for analyzing the tumor microvascular architecture at micron-scale resolution, magnetic resonance imaging (MRI) with its sub-millimeter resolution is useful for obtaining in vivo vascular data (e.g. tumor blood volume and vessel size index). However, integration of these microscopic and macroscopic angiogenesis data across spatial resolutions remains challenging. Here we demonstrate the feasibility of ‘multiscale’ angiogenesis imaging in a human breast cancer model, wherein we bridge the resolution gap between ex vivo μCT and in vivo MRI using intermediate resolution ex vivo MR microscopy (μMRI). To achieve this integration, we developed suitable vessel segmentation techniques for the ex vivo imaging data and co-registered the vascular data from all three imaging modalities. We showcase two applications of this multiscale, multi-modality imaging approach: (1) creation of co-registered maps of vascular volume from three independent imaging modalities, and (2) visualization of differences in tumor vasculature between viable and necrotic tumor regions by integrating μCT vascular data with tumor cellularity data obtained using diffusion-weighted MRI. Collectively, these results demonstrate the utility of ‘mesoscopic’ resolution μMRI for integrating macroscopic in vivo MRI data and microscopic μCT data. Although focused on the breast tumor xenograft vasculature, our imaging platform could be extended to include additional data types for a detailed characterization of the tumor microenvironment and computational systems biology applications. PMID:24719185

In situ hybridization at the electron microscope level: localization of transcripts on ultrathin sections of Lowicryl K4M-embedded tissue using biotinylated probes and protein A-gold complexes

PubMed Central

1986-01-01

A technique has been developed for localizing hybrids formed in situ on semi-thin and ultrathin sections of Lowicryl K4M-embedded tissue. Biotinylated dUTP (Bio-11-dUTP and/or Bio-16-dUTP) was incorporated into mitochondrial rDNA and small nuclear U1 probes by nick- translation. The probes were hybridized to sections of Drosophila ovaries and subsequently detected with an anti-biotin antibody and protein A-gold complex. On semi-thin sections, probe detection was achieved by amplification steps with anti-protein A antibody and protein A-gold with subsequent silver enhancement. At the electron microscope level, specific labeling was obtained over structures known to be the site of expression of the appropriate genes (i.e., either over mitochondria or over nuclei). The labeling pattern at the light microscope level (semi-thin sections) was consistent with that obtained at the electron microscope level. The described nonradioactive procedures for hybrid detection on Lowicryl K4M-embedded tissue sections offer several advantages: rapid signal detection: superior morphological preservation and spatial resolution; and signal-to-noise ratios equivalent to radiolabeling. PMID:3084498

Enabling low-noise null-point scanning thermal microscopy by the optimization of scanning thermal microscope probe through a rigorous theory of quantitative measurement.

PubMed

Hwang, Gwangseok; Chung, Jaehun; Kwon, Ohmyoung

2014-11-01

The application of conventional scanning thermal microscopy (SThM) is severely limited by three major problems: (i) distortion of the measured signal due to heat transfer through the air, (ii) the unknown and variable value of the tip-sample thermal contact resistance, and (iii) perturbation of the sample temperature due to the heat flux through the tip-sample thermal contact. Recently, we proposed null-point scanning thermal microscopy (NP SThM) as a way of overcoming these problems in principle by tracking the thermal equilibrium between the end of the SThM tip and the sample surface. However, in order to obtain high spatial resolution, which is the primary motivation for SThM, NP SThM requires an extremely sensitive SThM probe that can trace the vanishingly small heat flux through the tip-sample nano-thermal contact. Herein, we derive a relation between the spatial resolution and the design parameters of a SThM probe, optimize the thermal and electrical design, and develop a batch-fabrication process. We also quantitatively demonstrate significantly improved sensitivity, lower measurement noise, and higher spatial resolution of the fabricated SThM probes. By utilizing the exceptional performance of these fabricated probes, we show that NP SThM can be used to obtain a quantitative temperature profile with nanoscale resolution independent of the changing tip-sample thermal contact resistance and without perturbation of the sample temperature or distortion due to the heat transfer through the air.

Alpha particle spectroscopy using FNTD and SIM super-resolution microscopy.

PubMed

Kouwenberg, J J M; Kremers, G J; Slotman, J A; Wolterbeek, H T; Houtsmuller, A B; Denkova, A G; Bos, A J J

2018-06-01

Structured illumination microscopy (SIM) for the imaging of alpha particle tracks in fluorescent nuclear track detectors (FNTD) was evaluated and compared to confocal laser scanning microscopy (CLSM). FNTDs were irradiated with an external alpha source and imaged using both methodologies. SIM imaging resulted in improved resolution, without increase in scan time. Alpha particle energy estimation based on the track length, direction and intensity produced results in good agreement with the expected alpha particle energy distribution. A pronounced difference was seen in the spatial scattering of alpha particles in the detectors, where SIM showed an almost 50% reduction compared to CLSM. The improved resolution of SIM allows for more detailed studies of the tracks induced by ionising particles. The combination of SIM and FNTDs for alpha radiation paves the way for affordable and fast alpha spectroscopy and dosimetry. © 2018 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.

Self-interference 3D super-resolution microscopy for deep tissue investigations.

PubMed

Bon, Pierre; Linarès-Loyez, Jeanne; Feyeux, Maxime; Alessandri, Kevin; Lounis, Brahim; Nassoy, Pierre; Cognet, Laurent

2018-06-01

Fluorescence localization microscopy has achieved near-molecular resolution capable of revealing ultra-structures, with a broad range of applications, especially in cellular biology. However, it remains challenging to attain such resolution in three dimensions and inside biological tissues beyond the first cell layer. Here we introduce SELFI, a framework for 3D single-molecule localization within multicellular specimens and tissues. The approach relies on self-interference generated within the microscope's point spread function (PSF) to simultaneously encode equiphase and intensity fluorescence signals, which together provide the 3D position of an emitter. We combined SELFI with conventional localization microscopy to visualize F-actin 3D filament networks and reveal the spatial distribution of the transcription factor OCT4 in human induced pluripotent stem cells at depths up to 50 µm inside uncleared tissue spheroids. SELFI paves the way to nanoscale investigations of native cellular processes in intact tissues.

Confocal detection of Rayleigh scattering for residual stress measurement in chemically tempered glass

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

Hödemann, S., E-mail: siim.hodemann@ut.ee; Möls, P.; Kiisk, V.

2015-12-28

A new optical method is presented for evaluation of the stress profile in chemically tempered (chemically strengthened) glass based on confocal detection of scattered laser beam. Theoretically, a lateral resolution of 0.2 μm and a depth resolution of 0.6 μm could be achieved by using a confocal microscope with high-NA immersion objective. The stress profile in the 250 μm thick surface layer of chemically tempered lithium aluminosilicate glass was measured with a high spatial resolution to illustrate the capability of the method. The confocal method is validated using transmission photoelastic and Na{sup +} ion concentration profile measurement. Compositional influence on the stress-optic coefficientmore » is calculated and discussed. Our method opens up new possibilities for three-dimensional scattered light tomography of mechanical imaging in birefringent materials.« less

Nanoscale deformation analysis with high-resolution transmission electron microscopy and digital image correlation

DOE PAGES

Wang, Xueju; Pan, Zhipeng; Fan, Feifei; ...

2015-09-10

We present an application of the digital image correlation (DIC) method to high-resolution transmission electron microscopy (HRTEM) images for nanoscale deformation analysis. The combination of DIC and HRTEM offers both the ultrahigh spatial resolution and high displacement detection sensitivity that are not possible with other microscope-based DIC techniques. We demonstrate the accuracy and utility of the HRTEM-DIC technique through displacement and strain analysis on amorphous silicon. Two types of error sources resulting from the transmission electron microscopy (TEM) image noise and electromagnetic-lens distortions are quantitatively investigated via rigid-body translation experiments. The local and global DIC approaches are applied for themore » analysis of diffusion- and reaction-induced deformation fields in electrochemically lithiated amorphous silicon. As a result, the DIC technique coupled with HRTEM provides a new avenue for the deformation analysis of materials at the nanometer length scales.« less

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.

Overview of Athena Microscopic Imager Results

NASA Technical Reports Server (NTRS)

Herkenhoff, K.; Squyres, S.; Arvidson, R.; Bass, D.; Bell, J., III; Bertelsen, P.; Cabrol, N.; Ehlmann, B.; Farrand, W.; Gaddis, L.

2005-01-01

The Athena science payload on the Mars Exploration Rovers (MER) includes the Microscopic Imager (MI). The MI is a fixed-focus camera mounted on an extendable arm, the Instrument Deployment Device (IDD). The MI acquires images at a spatial resolution of 31 microns/pixel over a broad spectral range (400 - 700 nm). The MI uses the same electronics design as the other MER cameras but its optics yield a field of view of 32 32 mm across a 1024 1024 pixel CCD image. The MI acquires images using only solar or skylight illumination of the target surface. The MI science objectives, instrument design and calibration, operation, and data processing were described by Herkenhoff et al. Initial results of the MI experiment on both MER rovers (Spirit and Opportunity) have been published previously. Highlights of these and more recent results are described.

X-ray microanalysis in the scanning electron microscope.

PubMed

Roomans, Godfried M; Dragomir, Anca

2014-01-01

X-ray microanalysis conducted using the scanning electron microscope is a technique that allows the determination of chemical elements in bulk or semi-thick specimens. The lowest concentration of an element that can be detected is in the order of a few mmol/kg or a few hundred parts per million, and the smallest amount is in the order of 10(-18) g. The spatial resolution of the analysis depends on the thickness of the specimen. For biological specimen analysis, care must be taken to prevent displacement/loss of the element of interest (usually ions). Protocols are presented for the processing of frozen-hydrated and freeze-dried specimens, as well as for the analysis of small volumes of fluid, cell cultures, and other specimens. Aspects of qualitative and quantitative analysis are covered, including limitations of the technique.

X-ray microanalysis in the scanning electron microscope.

PubMed

Roomans, Godfried M; Dragomir, Anca

2007-01-01

X-ray microanalysis conducted using the scanning electron microscope is a technique that allows the determination of chemical elements in bulk or semithick specimens. The lowest concentration of an element that can be detected is in the order of a few mmol/kg or a few hundred parts per million, and the smallest amount is in the order of 10(-18) g. The spatial resolution of the analysis depends on the thickness of the specimen. For biological specimen analysis, care must be taken to prevent displacement/loss of the element of interest (usually ions). Protocols are presented for the processing of frozen-hydrated and freeze-dried specimens, as well as for the analysis of small volumes of fluid, cell cultures and other specimens. Aspects of qualitative and quantitative analysis are covered, including limitations of the technique.

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

The ALBA spectroscopic LEEM-PEEM experimental station: layout and performance

PubMed Central

Aballe, Lucia; Foerster, Michael; Pellegrin, Eric; Nicolas, Josep; Ferrer, Salvador

2015-01-01

The spectroscopic LEEM-PEEM experimental station at the CIRCE helical undulator beamline, which started user operation at the ALBA Synchrotron Light Facility in 2012, is presented. This station, based on an Elmitec LEEM III microscope with electron imaging energy analyzer, permits surfaces to be imaged with chemical, structural and magnetic sensitivity down to a lateral spatial resolution better than 20 nm with X-ray excited photoelectrons and 10 nm in LEEM and UV-PEEM modes. Rotation around the surface normal and application of electric and (weak) magnetic fields are possible in the microscope chamber. In situ surface preparation capabilities include ion sputtering, high-temperature flashing, exposure to gases, and metal evaporation with quick evaporator exchange. Results from experiments in a variety of fields and imaging modes will be presented in order to illustrate the ALBA XPEEM capabilities. PMID:25931092

Optical Antenna-Based Fluorescence Correlation Spectroscopy to Probe the Nanoscale Dynamics of Biological Membranes.

PubMed

Winkler, Pamina M; Regmi, Raju; Flauraud, Valentin; Brugger, Jürgen; Rigneault, Hervé; Wenger, Jérôme; García-Parajo, María F

2018-01-04

The plasma membrane of living cells is compartmentalized at multiple spatial scales ranging from the nano- to the mesoscale. This nonrandom organization is crucial for a large number of cellular functions. At the nanoscale, cell membranes organize into dynamic nanoassemblies enriched by cholesterol, sphingolipids, and certain types of proteins. Investigating these nanoassemblies known as lipid rafts is of paramount interest in fundamental cell biology. However, this goal requires simultaneous nanometer spatial precision and microsecond temporal resolution, which is beyond the reach of common microscopes. Optical antennas based on metallic nanostructures efficiently enhance and confine light into nanometer dimensions, breaching the diffraction limit of light. In this Perspective, we discuss recent progress combining optical antennas with fluorescence correlation spectroscopy (FCS) to monitor microsecond dynamics at nanoscale spatial dimensions. These new developments offer numerous opportunities to investigate lipid and protein dynamics in both mimetic and native biological membranes.

Development and characterization of monolithic multilayer Laue lens nanofocusing optics

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

Nazaretski, E.; Xu, W.; Bouet, N.

2016-06-27

We have developed an experimental approach to bond two independent linear Multilayer Laue Lenses (MLLs) together. A monolithic MLL structure was characterized using ptychography at 12 keV photon energy, and we demonstrated 12 nm and 24 nm focusing in horizontal and vertical directions, respectively. Fabrication of 2D MLL optics allows installation of these focusing elements in more conventional microscopes suitable for x-ray imaging using zone plates, and opens easier access to 2D imaging with high spatial resolution in the hard x-ray regime.

Development and characterization of monolithic multilayer Laue lens nanofocusing optics

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

Nazaretski, E.; Xu, W., E-mail: weihexu@bnl.gov; Bouet, N.

2016-06-27

We have developed an experimental approach to bond two independent linear Multilayer Laue Lenses (MLLs) together. A monolithic MLL structure was characterized using ptychography at 12 keV photon energy, and we demonstrated 12 nm and 24 nm focusing in horizontal and vertical directions, respectively. Fabrication of 2D MLL optics allows installation of these focusing elements in more conventional microscopes suitable for x-ray imaging using zone plates, and opens easier access to 2D imaging with high spatial resolution in the hard x-ray regime.

Organic chemical analysis on a microscopic scale using two-step laser desorption/laser ionization mass spectrometry

NASA Technical Reports Server (NTRS)

Kovalenko, L. J.; Philippoz, J.-M.; Bucenell, J. R.; Zenobi, R.; Zare, R. N.

1991-01-01

The distribution of PAHs in the Allende meteorite has been measured using two-step laser desorption and laser multiphoton-ionization mass spectrometry. This method enables in situ analysis (with a spatial resolution of 1 mm or better) of selected organic molecules. Results show that PAH concentrations are locally high compared to the average concentration found by analysis of pulverized samples, and are found primarily in the fine-grained matrix; no PAHs were detected in the interiors of individual chondrules at the detection limit (about 0.05 ppm).

Movement measurement of isolated skeletal muscle using imaging microscopy

NASA Astrophysics Data System (ADS)

Elias, David; Zepeda, Hugo; Leija, Lorenzo S.; Sossa, Humberto; de la Rosa, Jose I.

1997-05-01

An imaging-microscopy methodology to measure contraction movement in chemically stimulated crustacean skeletal muscle, whose movement speed is about 0.02 mm/s is presented. For this, a CCD camera coupled to a microscope and a high speed digital image acquisition system, allowing us to capture 960 images per second are used. The images are digitally processed in a PC and displayed in a video monitor. A maximal field of 0.198 X 0.198 mm2 and a spatial resolution of 3.5 micrometers are obtained.

Development and characterization of monolithic multilayer Laue lens nanofocusing optics

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

Nazaretski, E.; Xu, W.; Bouet, N.

In this study, we have developed an experimental approach to bond two independent linear Multilayer Laue Lenses (MLLs) together. A monolithic MLL structure was characterized using ptychography at 12 keV photon energy, and we demonstrated 12 nm and 24 nm focusing in horizontal and vertical directions, respectively. Fabrication of 2D MLL optics allows installation of these focusing elements in more conventional microscopes suitable for x-ray imaging using zone plates, and opens easier access to 2D imaging with high spatial resolution in the hard x-ray regime.

Development and characterization of monolithic multilayer Laue lens nanofocusing optics

DOE PAGES

Nazaretski, E.; Xu, W.; Bouet, N.; ...

2016-06-27

In this study, we have developed an experimental approach to bond two independent linear Multilayer Laue Lenses (MLLs) together. A monolithic MLL structure was characterized using ptychography at 12 keV photon energy, and we demonstrated 12 nm and 24 nm focusing in horizontal and vertical directions, respectively. Fabrication of 2D MLL optics allows installation of these focusing elements in more conventional microscopes suitable for x-ray imaging using zone plates, and opens easier access to 2D imaging with high spatial resolution in the hard x-ray regime.

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

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

Spatial-spectral blood cell classification with microscopic hyperspectral imagery

NASA Astrophysics Data System (ADS)

Ran, Qiong; Chang, Lan; Li, Wei; Xu, Xiaofeng

2017-10-01

Microscopic hyperspectral images provide a new way for blood cell examination. The hyperspectral imagery can greatly facilitate the classification of different blood cells. In this paper, the microscopic hyperspectral images are acquired by connecting the microscope and the hyperspectral imager, and then tested for blood cell classification. For combined use of the spectral and spatial information provided by hyperspectral images, a spatial-spectral classification method is improved from the classical extreme learning machine (ELM) by integrating spatial context into the image classification task with Markov random field (MRF) model. Comparisons are done among ELM, ELM-MRF, support vector machines(SVM) and SVMMRF methods. Results show the spatial-spectral classification methods(ELM-MRF, SVM-MRF) perform better than pixel-based methods(ELM, SVM), and the proposed ELM-MRF has higher precision and show more accurate location of cells.

Towards native-state imaging in biological context in the electron microscope

PubMed Central

Weston, Anne E.; Armer, Hannah E. J.

2009-01-01

Modern cell biology is reliant on light and fluorescence microscopy for analysis of cells, tissues and protein localisation. However, these powerful techniques are ultimately limited in resolution by the wavelength of light. Electron microscopes offer much greater resolution due to the shorter effective wavelength of electrons, allowing direct imaging of sub-cellular architecture. The harsh environment of the electron microscope chamber and the properties of the electron beam have led to complex chemical and mechanical preparation techniques, which distance biological samples from their native state and complicate data interpretation. Here we describe recent advances in sample preparation and instrumentation, which push the boundaries of high-resolution imaging. Cryopreparation, cryoelectron microscopy and environmental scanning electron microscopy strive to image samples in near native state. Advances in correlative microscopy and markers enable high-resolution localisation of proteins. Innovation in microscope design has pushed the boundaries of resolution to atomic scale, whilst automatic acquisition of high-resolution electron microscopy data through large volumes is finally able to place ultrastructure in biological context. PMID:19916039

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

PubMed

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

2011-04-07

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

Wave optics theory and 3-D deconvolution for the light field microscope

PubMed Central

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

2013-01-01

Light field microscopy is a new technique for high-speed volumetric imaging of weakly scattering or fluorescent specimens. It employs an array of microlenses to trade off spatial resolution against angular resolution, thereby allowing a 4-D light field to be captured using a single photographic exposure without the need for scanning. The recorded light field can then be used to computationally reconstruct a full volume. In this paper, we present an optical model for light field microscopy based on wave optics, instead of previously reported ray optics models. We also present a 3-D deconvolution method for light field microscopy that is able to reconstruct volumes at higher spatial resolution, and with better optical sectioning, than previously reported. To accomplish this, we take advantage of the dense spatio-angular sampling provided by a microlens array at axial positions away from the native object plane. This dense sampling permits us to decode aliasing present in the light field to reconstruct high-frequency information. We formulate our method as an inverse problem for reconstructing the 3-D volume, which we solve using a GPU-accelerated iterative algorithm. Theoretical limits on the depth-dependent lateral resolution of the reconstructed volumes are derived. We show that these limits are in good agreement with experimental results on a standard USAF 1951 resolution target. Finally, we present 3-D reconstructions of pollen grains that demonstrate the improvements in fidelity made possible by our method. PMID:24150383

Analytical SuperSTEM for extraterrestrial materials research

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

Bradley, J P; Dai, Z R

2009-09-08

Electron-beam studies of extraterrestrial materials with significantly improved spatial resolution, energy resolution and sensitivity are enabled using a 300 keV SuperSTEM scanning transmission electron microscope with a monochromator and two spherical aberration correctors. The improved technical capabilities enable analyses previously not possible. Mineral structures can be directly imaged and analyzed with single-atomic-column resolution, liquids and implanted gases can be detected, and UV-VIS optical properties can be measured. Detection limits for minor/trace elements in thin (<100 nm thick) specimens are improved such that quantitative measurements of some extend to the sub-500 ppm level. Electron energy-loss spectroscopy (EELS) can be carried outmore » with 0.10-0.20 eV energy resolution and atomic-scale spatial resolution such that variations in oxidation state from one atomic column to another can be detected. Petrographic mapping is extended down to the atomic scale using energy-dispersive x-ray spectroscopy (EDS) and energy-filtered transmission electron microscopy (EFTEM) imaging. Technical capabilities and examples of the applications of SuperSTEM to extraterrestrial materials are presented, including the UV spectral properties and organic carbon K-edge fine structure of carbonaceous matter in interplanetary dust particles (IDPs), x-ray elemental maps showing the nanometer-scale distribution of carbon within GEMS (glass with embedded metal and sulfides), the first detection and quantification of trace Ti in GEMS using EDS, and detection of molecular H{sub 2}O in vesicles and implanted H{sub 2} and He in irradiated mineral and glass grains.« less

In situ high-resolution thermal microscopy on integrated circuits.

PubMed

Zhuo, Guan-Yu; Su, Hai-Ching; Wang, Hsien-Yi; Chan, Ming-Che

2017-09-04

The miniaturization of metal tracks in integrated circuits (ICs) can cause abnormal heat dissipation, resulting in electrostatic discharge, overvoltage breakdown, and other unwanted issues. Unfortunately, locating areas of abnormal heat dissipation is limited either by the spatial resolution or imaging acquisition speed of current thermal analytical techniques. A rapid, non-contact approach to the thermal imaging of ICs with sub-μm resolution could help to alleviate this issue. In this work, based on the intensity of the temperature-dependent two-photon fluorescence (TPF) of Rhodamine 6G (R6G) material, we developed a novel fast and non-invasive thermal microscopy with a sub-μm resolution. Its application to the location of hotspots that may evolve into thermally induced defects in ICs was also demonstrated. To the best of our knowledge, this is the first study to present high-resolution 2D thermal microscopic images of ICs, showing the generation, propagation, and distribution of heat during its operation. According to the demonstrated results, this scheme has considerable potential for future in situ hotspot analysis during the optimization stage of IC development.

Co-Registered In Situ Secondary Electron and Mass Spectral Imaging on the Helium Ion Microscope Demonstrated Using Lithium Titanate and Magnesium Oxide Nanoparticles.

PubMed

Dowsett, D; Wirtz, T

2017-09-05

The development of a high resolution elemental imaging platform combining coregistered secondary ion mass spectrometry and high resolution secondary electron imaging is reported. The basic instrument setup and operation are discussed and in situ image correlation is demonstrated on a lithium titanate and magnesium oxide nanoparticle mixture. The instrument uses both helium and neon ion beams generated by a gas field ion source to irradiate the sample. Both secondary electrons and secondary ions may be detected. Secondary ion mass spectrometry (SIMS) is performed using an in-house developed double focusing magnetic sector spectrometer with parallel detection. Spatial resolutions of 10 nm have been obtained in SIMS mode. Both the secondary electron and SIMS image data are very surface sensitive and have approximately the same information depth. While the spatial resolutions are approximately a factor of 10 different, switching between the different images modes may be done in situ and extremely rapidly, allowing for simple imaging of the same region of interest and excellent coregistration of data sets. The ability to correlate mass spectral images on the 10 nm scale with secondary electron images on the nanometer scale in situ has the potential to provide a step change in our understanding of nanoscale phenomena in fields from materials science to life science.

Quantitative 3D high resolution transmission ultrasound tomography: creating clinically relevant images (Conference Presentation)

NASA Astrophysics Data System (ADS)

Wiskin, James; Klock, John; Iuanow, Elaine; Borup, Dave T.; Terry, Robin; Malik, Bilal H.; Lenox, Mark

2017-03-01

There has been a great deal of research into ultrasound tomography for breast imaging over the past 35 years. Few successful attempts have been made to reconstruct high-resolution images using transmission ultrasound. To this end, advances have been made in 2D and 3D algorithms that utilize either time of arrival or full wave data to reconstruct images with high spatial and contrast resolution suitable for clinical interpretation. The highest resolution and quantitative accuracy result from inverse scattering applied to full wave data in 3D. However, this has been prohibitively computationally expensive, meaning that full inverse scattering ultrasound tomography has not been considered clinically viable. Here we show the results of applying a nonlinear inverse scattering algorithm to 3D data in a clinically useful time frame. This method yields Quantitative Transmission (QT) ultrasound images with high spatial and contrast resolution. We reconstruct sound speeds for various 2D and 3D phantoms and verify these values with independent measurements. The data are fully 3D as is the reconstruction algorithm, with no 2D approximations. We show that 2D reconstruction algorithms can introduce artifacts into the QT breast image which are avoided by using a full 3D algorithm and data. We show high resolution gross and microscopic anatomic correlations comparing cadaveric breast QT images with MRI to establish imaging capability and accuracy. Finally, we show reconstructions of data from volunteers, as well as an objective visual grading analysis to confirm clinical imaging capability and accuracy.

High resolution imaging of intracellular oxygen concentration by phosphorescence lifetime

PubMed Central

Kurokawa, Hiromi; Ito, Hidehiro; Inoue, Mai; Tabata, Kenji; Sato, Yoshifumi; Yamagata, Kazuya; Kizaka-Kondoh, Shinae; Kadonosono, Tetsuya; Yano, Shigenobu; Inoue, Masahiro; Kamachi, Toshiaki

2015-01-01

Optical methods using phosphorescence quenching by oxygen are suitable for sequential monitoring and non-invasive measurements for oxygen concentration (OC) imaging within cells. Phosphorescence intensity measurement is widely used with phosphorescent dyes. These dyes are ubiquitously but heterogeneously distributed inside the whole cell. The distribution of phosphorescent dye is a major disadvantage in phosphorescence intensity measurement. We established OC imaging system for a single cell using phosphorescence lifetime and a laser scanning confocal microscope. This system had improved spatial resolution and reduced the measurement time with the high repetition rate of the laser. By the combination of ubiquitously distributed phosphorescent dye with this lifetime imaging microscope, we can visualize the OC inside the whole cell and spheroid. This system uses reversible phosphorescence quenching by oxygen, so it can measure successive OC changes from normoxia to anoxia. Lower regions of OC inside the cell colocalized with mitochondria. The time-dependent OC change in an insulin-producing cell line MIN6 by the glucose stimulation was successfully visualized. Assessing the detailed distribution and dynamics of OC inside cells achieved by the presented system will be useful to understanding a physiological and pathological oxygen metabolism. PMID:26065366

Cryo X-ray microscope with flat sample geometry for correlative fluorescence and nanoscale tomographic imaging.

PubMed

Schneider, Gerd; Guttmann, Peter; Rehbein, Stefan; Werner, Stephan; Follath, Rolf

2012-02-01

X-ray imaging offers a new 3-D view into cells. With its ability to penetrate whole hydrated cells it is ideally suited for pairing fluorescence light microscopy and nanoscale X-ray tomography. In this paper, we describe the X-ray optical set-up and the design of the cryo full-field transmission X-ray microscope (TXM) at the electron storage ring BESSY II. Compared to previous TXM set-ups with zone plate condenser monochromator, the new X-ray optical layout employs an undulator source, a spherical grating monochromator and an elliptically shaped glass capillary mirror as condenser. This set-up improves the spectral resolution by an order of magnitude. Furthermore, the partially coherent object illumination improves the contrast transfer of the microscope compared to incoherent conditions. With the new TXM, cells grown on flat support grids can be tilted perpendicular to the optical axis without any geometrical restrictions by the previously required pinhole for the zone plate monochromator close to the sample plane. We also developed an incorporated fluorescence light microscope which permits to record fluorescence, bright field and DIC images of cryogenic cells inside the TXM. For TXM tomography, imaging with multi-keV X-rays is a straightforward approach to increase the depth of focus. Under these conditions phase contrast imaging is necessary. For soft X-rays with shrinking depth of focus towards 10nm spatial resolution, thin optical sections through a thick specimen might be obtained by deconvolution X-ray microscopy. As alternative 3-D X-ray imaging techniques, the confocal cryo-STXM and the dual beam cryo-FIB/STXM with photoelectron detection are proposed. Copyright © 2012 Elsevier Inc. All rights reserved.

Upgrade of a Scanning Confocal Microscope to a Single-Beam Path STED Microscope

PubMed Central

Klauss, André; König, Marcelle; Hille, Carsten

2015-01-01

By overcoming the diffraction limit in light microscopy, super-resolution techniques, such as stimulated emission depletion (STED) microscopy, are experiencing an increasing impact on life sciences. High costs and technically demanding setups, however, may still hinder a wider distribution of this innovation in biomedical research laboratories. As far-field microscopy is the most widely employed microscopy modality in the life sciences, upgrading already existing systems seems to be an attractive option for achieving diffraction-unlimited fluorescence microscopy in a cost-effective manner. Here, we demonstrate the successful upgrade of a commercial time-resolved confocal fluorescence microscope to an easy-to-align STED microscope in the single-beam path layout, previously proposed as “easy-STED”, achieving lateral resolution < λ/10 corresponding to a five-fold improvement over a confocal modality. For this purpose, both the excitation and depletion laser beams pass through a commercially available segmented phase plate that creates the STED-doughnut light distribution in the focal plane, while leaving the excitation beam unaltered when implemented into the joint beam path. Diffraction-unlimited imaging of 20 nm-sized fluorescent beads as reference were achieved with the wavelength combination of 635 nm excitation and 766 nm depletion. To evaluate the STED performance in biological systems, we compared the popular phalloidin-coupled fluorescent dyes Atto647N and Abberior STAR635 by labeling F-actin filaments in vitro as well as through immunofluorescence recordings of microtubules in a complex epithelial tissue. Here, we applied a recently proposed deconvolution approach and showed that images obtained from time-gated pulsed STED microscopy may benefit concerning the signal-to-background ratio, from the joint deconvolution of sub-images with different spatial information which were extracted from offline time gating. PMID:26091552

Vector sensor for scanning SQUID microscopy

NASA Astrophysics Data System (ADS)

Dang, Vu The; Toji, Masaki; Thanh Huy, Ho; Miyajima, Shigeyuki; Shishido, Hiroaki; Hidaka, Mutsuo; Hayashi, Masahiko; Ishida, Takekazu

2017-07-01

We plan to build a novel 3-dimensional (3D) scanning SQUID microscope with high sensitivity and high spatial resolution. In the system, a vector sensor consists of three SQUID sensors and three pick-up coils realized on a single chip. Three pick-up coils are configured in orthogonal with each other to measure the magnetic field vector of X, Y, Z components. We fabricated some SQUID chips with one uniaxial pick-up coil or three vector pick-up coils and carried out fundamental measurements to reveal the basic characteristics. Josephson junctions (JJs) of sensors are designed to have the critical current density J c of 320 A/cm2, and the critical current I c becomes 12.5 μA for the 2.2μm × 2.2μm JJ. We carefully positioned the three pickup coils so as to keep them at the same height at the centers of all three X, Y and Z coils. This can be done by arranging them along single line parallel to a sample surface. With the aid of multilayer technology of Nb-based fabrication, we attempted to reduce an inner diameter of the pickup coils to enhance both sensitivity and spatial resolution. The method for improving a spatial resolution of a local magnetic field image is to employ an XYZ piezo-driven scanner for controlling the positions of the pick-up coils. The fundamental characteristics of our SQUID sensors confirmed the proper operation of our SQUID sensors and found a good agreement with our design parameters.

Multiplexed 3D FRET imaging in deep tissue of live embryos

PubMed Central

Zhao, Ming; Wan, Xiaoyang; Li, Yu; Zhou, Weibin; Peng, Leilei

2015-01-01

Current deep tissue microscopy techniques are mostly restricted to intensity mapping of fluorophores, which significantly limit their applications in investigating biochemical processes in vivo. We present a deep tissue multiplexed functional imaging method that probes multiple Förster resonant energy transfer (FRET) sensors in live embryos with high spatial resolution. The method simultaneously images fluorescence lifetimes in 3D with multiple excitation lasers. Through quantitative analysis of triple-channel intensity and lifetime images, we demonstrated that Ca2+ and cAMP levels of live embryos expressing dual FRET sensors can be monitored simultaneously at microscopic resolution. The method is compatible with a broad range of FRET sensors currently available for probing various cellular biochemical functions. It opens the door to imaging complex cellular circuitries in whole live organisms. PMID:26387920

Adapting a compact confocal microscope system to a two-photon excitation fluorescence imaging architecture.

PubMed

Diaspro, A; Corosu, M; Ramoino, P; Robello, M

1999-11-01

Within the framework of a national National Institute of Physics of Matter (INFM) project, we have realised a two-photon excitation (TPE) fluorescence microscope based on a new generation commercial confocal scanning head. The core of the architecture is a mode-locked Ti:Sapphire laser (Tsunami 3960, Spectra Physics Inc., Mountain View, CA) pumped by a high-power (5 W, 532 nm) laser (Millennia V, Spectra Physics Inc.) and an ultracompact confocal scanning head, Nikon PCM2000 (Nikon Instruments, Florence, Italy) using a single-pinhole design. Three-dimensional point-spread function has been measured to define spatial resolution performances. The TPE microscope has been used with a wide range of excitable fluorescent molecules (DAPI, Fura-2, Indo-1, DiOC(6)(3), fluoresceine, Texas red) covering a single photon spectral range from UV to green. An example is reported on 3D imaging of the helical structure of the sperm head of the Octopus Eledone cirrhosa labelled with an UV excitable dye, i.e., DAPI. The system can be easily switched for operating both in conventional and two-photon mode. Copyright 1999 Wiley-Liss, Inc.

Thermal radiation scanning tunnelling microscopy

NASA Astrophysics Data System (ADS)

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

2006-12-01

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

Single shot, three-dimensional fluorescence microscopy with a spatially rotating point spread function

PubMed Central

Wang, Zhaojun; Cai, Yanan; Liang, Yansheng; Zhou, Xing; Yan, Shaohui; Dan, Dan; Bianco, Piero R.; Lei, Ming; Yao, Baoli

2017-01-01

A wide-field fluorescence microscope with a double-helix point spread function (PSF) is constructed to obtain the specimen’s three-dimensional distribution with a single snapshot. Spiral-phase-based computer-generated holograms (CGHs) are adopted to make the depth-of-field of the microscope adjustable. The impact of system aberrations on the double-helix PSF at high numerical aperture is analyzed to reveal the necessity of the aberration correction. A modified cepstrum-based reconstruction scheme is promoted in accordance with properties of the new double-helix PSF. The extended depth-of-field images and the corresponding depth maps for both a simulated sample and a tilted section slice of bovine pulmonary artery endothelial (BPAE) cells are recovered, respectively, verifying that the depth-of-field is properly extended and the depth of the specimen can be estimated at a precision of 23.4nm. This three-dimensional fluorescence microscope with a framerate-rank time resolution is suitable for studying the fast developing process of thin and sparsely distributed micron-scale cells in extended depth-of-field. PMID:29296483

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

NASA Astrophysics Data System (ADS)

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

2017-10-01

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

Proton beam spatial distribution and Bragg peak imaging by photoluminescence of color centers in lithium fluoride crystals at the TOP-IMPLART linear accelerator

NASA Astrophysics Data System (ADS)

Piccinini, M.; Ronsivalle, C.; Ampollini, A.; Bazzano, G.; Picardi, L.; Nenzi, P.; Trinca, E.; Vadrucci, M.; Bonfigli, F.; Nichelatti, E.; Vincenti, M. A.; Montereali, R. M.

2017-11-01

Solid-state radiation detectors based on the photoluminescence of stable point defects in lithium fluoride crystals have been used for advanced diagnostics during the commissioning of the segment up to 27 MeV of the TOP-IMPLART proton linear accelerator for proton therapy applications, under development at ENEA C.R. Frascati, Italy. The LiF detectors high intrinsic spatial resolution and wide dynamic range allow obtaining two-dimensional images of the beam transverse intensity distribution and also identifying the Bragg peak position with micrometric precision by using a conventional optical fluorescence microscope. Results of the proton beam characterization, among which, the estimation of beam energy components and dynamics, are reported and discussed for different operating conditions of the accelerator.

Learning surface molecular structures via machine vision

DOE PAGES

Ziatdinov, Maxim; Maksov, Artem; Kalinin, Sergei V.

2017-08-10

Recent advances in high resolution scanning transmission electron and scanning probe microscopies have allowed researchers to perform measurements of materials structural parameters and functional properties in real space with a picometre precision. In many technologically relevant atomic and/or molecular systems, however, the information of interest is distributed spatially in a non-uniform manner and may have a complex multi-dimensional nature. One of the critical issues, therefore, lies in being able to accurately identify (‘read out’) all the individual building blocks in different atomic/molecular architectures, as well as more complex patterns that these blocks may form, on a scale of hundreds andmore » thousands of individual atomic/molecular units. Here we employ machine vision to read and recognize complex molecular assemblies on surfaces. Specifically, we combine Markov random field model and convolutional neural networks to classify structural and rotational states of all individual building blocks in molecular assembly on the metallic surface visualized in high-resolution scanning tunneling microscopy measurements. We show how the obtained full decoding of the system allows us to directly construct a pair density function—a centerpiece in analysis of disorder-property relationship paradigm—as well as to analyze spatial correlations between multiple order parameters at the nanoscale, and elucidate reaction pathway involving molecular conformation changes. Here, the method represents a significant shift in our way of analyzing atomic and/or molecular resolved microscopic images and can be applied to variety of other microscopic measurements of structural, electronic, and magnetic orders in different condensed matter systems.« less

Learning surface molecular structures via machine vision

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

Ziatdinov, Maxim; Maksov, Artem; Kalinin, Sergei V.

Recent advances in high resolution scanning transmission electron and scanning probe microscopies have allowed researchers to perform measurements of materials structural parameters and functional properties in real space with a picometre precision. In many technologically relevant atomic and/or molecular systems, however, the information of interest is distributed spatially in a non-uniform manner and may have a complex multi-dimensional nature. One of the critical issues, therefore, lies in being able to accurately identify (‘read out’) all the individual building blocks in different atomic/molecular architectures, as well as more complex patterns that these blocks may form, on a scale of hundreds andmore » thousands of individual atomic/molecular units. Here we employ machine vision to read and recognize complex molecular assemblies on surfaces. Specifically, we combine Markov random field model and convolutional neural networks to classify structural and rotational states of all individual building blocks in molecular assembly on the metallic surface visualized in high-resolution scanning tunneling microscopy measurements. We show how the obtained full decoding of the system allows us to directly construct a pair density function—a centerpiece in analysis of disorder-property relationship paradigm—as well as to analyze spatial correlations between multiple order parameters at the nanoscale, and elucidate reaction pathway involving molecular conformation changes. Here, the method represents a significant shift in our way of analyzing atomic and/or molecular resolved microscopic images and can be applied to variety of other microscopic measurements of structural, electronic, and magnetic orders in different condensed matter systems.« less

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

The National Ignition Facility modular Kirkpatrick-Baez microscope

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

Pickworth, L. A., E-mail: pickworth1@llnl.gov; Ayers, J.; Bell, P.

Current two-dimensional X-ray imaging at the National Ignition Facility (NIF) uses time resolved pinhole cameras with ∼10-25 μm pinholes. This method has limitations in the smallest resolvable features that can be imaged with reasonable photon statistics for inertial confinement fusion (ICF) applications. ICF sources have a broadband self-emission spectrum that causes the pinhole images obtained, through thin foil filters, to contain a similarly broadband spectrum complicating the interpretation of structure in the source. In order to study phenomena on the scale of ∼5 μm, such as dopant mix in the ICF capsule, a narrow energy band, higher spatial resolution microscopemore » system with improved signal/noise has been developed using X-ray optics. Utilizing grazing incidence mirrors in a Kirkpatrick-Baez microscope (KBM) configuration [P. Kirkpatrick and A. V. Baez, J. Opt. Soc. Am. 38, 766–774 (1948)], an X-ray microscope has been designed and fielded on NIF with four imaging channels. The KBM has ∼12 × magnification, <8 μm resolution, and higher throughput in comparison to similar pinhole systems. The first KBM mirrors are coated with a multilayer mirror to allow a “narrow band” energy response at 10.2 keV with ΔE ∼ 3 keV. By adjusting the mirror coating only, the energy response can be matched to the future experimental requirements. Several mirror packs have been commissioned and are interchangeable in the diagnostic snout.« less

Relative merits and limiting factors for x-ray and electron microscopy of thick, hydrated organic materials.

PubMed

Du, Ming; Jacobsen, Chris

2018-01-01

Electron and x-ray microscopes allow one to image the entire, unlabeled structure of hydrated materials at a resolution well beyond what visible light microscopes can achieve. However, both approaches involve ionizing radiation, so that radiation damage must be considered as one of the limits to imaging. Drawing upon earlier work, we describe here a unified approach to estimating the image contrast (and thus the required exposure and corresponding radiation dose) in both x-ray and electron microscopy. This approach accounts for factors such as plural and inelastic scattering, and (in electron microscopy) the use of energy filters to obtain so-called "zero loss" images. As expected, it shows that electron microscopy offers lower dose for specimens thinner than about 1 µm (such as for studies of macromolecules, viruses, bacteria and archaebacteria, and thin sectioned material), while x-ray microscopy offers superior characteristics for imaging thicker specimen such as whole eukaryotic cells, thick-sectioned tissues, and organs. The required radiation dose scales strongly as a function of the desired spatial resolution, allowing one to understand the limits of live and frozen hydrated specimen imaging. Finally, we consider the factors limiting x-ray microscopy of thicker materials, suggesting that specimens as thick as a whole mouse brain can be imaged with x-ray microscopes without significant image degradation should appropriate image reconstruction methods be identified. Copyright © 2017 Elsevier B.V. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

2015-08-01

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

Underwater microscopy for in situ studies of benthic ecosystems

NASA Astrophysics Data System (ADS)

Mullen, Andrew D.; Treibitz, Tali; Roberts, Paul L. D.; Kelly, Emily L. A.; Horwitz, Rael; Smith, Jennifer E.; Jaffe, Jules S.

2016-07-01

Microscopic-scale processes significantly influence benthic marine ecosystems such as coral reefs and kelp forests. Due to the ocean's complex and dynamic nature, it is most informative to study these processes in the natural environment yet it is inherently difficult. Here we present a system capable of non-invasively imaging seafloor environments and organisms in situ at nearly micrometre resolution. We overcome the challenges of underwater microscopy through the use of a long working distance microscopic objective, an electrically tunable lens and focused reflectance illumination. The diver-deployed instrument permits studies of both spatial and temporal processes such as the algal colonization and overgrowth of bleaching corals, as well as coral polyp behaviour and interspecific competition. By enabling in situ observations at previously unattainable scales, this instrument can provide important new insights into micro-scale processes in benthic ecosystems that shape observed patterns at much larger scales.

Graphene Nanoribbons Fabricated by Helium Ion microscope

NASA Astrophysics Data System (ADS)

Pickard, D.; Oezyilmaz, B.; Thong, J.; Loh, K. P.; Viswanathan, V.; Zhongkai, A.; Mathew, S.; Kundu, T.; Park, C.; Yi, Z.; Xu, X.; Zhang, K.; Tat, T. C.; Wang, H.; Venkatesan, T.; Botton, G.; Couillard, M.

2010-03-01

Graphene, a monolayer graphitic lattice of carbon atoms has tremendous promise for a variety of applications on account of the zero mass of electrons, high mobility and the sensitivity of transport to perturbations at the interface. Patterning graphene is an obvious challenge and mesoscopic devices based on graphene require high spatial resolution patterning that will induce as little damage as possible. We use a helium ion microscope with its 0.4nm spot size beam to directly write patterns on free standing graphene films. TEM images of the patterns reveal holes as small as 4 nm and ribbons with line widths as narrow as 3 nm. The images show recovery of the graphene lattice at a distance of about a nm from the patterned edge. The linewidths of the ribbon can be varied considerably in a controllable fashion over ribbon lengths of the order of microns. . .

Fano Description of Single-Hydrocarbon Fluorescence Excited by a Scanning Tunneling Microscope.

PubMed

Kröger, Jörg; Doppagne, Benjamin; Scheurer, Fabrice; Schull, Guillaume

2018-06-13

The detection of fluorescence with submolecular resolution enables the exploration of spatially varying photon yields and vibronic properties at the single-molecule level. By placing individual polycyclic aromatic hydrocarbon molecules into the plasmon cavity formed by the tip of a scanning tunneling microscope and a NaCl-covered Ag(111) surface, molecular light emission spectra are obtained that unravel vibrational progression. In addition, light spectra unveil a signature of the molecule even when the tunneling current is injected well separated from the molecular emitter. This signature exhibits a distance-dependent Fano profile that reflects the subtle interplay between inelastic tunneling electrons, the molecular exciton and localized plasmons in at-distance as well as on-molecule fluorescence. The presented findings open the path to luminescence of a different class of molecules than investigated before and contribute to the understanding of single-molecule luminescence at surfaces in a unified picture.

Electron-excited energy dispersive x-ray spectrometry in the variable pressure scanning electron microscope (EDS/VPSEM): it's not microanalysis anymore!

NASA Astrophysics Data System (ADS)

Newbury, Dale E.; Ritchie, Nicholas W. M.

2015-10-01

X-ray spectra suffer significantly degraded spatial resolution when measured in the variable-pressure scanning electron microscope (VPSEM, chamber pressure 1 Pa to 2500 Pa) as compared to highvacuum SEM (operating pressure < 10 mPa). Depending on the gas path length, electrons that are scattered hundreds of micrometers outside the focused beam can contribute 90% or more of the measured spectrum. Monte Carlo electron trajectory simulation, available in NIST DTSA-II, models the gas scattering and simulates mixed composition targets, e.g., particle on substrate. The impact of gas scattering at the major (C > 0.1 mass fraction), minor (0.01 <= C <= 0.1), and trace (C < 0.01) constituent levels can be estimated. NIST DTSA-II for Java-platforms is available free at: http://www.cstl.nist.gov/div837/837.02/epq/dtsa2/index.html).

An in-plane magnetic chiral dichroism approach for measurement of intrinsic magnetic signals using transmitted electrons

PubMed Central

Song, Dongsheng; Tavabi, Amir H.; Li, Zi-An; Kovács, András; Rusz, Ján; Huang, Wenting; Richter, Gunther; Dunin-Borkowski, Rafal E.; Zhu, Jing

2017-01-01

Electron energy-loss magnetic chiral dichroism is a powerful technique that allows the local magnetic properties of materials to be measured quantitatively with close-to-atomic spatial resolution and element specificity in the transmission electron microscope. Until now, the technique has been restricted to measurements of the magnetic circular dichroism signal in the electron beam direction. However, the intrinsic magnetization directions of thin samples are often oriented in the specimen plane, especially when they are examined in magnetic-field-free conditions in the transmission electron microscope. Here, we introduce an approach that allows in-plane magnetic signals to be measured using electron magnetic chiral dichroism by selecting a specific diffraction geometry. We compare experimental results recorded from a cobalt nanoplate with simulations to demonstrate that an electron magnetic chiral dichroism signal originating from in-plane magnetization can be detected successfully. PMID:28504267

Real-time Image Processing for Microscopy-based Label-free Imaging Flow Cytometry in a Microfluidic Chip.

PubMed

Heo, Young Jin; Lee, Donghyeon; Kang, Junsu; Lee, Keondo; Chung, Wan Kyun

2017-09-14

Imaging flow cytometry (IFC) is an emerging technology that acquires single-cell images at high-throughput for analysis of a cell population. Rich information that comes from high sensitivity and spatial resolution of a single-cell microscopic image is beneficial for single-cell analysis in various biological applications. In this paper, we present a fast image-processing pipeline (R-MOD: Real-time Moving Object Detector) based on deep learning for high-throughput microscopy-based label-free IFC in a microfluidic chip. The R-MOD pipeline acquires all single-cell images of cells in flow, and identifies the acquired images as a real-time process with minimum hardware that consists of a microscope and a high-speed camera. Experiments show that R-MOD has the fast and reliable accuracy (500 fps and 93.3% mAP), and is expected to be used as a powerful tool for biomedical and clinical applications.

Determining the phonon energy of highly oriented pyrolytic graphite by scanning tunneling microscope light emission spectroscopy

NASA Astrophysics Data System (ADS)

Uehara, Yoichi; Michimata, Junichi; Watanabe, Shota; Katano, Satoshi; Inaoka, Takeshi

2018-03-01

We have investigated the scanning tunneling microscope (STM) light emission spectra of isolated single Ag nanoparticles lying on highly oriented pyrolytic graphite (HOPG). The STM light emission spectra exhibited two types of spectral structures (step-like and periodic). Comparisons of the observed structures and theoretical predictions indicate that the phonon energy of the ZO mode of HOPG [M. Mohr et al., Phys. Rev. B 76, 035439 (2007)] can be determined from the energy difference between the cutoff of STM light emission and the step in the former structure, and from the period of the latter structure. Since the role of the Ag nanoparticles does not depend on the substrate materials, this method will enable the phonon energies of various materials to be measured by STM light emission spectroscopy. The spatial resolution is comparable to the lateral size of the individual Ag nanoparticles (that is, a few nm).

Glue-Free Stacked Luminescent Nanosheets Enable High-Resolution Ratiometric Temperature Mapping in Living Small Animals.

PubMed

Miyagawa, Takuya; Fujie, Toshinori; Ferdinandus; Vo Doan, Tat Thang; Sato, Hirotaka; Takeoka, Shinji

2016-12-14

In this paper, a microthermograph, temperature mapping with high spatial resolution, was established using luminescent molecules embedded ultrathin polymeric films (nanosheets), and demonstrated in a living small animal to map out and visualize temperature shift due to animal's muscular activity. Herein, we report super flexible and self-adhesive (no need of glue) nanothermosensor consisting of stacked two different polymeric nanosheets with thermosensitive (Eu-tris (dinaphthoylmethane)-bis-trioctylphosphine oxide: EuDT) and insensitive (Rhodamine 800) dyes being embedded. Such stacked nanosheets allow for the ratiometric thermometry, with which the undesired luminescence intensity shift due to focal drift or animal's z-axis displacement is eliminated and the desired intensity shift solely due to the temperature shift of the sample (living muscle) can be acquired. With the stacked luminescent nanosheets, we achieved the first-ever demonstration of video filming of chronologically changing temperature-shift distribution from the rest state to the active state of the muscles in the living animal. The polymer nanosheet engineering and in vivo microthermography presented in the paper are promising technologies to microscopically explore the heat production and heat transfer in living cells, tissues, and organisms with high spatial resolution beyond what existing thermometric technologies such as infrared thermography have ever achieved.

Quantitative Vectorial Magnetic Imaging of Multi Domain Rock Forming Minerals using Nitrogen-Vacancy Centers in Diamond

NASA Astrophysics Data System (ADS)

Shaar, R.; Farchi, E.; Farfurnik, D.; Ebert, Y.; Haim, G.; Bar-Gill, N.

2017-12-01

Magnetization in rock samples is crucial for paleomagnetometry research, as it harbors valuable geological information on long term processes, such as tectonic movements and the formation of oceans and continents. Nevertheless, current techniques are limited in their ability to measure high spatial resolution and high-sensitivity quantitative vectorial magnetic signatures from individual minerals and micrometer scale samples. As a result, our understanding of bulk rock magnetization is limited, specifically for the case of multi-domain minerals. In this work we use a newly developed nitrogen-vacancy magnetic microscope, capable of quantitative vectorial magnetic imaging with optical resolution. We demonstrate direct imaging of the vectorial magnetic field of a single, multi-domain dendritic magnetite, as well as the measurement and calculation of the weak magnetic moments of an individual grain on the micron scale. Our results were measured in a standoff distance of 3-10 μm, with 350 nm spatial resolution, magnetic sensitivity of 6 μT/√(Hz) and a field of view of 35 μm. The results presented here show the capabilities and the future potential of NV microscopy in measuring the magnetic signals of individual micrometer scale grains. These outcomes pave the way for future applications in paleomagnetometry, and for the fundamental understanding of magnetization in multi-domain samples.

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

Geostatistical analysis of 3D microCT images of porous media for stochastic upscaling of spatially variable reactive surfaces

NASA Astrophysics Data System (ADS)

De Lucia, Marco; Kühn, Michael

2015-04-01

The 3D imaging of porous media through micro tomography allows the characterization of porous space and mineral abundances with unprecedented resolution. Such images can be used to perform computational determination of permeability and to obtain a realistic measure of the mineral surfaces exposed to fluid flow and thus to chemical interactions. However, the volume of the plugs that can be analysed with such detail is in the order of 1 cm3, so that their representativity at a larger scale, i.e. as needed for reactive transport modelling at Darcy scale, is questionable at best. In fact, the fine scale heterogeneity (from plug to plug at few cm distance within the same core) would originate substantially different readings of the investigated properties. Therefore, a comprehensive approach including the spatial variability and heterogeneity at the micro- and plug scale needs to be adopted to gain full advantage from the high resolution images in view of the upscaling to Darcy scale. In the framework of the collaborative project H2STORE, micro-CT imaging of different core samples from potential H2-storage sites has been performed by partners at TU Clausthal and Jena University before and after treatment with H2/CO2 mixtures in pressurized autoclaves. We present here the workflow which has been implemented to extract the relevant features from the available data concerning the heterogeneity of the medium at the microscopic and plug scale and to correlate the observed chemical reactions and changes in the porous structure with the geometrical features of the medium. First, a multivariate indicator-based geostatistical model for the microscopic structure of the plugs has been built and fitted to the available images. This involved the implementation of exploratory analysis algorithms such as experimental indicator variograms and cross-variograms. The implemented methods are able to efficiently deal with images in the order of 10003 voxels making use of parallelization. Sequential Indicator Simulations are then employed to generate equi-probable realizations of microscopic structures with varying mineral proportions and porosity but constrained to the spatial variability observed in the plugs. The statistics computed on the ensemble of realizations (essentially the distribution of mineral reactive surfaces exposed to porous space) is integrated at a larger, Darcy scale. In a further step, the analysis of the microscopic changes in the plugs after exposure to reactive solution establishes the correlations betweens amount of chemical reactions and changes in the spatial models, thus deriving some effective correlations which can be injected into the reactive transport modelling. In this contribution, we demonstrate the implemented workflow on a series of images obtained from plugs from a german depleted gas field exposed to H2 and CO2-charged brines. The geostatistical evaluation of microscale variability of the porous media contributes to the upscaling of relevant variables and helps estimating - if not reducing - the uncertainty due to the heterogeneity across scales of the natural systems.

Design and analysis of multilayer x ray/XUV microscope

NASA Technical Reports Server (NTRS)

Shealy, David L.

1990-01-01

The design and analysis of a large number of normal incidence multilayer x ray microscopes based on the spherical mirror Schwarzschild configuration is examined. Design equations for the spherical mirror Schwarzschild microscopes are summarized and used to evaluate mirror parameters for microscopes with magnifications ranging from 2 to 50x. Ray tracing and diffraction analyses are carried out for many microscope configurations to determine image resolution as a function of system parameters. The results are summarized in three publication included herein. A preliminary study of advanced reflecting microscope configurations, where aspherics are used in place of the spherical microscope mirror elements, has indicated that the aspherical elements will improve off-axis image resolution and increase the effective field of view.

High axial resolution imaging system for large volume tissues using combination of inclined selective plane illumination and mechanical sectioning

PubMed Central

Zhang, Qi; Yang, Xiong; Hu, Qinglei; Bai, Ke; Yin, Fangfang; Li, Ning; Gang, Yadong; Wang, Xiaojun; Zeng, Shaoqun

2017-01-01

To resolve fine structures of biological systems like neurons, it is required to realize microscopic imaging with sufficient spatial resolution in three dimensional systems. With regular optical imaging systems, high lateral resolution is accessible while high axial resolution is hard to achieve in a large volume. We introduce an imaging system for high 3D resolution fluorescence imaging of large volume tissues. Selective plane illumination was adopted to provide high axial resolution. A scientific CMOS working in sub-array mode kept the imaging area in the sample surface, which restrained the adverse effect of aberrations caused by inclined illumination. Plastic embedding and precise mechanical sectioning extended the axial range and eliminated distortion during the whole imaging process. The combination of these techniques enabled 3D high resolution imaging of large tissues. Fluorescent bead imaging showed resolutions of 0.59 μm, 0.47μm, and 0.59 μm in the x, y, and z directions, respectively. Data acquired from the volume sample of brain tissue demonstrated the applicability of this imaging system. Imaging of different depths showed uniform performance where details could be recognized in either the near-soma area or terminal area, and fine structures of neurons could be seen in both the xy and xz sections. PMID:29296503

Intrinsic instability of aberration-corrected electron microscopes.

PubMed

Schramm, S M; van der Molen, S J; Tromp, R M

2012-10-19

Aberration-corrected microscopes with subatomic resolution will impact broad areas of science and technology. However, the experimentally observed lifetime of the corrected state is just a few minutes. Here we show that the corrected state is intrinsically unstable; the higher its quality, the more unstable it is. Analyzing the contrast transfer function near optimum correction, we define an "instability budget" which allows a rational trade-off between resolution and stability. Unless control systems are developed to overcome these challenges, intrinsic instability poses a fundamental limit to the resolution practically achievable in the electron microscope.

SU-E-T-98: Towards Cell Nucleus Microdosimetry: Construction of a Confocal Laser-Scanning Fluorescence Microscope to Readout Fluorescence Nuclear Track Detectors (FNTDs).

PubMed

McFadden, C; Bartz, J; Akselrod, M; Sawakuchi, G

2012-06-01

To construct a custom confocal laser scanning microscope (CLSM) capable of resolving individual proton tracks in the volume of an Al 2 O 3 :C,Mg fluorescent nuclear track detector (FNTD). The spatial resolution of the FNTD technique is at the sub-micrometer scale. Therefore the FNTD technique has the potential to perform radiation measurements at the cell nucleus scale. The crystal volume of an FNTD contains defects which become fluorescent F 2 + centers after trapping delta electrons from ionizing radiation. These centers have an absorption band centered at 620 nm and an emission band in the near infrared. Events of energy deposition in the crystal are read-out using a CLSM with sub-micrometer spatial resolution. Excitation light from a 635 nm laser is focused in the crystal volume by an objective lens. Fluorescence is collected back through the same path, filtered through a dichroic mirror, and focused through a small pinhole onto an avalanche photodiode. Lateral scanning of the focal point is performed with a scanning mirror galvanometer, and axial scanning is performed using a stepper-motor stage. Control of electronics and image acquisition was performed using a custom built LabVIEW VI and further image processing was done using Java. The system was used to scan FNTDs exposed to a 6 MV x-ray beam and an unexposed FNTD. Fluorescence images above the unexposed background were obtained at scan depths ranging from 5 - 10 micrometer below the crystal surface using a 100 micrometer pinhole size. Further work needs to be done to increase the resolution and the signal to noise ratio of the images so that energy deposition events may be identified more easily. Natural Sciences and Engineering Research Council of Canada. © 2012 American Association of Physicists in Medicine.

Programmable Illumination and High-Speed, Multi-Wavelength, Confocal Microscopy Using a Digital Micromirror

PubMed Central

Martial, Franck P.; Hartell, Nicholas A.

2012-01-01

Confocal microscopy is routinely used for high-resolution fluorescence imaging of biological specimens. Most standard confocal systems scan a laser across a specimen and collect emitted light passing through a single pinhole to produce an optical section of the sample. Sequential scanning on a point-by-point basis limits the speed of image acquisition and even the fastest commercial instruments struggle to resolve the temporal dynamics of rapid cellular events such as calcium signals. Various approaches have been introduced that increase the speed of confocal imaging. Nipkov disk microscopes, for example, use arrays of pinholes or slits on a spinning disk to achieve parallel scanning which significantly increases the speed of acquisition. Here we report the development of a microscope module that utilises a digital micromirror device as a spatial light modulator to provide programmable confocal optical sectioning with a single camera, at high spatial and axial resolution at speeds limited by the frame rate of the camera. The digital micromirror acts as a solid state Nipkov disk but with the added ability to change the pinholes size and separation and to control the light intensity on a mirror-by-mirror basis. The use of an arrangement of concave and convex mirrors in the emission pathway instead of lenses overcomes the astigmatism inherent with DMD devices, increases light collection efficiency and ensures image collection is achromatic so that images are perfectly aligned at different wavelengths. Combined with non-laser light sources, this allows low cost, high-speed, multi-wavelength image acquisition without the need for complex wavelength-dependent image alignment. The micromirror can also be used for programmable illumination allowing spatially defined photoactivation of fluorescent proteins. We demonstrate the use of this system for high-speed calcium imaging using both a single wavelength calcium indicator and a genetically encoded, ratiometric, calcium sensor. PMID:22937130

Programmable illumination and high-speed, multi-wavelength, confocal microscopy using a digital micromirror.

PubMed

Martial, Franck P; Hartell, Nicholas A

2012-01-01

Confocal microscopy is routinely used for high-resolution fluorescence imaging of biological specimens. Most standard confocal systems scan a laser across a specimen and collect emitted light passing through a single pinhole to produce an optical section of the sample. Sequential scanning on a point-by-point basis limits the speed of image acquisition and even the fastest commercial instruments struggle to resolve the temporal dynamics of rapid cellular events such as calcium signals. Various approaches have been introduced that increase the speed of confocal imaging. Nipkov disk microscopes, for example, use arrays of pinholes or slits on a spinning disk to achieve parallel scanning which significantly increases the speed of acquisition. Here we report the development of a microscope module that utilises a digital micromirror device as a spatial light modulator to provide programmable confocal optical sectioning with a single camera, at high spatial and axial resolution at speeds limited by the frame rate of the camera. The digital micromirror acts as a solid state Nipkov disk but with the added ability to change the pinholes size and separation and to control the light intensity on a mirror-by-mirror basis. The use of an arrangement of concave and convex mirrors in the emission pathway instead of lenses overcomes the astigmatism inherent with DMD devices, increases light collection efficiency and ensures image collection is achromatic so that images are perfectly aligned at different wavelengths. Combined with non-laser light sources, this allows low cost, high-speed, multi-wavelength image acquisition without the need for complex wavelength-dependent image alignment. The micromirror can also be used for programmable illumination allowing spatially defined photoactivation of fluorescent proteins. We demonstrate the use of this system for high-speed calcium imaging using both a single wavelength calcium indicator and a genetically encoded, ratiometric, calcium sensor.

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.

The nature and origin of interstellar diamond

NASA Technical Reports Server (NTRS)

Blake, David F.; Freund, Friedemann; Shipp, Ruth; Krishnan, Kannan F. M.; Echer, Charles J.

1988-01-01

The C-delta component of the Allende meteorite is a microscopic diamond some of whose properties seem in conflict with those expected of diamond. High spatial resolution analytical data are presented here which may help explain such results. Surface and interfacial carbon atoms in the component, which may comprise as much as 25 percent of the total, impart an 'amorphous' character to some spectral data. These data support the proposed high-pressure conversion of amorphous carbon and graphite into diamonds due to grain-grain collisions in the ISM, although a low-pressure mechanism of formation cannot be ruled out.

Resolving the multipolar scattering modes of a submicron particle using parametric indirect microscopic imaging

NASA Astrophysics Data System (ADS)

Ullah, Kaleem; Liu, Xuefeng; Krasnok, Alex; Habib, Muhammad; Song, Li; Garcia-Camara, Braulio

2018-07-01

In this work, we show the spatial distribution of the scattered electromagnetic field of dielectric particles by using a new super-resolution method based on polarization modulation. Applying this technique, we were able to resolve the multipolar distribution of a Cu2O particle with a radius of 450 nm. In addition, FDTD and Mie simulations have been carried out to validate and confirm the experimental results. The results are helpful to understand the resonant modes of dielectric submicron particles which have a broad range of potential applications, such as all-optical devices or nanoantennas.

Three-dimensional real-time imaging of bi-phasic flow through porous media

NASA Astrophysics Data System (ADS)

Sharma, Prerna; Aswathi, P.; Sane, Anit; Ghosh, Shankar; Bhattacharya, S.

2011-11-01

We present a scanning laser-sheet video imaging technique to image bi-phasic flow in three-dimensional porous media in real time with pore-scale spatial resolution, i.e., 35 μm and 500 μm for directions parallel and perpendicular to the flow, respectively. The technique is illustrated for the case of viscous fingering. Using suitable image processing protocols, both the morphology and the movement of the two-fluid interface, were quantitatively estimated. Furthermore, a macroscopic parameter such as the displacement efficiency obtained from a microscopic (pore-scale) analysis demonstrates the versatility and usefulness of the method.

Electrochemical wall shear rate microscopy of collapsing bubbles

NASA Astrophysics Data System (ADS)

Reuter, Fabian; Mettin, Robert

2018-06-01

An electrochemical high-speed wall shear raster microscope is presented. It involves chronoamperometric measurements on a microelectrode that is flush-mounted in a submerged test specimen. Wall shear rates are derived from the measured microelectrode signal by numerically solving a convection-diffusion equation with an optimization approach. This way, the unsteady wall shear rates from the collapse of a laser pulse seeded cavitation bubble close to a substrate are measured. By planar scanning, they are resolved in high spatial resolution. The wall shear rates are related to the bubble dynamics via synchronized high-speed imaging of the bubble shape.

Maximum: Recent Implementation and Application to the Study of Corrosion-Induced Microstructures in Thin Films of Aluminum-Copper Metallization.

NASA Astrophysics Data System (ADS)

Liang, Shoudeng

We describe the recent implementation of a synchrotron radiation based scanning soft X-ray photoemission microscope - MAXIMUM, and discuss its application to the investigation of corrosion-induced microstructures in Al-Cu-Si thin films. The microscope employs a Mo/Si multilayer-coated Schwarzschild objective to focus 95eV X-rays from an undulator beamline. The photoelectrons are energy-analyzed by a CMA, and the sample is rastered to produce an image. We have achieved 980A spatial and 250meV energy resolution. Recent addition of a sample preparation and transfer system to the microscope enables us to perform surface and materials studies under UHV conditions. Since the spatial resolution of the microscope is determined by the spot size of the focused X-rays, any electrostatic potential from surface charging will not affect the image quality. This allowed the study of highly insulating films with the use of an electron flood gun to compensate for spectral shifts. We have employed MAXIMUM to investigate corrosion -induced surface microstructures in the Al-Cu-Si thin films commonly utilized in VLSI metallization. Spectromicroscopy was performed to characterize the chemical species and their distribution on the film surface after corrosion under 85% relative humidity at 85^circ C. The experimental images demonstrated that Cu -rich precipitates were formed near the surface region beneath the oxide layer upon annealing. We also observed a correlation between the precipitates and the increased corrosion in the alloy film: the localized corrosion occurs only at those sites where precipitation has taken place. This implies that the surface oxide layer is modified by the underlying Cu-rich phase such that it loses protection against moisture. After pitting, the Cu-rich phase acts as a cathode to facilitate corrosion of the surrounding Cu-deficient Al matrix via galvanic action. The corrosion -induced microstructures show characteristic circular features in the micrographs of energy-specific photoelectrons from Cu 3d and O 2p valence bands. Such characteristic structures were observed only when the film was annealed below the solvus temperatures in the Al-Cu binary phase diagram, i.e., when a phase separation occurred. These results demonstrated the usefulness of spectromicroscopy in corrosion studies.

Future of Electron Scattering and Diffraction

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

Hall, Ernest; Stemmer, Susanne; Zheng, Haimei

2014-02-25

The ability to correlate the atomic- and nanoscale-structure of condensed matter with physical properties (e.g., mechanical, electrical, catalytic, and optical) and functionality forms the core of many disciplines. Directing and controlling materials at the quantum-, atomic-, and molecular-levels creates enormous challenges and opportunities across a wide spectrum of critical technologies, including those involving the generation and use of energy. The workshop identified next generation electron scattering and diffraction instruments that are uniquely positioned to address these grand challenges. The workshop participants identified four key areas where the next generation of such instrumentation would have major impact: A – Multidimensional Visualizationmore » of Real Materials B – Atomic-scale Molecular Processes C – Photonic Control of Emergence in Quantum Materials D – Evolving Interfaces, Nucleation, and Mass Transport Real materials are comprised of complex three-dimensional arrangements of atoms and defects that directly determine their potential for energy applications. Understanding real materials requires new capabilities for three-dimensional atomic scale tomography and spectroscopy of atomic and electronic structures with unprecedented sensitivity, and with simultaneous spatial and energy resolution. Many molecules are able to selectively and efficiently convert sunlight into other forms of energy, like heat and electric current, or store it in altered chemical bonds. Understanding and controlling such process at the atomic scale require unprecedented time resolution. One of the grand challenges in condensed matter physics is to understand, and ultimately control, emergent phenomena in novel quantum materials that necessitate developing a new generation of instruments that probe the interplay among spin, charge, orbital, and lattice degrees of freedom with intrinsic time- and length-scale resolutions. Molecules and soft matter require imaging and spectroscopy with high spatial resolution without damaging their structure. The strong interaction of electrons with matter allows high-energy electron pulses to gather structural information before a sample is damaged. Electron ScatteringImaging, diffraction, and spectroscopy are the fundamental capabilities of electron-scattering instruments. The DOE BES-funded TEAM (Transmission Electron Aberration-corrected Microscope) project achieved unprecedented sub-atomic spatial resolution in imaging through aberration-corrected transmission electron microscopy. To further advance electron scattering techniques that directly enable groundbreaking science, instrumentation must advance beyond traditional two-dimensional imaging. Advances in temporal resolution, recording the full phase and energy spaces, and improved spatial resolution constitute a new frontier in electron microscopy, and will directly address the BES Grand Challenges, such as to “control the emergent properties that arise from the complex correlations of atomic and electronic constituents” and the “hidden states” “very far away from equilibrium”. Ultrafast methods, such as the pump-probe approach, enable pathways toward understanding, and ultimately controlling, the chemical dynamics of molecular systems and the evolution of complexity in mesoscale and nanoscale systems. Central to understanding how to synthesize and exploit functional materials is having the ability to apply external stimuli (such as heat, light, a reactive flux, and an electrical bias) and to observe the resulting dynamic process in situ and in operando, and under the appropriate environment (e.g., not limited to UHV conditions). To enable revolutionary advances in electron scattering and science, the participants of the workshop recommended three major new instrumental developments: A. Atomic-Resolution Multi-Dimensional Transmission Electron Microscope: This instrument would provide quantitative information over the entire real space, momentum space, and energy space for visualizing dopants, interstitials, and light elements; for imaging localized vibrational modes and the motion of charged particles and vacancies; for correlating lattice, spin, orbital, and charge; and for determining the structure and molecular chemistry of organic and soft matter. The instrument will be uniquely suited to answer fundamental questions in condensed matter physics that require understanding the physical and electronic structure at the atomic scale. Key developments include stable cryogenic capabilities that will allow access to emergent electronic phases, as well as hard/soft interfaces and radiation- sensitive materials. B. Ultrafast Electron Diffraction and Microscopy Instrument: This instrument would be capable of nano-diffraction with 10 fs temporal resolution in stroboscopic mode, and better than 100 fs temporal resolution in single shot mode. The instrument would also achieve single- shot real-space imaging with a spatial/temporal resolution of 10 nm/10 ps, representing a thousand fold improvement over current microscopes. Such a capability would be complementary to x-ray free electron lasers due to the difference in the nature of electron and x-ray scattering, enabling space-time mapping of lattice vibrations and energy transport, facilitating the understanding of molecular dynamics of chemical reactions, the photonic control of emergence in quantum materials, and the dynamics of mesoscopic materials. C. Lab-In-Gap Dynamic Microscope: This instrument would enable quantitative measurements of materials structure, composition, and bonding evolution in technologically relevant environments, including liquids, gases and plasmas, thereby assuring the understanding of structure function relationship at the atomic scale with up to nanosecond temporal resolution. This instrument would employ a versatile, modular sample stage and holder geometry to allow the multi-modal (e.g., optical, thermal, mechanical, electrical, and electrochemical) probing of materials’ functionality in situ and in operando. The electron optics encompasses a pole piece that can accommodate the new stage, differential pumping, detectors, aberration correctors, and other electron optical elements for measurement of materials dynamics. To realize the proposed instruments in a timely fashion, BES should aggressively support research and development of complementary and enabling instruments, including new electron sources, advanced electron optics, new tunable specimen pumps and sample stages, and new detectors. The proposed instruments would have transformative impact on physics, chemistry, materials science, engineering« less

Coherence and diffraction limited resolution in microscopic OCT by a unified approach for the correction of dispersion and aberrations

NASA Astrophysics Data System (ADS)

Schulz-Hildebrandt, H.; Münter, Michael; Ahrens, M.; Spahr, H.; Hillmann, D.; König, P.; Hüttmann, G.

2018-03-01

Optical coherence tomography (OCT) images scattering tissues with 5 to 15 μm resolution. This is usually not sufficient for a distinction of cellular and subcellular structures. Increasing axial and lateral resolution and compensation of artifacts caused by dispersion and aberrations is required to achieve cellular and subcellular resolution. This includes defocus which limit the usable depth of field at high lateral resolution. OCT gives access the phase of the scattered light and hence correction of dispersion and aberrations is possible by numerical algorithms. Here we present a unified dispersion/aberration correction which is based on a polynomial parameterization of the phase error and an optimization of the image quality using Shannon's entropy. For validation, a supercontinuum light sources and a costume-made spectrometer with 400 nm bandwidth were combined with a high NA microscope objective in a setup for tissue and small animal imaging. Using this setup and computation corrections, volumetric imaging at 1.5 μm resolution is possible. Cellular and near cellular resolution is demonstrated in porcine cornea and the drosophila larva, when computational correction of dispersion and aberrations is used. Due to the excellent correction of the used microscope objective, defocus was the main contribution to the aberrations. In addition, higher aberrations caused by the sample itself were successfully corrected. Dispersion and aberrations are closely related artifacts in microscopic OCT imaging. Hence they can be corrected in the same way by optimization of the image quality. This way microscopic resolution is easily achieved in OCT imaging of static biological tissues.

Set-up of a high-resolution 300 mK atomic force microscope in an ultra-high vacuum compatible (3)He/10 T cryostat.

PubMed

von Allwörden, H; Ruschmeier, K; Köhler, A; Eelbo, T; Schwarz, A; Wiesendanger, R

2016-07-01

The design of an atomic force microscope with an all-fiber interferometric detection scheme capable of atomic resolution at about 500 mK is presented. The microscope body is connected to a small pumped (3)He reservoir with a base temperature of about 300 mK. The bakeable insert with the cooling stage can be moved from its measurement position inside the bore of a superconducting 10 T magnet into an ultra-high vacuum chamber, where the tip and sample can be exchanged in situ. Moreover, single atoms or molecules can be evaporated onto a cold substrate located inside the microscope. Two side chambers are equipped with standard surface preparation and surface analysis tools. The performance of the microscope at low temperatures is demonstrated by resolving single Co atoms on Mn/W(110) and by showing atomic resolution on NaCl(001).

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.

High-resolution electron microscope

NASA Technical Reports Server (NTRS)

Nathan, R.

1977-01-01

Employing scanning transmission electron microscope as interferometer, relative phases of diffraction maximums can be determined by analysis of dark field images. Synthetic aperture technique and Fourier-transform computer processing of amplitude and phase information provide high resolution images at approximately one angstrom.

Use of radiochromic film as a high-spatial resolution dosimeter by Raman spectroscopy

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

Mirza, Jamal Ahmad; Park, Hyeonsuk

Purpose: Due to increasing demand for high-spatial resolution dosimetry, radiochromic films have been investigated as potential candidates but are often limited by the scanning system, e.g., flatbed optical scanner. In this study, Raman spectroscopy in conjunction with a microscope was selected as an alternative method for high-spatial resolution dosimetry of radiochromic film. Methods: Unlaminated Gafchromic™ EBT3 films were irradiated with doses between 0 and 50 Gy using 6 MV x-rays of a clinical linear accelerator. Depth profiling from the surface of unlaminated film was performed to acquire the maximum Raman intensity peaks of C≡C and C=C stretching bands of diacetylenemore » polymer. The Raman mapping technique for a region of interest (200 × 200, 30 × 30 μm{sup 2}) was developed to reduce a large variation in a Raman spectrum produced with a sampling resolution of a few μm. The preprocessing of Raman spectra was carried out to determine a dosimetric relationship with the amount of diacetylene polymerization. Results: Due to partial diacetylene polymerization upon irradiation, two Raman peaks of C=C and C≡C stretching bands were observed around 1447 and 2060 cm{sup −1}, respectively. The maximum intensities of the two peaks were obtained by positioning a focused laser spot on the surface of unlaminated film. For the dose range of 0–50 Gy, the band heights of both C≡C and C=C peaks increase asymptotically with increasing doses and can be fit with an exponential function of two components. The relative standard deviation in Raman mapping was found to be less than ±5%. By using this technique, dose uniformity was found to be within ±2%. Conclusions: The Raman intensity for C=C and C≡C peaks increases with an increase in the amount of diacetylene polymerization due to an increase in dose. This study shows the potential of Raman spectroscopy as an alternative for absolute dosimetry verifications with a high-spatial resolution of a few μm, but these findings need to be further validated for the purpose of microdosimetry.« less

Progress toward an aberration-corrected low energy electron microscope for DNA sequencing and surface analysis.

PubMed

Mankos, Marian; Shadman, Khashayar; N'diaye, Alpha T; Schmid, Andreas K; Persson, Henrik H J; Davis, Ronald W

2012-11-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel imaging technique aimed at high resolution imaging of macromolecules, nanoparticles, and surfaces. MAD-LEEM combines three innovative electron-optical concepts in a single tool: a monochromator, a mirror aberration corrector, and dual electron beam illumination. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. The aberration corrector is needed to achieve subnanometer resolution at landing energies of a few hundred electronvolts. The dual flood illumination approach eliminates charging effects generated when a conventional, single-beam LEEM is used to image insulating specimens. The low landing energy of electrons in the range of 0 to a few hundred electronvolts is also critical for avoiding radiation damage, as high energy electrons with kilo-electron-volt kinetic energies cause irreversible damage to many specimens, in particular biological molecules. The performance of the key electron-optical components of MAD-LEEM, the aberration corrector combined with the objective lens and a magnetic beam separator, was simulated. Initial results indicate that an electrostatic electron mirror has negative spherical and chromatic aberration coefficients that can be tuned over a large parameter range. The negative aberrations generated by the electron mirror can be used to compensate the aberrations of the LEEM objective lens for a range of electron energies and provide a path to achieving subnanometer spatial resolution. First experimental results on characterizing DNA molecules immobilized on Au substrates in a LEEM are presented. Images obtained in a spin-polarized LEEM demonstrate that high contrast is achievable at low electron energies in the range of 1-10 eV and show that small changes in landing energy have a strong impact on the achievable contrast. The MAD-LEEM approach promises to significantly improve the performance of a LEEM for a wide range of applications in the biosciences, material sciences, and nanotechnology where nanometer scale resolution and analytical capabilities are required. In particular, the microscope has the potential of delivering images of unlabeled DNA strands with nucleotide-specific contrast. This simplifies specimen preparation and significantly eases the computational complexity needed to assemble the DNA sequence from individual reads.

Progress toward an aberration-corrected low energy electron microscope for DNA sequencing and surface analysis

PubMed Central

Mankos, Marian; Shadman, Khashayar; N'Diaye, Alpha T.; Schmid, Andreas K.; Persson, Henrik H. J.; Davis, Ronald W.

2012-01-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel imaging technique aimed at high resolution imaging of macromolecules, nanoparticles, and surfaces. MAD-LEEM combines three innovative electron–optical concepts in a single tool: a monochromator, a mirror aberration corrector, and dual electron beam illumination. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. The aberration corrector is needed to achieve subnanometer resolution at landing energies of a few hundred electronvolts. The dual flood illumination approach eliminates charging effects generated when a conventional, single-beam LEEM is used to image insulating specimens. The low landing energy of electrons in the range of 0 to a few hundred electronvolts is also critical for avoiding radiation damage, as high energy electrons with kilo-electron-volt kinetic energies cause irreversible damage to many specimens, in particular biological molecules. The performance of the key electron–optical components of MAD-LEEM, the aberration corrector combined with the objective lens and a magnetic beam separator, was simulated. Initial results indicate that an electrostatic electron mirror has negative spherical and chromatic aberration coefficients that can be tuned over a large parameter range. The negative aberrations generated by the electron mirror can be used to compensate the aberrations of the LEEM objective lens for a range of electron energies and provide a path to achieving subnanometer spatial resolution. First experimental results on characterizing DNA molecules immobilized on Au substrates in a LEEM are presented. Images obtained in a spin-polarized LEEM demonstrate that high contrast is achievable at low electron energies in the range of 1–10 eV and show that small changes in landing energy have a strong impact on the achievable contrast. The MAD-LEEM approach promises to significantly improve the performance of a LEEM for a wide range of applications in the biosciences, material sciences, and nanotechnology where nanometer scale resolution and analytical capabilities are required. In particular, the microscope has the potential of delivering images of unlabeled DNA strands with nucleotide-specific contrast. This simplifies specimen preparation and significantly eases the computational complexity needed to assemble the DNA sequence from individual reads. PMID:23847748

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

PubMed

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

2013-01-01

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

High-resolution scanning Hall probe microscopy

NASA Astrophysics Data System (ADS)

Hallen, Hans D.; Hess, H. F.; Chang, A. M.; Pfeiffer, Loren N.; West, Kenneth W.; Mitzi, David B.

1993-06-01

A high resolution scanning Hall probe microscope is used to spatially resolve vortices in high temperature superconducting Bi2Sr2CaCu2O8+(delta) crystals. We observe a partially ordered vortex lattice at several different applied magnetic fields and temperatures. At higher temperatures, a limited amount of vortex re-arrangement is observed, but most vortices remain fixed for periods long compared to the imaging time of several hours even at temperatures as high as 75 degree(s)K (the superconducting transition temperature for these crystals is approximately 84 degree(s)K). A measure of these local magnetic penetration depth can be obtained from a fit to the surface field of several neighboring vortices, and has been measured as a function of temperature. In particular, we have measured the zero temperature penetration depth and found it to be 275 +/- 40 nm.

eV-TEM: Transmission electron microscopy in a low energy cathode lens instrument.

PubMed

Geelen, Daniël; Thete, Aniket; Schaff, Oliver; Kaiser, Alexander; van der Molen, Sense Jan; Tromp, Rudolf

2015-12-01

We are developing a transmission electron microscope that operates at extremely low electron energies, 0-40 eV. We call this technique eV-TEM. Its feasibility is based on the fact that at very low electron energies the number of energy loss pathways decreases. Hence, the electron inelastic mean free path increases dramatically. eV-TEM will enable us to study elastic and inelastic interactions of electrons with thin samples. With the recent development of aberration correction in cathode lens instruments, a spatial resolution of a few nm appears within range, even for these very low electron energies. Such resolution will be highly relevant to study biological samples such as proteins and cell membranes. The low electron energies minimize adverse effects due to radiation damage. Copyright © 2015. Published by Elsevier B.V.

Neural Network for Image-to-Image Control of Optical Tweezers

NASA Technical Reports Server (NTRS)

Decker, Arthur J.; Anderson, Robert C.; Weiland, Kenneth E.; Wrbanek, Susan Y.

2004-01-01

A method is discussed for using neural networks to control optical tweezers. Neural-net outputs are combined with scaling and tiling to generate 480 by 480-pixel control patterns for a spatial light modulator (SLM). The SLM can be combined in various ways with a microscope to create movable tweezers traps with controllable profiles. The neural nets are intended to respond to scattered light from carbon and silicon carbide nanotube sensors. The nanotube sensors are to be held by the traps for manipulation and calibration. Scaling and tiling allow the 100 by 100-pixel maximum resolution of the neural-net software to be applied in stages to exploit the full 480 by 480-pixel resolution of the SLM. One of these stages is intended to create sensitive null detectors for detecting variations in the scattered light from the nanotube sensors.

Penetrating view of nano-structures in Aleochara verna spermatheca and flagellum by hard X-ray microscopy

NASA Astrophysics Data System (ADS)

Zhang, Kai; Li, De-E.; Hong, You-Li; Zhu, Pei-Ping; Yuan, Qing-Xi; Huang, Wan-Xia; Gao, Kun; Zhou, Hong-Zhang; Wu, Zi-Yu

2013-07-01

A penetrating view of the three-dimensional nanostructure of female spermatheca and male flagellum in the species Aleochara verna is obtained with 100-nm resolution using a hard X-ray microscope, which provides a fast noninvasive imaging technology for insect morphology. Through introducing Zernike phase contrast and heavy metal staining, images taken at 8 keV displayed sufficient contrast for observing nanoscale fine structures, such as the spermatheca cochleate duct and the subapex of the flagellum, which have some implications for the study of the sperm transfer process and genital evolution in insects. This work shows that both the spatial resolution and the contrast characteristic of hard X-ray microscopy are quite promising for insect morphology studies and, particularly, provide an attractive alternative to the destructive techniques used for investigating internal soft tissues.

Continuous Fluorescence Microphotolysis and Correlation Spectroscopy Using 4Pi Microscopy

PubMed Central

Arkhipov, Anton; Hüve, Jana; Kahms, Martin; Peters, Reiner; Schulten, Klaus

2007-01-01

Continuous fluorescence microphotolysis (CFM) and fluorescence correlation spectroscopy (FCS) permit measurement of molecular mobility and association reactions in single living cells. CFM and FCS complement each other ideally and can be realized using identical equipment. So far, the spatial resolution of CFM and FCS was restricted by the resolution of the light microscope to the micrometer scale. However, cellular functions generally occur on the nanometer scale. Here, we develop the theoretical and computational framework for CFM and FCS experiments using 4Pi microscopy, which features an axial resolution of ∼100 nm. The framework, taking the actual 4Pi point spread function of the instrument into account, was validated by measurements on model systems, employing 4Pi conditions or normal confocal conditions together with either single- or two-photon excitation. In all cases experimental data could be well fitted by computed curves for expected diffusion coefficients, even when the signal/noise ratio was small due to the small number of fluorophores involved. PMID:17704168

Experimental assessment and analysis of super-resolution in fluorescence microscopy based on multiple-point spread function fitting of spectrally demultiplexed images

NASA Astrophysics Data System (ADS)

Nishimura, Takahiro; Kimura, Hitoshi; Ogura, Yusuke; Tanida, Jun

2018-06-01

This paper presents an experimental assessment and analysis of super-resolution microscopy based on multiple-point spread function fitting of spectrally demultiplexed images using a designed DNA structure as a test target. For the purpose, a DNA structure was designed to have binding sites at a certain interval that is smaller than the diffraction limit. The structure was labeled with several types of quantum dots (QDs) to acquire their spatial information as spectrally encoded images. The obtained images are analyzed with a point spread function multifitting algorithm to determine the QD locations that indicate the binding site positions. The experimental results show that the labeled locations can be observed beyond the diffraction-limited resolution using three-colored fluorescence images that were obtained with a confocal fluorescence microscope. Numerical simulations show that labeling with eight types of QDs enables the positions aligned at 27.2-nm pitches on the DNA structure to be resolved with high accuracy.

Visualization of Nanoplasmonic Coupling to Molecular Orbital in Light Emission Induced by Tunneling Electrons.

PubMed

Yu, Arthur; Li, Shaowei; Wang, Hui; Chen, Siyu; Wu, Ruqian; Ho, W

2018-05-09

The coupling between localized plasmon and molecular orbital in the light emission from a metallic nanocavity has been directly detected and imaged with sub-0.1 nm resolution. The light emission intensity was enhanced when the energy difference between the tunneling electrons and the lowest unoccupied molecular orbital (LUMO) of an azulene molecule matches the energy of a plasmon mode of the nanocavity defined by the Ag-tip and Ag (110) substrate of a scanning tunneling microscope (STM). The spatially resolved image of the light emission intensity matches the spatial distribution of the LUMO obtained by scanning tunneling spectroscopy (STS) and density functional theory (DFT) calculations. Our results highlight the near-field coupling of a molecular orbital to the radiative decay of a plasmonic excitation in a confined nanoscale junction.

Multimodal hyperspectral optical microscopy

DOE PAGES

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

2017-09-02

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

Multimodal hyperspectral optical microscopy

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

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

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