Automatic analysis of digitized TV-images by a computer-driven optical microscope

International Nuclear Information System (INIS)

Rosa, G.; Di Bartolomeo, A.; Grella, G.; Romano, G.

1997-01-01

New methods of image analysis and three-dimensional pattern recognition were developed in order to perform the automatic scan of nuclear emulsion pellicles. An optical microscope, with a motorized stage, was equipped with a CCD camera and an image digitizer, and interfaced to a personal computer. Selected software routines inspired the design of a dedicated hardware processor. Fast operation, high efficiency and accuracy were achieved. First applications to high-energy physics experiments are reported. Further improvements are in progress, based on a high-resolution fast CCD camera and on programmable digital signal processors. Applications to other research fields are envisaged. (orig.)

Polarization resolved imaging with a reflection near-field optical microscope

DEFF Research Database (Denmark)

Bozhevolnyi, Sergey I.; Xiao, Mufei; Hvam, Jørn Märcher

1999-01-01

Using a rigorous microscopic point-dipole description of probe-sample interactions, we study imaging with a reflection scanning near-field optical microscope. Optical content, topographical artifacts, sensitivity window-i.e., the scale on which near-field optical images represent mainly optical...... configuration is preferable to the cross-linear one, since it ensures more isotropic (in the surface plane) near-field imaging of surface features. The numerical results are supported with experimental near-field images obtained by using a reflection microscope with an uncoated fiber tip....

Design of a normal incidence multilayer imaging X-ray microscope

Science.gov (United States)

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.

Imaging arrangement and microscope

Science.gov (United States)

Pertsinidis, Alexandros; Chu, Steven

2015-12-15

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

Remote Histology Learning from Static versus Dynamic Microscopic Images

Science.gov (United States)

Mione, Sylvia; Valcke, Martin; Cornelissen, Maria

2016-01-01

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

Quantitative imaging with a mobile phone microscope.

Directory of Open Access Journals (Sweden)

Arunan Skandarajah

Full Text Available 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.

Quantitative Imaging with a Mobile Phone Microscope

Science.gov (United States)

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

Microscopic validation of whole mouse micro-metastatic tumor imaging agents using cryo-imaging and sliding organ image registration

OpenAIRE

Liu, Yiqiao; Zhou, Bo; Qutaish, Mohammed; Wilson, David L.

2016-01-01

We created a metastasis imaging, analysis platform consisting of software and multi-spectral cryo-imaging system suitable for evaluating emerging imaging agents targeting micro-metastatic tumor. We analyzed CREKA-Gd in MRI, followed by cryo-imaging which repeatedly sectioned and tiled microscope images of the tissue block face, providing anatomical bright field and molecular fluorescence, enabling 3D microscopic imaging of the entire mouse with single metastatic cell sensitivity. To register ...

Microscopic hyperspectral imaging studies of normal and diabetic retina of rats

Institute of Scientific and Technical Information of China (English)

2008-01-01

A microscopic hyperspectral imager was developed based on the microscopic technology and the spectral imaging technology. Some microscopic hyperspectral images of retina sections of the normal, the diabetic, and the treated rats were collected by the new imager. Single-band images and pseudo-color images of each group were obtained and the typical transmittance spectrums were ex-tracted. The results showed that the transmittance of outer nuclear layer cells of the diabetic group was generally higher than that of the normal. A small absorption peak appeared near the 180th band in the spectrum of the diabetic group and this peak weakened or disappeared in the spectrum of the treated group. Our findings indicate that the microscopic hyperspectral images include wealthy information of retina sections which is helpful for the ophthalmologist to reveal the pathogenesis of diabetic reti-nopathy and explore the therapeutic effect of drugs.

Stitching Grid-wise Atomic Force Microscope Images

DEFF Research Database (Denmark)

Vestergaard, Mathias Zacho; Bengtson, Stefan Hein; Pedersen, Malte

2016-01-01

Atomic Force Microscopes (AFM) are able to capture images with a resolution in the nano metre scale. Due to this high resolution, the covered area per image is relatively small, which can be problematic when surveying a sample. A system able to stitch AFM images has been developed to solve this p...

On the Progress of Scanning Transmission Electron Microscopy (STEM) Imaging in a Scanning Electron Microscope.

Science.gov (United States)

Sun, Cheng; Müller, Erich; Meffert, Matthias; Gerthsen, Dagmar

2018-04-01

Transmission electron microscopy (TEM) with low-energy electrons has been recognized as an important addition to the family of electron microscopies as it may avoid knock-on damage and increase the contrast of weakly scattering objects. Scanning electron microscopes (SEMs) are well suited for low-energy electron microscopy with maximum electron energies of 30 keV, but they are mainly used for topography imaging of bulk samples. Implementation of a scanning transmission electron microscopy (STEM) detector and a charge-coupled-device camera for the acquisition of on-axis transmission electron diffraction (TED) patterns, in combination with recent resolution improvements, make SEMs highly interesting for structure analysis of some electron-transparent specimens which are traditionally investigated by TEM. A new aspect is correlative SEM, STEM, and TED imaging from the same specimen region in a SEM which leads to a wealth of information. Simultaneous image acquisition gives information on surface topography, inner structure including crystal defects and qualitative material contrast. Lattice-fringe resolution is obtained in bright-field STEM imaging. The benefits of correlative SEM/STEM/TED imaging in a SEM are exemplified by structure analyses from representative sample classes such as nanoparticulates and bulk materials.

Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.

Science.gov (United States)

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).

Laser speckle contrast imaging using light field microscope approach

Science.gov (United States)

Ma, Xiaohui; Wang, Anting; Ma, Fenghua; Wang, Zi; Ming, Hai

2018-01-01

In this paper, a laser speckle contrast imaging (LSCI) system using light field (LF) microscope approach is proposed. As far as we known, it is first time to combine LSCI with LF. To verify this idea, a prototype consists of a modified LF microscope imaging system and an experimental device was built. A commercially used Lytro camera was modified for microscope imaging. Hollow glass tubes with different depth fixed in glass dish were used to simulate the vessels in brain and test the performance of the system. Compared with conventional LSCI, three new functions can be realized by using our system, which include refocusing, extending the depth of field (DOF) and gathering 3D information. Experiments show that the principle is feasible and the proposed system works well.

Microscopic imaging through turbid media Monte Carlo modeling and applications

CERN Document Server

Gu, Min; Deng, Xiaoyuan

2015-01-01

This book provides a systematic introduction to the principles of microscopic imaging through tissue-like turbid media in terms of Monte-Carlo simulation. It describes various gating mechanisms based on the physical differences between the unscattered and scattered photons and method for microscopic image reconstruction, using the concept of the effective point spread function. Imaging an object embedded in a turbid medium is a challenging problem in physics as well as in biophotonics. A turbid medium surrounding an object under inspection causes multiple scattering, which degrades the contrast, resolution and signal-to-noise ratio. Biological tissues are typically turbid media. Microscopic imaging through a tissue-like turbid medium can provide higher resolution than transillumination imaging in which no objective is used. This book serves as a valuable reference for engineers and scientists working on microscopy of tissue turbid media.

Lateral resolution testing of a novel developed confocal microscopic imaging system

Science.gov (United States)

Zhang, Xin; Zhang, Yunhai; Chang, Jian; Huang, Wei; Xue, Xiaojun; Xiao, Yun

2015-10-01

Laser scanning confocal microscope has been widely used in biology, medicine and material science owing to its advantages of high resolution and tomographic imaging. Based on a set of confirmatory experiments and system design, a novel confocal microscopic imaging system is developed. The system is composed of a conventional fluorescence microscope and a confocal scanning unit. In the scanning unit a laser beam coupling module provides four different wavelengths 405nm 488nm 561nm and 638nm which can excite a variety of dyes. The system works in spot-to-spot scanning mode with a two-dimensional galvanometer. A 50 microns pinhole is used to guarantee that stray light is blocked and only the fluorescence signal from the focal point can be received . The three-channel spectral splitter is used to perform fluorescence imaging at three different working wavelengths simultaneously. The rat kidney tissue slice is imaged using the developed confocal microscopic imaging system. Nucleues labeled by DAPI and kidney spherule curved pipe labeled by Alexa Fluor 488 can be imaged clearly and respectively, realizing the distinction between the different components of mouse kidney tissue. The three-dimensional tomographic imaging of mouse kidney tissue is reconstructed by several two-dimensional images obtained in different depths. At last the resolution of the confocal microscopic imaging system is tested quantitatively. The experimental result shows that the system can achieve lateral resolution priority to 230nm.

Imaging differential polarization microscope with electronic readout

International Nuclear Information System (INIS)

Mickols, W.; Tinoco, I.; Katz, J.E.; Maestre, M.F.; Bustamante, C.

1985-01-01

A differential polarization microscope forms two images: one of the transmitted intensity and the other due to the change in intensity between images formed when different polarizations of light are used. The interpretation of these images for linear dichroism and circular dichroism are described. The design constraints on the data acquisition systems and the polarization modulation are described. The advantage of imaging several biological systems which contain optically anisotropic structures are described

Performance evaluation of image segmentation algorithms on microscopic image data

Czech Academy of Sciences Publication Activity Database

Beneš, Miroslav; Zitová, Barbara

2015-01-01

Roč. 275, č. 1 (2015), s. 65-85 ISSN 0022-2720 R&D Projects: GA ČR GAP103/12/2211 Institutional support: RVO:67985556 Keywords : image segmentation * performance evaluation * microscopic images Subject RIV: JC - Computer Hardware ; Software Impact factor: 2.136, year: 2015 http://library.utia.cas.cz/separaty/2014/ZOI/zitova-0434809-DOI.pdf

Remote laboratory for phase-aided 3D microscopic imaging and metrology

Science.gov (United States)

Wang, Meng; Yin, Yongkai; Liu, Zeyi; He, Wenqi; Li, Boqun; Peng, Xiang

2014-05-01

In this paper, the establishment of a remote laboratory for phase-aided 3D microscopic imaging and metrology is presented. Proposed remote laboratory consists of three major components, including the network-based infrastructure for remote control and data management, the identity verification scheme for user authentication and management, and the local experimental system for phase-aided 3D microscopic imaging and metrology. The virtual network computer (VNC) is introduced to remotely control the 3D microscopic imaging system. Data storage and management are handled through the open source project eSciDoc. Considering the security of remote laboratory, the fingerprint is used for authentication with an optical joint transform correlation (JTC) system. The phase-aided fringe projection 3D microscope (FP-3DM), which can be remotely controlled, is employed to achieve the 3D imaging and metrology of micro objects.

Development and applications of the positron microscope

International Nuclear Information System (INIS)

1991-01-01

Progress on the positron microscope during the past year has been steady, and we currently project that initial microscope images can be collected during mid to late summer of 1992. Work during the year has mainly been divided among four areas of effort: hardware construction; power supply and control system development; radioactive source fabrication; and planning of initial experimental projects. Details of progress in these areas will be given below. An initial optical design of the microscope was completed during 1990, but during the past year, significant improvements have been made to this design, and several limiting cases of microscope performance have been evaluated. The results of these evaluations have been extremely encouraging, giving us strong indications that the optical performance of the microscope will be better than originally anticipated. In particular, we should be able to explore ultimate performance capabilities of positron microscopy using our currently planned optical system, with improvements only in the image detector system, and the positron-source/moderator configuration. We should be able to study imaging reemission microscopy with resolutions approaching 10 Angstrom and be able to produce beam spots for rastered microscope work with diameters below the 1000 Angstrom diffusion limit. Because of these exciting new possibilities, we have decided to upgrade several microscope subsystems to levels consistent with ultimate performance earlier in our construction schedule than we had previously intended. In particular, alignment facilities in the optical system, vibration isolation, and power supply and control system flexibility have all been upgraded in their design over the past year

In vivo cellular imaging with microscopes enabled by MEMS scanners

Science.gov (United States)

Ra, Hyejun

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

Imaging C. elegans embryos using an epifluorescent microscope and open source software.

Science.gov (United States)

Verbrugghe, Koen J C; Chan, Raymond C

2011-03-24

Cellular processes, such as chromosome assembly, segregation and cytokinesis,are inherently dynamic. Time-lapse imaging of living cells, using fluorescent-labeled reporter proteins or differential interference contrast (DIC) microscopy, allows for the examination of the temporal progression of these dynamic events which is otherwise inferred from analysis of fixed samples(1,2). Moreover, the study of the developmental regulations of cellular processes necessitates conducting time-lapse experiments on an intact organism during development. The Caenorhabiditis elegans embryo is light-transparent and has a rapid, invariant developmental program with a known cell lineage(3), thus providing an ideal experiment model for studying questions in cell biology(4,5)and development(6-9). C. elegans is amendable to genetic manipulation by forward genetics (based on random mutagenesis(10,11)) and reverse genetics to target specific genes (based on RNAi-mediated interference and targeted mutagenesis(12-15)). In addition, transgenic animals can be readily created to express fluorescently tagged proteins or reporters(16,17). These traits combine to make it easy to identify the genetic pathways regulating fundamental cellular and developmental processes in vivo(18-21). In this protocol we present methods for live imaging of C. elegans embryos using DIC optics or GFP fluorescence on a compound epifluorescent microscope. We demonstrate the ease with which readily available microscopes, typically used for fixed sample imaging, can also be applied for time-lapse analysis using open-source software to automate the imaging process.

Sensing of Streptococcus mutans by microscopic imaging ellipsometry

Science.gov (United States)

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

2017-05-01

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

3D widefield light microscope image reconstruction without dyes

Science.gov (United States)

Larkin, S.; Larson, J.; Holmes, C.; Vaicik, M.; Turturro, M.; Jurkevich, A.; Sinha, S.; Ezashi, T.; Papavasiliou, G.; Brey, E.; Holmes, T.

2015-03-01

3D image reconstruction using light microscope modalities without exogenous contrast agents is proposed and investigated as an approach to produce 3D images of biological samples for live imaging applications. Multimodality and multispectral imaging, used in concert with this 3D optical sectioning approach is also proposed as a way to further produce contrast that could be specific to components in the sample. The methods avoid usage of contrast agents. Contrast agents, such as fluorescent or absorbing dyes, can be toxic to cells or alter cell behavior. Current modes of producing 3D image sets from a light microscope, such as 3D deconvolution algorithms and confocal microscopy generally require contrast agents. Zernike phase contrast (ZPC), transmitted light brightfield (TLB), darkfield microscopy and others can produce contrast without dyes. Some of these modalities have not previously benefitted from 3D image reconstruction algorithms, however. The 3D image reconstruction algorithm is based on an underlying physical model of scattering potential, expressed as the sample's 3D absorption and phase quantities. The algorithm is based upon optimizing an objective function - the I-divergence - while solving for the 3D absorption and phase quantities. Unlike typical deconvolution algorithms, each microscope modality, such as ZPC or TLB, produces two output image sets instead of one. Contrast in the displayed image and 3D renderings is further enabled by treating the multispectral/multimodal data as a feature set in a mathematical formulation that uses the principal component method of statistics.

Three-dimensional phase-contrast X-ray microtomography with scanning–imaging X-ray microscope optics

International Nuclear Information System (INIS)

Takeuchi, Akihisa; Uesugi, Kentaro; Suzuki, Yoshio

2013-01-01

A novel three-dimensional X-ray microtomographic micro-imaging system which enables simultaneous measurement of differential phase contrast and absorption contrast has been developed. The optical system consists of a scanning microscope with one-dimensional focusing device and an imaging microscope with one-dimensional objective. A three-dimensional (3D) X-ray tomographic micro-imaging system has been developed. The optical system is based on a scanning–imaging X-ray microscope (SIXM) optics, which is a hybrid system consisting of a scanning microscope optics with a one-dimensional (1D) focusing (line-focusing) device and an imaging microscope optics with a 1D objective. In the SIXM system, each 1D dataset of a two-dimensional (2D) image is recorded independently. An object is illuminated with a line-focused beam. Positional information of the region illuminated by the line-focused beam is recorded with the 1D imaging microscope optics as line-profile data. By scanning the object with the line focus, 2D image data are obtained. In the same manner as for a scanning microscope optics with a multi-pixel detector, imaging modes such as phase contrast and absorption contrast can be arbitrarily configured after the image data acquisition. By combining a tomographic scan method and the SIXM system, quantitative 3D imaging is performed. Results of a feasibility study of the SIXM for 3D imaging are shown

High-resolution, high-throughput imaging with a multibeam scanning electron microscope.

Science.gov (United States)

Eberle, A L; Mikula, S; Schalek, R; Lichtman, J; Knothe Tate, M L; Zeidler, D

2015-08-01

Electron-electron interactions and detector bandwidth limit the maximal imaging speed of single-beam scanning electron microscopes. We use multiple electron beams in a single column and detect secondary electrons in parallel to increase the imaging speed by close to two orders of magnitude and demonstrate imaging for a variety of samples ranging from biological brain tissue to semiconductor wafers. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

A Simple Metric for Determining Resolution in Optical, Ion, and Electron Microscope Images.

Science.gov (United States)

Curtin, Alexandra E; Skinner, Ryan; Sanders, Aric W

2015-06-01

A resolution metric intended for resolution analysis of arbitrary spatially calibrated images is presented. By fitting a simple sigmoidal function to pixel intensities across slices of an image taken perpendicular to light-dark edges, the mean distance over which the light-dark transition occurs can be determined. A fixed multiple of this characteristic distance is then reported as the image resolution. The prefactor is determined by analysis of scanning transmission electron microscope high-angle annular dark field images of Si. This metric has been applied to optical, scanning electron microscope, and helium ion microscope images. This method provides quantitative feedback about image resolution, independent of the tool on which the data were collected. In addition, our analysis provides a nonarbitrary and self-consistent framework that any end user can utilize to evaluate the resolution of multiple microscopes from any vendor using the same metric.

Soft X-ray imaging with axisymmetry microscope and electronic readout

International Nuclear Information System (INIS)

Sauneuf, A.; Cavailler, C.; Henry, Ph.; Launspach, J.; Mascureau, J. de; Rostaing, M.

1984-11-01

An axisymmetric microscope with 10 X magnification has been constructed; its resolution has been measured using severals grids, backlighted by an X-ray source and found to be near 25 μm. So it could be used to make images of laser driven plasmas in the soft X-ray region. In order to see rapidly those images we have associated it with a new detector. It is a small image converter tube with a soft X-ray photocathode and a P20 phosphor deposited on an optic fiber plate. The electronic image appearing on the screen is read by a CCD working in the spectral range. An electronic image readout chain, which is identical to those we use with streak cameras, then processes automatically and immediatly the images given by the microscope

Dual-mode optical microscope based on single-pixel imaging

Science.gov (United States)

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

2016-07-01

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

Simulation of high-resolution X-ray microscopic images for improved alignment

International Nuclear Information System (INIS)

Song Xiangxia; Zhang Xiaobo; Liu Gang; Cheng Xianchao; Li Wenjie; Guan Yong; Liu Ying; Xiong Ying; Tian Yangchao

2011-01-01

The introduction of precision optical elements to X-ray microscopes necessitates fine realignment to achieve optimal high-resolution imaging. In this paper, we demonstrate a numerical method for simulating image formation that facilitates alignment of the source, condenser, objective lens, and CCD camera. This algorithm, based on ray-tracing and Rayleigh-Sommerfeld diffraction theory, is applied to simulate the X-ray microscope beamline U7A of National Synchrotron Radiation Laboratory (NSRL). The simulations and imaging experiments show that the algorithm is useful for guiding experimental adjustments. Our alignment simulation method is an essential tool for the transmission X-ray microscope (TXM) with optical elements and may also be useful for the alignment of optical components in other modes of microscopy.

The plant virus microscope image registration method based on mismatches removing.

Science.gov (United States)

Wei, Lifang; Zhou, Shucheng; Dong, Heng; Mao, Qianzhuo; Lin, Jiaxiang; Chen, Riqing

2016-01-01

The electron microscopy is one of the major means to observe the virus. The view of virus microscope images is limited by making specimen and the size of the camera's view field. To solve this problem, the virus sample is produced into multi-slice for information fusion and image registration techniques are applied to obtain large field and whole sections. Image registration techniques have been developed in the past decades for increasing the camera's field of view. Nevertheless, these approaches typically work in batch mode and rely on motorized microscopes. Alternatively, the methods are conceived just to provide visually pleasant registration for high overlap ratio image sequence. This work presents a method for virus microscope image registration acquired with detailed visual information and subpixel accuracy, even when overlap ratio of image sequence is 10% or less. The method proposed focus on the correspondence set and interimage transformation. A mismatch removal strategy is proposed by the spatial consistency and the components of keypoint to enrich the correspondence set. And the translation model parameter as well as tonal inhomogeneities is corrected by the hierarchical estimation and model select. In the experiments performed, we tested different registration approaches and virus images, confirming that the translation model is not always stationary, despite the fact that the images of the sample come from the same sequence. The mismatch removal strategy makes building registration of virus microscope images at subpixel accuracy easier and optional parameters for building registration according to the hierarchical estimation and model select strategies make the proposed method high precision and reliable for low overlap ratio image sequence. Copyright © 2015 Elsevier Ltd. All rights reserved.

Scanning tunneling microscope for magneto-optical imaging

NARCIS (Netherlands)

Prins, M.W.J.; Groeneveld, R.H.M.; Abraham, D.L.; Schad, R.; Kempen, van H.; Kesteren, van H.W.

1996-01-01

Images of magnetic bits written in a Pt/Co multilayer are presented. Using photosensitive semiconducting tips in a scanning tunneling microscope the surface topography as well as the polarization-dependent optical transmission are measured. Magnetic contrast is achieved by detection of the Faraday

Microdose fluorescence imaging of ABY-029 on an operating microscope adapted by custom illumination and imaging modules

OpenAIRE

Elliott, Jonathan T.; Dsouza, Alisha V.; Marra, Kayla; Pogue, Brian W.; Roberts, David W.; Paulsen, Keith D.

2016-01-01

Fluorescence guided surgery has the potential to positively impact surgical oncology; current operating microscopes and stand-alone imaging systems are too insensitive or too cumbersome to maximally take advantage of new tumor-specific agents developed through the microdose pathway. To this end, a custom-built illumination and imaging module enabling picomolar-sensitive near-infrared fluorescence imaging on a commercial operating microscope is described. The limits of detection and system spe...

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

Science.gov (United States)

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

2018-03-01

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

Image enhancement of x-ray microscope using frequency spectrum analysis

International Nuclear Information System (INIS)

Li Wenjie; Chen Jie; Tian Jinping; Zhang Xiaobo; Liu Gang; Tian Yangchao; Liu Yijin; Wu Ziyu

2009-01-01

We demonstrate a new method for x-ray microscope image enhancement using frequency spectrum analysis. Fine sample characteristics are well enhanced with homogeneous visibility and better contrast from single image. This method is easy to implement and really helps to improve the quality of image taken by our imaging system.

Image enhancement of x-ray microscope using frequency spectrum analysis

Energy Technology Data Exchange (ETDEWEB)

Li Wenjie; Chen Jie; Tian Jinping; Zhang Xiaobo; Liu Gang; Tian Yangchao [National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui 230029 (China); Liu Yijin; Wu Ziyu, E-mail: wuzy@ihep.ac.c, E-mail: ychtian@ustc.edu.c [Institute of High Energy Physics, Chinese Academy of Science, Beijing 100049 (China)

2009-09-01

We demonstrate a new method for x-ray microscope image enhancement using frequency spectrum analysis. Fine sample characteristics are well enhanced with homogeneous visibility and better contrast from single image. This method is easy to implement and really helps to improve the quality of image taken by our imaging system.

Optical modeling of Fresnel zoneplate microscopes

International Nuclear Information System (INIS)

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

2011-01-01

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

Morphometric Evaluation of Preeclamptic Placenta Using Light Microscopic Images

Directory of Open Access Journals (Sweden)

Rashmi Mukherjee

2014-01-01

Full Text Available Deficient trophoblast invasion and anomalies in placental development generally lead to preeclampsia (PE but the inter-relationship between placental function and morphology in PE still remains unknown. The aim of this study was to evaluate the morphometric features of placental villi and capillaries in preeclamptic and normal placentae. The study included light microscopic images of placental tissue sections of 40 preeclamptic and 35 normotensive pregnant women. Preprocessing and segmentation of these images were performed to characterize the villi and capillaries. Fisher’s linear discriminant analysis (FLDA, hierarchical cluster analysis (HCA, and principal component analysis (PCA were applied to identify the most significant placental (morphometric features from microscopic images. A total of 10 morphometric features were extracted, of which the villous parameters were significantly altered in PE. FLDA identified 5 highly significant morphometric features (>90% overall discrimination accuracy. Two large subclusters were clearly visible in HCA based dendrogram. PCA returned three most significant principal components cumulatively explaining 98.4% of the total variance based on these 5 significant features. Hence, quantitative microscopic evaluation revealed that placental morphometry plays an important role in characterizing PE, where the villous is the major component that is affected.

A New Multichannel Spectral Imaging Laser Scanning Confocal Microscope

Directory of Open Access Journals (Sweden)

Yunhai Zhang

2013-01-01

Full Text Available We have developed a new multichannel spectral imaging laser scanning confocal microscope for effective detection of multiple fluorescent labeling in the research of biological tissues. In this paper, the design and key technologies of the system are introduced. Representative results on confocal imaging, 3-dimensional sectioning imaging, and spectral imaging are demonstrated. The results indicated that the system is applicable to multiple fluorescent labeling in biological experiments.

Real-time image processing and control interface for remote operation of a microscope

Science.gov (United States)

Leng, Hesong; Wilder, Joseph

1999-08-01

A real-time image processing and control interface for remote operation of a microscope is presented in this paper. The system has achieved real-time color image display for 640 X 480 pixel images. Multi-resolution image representation can be provided for efficient transmission through the network. Through the control interface the computer can communicate with the programmable microscope via the RS232 serial ports. By choosing one of three scanning patterns, a sequence of images can be saved as BMP or PGM files to record information on an entire microscope slide. The system will be used by medical and graduate students at the University of Medicine and Dentistry of New Jersey for distance learning. It can be used in many network-based telepathology applications.

Thermal diffusivity imaging with the thermal lens microscope.

Science.gov (United States)

Dada, Oluwatosin O; Feist, Peter E; Dovichi, Norman J

2011-12-01

A coaxial thermal lens microscope was used to generate images based on both the absorbance and thermal diffusivity of histological samples. A pump beam was modulated at frequencies ranging from 50 kHz to 5 MHz using an acousto-optic modulator. The pump and a CW probe beam were combined with a dichroic mirror, directed into an inverted microscope, and focused onto the specimen. The change in the transmitted probe beam's center intensity was detected with a photodiode. The photodiode's signal and a reference signal from the modulator were sent to a high-speed lock-in amplifier. The in-phase and quadrature signals were recorded as a sample was translated through the focused beams and used to generate images based on the amplitude and phase of the lock-in amplifier's signal. The amplitude is related to the absorbance and the phase is related to the thermal diffusivity of the sample. Thin sections of stained liver and bone tissues were imaged; the contrast and signal-to-noise ratio of the phase image was highest at frequencies from 0.1-1 MHz and dropped at higher frequencies. The spatial resolution was 2.5 μm for both amplitude and phase images, limited by the pump beam spot size. © 2011 Optical Society of America

Development of the water window imaging x-ray microscope

International Nuclear Information System (INIS)

Hoover, R.B.; Shealy, D.L.; Baker, P.C.; Barbee, T.W. Jr.; Walker, A.B.C. Jr.

1991-01-01

This paper reports on the Water Window Imaging X-ray Microscopy which is currently being developed by a consortium from the Marshall Space Flight Center, the University of Alabama at Birmingham, Baker Consulting, the Lawrence Livermore National Laboratory, and Stanford University. The high quality solar images achieved during the Stanford/MSFC/LLNL Rocket X-ray Spectroheliograph flight conclusively established that excellent imaging could be obtained with doubly reflecting multilayer optical systems. Theoretical studies carried out as part of the MSFC X-ray Microscopy Program, demonstrated that high quality, high resolution multilayer x-ray imaging microscopes could be achieved with spherical optics in the Schwarzschild configuration and with Aspherical optical systems. Advanced Flow Polishing methods have been used to fabricate substrates for multilayer optics. On hemlite grade Sapphire, the authors have achieved microscopy mirror substrates on concave and convex spherical surfaces with 0.5 Angstrom rms surface smoothness, as measured by the Zygo profilometer. In this paper the authors report on the current status of fabrication and testing of the optical and mechanical subsystems for the Water Window Imaging X-ray Microscope

The importance of radiographic imaging in the microscopic assessment of bone tumors

International Nuclear Information System (INIS)

Larousserie, F.; Kreshak, J.; Gambarotti, M.; Alberghini, M.; Vanel, D.

2013-01-01

Introduction: Primary bone tumors are rare and require a multidisciplinary approach. Diagnosis involves primarily the radiologist and the pathologist. Bone lesions are often heterogeneous and the microscopic diagnostic component(s) may be in the minority, especially on core needle biopsies. Reactive processes, benign, and malignant tumors may have similar microscopic aspects. For these challenging cases, the correlation of microscopic and radiologic information is critical, or diagnostic mistakes may be made with severe clinical consequences for the patient. The purpose of this article is to explain how pathologists can best use imaging studies to improve the diagnostic accuracy of bone lesions. Diagnosis: Many bone lesions are microscopically and/or radiographically heterogeneous, especially those with both lytic and matrix components. Final diagnosis may require specific microscopic diagnostic features that may be present in the lesion, but not the biopsy specimen. A review of the imaging helps assess if sampling was adequate. The existence of a pre-existing bone lesion, syndrome (such as Ollier disease or multiple hereditary exostosis), or oncologic history may be of crucial importance. Finally, imaging information is very useful for the pathologist to perform accurate local and regional staging during gross examination. Conclusion: Close teamwork between pathologists, radiologists, and clinicians is of utmost importance in the evaluation and management of bone tumors. These lesions can be very difficult to interpret microscopically; imaging studies therefore play a crucial role in their accurate diagnosis

The importance of radiographic imaging in the microscopic assessment of bone tumors

Energy Technology Data Exchange (ETDEWEB)

Larousserie, F., E-mail: frederique.larousserie@cch.aphp.fr [Université Paris Descartes, Sorbonne Paris Cité, Paris (France); Department of pathology, Rizzoli Institute, Bologna (Italy); Kreshak, J.; Gambarotti, M.; Alberghini, M.; Vanel, D. [Department of pathology, Rizzoli Institute, Bologna (Italy)

2013-12-01

Introduction: Primary bone tumors are rare and require a multidisciplinary approach. Diagnosis involves primarily the radiologist and the pathologist. Bone lesions are often heterogeneous and the microscopic diagnostic component(s) may be in the minority, especially on core needle biopsies. Reactive processes, benign, and malignant tumors may have similar microscopic aspects. For these challenging cases, the correlation of microscopic and radiologic information is critical, or diagnostic mistakes may be made with severe clinical consequences for the patient. The purpose of this article is to explain how pathologists can best use imaging studies to improve the diagnostic accuracy of bone lesions. Diagnosis: Many bone lesions are microscopically and/or radiographically heterogeneous, especially those with both lytic and matrix components. Final diagnosis may require specific microscopic diagnostic features that may be present in the lesion, but not the biopsy specimen. A review of the imaging helps assess if sampling was adequate. The existence of a pre-existing bone lesion, syndrome (such as Ollier disease or multiple hereditary exostosis), or oncologic history may be of crucial importance. Finally, imaging information is very useful for the pathologist to perform accurate local and regional staging during gross examination. Conclusion: Close teamwork between pathologists, radiologists, and clinicians is of utmost importance in the evaluation and management of bone tumors. These lesions can be very difficult to interpret microscopically; imaging studies therefore play a crucial role in their accurate diagnosis.

A portable fluorescence microscopic imaging system for cholecystectomy

Science.gov (United States)

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.

DHM (Digital Holography Microscope) for imaging cells

International Nuclear Information System (INIS)

Emery, Yves; Cuche, Etienne; Colomb, Tristan; Depeursinge, Christian; Rappaz, Benjamin; Marquet, Pierre; Magistretti, Pierre

2007-01-01

Light interaction with a sample modifies both intensity and phase of the illuminating wave. Any available supports for image recording are only sensitive to intensity, but Denis Gabor [P. Marquet, B. Rappaz, P. Magistretti, et. al. Digital Holography for quantitative phase-contrast imaging, Optics Letters, 30, 5, pp 291-93 (2005)] invented in 1948 a way to encode the phase as an intensity variation: the h ologram . Digital Holographic Microscopy (DHM) [D. Gabor, A new microscopic principle, Nature, 1948] implements digitally this powerful hologram. Characterization of various pollen grains and of morphology changes of neurones associated with hypotonic shock demonstrates the potential of DHM for imaging cells

Chemical imaging of structured SAMs with a novel SFG microscope

Science.gov (United States)

Hoffmann, Dominik M. P.; Kuhnke, Klaus; Kern, Klaus

2002-11-01

We present a newly developed microscope for sum frequency generation (SFG) imaging of opaque and reflecting interfaces. The sample is viewed at an angle of 60° with respect to the surface normal in order to increase the collected SFG intensity. Our setup is designed to keep the whole field of view (FOV) in focus and to compensate for the distortion usually related to oblique imaging by means of a blazed grating. The separation of the SFG intensity and the reflected visible beam is accomplished by a suitable combination of spectral filters. The sum frequency microscope (SFM) is capable of in-situ chemically selective imaging by tuning the IR-beam to vibrational transitions of the respective molecules. The SFM is applied to imaging of structured self-assembled monolayers (SAM) of thiol molecules on a gold surface.

Interferometric and optical tests of water window imaging x ray microscopes

Science.gov (United States)

Johnson, R. Barry

1993-01-01

Interferometric tests of Schwarzchild X-ray Microscope are performed to evaluate the optical properties and alignment of the components. Photographic measurements of the spatial resolution, focal properties, and vignetting characteristics of the prototype Water Window Imaging X-ray Microscope are made and analyzed.

Live Imaging of Shoot Meristems on an Inverted Confocal Microscope Using an Objective Lens Inverter Attachment

Science.gov (United States)

Nimchuk, Zachary L.; Perdue, Tony D.

2017-01-01

Live imaging of above ground meristems can lead to new insights in plant development not possible from static imaging of fixed tissue. The use of an upright confocal microscope offers several technical and biological advantages for live imaging floral or shoot meristems. However, many departments and core facilities possess only inverted confocal microscopes and lack the funding for an additional upright confocal microscope. Here we show that imaging of living apical meristems can be performed on existing inverted confocal microscopes with the use of an affordable and detachable InverterScope accessory. PMID:28579995

Live Imaging of Shoot Meristems on an Inverted Confocal Microscope Using an Objective Lens Inverter Attachment.

Science.gov (United States)

Nimchuk, Zachary L; Perdue, Tony D

2017-01-01

Live imaging of above ground meristems can lead to new insights in plant development not possible from static imaging of fixed tissue. The use of an upright confocal microscope offers several technical and biological advantages for live imaging floral or shoot meristems. However, many departments and core facilities possess only inverted confocal microscopes and lack the funding for an additional upright confocal microscope. Here we show that imaging of living apical meristems can be performed on existing inverted confocal microscopes with the use of an affordable and detachable InverterScope accessory.

Dual-mode optical microscope based on single-pixel imaging

OpenAIRE

Rodríguez Jiménez, Angel David; Clemente Pesudo, Pedro Javier; Tajahuerce, Enrique; Lancis Sáez, Jesús

2016-01-01

We demonstrate an inverted microscope that can image specimens in both reflection and transmission modes simultaneously with a single light source. The microscope utilizes a digital micromirror device (DMD) for patterned illumination altogether with two single-pixel photosensors for efficient light detection. The system, a scan-less device with no moving parts, works by sequential projection of a set of binary intensity patterns onto the sample that are codified onto a modified commercial DMD...

Magnetoacoustic microscopic imaging of conductive objects and nanoparticles distribution

Science.gov (United States)

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.

Electric field effects in scanning tunneling microscope imaging

DEFF Research Database (Denmark)

Stokbro, Kurt; Quaade, Ulrich; Grey, Francois

1998-01-01

We present a high-voltage extension of the Tersoff-Hamann theory of scanning tunneling microscope (STM) images, which includes the effect of the electric field between the tip and the sample. The theoretical model is based on first-principles electronic structure calculations and has no adjustable...... parameters. We use the method to calculate theoretical STM images of the monohydrate Si(100)-H(2x1) surface with missing hydrogen defects at -2V and find an enhanced corrugation due to the electric field, in good agreement with experimental images....

Design of an imaging microscope for soft X-ray applications

Science.gov (United States)

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.

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

Science.gov (United States)

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

A new clustering algorithm for scanning electron microscope images

Science.gov (United States)

Yousef, Amr; Duraisamy, Prakash; Karim, Mohammad

2016-04-01

A scanning electron microscope (SEM) is a type of electron microscope that produces images of a sample by scanning it with a focused beam of electrons. The electrons interact with the sample atoms, producing various signals that are collected by detectors. The gathered signals contain information about the sample's surface topography and composition. The electron beam is generally scanned in a raster scan pattern, and the beam's position is combined with the detected signal to produce an image. The most common configuration for an SEM produces a single value per pixel, with the results usually rendered as grayscale images. The captured images may be produced with insufficient brightness, anomalous contrast, jagged edges, and poor quality due to low signal-to-noise ratio, grained topography and poor surface details. The segmentation of the SEM images is a tackling problems in the presence of the previously mentioned distortions. In this paper, we are stressing on the clustering of these type of images. In that sense, we evaluate the performance of the well-known unsupervised clustering and classification techniques such as connectivity based clustering (hierarchical clustering), centroid-based clustering, distribution-based clustering and density-based clustering. Furthermore, we propose a new spatial fuzzy clustering technique that works efficiently on this type of images and compare its results against these regular techniques in terms of clustering validation metrics.

Assessing microscope image focus quality with deep learning.

Science.gov (United States)

Yang, Samuel J; Berndl, Marc; Michael Ando, D; Barch, Mariya; Narayanaswamy, Arunachalam; Christiansen, Eric; Hoyer, Stephan; Roat, Chris; Hung, Jane; Rueden, Curtis T; Shankar, Asim; Finkbeiner, Steven; Nelson, Philip

2018-03-15

Large image datasets acquired on automated microscopes typically have some fraction of low quality, out-of-focus images, despite the use of hardware autofocus systems. Identification of these images using automated image analysis with high accuracy is important for obtaining a clean, unbiased image dataset. Complicating this task is the fact that image focus quality is only well-defined in foreground regions of images, and as a result, most previous approaches only enable a computation of the relative difference in quality between two or more images, rather than an absolute measure of quality. We present a deep neural network model capable of predicting an absolute measure of image focus on a single image in isolation, without any user-specified parameters. The model operates at the image-patch level, and also outputs a measure of prediction certainty, enabling interpretable predictions. The model was trained on only 384 in-focus Hoechst (nuclei) stain images of U2OS cells, which were synthetically defocused to one of 11 absolute defocus levels during training. The trained model can generalize on previously unseen real Hoechst stain images, identifying the absolute image focus to within one defocus level (approximately 3 pixel blur diameter difference) with 95% accuracy. On a simpler binary in/out-of-focus classification task, the trained model outperforms previous approaches on both Hoechst and Phalloidin (actin) stain images (F-scores of 0.89 and 0.86, respectively over 0.84 and 0.83), despite only having been presented Hoechst stain images during training. Lastly, we observe qualitatively that the model generalizes to two additional stains, Hoechst and Tubulin, of an unseen cell type (Human MCF-7) acquired on a different instrument. Our deep neural network enables classification of out-of-focus microscope images with both higher accuracy and greater precision than previous approaches via interpretable patch-level focus and certainty predictions. The use of

Development of Scanning-Imaging X-Ray Microscope for Quantitative Three-Dimensional Phase Contrast Microimaging

International Nuclear Information System (INIS)

Takeuchi, Akihisa; Suzuki, Yoshio; Uesugi, Kentaro

2013-01-01

A novel x-ray microscope system has been developed for the purpose of quantitative and sensitive three-dimensional (3D) phase-contrast x-ray microimaging. The optical system is a hybrid that consists of a scanning microscope optics with a one-dimensional (1D) focusing (line-focusing) device and an imaging microscope optics with a 1D objective. These two optics are orthogonally arranged regarding their common optical axis. Each is used for forming each dimension of two-dimensional (2D) image. The same data acquisition process as that of the scanning microscope system enables quantitative and sensitive x-ray imaging such as phase contrast and absorption contrast. Because a 2D image is measured with only 1D translation scan, much shorter measurement time than that of conventional scanning optics has been realized. By combining a computed tomography (CT) technique, some 3D CT application examples are demonstrated

The development of a high speed 3D 2-photon microscope for neuroscience

OpenAIRE

Kirkby, P. A.

2010-01-01

The progress of neuroscience is limited by the instrumentation available to it for studying the brain. At present, there is a serious instrumentation gap between functional Magnetic Resonance Imaging (fMRI) of whole brains and the microscopic scale functional imaging possible with today’s optical microscopes and electrophysiology techniques, such as patch clamping of individual neurons. This thesis describes the development of a new extension to optical microscopy that enabl...

Characteristics of different frequency ranges in scanning electron microscope images

International Nuclear Information System (INIS)

Sim, K. S.; Nia, M. E.; Tan, T. L.; Tso, C. P.; Ee, C. S.

2015-01-01

We demonstrate a new approach to characterize the frequency range in general scanning electron microscope (SEM) images. First, pure frequency images are generated from low frequency to high frequency, and then, the magnification of each type of frequency image is implemented. By comparing the edge percentage of the SEM image to the self-generated frequency images, we can define the frequency ranges of the SEM images. Characterization of frequency ranges of SEM images benefits further processing and analysis of those SEM images, such as in noise filtering and contrast enhancement

Characteristics of different frequency ranges in scanning electron microscope images

Energy Technology Data Exchange (ETDEWEB)

Sim, K. S., E-mail: kssim@mmu.edu.my; Nia, M. E.; Tan, T. L.; Tso, C. P.; Ee, C. S. [Faculty of Engineering and Technology, Multimedia University, 75450 Melaka (Malaysia)

2015-07-22

We demonstrate a new approach to characterize the frequency range in general scanning electron microscope (SEM) images. First, pure frequency images are generated from low frequency to high frequency, and then, the magnification of each type of frequency image is implemented. By comparing the edge percentage of the SEM image to the self-generated frequency images, we can define the frequency ranges of the SEM images. Characterization of frequency ranges of SEM images benefits further processing and analysis of those SEM images, such as in noise filtering and contrast enhancement.

Single-cell magnetic imaging using a quantum diamond microscope.

Science.gov (United States)

Glenn, D R; Lee, K; Park, H; Weissleder, R; Yacoby, A; Lukin, M D; Lee, H; Walsworth, R L; Connolly, C B

2015-08-01

We apply a quantum diamond microscope for detection and imaging of immunomagnetically labeled cells. This instrument uses nitrogen-vacancy (NV) centers in diamond for correlated magnetic and fluorescence imaging. Our device provides single-cell resolution and a field of view (∼1 mm(2)) two orders of magnitude larger than that of previous NV imaging technologies, enabling practical applications. To illustrate, we quantified cancer biomarkers expressed by rare tumor cells in a large population of healthy cells.

Adaptation of commercial microscopes for advanced imaging applications

Science.gov (United States)

Brideau, Craig; Poon, Kelvin; Stys, Peter

2015-03-01

Today's commercially available microscopes offer a wide array of options to accommodate common imaging experiments. Occasionally, an experimental goal will require an unusual light source, filter, or even irregular sample that is not compatible with existing equipment. In these situations the ability to modify an existing microscopy platform with custom accessories can greatly extend its utility and allow for experiments not possible with stock equipment. Light source conditioning/manipulation such as polarization, beam diameter or even custom source filtering can easily be added with bulk components. Custom and after-market detectors can be added to external ports using optical construction hardware and adapters. This paper will present various examples of modifications carried out on commercial microscopes to address both atypical imaging modalities and research needs. Violet and near-ultraviolet source adaptation, custom detection filtering, and laser beam conditioning and control modifications will be demonstrated. The availability of basic `building block' parts will be discussed with respect to user safety, construction strategies, and ease of use.

Laser based imaging of time depending microscopic scenes with strong light emission

Science.gov (United States)

Hahlweg, Cornelius; Wilhelm, Eugen; Rothe, Hendrik

2011-10-01

Investigating volume scatterometry methods based on short range LIDAR devices for non-static objects we achieved interesting results aside the intended micro-LIDAR: the high speed camera recording of the illuminated scene of an exploding wire -intended for Doppler LIDAR tests - delivered a very effective method of observing details of objects with extremely strong light emission. As a side effect a schlieren movie is gathered without any special effort. The fact that microscopic features of short time processes with high emission and material flow might be imaged without endangering valuable equipment makes this technique at least as interesting as the intended one. So we decided to present our results - including latest video and photo material - instead of a more theoretical paper on our progress concerning the primary goal.

Hard X-ray Microscopic Imaging Of Human Breast Tissues

Science.gov (United States)

Park, Sung H.; Kim, Hong T.; Kim, Jong K.; Jheon, Sang H.; Youn, Hwa S.

2007-01-01

X-ray microscopy with synchrotron radiation will be a useful tool for innovation of x-ray imaging in clinical and laboratory settings. It helps us observe detailed internal structure of material samples non-invasively in air. And, it also has the potential to solve some tough problems of conventional breast imaging if it could evaluate various conditions of breast tissue effectively. A new hard x-ray microscope with a spatial resolution better than 100 nm was installed at Pohang Light Source, a third generation synchrotron radiation facility in Pohang, Korea. The x-ray energy was set at 6.95 keV, and the x-ray beam was monochromatized by W/B4C monochromator. Condenser and objective zone plates were used as x-ray lenses. Zernike phase plate next to condenser zone plate was introduced for improved contrast imaging. The image of a sample was magnified 30 times by objective zone plate and 20 times by microscope objective, respectively. After additional 10 times digital magnification, the total magnifying power was up to 6000 times in the end. Phase contrast synchrotron images of 10-μm-thick female breast tissue of the normal, fibroadenoma, fibrocystic change and carcinoma cases were obtained. By phase contrast imaging, hard x-rays enable us to observe many structures of breast tissue without sample preparations such as staining or fixation.

Hard X-ray Microscopic Imaging Of Human Breast Tissues

International Nuclear Information System (INIS)

Park, Sung H.; Kim, Hong T.; Kim, Jong K.; Jheon, Sang H.; Youn, Hwa S.

2007-01-01

X-ray microscopy with synchrotron radiation will be a useful tool for innovation of x-ray imaging in clinical and laboratory settings. It helps us observe detailed internal structure of material samples non-invasively in air. And, it also has the potential to solve some tough problems of conventional breast imaging if it could evaluate various conditions of breast tissue effectively. A new hard x-ray microscope with a spatial resolution better than 100 nm was installed at Pohang Light Source, a third generation synchrotron radiation facility in Pohang, Korea. The x-ray energy was set at 6.95 keV, and the x-ray beam was monochromatized by W/B4C monochromator. Condenser and objective zone plates were used as x-ray lenses. Zernike phase plate next to condenser zone plate was introduced for improved contrast imaging. The image of a sample was magnified 30 times by objective zone plate and 20 times by microscope objective, respectively. After additional 10 times digital magnification, the total magnifying power was up to 6000 times in the end. Phase contrast synchrotron images of 10-μm-thick female breast tissue of the normal, fibroadenoma, fibrocystic change and carcinoma cases were obtained. By phase contrast imaging, hard x-rays enable us to observe many structures of breast tissue without sample preparations such as staining or fixation

Improvements in low-cost label-free QPI microscope for live cell imaging

Science.gov (United States)

Seniya, C.; Towers, C. E.; Towers, D. P.

2017-07-01

This paper reports an improvement in the development of a low-cost QPI microscope offering new capabilities in term of phase measurement accuracy for label-free live samples in the longer term (i.e., hours to days). The spatially separated scattered and non-scattered image light fields are reshaped in the Fourier plane and modulated to form an interference image at a CCD camera. The apertures that enable these two beams to be generated have been optimised by means of laser-cut apertures placed on the mirrors of a Michelson interferometer and has improved the phase measuring and reconstruction capability of the QPI microscope. The microscope was tested with transparent onion cells as an object of interest.

Towards native-state imaging in biological context in the electron microscope

Science.gov (United States)

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

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

Science.gov (United States)

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

First images from the Stanford tabletop scanning soft x-ray microscope

International Nuclear Information System (INIS)

Trail, J.A.; Byer, R.L.

1988-01-01

The authors have constructed a scanning soft x-ray microscope which uses a laser-produced plasma as the soft x-ray source and normal incidence multilayer coated mirrors in a Schwarzschild configuration as the focusing optics. The microscope operates at a wavelength of 140 angstrom, has a spatial resolution of 0.5 μm, and has a soft x-ray photon flux through the focus of 10 4 s -1 when operated with only 170 mW of average laser power. The microscope is compact; the complete system, including the laser, fits on a single optical table. In this paper they describe the microscope and present images of metallic microstructures

A simple water-immersion condenser for imaging living brain slices on an inverted microscope.

Science.gov (United States)

Prusky, G T

1997-09-05

Due to some physical limitations of conventional condensers, inverted compound microscopes are not optimally suited for imaging living brain slices with transmitted light. Herein is described a simple device that converts an inverted microscope into an effective tool for this application by utilizing an objective as a condenser. The device is mounted on a microscope in place of the condenser, is threaded to accept a water immersion objective, and has a slot for a differential interference contrast (DIC) slider. When combined with infrared video techniques, this device allows an inverted microscope to effectively image living cells within thick brain slices in an open perfusion chamber.

Confocal multispot microscope for fast and deep imaging in semicleared tissues

Science.gov (United States)

Adam, Marie-Pierre; Müllenbroich, Marie Caroline; Di Giovanna, Antonino Paolo; Alfieri, Domenico; Silvestri, Ludovico; Sacconi, Leonardo; Pavone, Francesco Saverio

2018-02-01

Although perfectly transparent specimens are imaged faster with light-sheet microscopy, less transparent samples are often imaged with two-photon microscopy leveraging its robustness to scattering; however, at the price of increased acquisition times. Clearing methods that are capable of rendering strongly scattering samples such as brain tissue perfectly transparent specimens are often complex, costly, and time intensive, even though for many applications a slightly lower level of tissue transparency is sufficient and easily achieved with simpler and faster methods. Here, we present a microscope type that has been geared toward the imaging of semicleared tissue by combining multispot two-photon excitation with rolling shutter wide-field detection to image deep and fast inside semicleared mouse brain. We present a theoretical and experimental evaluation of the point spread function and contrast as a function of shutter size. Finally, we demonstrate microscope performance in fixed brain slices by imaging dendritic spines up to 400-μm deep.

Microdose fluorescence imaging of ABY-029 on an operating microscope adapted by custom illumination and imaging modules.

Science.gov (United States)

Elliott, Jonathan T; Dsouza, Alisha V; Marra, Kayla; Pogue, Brian W; Roberts, David W; Paulsen, Keith D

2016-09-01

Fluorescence guided surgery has the potential to positively impact surgical oncology; current operating microscopes and stand-alone imaging systems are too insensitive or too cumbersome to maximally take advantage of new tumor-specific agents developed through the microdose pathway. To this end, a custom-built illumination and imaging module enabling picomolar-sensitive near-infrared fluorescence imaging on a commercial operating microscope is described. The limits of detection and system specifications are characterized, and in vivo efficacy of the system in detecting ABY-029 is evaluated in a rat orthotopic glioma model following microdose injections, showing the suitability of the device for microdose phase 0 clinical trials.

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

Science.gov (United States)

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

Fast processing of microscopic images using object-based extended depth of field.

Science.gov (United States)

Intarapanich, Apichart; Kaewkamnerd, Saowaluck; Pannarut, Montri; Shaw, Philip J; Tongsima, Sissades

2016-12-22

Microscopic analysis requires that foreground objects of interest, e.g. cells, are in focus. In a typical microscopic specimen, the foreground objects may lie on different depths of field necessitating capture of multiple images taken at different focal planes. The extended depth of field (EDoF) technique is a computational method for merging images from different depths of field into a composite image with all foreground objects in focus. Composite images generated by EDoF can be applied in automated image processing and pattern recognition systems. However, current algorithms for EDoF are computationally intensive and impractical, especially for applications such as medical diagnosis where rapid sample turnaround is important. Since foreground objects typically constitute a minor part of an image, the EDoF technique could be made to work much faster if only foreground regions are processed to make the composite image. We propose a novel algorithm called object-based extended depths of field (OEDoF) to address this issue. The OEDoF algorithm consists of four major modules: 1) color conversion, 2) object region identification, 3) good contrast pixel identification and 4) detail merging. First, the algorithm employs color conversion to enhance contrast followed by identification of foreground pixels. A composite image is constructed using only these foreground pixels, which dramatically reduces the computational time. We used 250 images obtained from 45 specimens of confirmed malaria infections to test our proposed algorithm. The resulting composite images with all in-focus objects were produced using the proposed OEDoF algorithm. We measured the performance of OEDoF in terms of image clarity (quality) and processing time. The features of interest selected by the OEDoF algorithm are comparable in quality with equivalent regions in images processed by the state-of-the-art complex wavelet EDoF algorithm; however, OEDoF required four times less processing time. This

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

Science.gov (United States)

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.

Direct microscopic image and measurement of the atomization process of a port fuel injector

International Nuclear Information System (INIS)

Esmail, Mohamed; Kawahara, Nobuyuki; Tomita, Eiji; Sumida, Mamoru

2010-01-01

The main objective of this study is to observe and investigate the phenomena of atomization, i.e. the fuel break-up process very close to the nozzle exit of a practical port fuel injector (PFI). In order to achieve this objective, direct microscopic images of the atomization process were obtained using an ultra-high-speed video camera that could record 102 frames at rates of up to 1 Mfps, coupled with a long-distance microscope and Barlow lens. The experiments were carried out using a PFI in a closed chamber at atmospheric pressure. Time-series images of the spray behaviour were obtained with a high temporal resolution using backlighting. The direct microscopic images of a liquid column break-up were compared with experimental results from laser-induced exciplex fluorescence (LIEF), and the wavelength obtained from the experimental results compared with that predicated from the Kelvin–Helmholtz break-up model. The droplet size diameters from a ligament break-up were compared with results predicated from Weber's analysis. Furthermore, experimental results of the mean droplet diameter from a direct microscopic image were compared with the results obtained from phase Doppler anemometry (PDA) experimental results. Three conclusions were obtained from this study. The atomization processes and detailed characterizations of the break-up of a liquid column were identified; the direct microscopic image results were in good agreement with the results obtained from LIEF, experimental results of the wavelength were in good agreement with those from the Kelvin–Helmholtz break-up model. The break-up process of liquid ligaments into droplets was investigated, and Weber's analysis of the predicated droplet diameter from ligament break-up was found to be applicable only at larger wavelengths. Finally, the direct microscopic image method and PDA method give qualitatively similar trends for droplet size distribution and quantitatively similar values of Sauter mean diameter

Direct microscopic image and measurement of the atomization process of a port fuel injector

Science.gov (United States)

Esmail, Mohamed; Kawahara, Nobuyuki; Tomita, Eiji; Sumida, Mamoru

2010-07-01

The main objective of this study is to observe and investigate the phenomena of atomization, i.e. the fuel break-up process very close to the nozzle exit of a practical port fuel injector (PFI). In order to achieve this objective, direct microscopic images of the atomization process were obtained using an ultra-high-speed video camera that could record 102 frames at rates of up to 1 Mfps, coupled with a long-distance microscope and Barlow lens. The experiments were carried out using a PFI in a closed chamber at atmospheric pressure. Time-series images of the spray behaviour were obtained with a high temporal resolution using backlighting. The direct microscopic images of a liquid column break-up were compared with experimental results from laser-induced exciplex fluorescence (LIEF), and the wavelength obtained from the experimental results compared with that predicated from the Kelvin-Helmholtz break-up model. The droplet size diameters from a ligament break-up were compared with results predicated from Weber's analysis. Furthermore, experimental results of the mean droplet diameter from a direct microscopic image were compared with the results obtained from phase Doppler anemometry (PDA) experimental results. Three conclusions were obtained from this study. The atomization processes and detailed characterizations of the break-up of a liquid column were identified; the direct microscopic image results were in good agreement with the results obtained from LIEF, experimental results of the wavelength were in good agreement with those from the Kelvin-Helmholtz break-up model. The break-up process of liquid ligaments into droplets was investigated, and Weber's analysis of the predicated droplet diameter from ligament break-up was found to be applicable only at larger wavelengths. Finally, the direct microscopic image method and PDA method give qualitatively similar trends for droplet size distribution and quantitatively similar values of Sauter mean diameter.

Excitation-scanning hyperspectral imaging system for microscopic and endoscopic applications

Science.gov (United States)

Mayes, Sam A.; Leavesley, Silas J.; Rich, Thomas C.

2016-04-01

Current microscopic and endoscopic technologies for cancer screening utilize white-light illumination sources. Hyper-spectral imaging has been shown to improve sensitivity while retaining specificity when compared to white-light imaging in both microscopy and in vivo imaging. However, hyperspectral imaging methods have historically suffered from slow acquisition times due to the narrow bandwidth of spectral filters. Often minutes are required to gather a full image stack. We have developed a novel approach called excitation-scanning hyperspectral imaging that provides 2-3 orders of magnitude increased signal strength. This reduces acquisition times significantly, allowing for live video acquisition. Here, we describe a preliminary prototype excitation-scanning hyperspectral imaging system that can be coupled with endoscopes or microscopes for hyperspectral imaging of tissues and cells. Our system is comprised of three subsystems: illumination, transmission, and imaging. The illumination subsystem employs light-emitting diode arrays to illuminate at different wavelengths. The transmission subsystem utilizes a unique geometry of optics and a liquid light guide. Software controls allow us to interface with and control the subsystems and components. Digital and analog signals are used to coordinate wavelength intensity, cycling and camera triggering. Testing of the system shows it can cycle 16 wavelengths at as fast as 1 ms per cycle. Additionally, more than 18% of the light transmits through the system. Our setup should allow for hyperspectral imaging of tissue and cells in real time.

Microscopic oxygen imaging based on fluorescein bleaching efficiency measurements

DEFF Research Database (Denmark)

Beutler, Martin; Heisterkamp, Ines M.; Piltz, Bastian

2014-01-01

by a charge-coupled-device (ccd) camera mounted on a fluorescence microscope allowed a pixelwise estimation of the ratio function in a microscopic image. Use of a microsensor and oxygen-consuming bacteria in a sample chamber enabled the calibration of the system for quantification of absolute oxygen......Photobleaching of the fluorophore fluorescein in an aqueous solution is dependent on the oxygen concentration. Therefore, the time-dependent bleaching behavior can be used to measure of dissolved oxygen concentrations. The method can be combined with epi-fluorescence microscopy. The molecular...... states of the fluorophore can be expressed by a three-state energy model. This leads to a set of differential equations which describe the photobleaching behavior of fluorescein. The numerical solution of these equations shows that in a conventional wide-field fluorescence microscope, the fluorescence...

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

Science.gov (United States)

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

2010-12-10

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

Recent progress in medical imaging technology

International Nuclear Information System (INIS)

Endo, Masahiro

2004-01-01

Medical imaging is name of methods for diagnosis and therapy, which make visible with physical media such as X-ray, structures and functions of man's inside those are usually invisible. These methods are classified by the physical media into ultrasound imaging, magnetic resonance imaging, nuclear medicine imaging and X-ray imaging etc. Having characteristics different from one another, these are used complementarily in medical fields though in some case being competitive. Medical imaging is supported by highly progressed technology, which is called medical imaging technology. This paper describes a survey of recent progress of medical imaging technology in magnetic resonance imaging, nuclear medicine imaging and X-ray imaging. (author)

A super-oscillatory lens optical microscope for subwavelength imaging.

Science.gov (United States)

Rogers, Edward T F; Lindberg, Jari; Roy, Tapashree; Savo, Salvatore; Chad, John E; Dennis, Mark R; Zheludev, Nikolay I

2012-03-25

The past decade has seen an intensive effort to achieve optical imaging resolution beyond the diffraction limit. Apart from the Pendry-Veselago negative index superlens, implementation of which in optics faces challenges of losses and as yet unattainable fabrication finesse, other super-resolution approaches necessitate the lens either to be in the near proximity of the object or manufactured on it, or work only for a narrow class of samples, such as intensely luminescent or sparse objects. Here we report a new super-resolution microscope for optical imaging that beats the diffraction limit of conventional instruments and the recently demonstrated near-field optical superlens and hyperlens. This non-invasive subwavelength imaging paradigm uses a binary amplitude mask for direct focusing of laser light into a subwavelength spot in the post-evanescent field by precisely tailoring the interference of a large number of beams diffracted from a nanostructured mask. The new technology, which--in principle--has no physical limits on resolution, could be universally used for imaging at any wavelength and does not depend on the luminescence of the object, which can be tens of micrometres away from the mask. It has been implemented as a straightforward modification of a conventional microscope showing resolution better than λ/6.

Imaging properties of the mesooptical Fourier transform microscope for nuclear research emulsion

International Nuclear Information System (INIS)

Bencze, Gy.L.; Soroko, L.M.

1987-01-01

The optical signal transformation in the Mesooptical Fourier Transform Microscope (MFTM) for nuclear emulsion is treated in terms of Fourier Optics. A continuous conversion of the traditional optical microscope into the MFTM is followed. The images of dot-like and straight line objects given by the MFTM are discussed

Structure Identification in High-Resolution Transmission Electron Microscopic Images

DEFF Research Database (Denmark)

Vestergaard, Jacob Schack; Kling, Jens; Dahl, Anders Bjorholm

2014-01-01

A connection between microscopic structure and macroscopic properties is expected for almost all material systems. High-resolution transmission electron microscopy is a technique offering insight into the atomic structure, but the analysis of large image series can be time consuming. The present ...

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

Science.gov (United States)

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

2018-05-01

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

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

Science.gov (United States)

Marshall, F. J.; Bahr, R. E.; Goncharov, V. N.; Glebov, V. Yu.; Peng, B.; Regan, S. P.; Sangster, T. C.; Stoeckl, C.

2017-09-01

A 16-image Kirkpatrick-Baez (KB)-type x-ray microscope consisting of compact KB mirrors [F. J. Marshall, Rev. Sci. Instrum. 83, 10E518 (2012)] 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-μm is achieved with this combination in a 400-μm region of laser-plasma x-ray emission in the 2- 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 [T. R. Boehly et al., Opt. Commun. 133, 495 (1997)]. The unprecedented time and spatial resolutions 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. 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 [Regan et al., Phys. Rev. Lett. 117, 025001 (2016)].

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

Science.gov (United States)

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

2018-06-01

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

Multi-technology Integration Based on Low-contrast Microscopic Image Enhancement

Directory of Open Access Journals (Sweden)

Haoge Ma

2014-01-01

Full Text Available Microscopic image enhancement is an important issue of image processing technique, which is used to improve the visual quality of image. This paper describes a novel multi resolution image segmentation algorithm for low DOF images. The algorithm is designed to separate a sharply focused object of interest from other foreground or background objects. The algorithm is fully automatic in that all parameters are image in dependent. A multiscale-approach based on high frequency wavelet coefficients and their statistics is used to perform context dependent classification of individual blocks of the image. Compared with the state of the art algorithms, this new algorithm provides better accuracy at higher speed.

Progress in element analysis on a high-voltage electron microscope

International Nuclear Information System (INIS)

Tivol, W.F.; Barnard, D.; Guha, T.

1985-01-01

X-Ray microprobe (XMA) and electron energy-loss (EELS) spectrometers have been installed on the high-voltage electron microscope (HVEM). The probe size has been measured and background reduction is in progress for XMA and EELS as are improvements in electron optics for EELS and sensitivity measurements. XMA is currently useful for qualitative analysis and has been used by several investigators from our laboratory and outside laboratories. However, EELS background levels are still too high for meaningful results to be obtained. Standards suitable for biological specimens are being measured, and a library for quantitative analysis is being compiled

Quantitatively characterizing the microstructural features of breast ductal carcinoma tissues in different progression stages by Mueller matrix microscope.

Science.gov (United States)

Dong, Yang; Qi, Ji; He, Honghui; He, Chao; Liu, Shaoxiong; Wu, Jian; Elson, Daniel S; Ma, Hui

2017-08-01

Polarization imaging has been recognized as a potentially powerful technique for probing the microstructural information and optical properties of complex biological specimens. Recently, we have reported a Mueller matrix microscope by adding the polarization state generator and analyzer (PSG and PSA) to a commercial transmission-light microscope, and applied it to differentiate human liver and cervical cancerous tissues with fibrosis. In this paper, we apply the Mueller matrix microscope for quantitative detection of human breast ductal carcinoma samples at different stages. The Mueller matrix polar decomposition and transformation parameters of the breast ductal tissues in different regions and at different stages are calculated and analyzed. For more quantitative comparisons, several widely-used image texture feature parameters are also calculated to characterize the difference in the polarimetric images. The experimental results indicate that the Mueller matrix microscope and the polarization parameters can facilitate the quantitative detection of breast ductal carcinoma tissues at different stages.

Fine-grained leukocyte classification with deep residual learning for microscopic images.

Science.gov (United States)

Qin, Feiwei; Gao, Nannan; Peng, Yong; Wu, Zizhao; Shen, Shuying; Grudtsin, Artur

2018-08-01

Leukocyte classification and cytometry have wide applications in medical domain, previous researches usually exploit machine learning techniques to classify leukocytes automatically. However, constrained by the past development of machine learning techniques, for example, extracting distinctive features from raw microscopic images are difficult, the widely used SVM classifier only has relative few parameters to tune, these methods cannot efficiently handle fine-grained classification cases when the white blood cells have up to 40 categories. Based on deep learning theory, a systematic study is conducted on finer leukocyte classification in this paper. A deep residual neural network based leukocyte classifier is constructed at first, which can imitate the domain expert's cell recognition process, and extract salient features robustly and automatically. Then the deep neural network classifier's topology is adjusted according to the prior knowledge of white blood cell test. After that the microscopic image dataset with almost one hundred thousand labeled leukocytes belonging to 40 categories is built, and combined training strategies are adopted to make the designed classifier has good generalization ability. The proposed deep residual neural network based classifier was tested on microscopic image dataset with 40 leukocyte categories. It achieves top-1 accuracy of 77.80%, top-5 accuracy of 98.75% during the training procedure. The average accuracy on the test set is nearly 76.84%. This paper presents a fine-grained leukocyte classification method for microscopic images, based on deep residual learning theory and medical domain knowledge. Experimental results validate the feasibility and effectiveness of our approach. Extended experiments support that the fine-grained leukocyte classifier could be used in real medical applications, assist doctors in diagnosing diseases, reduce human power significantly. Copyright © 2018 Elsevier B.V. All rights reserved.

Active Mask Segmentation of Fluorescence Microscope Images

OpenAIRE

Srinivasa, Gowri; Fickus, Matthew C.; Guo, Yusong; Linstedt, Adam D.; Kovačević, Jelena

2009-01-01

We propose a new active mask algorithm for the segmentation of fluorescence microscope images of punctate patterns. It combines the (a) flexibility offered by active-contour methods, (b) speed offered by multiresolution methods, (c) smoothing offered by multiscale methods, and (d) statistical modeling offered by region-growing methods into a fast and accurate segmentation tool. The framework moves from the idea of the “contour” to that of “inside and outside”, or, masks, allowing for easy mul...

Atomic Force Microscope for Imaging and Spectroscopy

Science.gov (United States)

Pike, W. T.; Hecht, M. H.; Anderson, M. S.; Akiyama, T.; Gautsch, S.; deRooij, N. F.; Staufer, U.; Niedermann, Ph.; Howald, L.; Mueller, D.

2000-01-01

We have developed, built, and tested an atomic force microscope (AFM) for extraterrestrial applications incorporating a micromachined tip array to allow for probe replacement. It is part of a microscopy station originally intended for NASA's 2001 Mars lander to identify the size, distribution, and shape of Martian dust and soil particles. As well as imaging topographically down to nanometer resolution, this instrument can be used to reveal chemical information and perform infrared and Raman spectroscopy at unprecedented resolution.

Foucault imaging by using non-dedicated transmission electron microscope

International Nuclear Information System (INIS)

Taniguchi, Yoshifumi; Matsumoto, Hiroaki; Harada, Ken

2012-01-01

An electron optical system for observing Foucault images was constructed using a conventional transmission electron microscope without any special equipment for Lorentz microscopy. The objective lens was switched off and an electron beam was converged by a condenser optical system to the crossover on the selected area aperture plane. The selected area aperture was used as an objective aperture to select the deflected beam for Foucault mode, and the successive image-forming lenses were controlled for observation of the specimen images. The irradiation area on the specimen was controlled by selecting the appropriate diameter of the condenser aperture.

Foucault imaging by using non-dedicated transmission electron microscope

Science.gov (United States)

Taniguchi, Yoshifumi; Matsumoto, Hiroaki; Harada, Ken

2012-08-01

An electron optical system for observing Foucault images was constructed using a conventional transmission electron microscope without any special equipment for Lorentz microscopy. The objective lens was switched off and an electron beam was converged by a condenser optical system to the crossover on the selected area aperture plane. The selected area aperture was used as an objective aperture to select the deflected beam for Foucault mode, and the successive image-forming lenses were controlled for observation of the specimen images. The irradiation area on the specimen was controlled by selecting the appropriate diameter of the condenser aperture.

Foucault imaging by using non-dedicated transmission electron microscope

Energy Technology Data Exchange (ETDEWEB)

Taniguchi, Yoshifumi [Science and Medical Systems Business Group, Hitachi High-Technologies Corp., Ichige, Hitachinaka, Ibaraki 312-8504 (Japan); Matsumoto, Hiroaki [Corporate Manufacturing Strategy Group, Hitachi High-Technologies Corp., Ishikawa-cho, Hitachinaka, Ibaraki 312-1991 (Japan); Harada, Ken [Central Research Laboratory, Hitachi Ltd., Hatoyama, Saitama 350-0395 (Japan)

2012-08-27

An electron optical system for observing Foucault images was constructed using a conventional transmission electron microscope without any special equipment for Lorentz microscopy. The objective lens was switched off and an electron beam was converged by a condenser optical system to the crossover on the selected area aperture plane. The selected area aperture was used as an objective aperture to select the deflected beam for Foucault mode, and the successive image-forming lenses were controlled for observation of the specimen images. The irradiation area on the specimen was controlled by selecting the appropriate diameter of the condenser aperture.

Performance limitations of imaging microscopes for soft x-ray applications

International Nuclear Information System (INIS)

Lewotsky, K.L.; Kotha, A.; Harvey, J.E.

1993-01-01

Recent advances in the fabrication of nanometer-scale multilayer structures have yielded high-reflectance mirrors operating at near-normal incidence for soft X-ray wavelengths. These developments have stimulated renewed interest in high-resolution soft X-ray microscopy. The design of a Schwarzschild imaging microscope for soft X-ray applications has been reported by Hoover and Shealy. Based upon a geometrical ray-trace analysis of the residual design errors, diffraction-limited performance at a wavelength of 100 angstrom was predicted over an object size (diameter) of 0.4 mm. In this paper the authors expand upon the previous analysis of the Schwarzschild X-ray microscope design by determining the total image degradation due to diffraction, geometrical aberrations, alignment errors, and realistic assumptions concerning optical fabrication errors. NASA's Optical Surface Analysis Code (OSAC) is used to model the image degradation effects of residual surface irregularities over the entire range of relevant spatial frequencies. This includes small angle scattering effects due to mid spatial frequency surface errors falling between the traditional figure and finish specifications. Performance predictions are presented parametrically to provide some insight into the optical fabrication and alignment tolerances necessary to meet a particular image quality requirement

Ghost microscope imaging system from the perspective of coherent-mode representation

Science.gov (United States)

Shen, Qian; Bai, Yanfeng; Shi, Xiaohui; Nan, Suqin; Qu, Lijie; Li, Hengxing; Fu, Xiquan

2018-03-01

The coherent-mode representation theory of partially coherent fields is firstly used to analyze a two-arm ghost microscope imaging system. It is shown that imaging quality of the generated images depend crucially on the distribution of the decomposition coefficients of the object imaged when the light source is fixed. This theory is also suitable for demonstrating the effects from the distance the object is moved away from the original plane on imaging quality. Our results are verified theoretically and experimentally.

Content Progressive Coding of Limited Bits/pixel Images

DEFF Research Database (Denmark)

Jensen, Ole Riis; Forchhammer, Søren

1999-01-01

A new lossless context based method for content progressive coding of limited bits/pixel images is proposed. Progressive coding is achieved by separating the image into contelnt layers. Digital maps are compressed up to 3 times better than GIF.......A new lossless context based method for content progressive coding of limited bits/pixel images is proposed. Progressive coding is achieved by separating the image into contelnt layers. Digital maps are compressed up to 3 times better than GIF....

Nanoscale X-Ray Microscopic Imaging of Mammalian Mineralized Tissue

OpenAIRE

Andrews, Joy C.; Almeida, Eduardo; van der Meulen, Marjolein C.H.; Alwood, Joshua S.; Lee, Chialing; Liu, Yijin; Chen, Jie; Meirer, Florian; Feser, Michael; Gelb, Jeff; Rudati, Juana; Tkachuk, Andrei; Yun, Wenbing; Pianetta, Piero

2010-01-01

A novel hard transmission X-ray microscope (TXM) at the Stanford Synchrotron Radiation Light-source operating from 5 to 15 keV X-ray energy with 14 to 30 µm2 field of view has been used for high-resolution (30–40 nm) imaging and density quantification of mineralized tissue. TXM is uniquely suited for imaging of internal cellular structures and networks in mammalian mineralized tissues using relatively thick (50 µm), untreated samples that preserve tissue micro- and nanostructure. To test this...

Use of an axisymmetric microscope with electronic readout for collecting soft X-ray images

International Nuclear Information System (INIS)

Cavailler, C.; Henry, P.; Launspach, J.; De Mascureau, J.; Millerioux, M.; Rostaing, M.; Sauneuf, R.

1984-08-01

The axisymmetric microscope, first discussed by Wolter, provides high resolution and sensitivity for investigating the soft X-ray emission of laser-driven plasmas. Such a device having a 10 X magnification has been constructed. We present a comparison between the images of laser-driven plasmas given by this microscope and by a 10 X pinhole camera. Until now these images were recorded on X-ray film. We have shown that film could be replaced by C.C.D. in a pinhole camera when the photon energy lies within the 1-10 keV range. Below 1 keV the quantum yield is too low so we have used an image converter tube made by RTC. It is a diode-inverter tube with a soft X-ray photocathode and a P20 phosphor deposited on an optic fiber plate. The electronic image appearing on the screen is read by a C.C.D. working in the visible spectral fields. An electronic image readout chain, which is identical to those associated with streak cameras, then processes automatically and immediately the images given by the microscope [fr

Atomic force microscopic imaging of Acanthamoeba castellanii and Balamuthia mandrillaris trophozoites and cysts.

Science.gov (United States)

Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Ateeq, Muhammad; Raza Shah, Muhammad; Kulsoom, Huma; Khan, Naveed Ahmed

2015-01-01

Light microscopy and electron microscopy have been successfully used in the study of microbes, as well as free-living protists. Unlike light microscopy, which enables us to observe living organisms or the electron microscope which provides a two-dimensional image, atomic force microscopy provides a three-dimensional surface profile. Here, we observed two free-living amoebae, Acanthamoeba castellanii and Balamuthia mandrillaris under the phase contrast inverted microscope, transmission electron microscope and atomic force microscope. Although light microscopy was of lower magnification, it revealed functional biology of live amoebae such as motility and osmoregulation using contractile vacuoles of the trophozoite stage, but it is of limited value in defining the cyst stage. In contrast, transmission electron microscopy showed significantly greater magnification and resolution to reveal the ultra-structural features of trophozoites and cysts including intracellular organelles and cyst wall characteristics but it only produced a snapshot in time of a dead amoeba cell. Atomic force microscopy produced three-dimensional images providing detailed topographic description of shape and surface, phase imaging measuring boundary stiffness, and amplitude measurements including width, height and length of A. castellanii and B. mandrillaris trophozoites and cysts. These results demonstrate the importance of the application of various microscopic methods in the biological and structural characterization of the whole cell, ultra-structural features, as well as surface components and cytoskeleton of protist pathogens. © 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.

The Reaction Microscope: Imaging and Pulse Shaping Control in Photodynamics

NARCIS (Netherlands)

Vredenborg, A.; Lehmann, C.S.; Irimia, D.; Roeterdink, W.; Janssen, M.H.M.

2011-01-01

Herein, we review the current capabilities and potential of advanced single-particle imaging techniques to study photodynamics in isolated molecules. These reaction microscopes are able to measure the full three-dimensional energy and angular distribution of (correlated) particles such as electrons

A multiphoton laser scanning microscope setup for transcranial in vivo brain imaging on mice

Science.gov (United States)

Nase, Gabriele; Helm, P. Johannes; Reppen, Trond; Ottersen, Ole Petter

2005-12-01

We describe a multiphoton laser scanning microscope setup for transcranial in vivo brain imaging in mice. The modular system is based on a modified industrial standard Confocal Scanning Laser Microscope (CSLM) and is assembled mainly from commercially available components. A special multifunctional stage, which is optimized for both laser scanning microscopic observation and preparative animal surgery, has been developed and built. The detection unit includes a highly efficient photomultiplier tube installed in a Peltier-cooled thermal box shielding the detector from changes in room temperature and from distortions caused by external electromagnetic fields. The images are recorded using a 12-bit analog-to-digital converter. Depending on the characteristics of the staining, individual nerve cells can be imaged down to at least 100μm below the intact cranium and down to at least 200μm below the opened cranium.

Evaluating EUV mask pattern imaging with two EUV microscopes

International Nuclear Information System (INIS)

Goldberg, Kenneth A.; Takase, Kei; Naulleau, Patrick P.; Han, Hakseung; Barty, Anton; Kinoshita, Hiroo; Hamamoto, Kazuhiro

2008-01-01

Aerial image measurement plays a key role in the development of patterned reticles for each generation of lithography. Studying the field transmitted (reflected) from EUV masks provides detailed information about potential disruptions caused by mask defects, and the performance of defect repair strategies, without the complications of photoresist imaging. Furthermore, by measuring the continuously varying intensity distribution instead of a thresholded, binary resist image, aerial image measurement can be used as feedback to improve mask and lithography system modeling methods. Interest in EUV, at-wavelength, aerial image measurement lead to the creation of several research tools worldwide. These tools are used in advanced mask development work, and in the evaluation of the need for commercial at-wavelength inspection tools. They describe performance measurements of two such tools, inspecting the same EUV mask in a series of benchmarking tests that includes brightfield and darkfield patterns. One tool is the SEMATECH Berkeley Actinic Inspection Tool (AIT) operating on a bending magnet beamline at Lawrence Berkeley National Laboratory's Advanced Light Source. The AIT features an EUV Fresnel zoneplate microscope that emulates the numerical aperture of a 0.25-NA stepper, and projects the aerial image directly onto a CCD camera, with 700x magnification. The second tool is an EUV microscope (EUVM) operating at the NewSUBARU synchrotron in Hyogo, Japan. The NewSUBARU tool projects the aerial image using a reflective, 30x Schwarzschild objective lens, followed by a 10-200x x-ray zooming tube. The illumination conditions and the imaging etendue are different for the two tools. The benchmarking measurements were used to determine many imaging and performance properties of the tools, including resolution, modulation transfer function (MTF), aberration magnitude, aberration field-dependence (including focal-plane tilt), illumination uniformity, line-edge roughness, and flare

Spectro-refractometry of individual microscopic objects using swept-source quantitative phase imaging.

Science.gov (United States)

Jung, Jae-Hwang; Jang, Jaeduck; Park, Yongkeun

2013-11-05

We present a novel spectroscopic quantitative phase imaging technique with a wavelength swept-source, referred to as swept-source diffraction phase microscopy (ssDPM), for quantifying the optical dispersion of microscopic individual samples. Employing the swept-source and the principle of common-path interferometry, ssDPM measures the multispectral full-field quantitative phase imaging and spectroscopic microrefractometry of transparent microscopic samples in the visible spectrum with a wavelength range of 450-750 nm and a spectral resolution of less than 8 nm. With unprecedented precision and sensitivity, we demonstrate the quantitative spectroscopic microrefractometry of individual polystyrene beads, 30% bovine serum albumin solution, and healthy human red blood cells.

Computer processing of microscopic images of bacteria : morphometry and fluorimetry

NARCIS (Netherlands)

Wilkinson, Michael H.F.; Jansen, Gijsbert J.; Waaij, Dirk van der

1994-01-01

Several techniques that use computer analysis of microscopic images have been developed to study the complicated microbial flora in the human intestine, including measuring the shape and fluorescence intensity of bacteria. These techniques allow rapid assessment of changes in the intestinal flora

Image alignment for tomography reconstruction from synchrotron X-ray microscopic images.

Directory of Open Access Journals (Sweden)

Chang-Chieh Cheng

Full Text Available A synchrotron X-ray microscope is a powerful imaging apparatus for taking high-resolution and high-contrast X-ray images of nanoscale objects. A sufficient number of X-ray projection images from different angles is required for constructing 3D volume images of an object. Because a synchrotron light source is immobile, a rotational object holder is required for tomography. At a resolution of 10 nm per pixel, the vibration of the holder caused by rotating the object cannot be disregarded if tomographic images are to be reconstructed accurately. This paper presents a computer method to compensate for the vibration of the rotational holder by aligning neighboring X-ray images. This alignment process involves two steps. The first step is to match the "projected feature points" in the sequence of images. The matched projected feature points in the x-θ plane should form a set of sine-shaped loci. The second step is to fit the loci to a set of sine waves to compute the parameters required for alignment. The experimental results show that the proposed method outperforms two previously proposed methods, Xradia and SPIDER. The developed software system can be downloaded from the URL, http://www.cs.nctu.edu.tw/~chengchc/SCTA or http://goo.gl/s4AMx.

Image alignment for tomography reconstruction from synchrotron X-ray microscopic images.

Science.gov (United States)

Cheng, Chang-Chieh; Chien, Chia-Chi; Chen, Hsiang-Hsin; Hwu, Yeukuang; Ching, Yu-Tai

2014-01-01

A synchrotron X-ray microscope is a powerful imaging apparatus for taking high-resolution and high-contrast X-ray images of nanoscale objects. A sufficient number of X-ray projection images from different angles is required for constructing 3D volume images of an object. Because a synchrotron light source is immobile, a rotational object holder is required for tomography. At a resolution of 10 nm per pixel, the vibration of the holder caused by rotating the object cannot be disregarded if tomographic images are to be reconstructed accurately. This paper presents a computer method to compensate for the vibration of the rotational holder by aligning neighboring X-ray images. This alignment process involves two steps. The first step is to match the "projected feature points" in the sequence of images. The matched projected feature points in the x-θ plane should form a set of sine-shaped loci. The second step is to fit the loci to a set of sine waves to compute the parameters required for alignment. The experimental results show that the proposed method outperforms two previously proposed methods, Xradia and SPIDER. The developed software system can be downloaded from the URL, http://www.cs.nctu.edu.tw/~chengchc/SCTA or http://goo.gl/s4AMx.

Inverted light-sheet microscope for imaging mouse pre-implantation development.

Science.gov (United States)

Strnad, Petr; Gunther, Stefan; Reichmann, Judith; Krzic, Uros; Balazs, Balint; de Medeiros, Gustavo; Norlin, Nils; Hiiragi, Takashi; Hufnagel, Lars; Ellenberg, Jan

2016-02-01

Despite its importance for understanding human infertility and congenital diseases, early mammalian development has remained inaccessible to in toto imaging. We developed an inverted light-sheet microscope that enabled us to image mouse embryos from zygote to blastocyst, computationally track all cells and reconstruct a complete lineage tree of mouse pre-implantation development. We used this unique data set to show that the first cell fate specification occurs at the 16-cell stage.

Microscopic validation of whole mouse micro-metastatic tumor imaging agents using cryo-imaging and sliding organ image registration

Science.gov (United States)

Liu, Yiqiao; Zhou, Bo; Qutaish, Mohammed; Wilson, David L.

2016-03-01

We created a metastasis imaging, analysis platform consisting of software and multi-spectral cryo-imaging system suitable for evaluating emerging imaging agents targeting micro-metastatic tumor. We analyzed CREKA-Gd in MRI, followed by cryo-imaging which repeatedly sectioned and tiled microscope images of the tissue block face, providing anatomical bright field and molecular fluorescence, enabling 3D microscopic imaging of the entire mouse with single metastatic cell sensitivity. To register MRI volumes to the cryo bright field reference, we used our standard mutual information, non-rigid registration which proceeded: preprocess --> affine --> B-spline non-rigid 3D registration. In this report, we created two modified approaches: mask where we registered locally over a smaller rectangular solid, and sliding organ. Briefly, in sliding organ, we segmented the organ, registered the organ and body volumes separately and combined results. Though sliding organ required manual annotation, it provided the best result as a standard to measure other registration methods. Regularization parameters for standard and mask methods were optimized in a grid search. Evaluations consisted of DICE, and visual scoring of a checkerboard display. Standard had accuracy of 2 voxels in all regions except near the kidney, where there were 5 voxels sliding. After mask and sliding organ correction, kidneys sliding were within 2 voxels, and Dice overlap increased 4%-10% in mask compared to standard. Mask generated comparable results with sliding organ and allowed a semi-automatic process.

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

Science.gov (United States)

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

2016-01-01

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

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

Directory of Open Access Journals (Sweden)

Md Mehedi Hasan

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

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

Science.gov (United States)

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

2016-01-01

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

Super-resolution for everybody: An image processing workflow to obtain high-resolution images with a standard confocal microscope.

Science.gov (United States)

Lam, France; Cladière, Damien; Guillaume, Cyndélia; Wassmann, Katja; Bolte, Susanne

2017-02-15

In the presented work we aimed at improving confocal imaging to obtain highest possible resolution in thick biological samples, such as the mouse oocyte. We therefore developed an image processing workflow that allows improving the lateral and axial resolution of a standard confocal microscope. Our workflow comprises refractive index matching, the optimization of microscope hardware parameters and image restoration by deconvolution. We compare two different deconvolution algorithms, evaluate the necessity of denoising and establish the optimal image restoration procedure. We validate our workflow by imaging sub resolution fluorescent beads and measuring the maximum lateral and axial resolution of the confocal system. Subsequently, we apply the parameters to the imaging and data restoration of fluorescently labelled meiotic spindles of mouse oocytes. We measure a resolution increase of approximately 2-fold in the lateral and 3-fold in the axial direction throughout a depth of 60μm. This demonstrates that with our optimized workflow we reach a resolution that is comparable to 3D-SIM-imaging, but with better depth penetration for confocal images of beads and the biological sample. Copyright © 2016 Elsevier Inc. All rights reserved.

Open-dish incubator for live cell imaging with an inverted microscope.

Science.gov (United States)

Heidemann, Steven R; Lamoureux, Phillip; Ngo, Kha; Reynolds, Matthew; Buxbaum, Robert E

2003-10-01

Here we describe the design and fabrication of an inexpensive cell culture incubator for the stage of an inverted light microscope for use in live cell imaging. This device maintains the temperature of the cell culture at 37 degrees C with great stability and, after reaching equilibrium, provides focal stability of an image for 20-25 min with oil-immersion lenses. We describe two versions of the incubator: one for use with standard 60-mm plastic culture dishes, and the other version for imaging of cells on glass coverslips. Either can be made for less than $400. Most components are widely available commercially, and it requires only simple wiring and 3 h to assemble. Although the device is generally useful for live cell imaging on an inverted microscope, it is particularly suitable for work in which instruments are introduced into the culture, such as electrophysiology or micromanipulation. The design is based on the principle that control performance is limited by the lag time between detection and response. The key element of the design is a heated, temperature-controlled aluminum ring serving as a mini-incubator surrounding the culture vessel. For this reason, we call our design a "ringcubator."

Martian Microscope

Science.gov (United States)

2004-01-01

The microscopic imager (circular device in center) is in clear view above the surface at Meridiani Planum, Mars, in this approximate true-color image taken by the panoramic camera on the Mars Exploration Rover Opportunity. The image was taken on the 9th sol of the rover's journey. The microscopic imager is located on the rover's instrument deployment device, or arm. The arrow is pointing to the lens of the instrument. Note the dust cover, which flips out to the left of the lens, is open. This approximated color image was created using the camera's violet and infrared filters as blue and red.

Active mask segmentation of fluorescence microscope images.

Science.gov (United States)

Srinivasa, Gowri; Fickus, Matthew C; Guo, Yusong; Linstedt, Adam D; Kovacević, Jelena

2009-08-01

We propose a new active mask algorithm for the segmentation of fluorescence microscope images of punctate patterns. It combines the (a) flexibility offered by active-contour methods, (b) speed offered by multiresolution methods, (c) smoothing offered by multiscale methods, and (d) statistical modeling offered by region-growing methods into a fast and accurate segmentation tool. The framework moves from the idea of the "contour" to that of "inside and outside," or masks, allowing for easy multidimensional segmentation. It adapts to the topology of the image through the use of multiple masks. The algorithm is almost invariant under initialization, allowing for random initialization, and uses a few easily tunable parameters. Experiments show that the active mask algorithm matches the ground truth well and outperforms the algorithm widely used in fluorescence microscopy, seeded watershed, both qualitatively, as well as quantitatively.

An image processing approach to analyze morphological features of microscopic images of muscle fibers.

Science.gov (United States)

Comin, Cesar Henrique; Xu, Xiaoyin; Wang, Yaming; Costa, Luciano da Fontoura; Yang, Zhong

2014-12-01

We present an image processing approach to automatically analyze duo-channel microscopic images of muscular fiber nuclei and cytoplasm. Nuclei and cytoplasm play a critical role in determining the health and functioning of muscular fibers as changes of nuclei and cytoplasm manifest in many diseases such as muscular dystrophy and hypertrophy. Quantitative evaluation of muscle fiber nuclei and cytoplasm thus is of great importance to researchers in musculoskeletal studies. The proposed computational approach consists of steps of image processing to segment and delineate cytoplasm and identify nuclei in two-channel images. Morphological operations like skeletonization is applied to extract the length of cytoplasm for quantification. We tested the approach on real images and found that it can achieve high accuracy, objectivity, and robustness. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

Science.gov (United States)

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

2018-02-28

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

New method for thickness determination and microscopic imaging of graphene-like two-dimensional materials

International Nuclear Information System (INIS)

Qin Xudong; Chen Yonghai; Liu Yu; Zhu Laipan; Li Yuan; Wu Qing; Huang Wei

2016-01-01

We employed the microscopic reflectance difference spectroscopy (micro-RDS) to determine the layer-number and microscopically image the surface topography of graphene and MoS 2 samples. The contrast image shows the efficiency and reliability of this new clipping technique. As a low-cost, quantifiable, no-contact and non-destructive method, it is not concerned with the characteristic signal of certain materials and can be applied to arbitrary substrates. Therefore it is a perfect candidate for characterizing the thickness of graphene-like two-dimensional materials. (paper)

Modulus design multiwavelength polarization microscope for transmission Mueller matrix imaging.

Science.gov (United States)

Zhou, Jialing; He, Honghui; Chen, Zhenhua; Wang, Ye; Ma, Hui

2018-01-01

We have developed a polarization microscope based on a commercial transmission microscope. We replace the halogen light source by a collimated LED light source module of six different colors. We use achromatic polarized optical elements that can cover the six different wavelength ranges in the polarization state generator (PSG) and polarization state analyzer (PSA) modules. The dual-rotating wave plate method is used to measure the Mueller matrix of samples, which requires the simultaneous rotation of the two quarter-wave plates in both PSG and PSA at certain angular steps. A scientific CCD detector is used as the image receiving module. A LabView-based software is developed to control the rotation angels of the wave plates and the exposure time of the detector to allow the system to run fully automatically in preprogrammed schedules. Standard samples, such as air, polarizers, and quarter-wave plates, are used to calibrate the intrinsic Mueller matrix of optical components, such as the objectives, using the eigenvalue calibration method. Errors due to the images walk-off in the PSA are studied. Errors in the Mueller matrices are below 0.01 using air and polarizer as standard samples. Data analysis based on Mueller matrix transformation and Mueller matrix polarization decomposition is used to demonstrate the potential application of this microscope in pathological diagnosis. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Microscope self-calibration based on micro laser line imaging and soft computing algorithms

Science.gov (United States)

Apolinar Muñoz Rodríguez, J.

2018-06-01

A technique to perform microscope self-calibration via micro laser line and soft computing algorithms is presented. In this technique, the microscope vision parameters are computed by means of soft computing algorithms based on laser line projection. To implement the self-calibration, a microscope vision system is constructed by means of a CCD camera and a 38 μm laser line. From this arrangement, the microscope vision parameters are represented via Bezier approximation networks, which are accomplished through the laser line position. In this procedure, a genetic algorithm determines the microscope vision parameters by means of laser line imaging. Also, the approximation networks compute the three-dimensional vision by means of the laser line position. Additionally, the soft computing algorithms re-calibrate the vision parameters when the microscope vision system is modified during the vision task. The proposed self-calibration improves accuracy of the traditional microscope calibration, which is accomplished via external references to the microscope system. The capability of the self-calibration based on soft computing algorithms is determined by means of the calibration accuracy and the micro-scale measurement error. This contribution is corroborated by an evaluation based on the accuracy of the traditional microscope calibration.

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

Science.gov (United States)

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

2013-01-01

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

Microscopic study of rock for estimating long-term behavior

International Nuclear Information System (INIS)

Ichikawa, Yasuaki

2002-03-01

Micro-structure of rock plays a essential role for their long-term behavior. For elucidating long-term characteristics of granite we here present the followings: 1) Conforcal Laser Scanning Microscope (LSM) observation of joint surfaces of granite and Fourier analysis, 2) characterization of the mechanism of microcrack initiation and propagation observed by stereoscopic microscope under uniaxial/triaxial compression and relaxation tests, 3) observation of microcrack initiation and propagation by LSM under uniaxial compression, and 4) a viscoelastic homogenization theory to predict the long-term behavior of micro/macro-level stress for granite. Rock image processing and analysis become a fundamental procedure to determine rock surface discontinuities. But the complexity of rock surface discontinuities seems beyond the manual image processing method. In Chapter 2 a Conforcal Laser Scanning Microscope that can acquire three-dimensional images is introduced to observe the rock roughness of a discontinuity. Then, scanning three-dimensional images are changed its data form in order to adapt various image analysis programs, and granitic rock roughness of discontinuities are displayed by graphic images. For example, these datas are analyzed by Discrete Fourier Transformation (DFT) program and Inverse Discrete Fourier Transformation (IDFT) program. Microcrack generation and propagation play an essential role to predict the long-term behavior of rock. In Chapter 3 a progressive development of cracking in granite is revealed by using stereoscopic microscope under triaxial compression condition and by using LSM under uniaxial compression condition. With a viscoelastic theory applied in homogenization method, we can calculate macro behavior of medium influenced by its micro structure by analyse the long-term time-dependent behavior of granite under the same condition to the relaxation experiment. (author)

On the detection of early osteoarthritis by quantitative microscopic imaging

Science.gov (United States)

Mittelstaedt, Daniel John

measurements. These studies demonstrate the ability to use two quantitative microscopic imaging techniques, microCT and microMRI, to detect microscopic changes in collagen and GAG from healthy, biochemically degraded, and early OA cartilage. The capability for microscopic imaging to detect alterations at the earliest stages of OA will ultimately improve the understanding of degradation and may help aid in the detection for the prevention of disease and repair of damaged cartilage.

Microscope image based fully automated stomata detection and pore measurement method for grapevines

Directory of Open Access Journals (Sweden)

Hiranya Jayakody

2017-11-01

Full Text Available Abstract Background Stomatal behavior in grapevines has been identified as a good indicator of the water stress level and overall health of the plant. Microscope images are often used to analyze stomatal behavior in plants. However, most of the current approaches involve manual measurement of stomatal features. The main aim of this research is to develop a fully automated stomata detection and pore measurement method for grapevines, taking microscope images as the input. The proposed approach, which employs machine learning and image processing techniques, can outperform available manual and semi-automatic methods used to identify and estimate stomatal morphological features. Results First, a cascade object detection learning algorithm is developed to correctly identify multiple stomata in a large microscopic image. Once the regions of interest which contain stomata are identified and extracted, a combination of image processing techniques are applied to estimate the pore dimensions of the stomata. The stomata detection approach was compared with an existing fully automated template matching technique and a semi-automatic maximum stable extremal regions approach, with the proposed method clearly surpassing the performance of the existing techniques with a precision of 91.68% and an F1-score of 0.85. Next, the morphological features of the detected stomata were measured. Contrary to existing approaches, the proposed image segmentation and skeletonization method allows us to estimate the pore dimensions even in cases where the stomatal pore boundary is only partially visible in the microscope image. A test conducted using 1267 images of stomata showed that the segmentation and skeletonization approach was able to correctly identify the stoma opening 86.27% of the time. Further comparisons made with manually traced stoma openings indicated that the proposed method is able to estimate stomata morphological features with accuracies of 89.03% for area

Early skin tumor detection from microscopic images through image processing

International Nuclear Information System (INIS)

Siddiqi, A.A.; Narejo, G.B.; Khan, A.M.

2017-01-01

The research is done to provide appropriate detection technique for skin tumor detection. The work is done by using the image processing toolbox of MATLAB. Skin tumors are unwanted skin growth with different causes and varying extent of malignant cells. It is a syndrome in which skin cells mislay the ability to divide and grow normally. Early detection of tumor is the most important factor affecting the endurance of a patient. Studying the pattern of the skin cells is the fundamental problem in medical image analysis. The study of skin tumor has been of great interest to the researchers. DIP (Digital Image Processing) allows the use of much more complex algorithms for image processing, and hence, can offer both more sophisticated performance at simple task, and the implementation of methods which would be impossibly by analog means. It allows much wider range of algorithms to be applied to the input data and can avoid problems such as build up of noise and signal distortion during processing. The study shows that few works has been done on cellular scale for the images of skin. This research allows few checks for the early detection of skin tumor using microscopic images after testing and observing various algorithms. After analytical evaluation the result has been observed that the proposed checks are time efficient techniques and appropriate for the tumor detection. The algorithm applied provides promising results in lesser time with accuracy. The GUI (Graphical User Interface) that is generated for the algorithm makes the system user friendly. (author)

Cryogenic immersion microscope

Science.gov (United States)

Le Gros, Mark; Larabell, Carolyn A.

2010-12-14

A cryogenic immersion microscope whose objective lens is at least partially in contact with a liquid reservoir of a cryogenic liquid, in which reservoir a sample of interest is immersed is disclosed. When the cryogenic liquid has an index of refraction that reduces refraction at interfaces between the lens and the sample, overall resolution and image quality are improved. A combination of an immersion microscope and x-ray microscope, suitable for imaging at cryogenic temperatures is also disclosed.

Tissue imaging with a stigmatic mass microscope using laser desorption/ionization

Science.gov (United States)

Awazu, Kunio; Hazama, Hisanao; Hamanaka, Tomonori; Aoki, Jun; Toyoda, Michisato; Naito, Yasuhide

2012-03-01

A novel stigmatic mass microscope using laser desorption/ionization and a multi-turn time-of-flight mass spectrometer, MULTUM-IMG, has been developed. Stigmatic ion images of crystal violet masked by a fine square mesh grid with a 12.7 μm pitch were clearly observed, and the estimated spatial resolution was about 3 μm in the linear mode with a 20-fold ion optical magnification. Tissue sections of a brain and eyes of a mouse stained with crystal violet and methylene blue were observed in the linear mode, and the stigmatic total ion images of crystal violet and methylene blue agreed well with the optical photomicrograph of the same sections. Especially, the fine structure in the cornea tissue was clearly observed with a spatial resolution in the range of micrometers. Although the total measurement time of the stigmatic ion image for the whole-eye section was about 59 minutes using a laser with a 10 Hz repetition rate, the measurement time could be reduced to about 35 s using a laser with a 1 kHz repetition rate and automation of measurements. The stigmatic mass microscope developed in this research should be suitable for high-spatial resolution and high-throughput imaging mass spectrometry for pathology, pharmacokinetics, and so on.

A universal fluid cell for the imaging of biological specimens in the atomic force microscope.

Science.gov (United States)

Kasas, Sandor; Radotic, Ksenja; Longo, Giovanni; Saha, Bashkar; Alonso-Sarduy, Livan; Dietler, Giovanni; Roduit, Charles

2013-04-01

Recently, atomic force microscope (AFM) manufacturers have begun producing instruments specifically designed to image biological specimens. In most instances, they are integrated with an inverted optical microscope, which permits concurrent optical and AFM imaging. An important component of the set-up is the imaging chamber, whose design determines the nature of the experiments that can be conducted. Many different imaging chamber designs are available, usually designed to optimize a single parameter, such as the dimensions of the substrate or the volume of fluid that can be used throughout the experiment. In this report, we present a universal fluid cell, which simultaneously optimizes all of the parameters that are important for the imaging of biological specimens in the AFM. This novel imaging chamber has been successfully tested using mammalian, plant, and microbial cells. Copyright © 2013 Wiley Periodicals, Inc.

Annular dark field electron microscope images with better than 2 A resolution at 100 kV

International Nuclear Information System (INIS)

Shin, D.H.; Kirkland, E.J.; Silcox, J.

1989-01-01

High-resolution scanning transmission electron microscope (STEM) images in the annular dark field (ADF) imaging mode approaching the theoretical point-to-point resolution limit are presented. The ADF images were obtained from a high T c superconducting YBa 2 Cu 3 O 7-x thin-film specimen at 100 kV. The 1.9 A resolution lattice image, which is the smallest lattice spacing in the specimen, corresponds to the minimum resolvable spatial frequency with 5% contrast in the contrast transfer function for annular dark field, and is smaller than the resolution limit given by the Rayleigh criterion. This demonstrates that STEM ADF imaging can have a resolution approximately 40% better than that of the bright field conventional transmission electron microscope (CTEM) imaging at Scherzer condition

Quantification of incisal tooth wear in upper anterior teeth: conventional vs new method using toolmakers microscope and a three-dimensional measuring technique.

Science.gov (United States)

Al-Omiri, Mahmoud K; Sghaireen, Mohd G; Alzarea, Bader K; Lynch, Edward

2013-12-01

This study aimed to quantify tooth wear in upper anterior teeth using a new CAD-CAM Laser scanning machine, tool maker microscope and conventional tooth wear index. Fifty participants (25 males and 25 females, mean age = 25 ± 4 years) were assessed for incisal tooth wear of upper anterior teeth using Smith and Knight clinical tooth wear index (TWI) on two occasions, the study baseline and 1 year later. Stone dies for each tooth were prepared and scanned using the CAD-CAM Laser Cercon System. Scanned images were printed and examined under a toolmaker microscope to quantify tooth wear and then the dies were directly assessed under the microscope to measure tooth wear. The Wilcoxon Signed Ranks Test was used to analyze the data. TWI scores for incisal edges were 0-3 and were similar at both occasions. Score 4 was not detected. Wear values measured by directly assessing the dies under the toolmaker microscope (range = 113 - 150 μm, mean = 130 ± 20 μm) were significantly more than those measured from Cercon Digital Machine images (range=52-80 μm, mean = 68 ± 23 μm) and both showed significant differences between the two occasions. Wear progression in upper anterior teeth was effectively detected by directly measuring the dies or the images of dies under toolmaker microscope. Measuring the dies of worn dentition directly under tool maker microscope enabled detection of wear progression more accurately than measuring die images obtained with Cercon Digital Machine. Conventional method was the least sensitive for tooth wear quantification and was unable to identify wear progression in most cases. Copyright © 2013 Elsevier Ltd. All rights reserved.

X-ray generation by femtosecond laser pulses and its application to soft X-ray imaging microscope

International Nuclear Information System (INIS)

Ikeda, Kenichi; Kotaki, Hideyuki; Nakajima, Kazuhisa

2002-01-01

We have developed laser-produced plasma X-ray sources using femtosecond laser pulses at 10Hz repetition rate in a table-top size in order to investigate basic mechanism of X-ray emission from laser-matter interactions and its application to a X-ray microscope. In a soft X-ray region over 5 nm wavelength, laser-plasma X-ray emission from a solid target achieved an intense flux of photons of the order of 1011 photons/rad per pulse with duration of a few 100 ps, which is intense enough to make a clear imaging in a short time exposure. As an application of laser-produced plasma X-ray source, we have developed a soft X-ray imaging microscope operating in the wavelength range around 14 nm. The microscope consists of a cylindrically ellipsoidal condenser mirror and a Schwarzshird objective mirror with highly-reflective multilayers. We report preliminary results of performance tests of the soft X-ray imaging microscope with a compact laser-produced plasma X-ray source

Extended morphological processing: a practical method for automatic spot detection of biological markers from microscopic images.

Science.gov (United States)

Kimori, Yoshitaka; Baba, Norio; Morone, Nobuhiro

2010-07-08

A reliable extraction technique for resolving multiple spots in light or electron microscopic images is essential in investigations of the spatial distribution and dynamics of specific proteins inside cells and tissues. Currently, automatic spot extraction and characterization in complex microscopic images poses many challenges to conventional image processing methods. A new method to extract closely located, small target spots from biological images is proposed. This method starts with a simple but practical operation based on the extended morphological top-hat transformation to subtract an uneven background. The core of our novel approach is the following: first, the original image is rotated in an arbitrary direction and each rotated image is opened with a single straight line-segment structuring element. Second, the opened images are unified and then subtracted from the original image. To evaluate these procedures, model images of simulated spots with closely located targets were created and the efficacy of our method was compared to that of conventional morphological filtering methods. The results showed the better performance of our method. The spots of real microscope images can be quantified to confirm that the method is applicable in a given practice. Our method achieved effective spot extraction under various image conditions, including aggregated target spots, poor signal-to-noise ratio, and large variations in the background intensity. Furthermore, it has no restrictions with respect to the shape of the extracted spots. The features of our method allow its broad application in biological and biomedical image information analysis.

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

Science.gov (United States)

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.

Analytical electron microscope based on scanning transmission electron microscope with wavelength dispersive x-ray spectroscopy to realize highly sensitive elemental imaging especially for light elements

International Nuclear Information System (INIS)

Koguchi, Masanari; Tsuneta, Ruriko; Anan, Yoshihiro; Nakamae, Koji

2017-01-01

An analytical electron microscope based on the scanning transmission electron microscope with wavelength dispersive x-ray spectroscopy (STEM-WDX) to realize highly sensitive elemental imaging especially for light elements has been developed. In this study, a large-solid-angle multi-capillary x-rays lens with a focal length of 5 mm, long-time data acquisition (e.g. longer than 26 h), and a drift-free system made it possible to visualize boron-dopant images in a Si substrate at a detection limit of 0.2 atomic percent. (paper)

Atomic force microscope with integrated optical microscope for biological applications

OpenAIRE

Putman, Constant A.J.; Putman, C.A.J.; van der Werf, Kees; de Grooth, B.G.; van Hulst, N.F.; Segerink, Franciscus B.; Greve, Jan

1992-01-01

Since atomic force microscopy (AFM) is capable of imaging nonconducting surfaces, the technique holds great promises for high‐resolution imaging of biological specimens. A disadvantage of most AFMs is the fact that the relatively large sample surface has to be scanned multiple times to pinpoint a specific biological object of interest. Here an AFM is presented which has an incorporated inverted optical microscope. The optical image from the optical microscope is not obscured by the cantilever...

STM-SQUID probe microscope

International Nuclear Information System (INIS)

Hayashi, Tadayuki; Tachiki, Minoru; Itozaki, Hideo

2007-01-01

We have developed a STM-SQUID probe microscope. A high T C SQUID probe microscope was combined with a scanning tunneling microscope for investigation of samples at room temperature in air. A high permeability probe needle was used as a magnetic flux guide to improve the spatial resolution. The probe with tip radius of less than 100 nm was prepared by microelectropolishing. The probe was also used as a scanning tunneling microscope tip. Topography of the sample surface could be measured by the scanning tunneling microscope with high spatial resolution prior to observation by SQUID microscopy. The SQUID probe microscope image could be observed while keeping the distance from the sample surface to the probe tip constant. We observed a topographic image and a magnetic image of Ni fine pattern and also a magnetically recorded hard disk. Furthermore we have investigated a sample vibration method of the static magnetic field emanating from a sample with the aim of achieving a higher signal-to-noise (S/N) ratio

Adaptive and automatic red blood cell counting method based on microscopic hyperspectral imaging technology

Science.gov (United States)

Liu, Xi; Zhou, Mei; Qiu, Song; Sun, Li; Liu, Hongying; Li, Qingli; Wang, Yiting

2017-12-01

Red blood cell counting, as a routine examination, plays an important role in medical diagnoses. Although automated hematology analyzers are widely used, manual microscopic examination by a hematologist or pathologist is still unavoidable, which is time-consuming and error-prone. This paper proposes a full-automatic red blood cell counting method which is based on microscopic hyperspectral imaging of blood smears and combines spatial and spectral information to achieve high precision. The acquired hyperspectral image data of the blood smear in the visible and near-infrared spectral range are firstly preprocessed, and then a quadratic blind linear unmixing algorithm is used to get endmember abundance images. Based on mathematical morphological operation and an adaptive Otsu’s method, a binaryzation process is performed on the abundance images. Finally, the connected component labeling algorithm with magnification-based parameter setting is applied to automatically select the binary images of red blood cell cytoplasm. Experimental results show that the proposed method can perform well and has potential for clinical applications.

Microscopic image processing system for measuring nonuniform film thickness profiles: Image scanning ellipsometry

International Nuclear Information System (INIS)

Liu, A.H.; Plawsky, J.L.; Wayner, P.C. Jr.

1993-01-01

The long-term objective of this research program is to determine the stability and heat transfer characteristics of evaporating thin films. The current objective is to develop and use a microscopic image-processing system (IPS) which has two parts: an image analyzing interferometer (IAI) and an image scanning ellipsometer (ISE). The primary purpose of this paper is to present the basic concept of ISE, which is a novel technique to measure the two dimensional thickness profile of a non-uniform, thin film, from several nm up to several μm, in a steady state as well as in a transient state. It is a full-field imaging technique which can study every point on the surface simultaneously with high spatial resolution and thickness sensitivity, i.e., it can measure and map the 2-D film thickness profile. The ISE was tested by measuring the thickness profile and the refractive index of a nonuniform solid film

A transmission positron microscope and a scanning positron microscope being built at KEK, Japan

International Nuclear Information System (INIS)

Doyama, M.; Inoue, M.; Kogure, Y.; Kurihara, T.; Yagishita, A.; Shidara, T.; Nakahara, K.; Hayashi, Y.; Yoshiie, T.

2001-01-01

This paper reports the plans of positron microscopes being built at KEK (High Energy Accelerator Research Organization), Tsukuba, Japan improving used electron microscopes. The kinetic energies of positron produced by accelerators or by nuclear decays have not a unique value but show a spread over in a wide range. Positron beam will be guided near electron microscopes, a transmission electron microscope (JEM100S) and a scanning electron microscope (JSM25S). Positrons are slowed down by a tungsten foil, accelerated and focused on a nickel sheet. The monochromatic focused beam will be injected into an electron microscope. The focusing of positrons and electrons is achieved by magnetic system of the electron microscopes. Imaging plates are used to record positron images for the transmission electron microscope. (orig.)

Proper alignment of the microscope.

Science.gov (United States)

Rottenfusser, Rudi

2013-01-01

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

Transmission environmental scanning electron microscope with scintillation gaseous detection device

International Nuclear Information System (INIS)

Danilatos, Gerasimos; Kollia, Mary; Dracopoulos, Vassileios

2015-01-01

A transmission environmental scanning electron microscope with use of a scintillation gaseous detection device has been implemented. This corresponds to a transmission scanning electron microscope but with addition of a gaseous environment acting both as environmental and detection medium. A commercial type of low vacuum machine has been employed together with appropriate modifications to the detection configuration. This involves controlled screening of various emitted signals in conjunction with a scintillation gaseous detection device already provided with the machine for regular surface imaging. Dark field and bright field imaging has been obtained along with other detection conditions. With a progressive series of modifications and tests, the theory and practice of a novel type of microscopy is briefly shown now ushering further significant improvements and developments in electron microscopy as a whole. - Highlights: • Novel scanning transmission electron microscopy (STEM) with an environmental scanning electron microscope (ESEM) called TESEM. • Use of the gaseous detection device (GDD) in scintillation mode that allows high resolution bright and dark field imaging in the TESEM. • Novel approach towards a unification of both vacuum and environmental conditions in both bulk/surface and transmission mode of electron microscopy

Atomic imaging of an InSe single-crystal surface with atomic force microscope

OpenAIRE

Uosaki, Kohei; Koinuma, Michio

1993-01-01

The atomic force microscope was employed to observed in air the surface atomic structure of InSe, one of III-VI compound semiconductors with layered structures. Atomic arrangements were observed in both n-type and p-type materials. The observed structures are in good agreement with those expected from bulk crystal structures. The atomic images became less clear by repeating the imaging process. Wide area imaging after the imaging of small area clearly showed that a mound was created at the sp...

Spatio-temporal imaging of voltage pulses with an ultrafast scanning tunneling microscope

DEFF Research Database (Denmark)

Jensen, Jacob Riis; Keil, Ulrich Dieter Felix; Hvam, Jørn Märcher

1997-01-01

Measurements on an ultrafast scanning tunneling microscope with simultaneous spatial and temporal resolution are presented. We show images of picosecond pulses propagating on a coplanar waveguide and resolve their mode structures. The influence of transmission line discontinuities on the mode...

Detection of nuclei in 4D Nomarski DIC microscope images of early Caenorhabditis elegans embryos using local image entropy and object tracking

Directory of Open Access Journals (Sweden)

Hamahashi Shugo

2005-05-01

Full Text Available Abstract Background The ability to detect nuclei in embryos is essential for studying the development of multicellular organisms. A system of automated nuclear detection has already been tested on a set of four-dimensional (4D Nomarski differential interference contrast (DIC microscope images of Caenorhabditis elegans embryos. However, the system needed laborious hand-tuning of its parameters every time a new image set was used. It could not detect nuclei in the process of cell division, and could detect nuclei only from the two- to eight-cell stages. Results We developed a system that automates the detection of nuclei in a set of 4D DIC microscope images of C. elegans embryos. Local image entropy is used to produce regions of the images that have the image texture of the nucleus. From these regions, those that actually detect nuclei are manually selected at the first and last time points of the image set, and an object-tracking algorithm then selects regions that detect nuclei in between the first and last time points. The use of local image entropy makes the system applicable to multiple image sets without the need to change its parameter values. The use of an object-tracking algorithm enables the system to detect nuclei in the process of cell division. The system detected nuclei with high sensitivity and specificity from the one- to 24-cell stages. Conclusion A combination of local image entropy and an object-tracking algorithm enabled highly objective and productive detection of nuclei in a set of 4D DIC microscope images of C. elegans embryos. The system will facilitate genomic and computational analyses of C. elegans embryos.

Imaging manifestations of progressive multifocal leukoencephalopathy

International Nuclear Information System (INIS)

Shah, R.; Bag, A.K.; Chapman, P.R.; Cure, J.K.

2010-01-01

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease caused by reactivation of JC virus in immunosuppressed patients. The diagnosis is usually suggested on imaging and confirmed by cerebrospinal fluid polymerase chain reaction (PCR) for JC virus DNA. In this article, we review the imaging manifestations of PML on computed tomography (CT), magnetic resonance imaging (MRI), diffusion-weighted imaging (DWI), diffusion tensor imaging (DTI), MR spectroscopy, single photon-emission computed tomography (SPECT) and positron-emission tomography (PET), and outline the role of imaging in follow-up and prognostication.

Imaging of surface plasmon polariton interference using phase-sensitive scanning tunneling microscope

NARCIS (Netherlands)

Jose, J.; Segerink, Franciscus B.; Korterik, Jeroen P.; Herek, Jennifer Lynn; Offerhaus, Herman L.

2011-01-01

We report the surface plasmon polariton interference, generated via a ‘buried’ gold grating, and imaged using a phase-sensitive Photon Scanning Tunneling Microscope (PSTM). The phase-resolved PSTM measurement unravels the complex surface plasmon polariton interference fields at the gold-air

Characterization of platelet adhesion under flow using microscopic image sequence analysis.

Science.gov (United States)

Machin, M; Santomaso, A; Cozzi, M R; Battiston, M; Mazzuccato, M; De Marco, L; Canu, P

2005-07-01

A method for quantitative analysis of platelet deposition under flow is discussed here. The model system is based upon perfusion of blood platelets over an adhesive substrate immobilized on a glass coverslip acting as the lower surface of a rectangular flow chamber. The perfusion apparatus is mounted onto an inverted microscope equipped with epifluorescent illumination and intensified CCD video camera. Characterization is based on information obtained from a specific image analysis method applied to continuous sequences of microscopical images. Platelet recognition across the sequence of images is based on a time-dependent, bidimensional, gaussian-like pdf. Once a platelet is located,the variation of its position and shape as a function of time (i.e., the platelet history) can be determined. Analyzing the history we can establish if the platelet is moving on the surface, the frequency of this movement and the distance traveled before its resumes the velocity of a non-interacting cell. Therefore, we can determine how long the adhesion would last which is correlated to the resistance of the platelet-substrate bond. This algorithm enables the dynamic quantification of trajectories, as well as residence times, arrest and release frequencies for a high numbers of platelets at the same time. Statistically significant conclusions on platelet-surface interactions can then be obtained. An image analysis tool of this kind can dramatically help the investigation and characterization of the thrombogenic properties of artificial surfaces such as those used in artificial organs and biomedical devices.

The optics of microscope image formation.

Science.gov (United States)

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.

Transmission environmental scanning electron microscope with scintillation gaseous detection device.

Science.gov (United States)

Danilatos, Gerasimos; Kollia, Mary; Dracopoulos, Vassileios

2015-03-01

A transmission environmental scanning electron microscope with use of a scintillation gaseous detection device has been implemented. This corresponds to a transmission scanning electron microscope but with addition of a gaseous environment acting both as environmental and detection medium. A commercial type of low vacuum machine has been employed together with appropriate modifications to the detection configuration. This involves controlled screening of various emitted signals in conjunction with a scintillation gaseous detection device already provided with the machine for regular surface imaging. Dark field and bright field imaging has been obtained along with other detection conditions. With a progressive series of modifications and tests, the theory and practice of a novel type of microscopy is briefly shown now ushering further significant improvements and developments in electron microscopy as a whole. Copyright © 2014 Elsevier B.V. All rights reserved.

Rapid identification of salmonella serotypes with stereo and hyperspectral microscope imaging Methods

Science.gov (United States)

The hyperspectral microscope imaging (HMI) method can reduce detection time within 8 hours including incubation process. The early and rapid detection with this method in conjunction with the high throughput capabilities makes HMI method a prime candidate for implementation for the food industry. Th...

High resolution imaging of dielectric surfaces with an evanescent field optical microscope

NARCIS (Netherlands)

van Hulst, N.F.; Segerink, Franciscus B.; Bölger, B.

1992-01-01

An evanescent field optical microscope (EFOM) is presented which employs frustrated total internal reflection o­n a localized scale by scanning a dielectric tip in close proximity to a sample surface. High resolution images of dielectric gratings and spheres containing both topographic and

Combined macroscopic and microscopic approach to the fracture of metals. Technical progress report

International Nuclear Information System (INIS)

Asaro, R.J.; Gurland, J.; Needleman, A.; Rice, R.J.

1979-06-01

Progress is reported on microscopic fracture mechanisms, including studies of void and crack initiation in steels in the absence and presence of hydrogen, the effects of hydrogen on ductile fracture in medium and high carbon steels; elastic--plastic crack growth including the quasi-stable growth of cracks in ductile solids under increasing load and conditions of instability; and elevated temperature rupture including analysis of the stress field near a crack tip in an elastic-nonlinear viscous material under tensile load as well as the processes of diffusion, and cavitation of grain boundaries in plastically creeping materials

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

Science.gov (United States)

2017-09-14

with a Light Field Microscope Gordon E. Lott Follow this and additional works at: https://scholar.afit.edu/etd Part of the Atomic, Molecular and......https://scholar.afit.edu/etd/774 THREE-DIMENSIONAL IMAGING OF COLD ATOMS IN A MAGNETO-OPTICAL TRAP WITH A LIGHT FIELD MICROSCOPE DISSERTATION Gordon E

Advances in imaging and electron physics the scanning transmission electron microscope

CERN Document Server

Hawkes, Peter W

2009-01-01

Advances in Imaging and Electron Physics merges two long-running serials--Advances in Electronics and Electron Physics and Advances in Optical and Electron Microscopy. This series features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science and digital image processing, electromagnetic wave propagation, electron microscopy, and the computing methods used in all these domains.  This particular volume presents several timely articles on the scanning transmission electron microscope. Updated with contributions from leading international scholars and industry experts Discusses hot topic areas and presents current and future research trends Provides an invaluable reference and guide for physicists, engineers and mathematicians.

Scanning Microscopes Using X Rays and Microchannels

Science.gov (United States)

Wang, Yu

2003-01-01

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

Design of an in-vivo microscope to characterize cataracts in human eyes

Science.gov (United States)

Feng, Chen; Ahmad, Anees

1996-11-01

The design of a compact contact microscope, which can be used in-vivo to study the cataracts in human eyes is presented. This microscope has the capability to evaluate the changes in the optical density within the eye lens itself, and thus enabling an examiner to ascertain the progression of a cataractous change or at least the optical changes associated with cataractous development. The microscope has a variable focal length so it can be focused at any depth through the entire thickness of the eye lens. A separate small objective lens is spring loaded against the cornea (like a tonometer tip) so that the natural eye movements can occur safely during the examination. The distance between the objective lens and rest of the optics is variable to accommodate the movements of the eye due to pulse or breathing without affecting the image quality of the instrument. The layer by layer images can be captured on a CCD camera and stored in the computer. The reconstruction software can quantitatively display the characteristics of the cataracts, such as the location, size, and density. The optical design and performance results for the microscope are presented. The optomechanical design features of the microscope are also discussed.

Garnet film rotator applied in polarizing microscope for domain image modulation (abstract)

Science.gov (United States)

Wakabayashi, K.; Numata, T.; Inokuchi, S.

1991-04-01

A garnet film polarization rotator placed before the analyzer in a polarizing microscope was investigated to obtain the difference image of a positive and a negative one of magnetic domain in real time along with an image processor. In the difference image, a nonmagnetic image can be reduced and hence the weak magnetic contrast enhanced. Theoretical calculation of S/N and contrast C of the domain image as a function of the rotation shows they take maxima at the rotation angle of 2.6° and 0.1°, respectively, with the extinction ratio of e=4×10-6 of a polarizing microscope. Thus, since the thickness of the garnet film required is 1 μm or so, the absorption by the garnet rotator does not bring a serious problem even in a visible region for the domain observation. The optimum rotation of the rotator for a high quality observation was obtained by a quantitative study of images obtained experimentally as well as by a visual evaluation. A magnetically unsaturated garnet film with perpendicular magnetization (i.e., multidomain) was employed as a rotator, in which the polarization rotation angle θm of the undeflected beam with respect to the light diffraction could be continuously varied by an applied magnetic field. The dependences of S/N and C on θm were measured, resulting in a well agreement between the measured and the calculated. The visually best image was obtained at θm=0.5° which made the product of S/N and C maximum. The domain image of the Kerr rotation angle of θk=0.22° was observed in S/N=47 dB and C=0.4 when Ar+ laser (λ=515 nm) of tenths of a watt was employed as a light source. Since the domain image with 47 dB S/N does not need an image summation for a noise reduction, a garnet film rotator makes it possible to invert the contrast of a domain image in a real time for an improved domain observation.

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

Science.gov (United States)

Demos,; Stavros, G [Livermore, CA

2011-05-17

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

An image processing pipeline to detect and segment nuclei in muscle fiber microscopic images.

Science.gov (United States)

Guo, Yanen; Xu, Xiaoyin; Wang, Yuanyuan; Wang, Yaming; Xia, Shunren; Yang, Zhong

2014-08-01

Muscle fiber images play an important role in the medical diagnosis and treatment of many muscular diseases. The number of nuclei in skeletal muscle fiber images is a key bio-marker of the diagnosis of muscular dystrophy. In nuclei segmentation one primary challenge is to correctly separate the clustered nuclei. In this article, we developed an image processing pipeline to automatically detect, segment, and analyze nuclei in microscopic image of muscle fibers. The pipeline consists of image pre-processing, identification of isolated nuclei, identification and segmentation of clustered nuclei, and quantitative analysis. Nuclei are initially extracted from background by using local Otsu's threshold. Based on analysis of morphological features of the isolated nuclei, including their areas, compactness, and major axis lengths, a Bayesian network is trained and applied to identify isolated nuclei from clustered nuclei and artifacts in all the images. Then a two-step refined watershed algorithm is applied to segment clustered nuclei. After segmentation, the nuclei can be quantified for statistical analysis. Comparing the segmented results with those of manual analysis and an existing technique, we find that our proposed image processing pipeline achieves good performance with high accuracy and precision. The presented image processing pipeline can therefore help biologists increase their throughput and objectivity in analyzing large numbers of nuclei in muscle fiber images. © 2014 Wiley Periodicals, Inc.

Switched capacitor charge pump used for low-distortion imaging in atomic force microscope.

Science.gov (United States)

Zhang, Jie; Zhang, Lian Sheng; Feng, Zhi Hua

2015-01-01

The switched capacitor charge pump (SCCP) is an effective method of linearizing charges on piezoelectric actuators and therefore constitute a significant approach to nano-positioning. In this work, it was for the first time implemented in an atomic force microscope for low-distortion imaging. Experimental results showed that the image quality was improved evidently under the SCCP drive compared with that under traditional linear voltage drive. © Wiley Periodicals, Inc.

Assessing various Infrared (IR) microscopic imaging techniques for post-mortem interval evaluation of human skeletal remains

Science.gov (United States)

Roider, Clemens; Ritsch-Marte, Monika; Pemberger, Nadin; Cemper-Kiesslich, Jan; Hatzer-Grubwieser, Petra; Parson, Walther; Pallua, Johannes Dominikus

2017-01-01

Due to the influence of many environmental processes, a precise determination of the post-mortem interval (PMI) of skeletal remains is known to be very complicated. Although methods for the investigation of the PMI exist, there still remains much room for improvement. In this study the applicability of infrared (IR) microscopic imaging techniques such as reflection-, ATR- and Raman- microscopic imaging for the estimation of the PMI of human skeletal remains was tested. PMI specific features were identified and visualized by overlaying IR imaging data with morphological tissue structures obtained using light microscopy to differentiate between forensic and archaeological bone samples. ATR and reflection spectra revealed that a more prominent peak at 1042 cm-1 (an indicator for bone mineralization) was observable in archeological bone material when compared with forensic samples. Moreover, in the case of the archaeological bone material, a reduction in the levels of phospholipids, proteins, nucleic acid sugars, complex carbohydrates as well as amorphous or fully hydrated sugars was detectable at (reciprocal wavelengths/energies) between 3000 cm-1 to 2800 cm-1. Raman spectra illustrated a similar picture with less ν2PO43−at 450 cm-1 and ν4PO43− from 590 cm-1 to 584 cm-1, amide III at 1272 cm-1 and protein CH2 deformation at 1446 cm-1 in archeological bone material/samples/sources. A semi-quantitative determination of various distributions of biomolecules by chemi-maps of reflection- and ATR- methods revealed that there were less carbohydrates and complex carbohydrates as well as amorphous or fully hydrated sugars in archaeological samples compared with forensic bone samples. Raman- microscopic imaging data showed a reduction in B-type carbonate and protein α-helices after a PMI of 3 years. The calculated mineral content ratio and the organic to mineral ratio displayed that the mineral content ratio increases, while the organic to mineral ratio decreases with

Assessing various Infrared (IR microscopic imaging techniques for post-mortem interval evaluation of human skeletal remains.

Directory of Open Access Journals (Sweden)

Claudia Woess

Full Text Available Due to the influence of many environmental processes, a precise determination of the post-mortem interval (PMI of skeletal remains is known to be very complicated. Although methods for the investigation of the PMI exist, there still remains much room for improvement. In this study the applicability of infrared (IR microscopic imaging techniques such as reflection-, ATR- and Raman- microscopic imaging for the estimation of the PMI of human skeletal remains was tested. PMI specific features were identified and visualized by overlaying IR imaging data with morphological tissue structures obtained using light microscopy to differentiate between forensic and archaeological bone samples. ATR and reflection spectra revealed that a more prominent peak at 1042 cm-1 (an indicator for bone mineralization was observable in archeological bone material when compared with forensic samples. Moreover, in the case of the archaeological bone material, a reduction in the levels of phospholipids, proteins, nucleic acid sugars, complex carbohydrates as well as amorphous or fully hydrated sugars was detectable at (reciprocal wavelengths/energies between 3000 cm-1 to 2800 cm-1. Raman spectra illustrated a similar picture with less ν2PO43-at 450 cm-1 and ν4PO43- from 590 cm-1 to 584 cm-1, amide III at 1272 cm-1 and protein CH2 deformation at 1446 cm-1 in archeological bone material/samples/sources. A semi-quantitative determination of various distributions of biomolecules by chemi-maps of reflection- and ATR- methods revealed that there were less carbohydrates and complex carbohydrates as well as amorphous or fully hydrated sugars in archaeological samples compared with forensic bone samples. Raman- microscopic imaging data showed a reduction in B-type carbonate and protein α-helices after a PMI of 3 years. The calculated mineral content ratio and the organic to mineral ratio displayed that the mineral content ratio increases, while the organic to mineral ratio

Assessing various Infrared (IR) microscopic imaging techniques for post-mortem interval evaluation of human skeletal remains.

Science.gov (United States)

Woess, Claudia; Unterberger, Seraphin Hubert; Roider, Clemens; Ritsch-Marte, Monika; Pemberger, Nadin; Cemper-Kiesslich, Jan; Hatzer-Grubwieser, Petra; Parson, Walther; Pallua, Johannes Dominikus

2017-01-01

Due to the influence of many environmental processes, a precise determination of the post-mortem interval (PMI) of skeletal remains is known to be very complicated. Although methods for the investigation of the PMI exist, there still remains much room for improvement. In this study the applicability of infrared (IR) microscopic imaging techniques such as reflection-, ATR- and Raman- microscopic imaging for the estimation of the PMI of human skeletal remains was tested. PMI specific features were identified and visualized by overlaying IR imaging data with morphological tissue structures obtained using light microscopy to differentiate between forensic and archaeological bone samples. ATR and reflection spectra revealed that a more prominent peak at 1042 cm-1 (an indicator for bone mineralization) was observable in archeological bone material when compared with forensic samples. Moreover, in the case of the archaeological bone material, a reduction in the levels of phospholipids, proteins, nucleic acid sugars, complex carbohydrates as well as amorphous or fully hydrated sugars was detectable at (reciprocal wavelengths/energies) between 3000 cm-1 to 2800 cm-1. Raman spectra illustrated a similar picture with less ν2PO43-at 450 cm-1 and ν4PO43- from 590 cm-1 to 584 cm-1, amide III at 1272 cm-1 and protein CH2 deformation at 1446 cm-1 in archeological bone material/samples/sources. A semi-quantitative determination of various distributions of biomolecules by chemi-maps of reflection- and ATR- methods revealed that there were less carbohydrates and complex carbohydrates as well as amorphous or fully hydrated sugars in archaeological samples compared with forensic bone samples. Raman- microscopic imaging data showed a reduction in B-type carbonate and protein α-helices after a PMI of 3 years. The calculated mineral content ratio and the organic to mineral ratio displayed that the mineral content ratio increases, while the organic to mineral ratio decreases with time

The system of digital-image optical microscope in semiconductor particle detector development

International Nuclear Information System (INIS)

Han Lixiang; Li Zhankui; Jin Genming; Wang Zhusheng; Xiao Guoqing

2009-01-01

Optical microscopic detection is very important in the process of semiconductor particle detector development. A system of digital-image optical microscope has been constructed with rather low price, which performance is comparable with the moderate-level imports. The system mounts powerful dry objective, and a 2μm resolution could be achieved. Observations with bright and dark field, polarized light,and interference light can be carried out on it. The system have large area on-line monitor,and the photographic device can be controlled by PC. It can be used in the control of defects and contaminations, pattern test, identification of crystal backing, inspection of the smoothness and the flatness of the crystal surface. It can also be used in some precise procedures, such as test, assembly, packaging and repairing. The quality of the bond could be examined by observing the appearance of the bond point and the microscopic structure of the solder. The surface fluctuation can be precisely measured under the microscope with the technology of multi-beam interference. In the article, the application of this system for semiconductor particle detector development has been illustrated, and the construction information has been described in detail. (authors)

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

Science.gov (United States)

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.

Computer Aided Solution for Automatic Segmenting and Measurements of Blood Leucocytes Using Static Microscope Images.

Science.gov (United States)

Abdulhay, Enas; Mohammed, Mazin Abed; Ibrahim, Dheyaa Ahmed; Arunkumar, N; Venkatraman, V

2018-02-17

Blood leucocytes segmentation in medical images is viewed as difficult process due to the variability of blood cells concerning their shape and size and the difficulty towards determining location of Blood Leucocytes. Physical analysis of blood tests to recognize leukocytes is tedious, time-consuming and liable to error because of the various morphological components of the cells. Segmentation of medical imagery has been considered as a difficult task because of complexity of images, and also due to the non-availability of leucocytes models which entirely captures the probable shapes in each structures and also incorporate cell overlapping, the expansive variety of the blood cells concerning their shape and size, various elements influencing the outer appearance of the blood leucocytes, and low Static Microscope Image disparity from extra issues outcoming about because of noise. We suggest a strategy towards segmentation of blood leucocytes using static microscope images which is a resultant of three prevailing systems of computer vision fiction: enhancing the image, Support vector machine for segmenting the image, and filtering out non ROI (region of interest) on the basis of Local binary patterns and texture features. Every one of these strategies are modified for blood leucocytes division issue, in this manner the subsequent techniques are very vigorous when compared with its individual segments. Eventually, we assess framework based by compare the outcome and manual division. The findings outcome from this study have shown a new approach that automatically segments the blood leucocytes and identify it from a static microscope images. Initially, the method uses a trainable segmentation procedure and trained support vector machine classifier to accurately identify the position of the ROI. After that, filtering out non ROI have proposed based on histogram analysis to avoid the non ROI and chose the right object. Finally, identify the blood leucocytes type using

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

Science.gov (United States)

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.

Helium Ion Microscope: A New Tool for Sub-nanometer Imaging of Soft Materials

Science.gov (United States)

Shutthanandan, V.; Arey, B.; Smallwood, C. R.; Evans, J. E.

2017-12-01

High-resolution inspection of surface details is needed in many biological and environmental researches to understand the Soil organic material (SOM)-mineral interactions along with identifying microbial communities and their interactions. SOM shares many imaging characteristics with biological samples and getting true surface details from these materials are challenging since they consist of low atomic number materials. FE-SEM imaging is the main imagining technique used to image these materials in the past. These SEM images often show loss of resolution and increase noise due to beam damage and charging issues. Newly developed Helium Ion Microscope (HIM), on the other hand can overcome these difficulties and give very fine details. HIM is very similar to scanning electron microscopy (SEM) but instead of using electrons as a probe beam, HIM uses helium ions with energy ranges from 5 to 40 keV. HIM offers a series of advantages compared to SEM such as nanometer and sub-nanometer image resolutions (about 0.35 nm), detailed surface topography, high surface sensitivity, low Z material imaging (especially for polymers and biological samples), high image contrast, and large depth of field. In addition, HIM also has the ability to image insulating materials without any conductive coatings so that surface details are not modified. In this presentation, several scientific applications across biology and geochemistry will be presented to highlight the effectiveness of this powerful microscope. Acknowledgements: Research was performed using the Environmental Molecular Sciences Laboratory (EMSL), a national scientific user facility sponsored by the Department of Energy's Office of Biological and Environmental Research and located at PNNL. Work was supported by DOE-BER Mesoscale to Molecules Bioimaging Project FWP# 66382.

Noise removal in extended depth of field microscope images through nonlinear signal processing.

Science.gov (United States)

Zahreddine, Ramzi N; Cormack, Robert H; Cogswell, Carol J

2013-04-01

Extended depth of field (EDF) microscopy, achieved through computational optics, allows for real-time 3D imaging of live cell dynamics. EDF is achieved through a combination of point spread function engineering and digital image processing. A linear Wiener filter has been conventionally used to deconvolve the image, but it suffers from high frequency noise amplification and processing artifacts. A nonlinear processing scheme is proposed which extends the depth of field while minimizing background noise. The nonlinear filter is generated via a training algorithm and an iterative optimizer. Biological microscope images processed with the nonlinear filter show a significant improvement in image quality and signal-to-noise ratio over the conventional linear filter.

Label-free cellular structure imaging with 82 nm lateral resolution using an electron-beam excitation-assisted optical microscope.

Science.gov (United States)

Fukuta, Masahiro; Masuda, Yuriko; Inami, Wataru; Kawata, Yoshimasa

2016-07-25

We present label-free and high spatial-resolution imaging for specific cellular structures using an electron-beam excitation-assisted optical microscope (EXA microscope). Images of the actin filament and mitochondria of stained HeLa cells, obtained by fluorescence and EXA microscopy, were compared to identify cellular structures. Based on these results, we demonstrated the feasibility of identifying label-free cellular structures at a spatial resolution of 82 nm. Using numerical analysis, we calculated the imaging depth region and determined the spot size of a cathodoluminescent (CL) light source to be 83 nm at the membrane surface.

Imaging of a large collection of human embryo using a super-parallel MR microscope

International Nuclear Information System (INIS)

Matsuda, Yoshimasa; Ono, Shinya; Otake, Yosuke; Handa, Shinya; Kose, Katsumi; Haishi, Tomoyuki; Yamada, Shigeto; Uwabe, Chikako; Shiota, Kohei

2007-01-01

Using 4 and 8-channel super-parallel magnetic resonance (MR) microscopes with a horizontal bore 2.34T superconducting magnet developed for 3-dimensional MR microscopy of the large Kyoto Collection of Human Embryos, we acquired T 1 -weighted 3D images of 1204 embryos at a spatial resolution of (40 μm) 3 to (150 μm) 3 in about 2 years. Similarity of image contrast between the T 1 -weighted images and stained anatomical sections indicated that T 1 -weighted 3D images could be used for an anatomical 3D image database for human embryology. (author)

The virtual microscopy database-sharing digital microscope images for research and education.

Science.gov (United States)

Lee, Lisa M J; Goldman, Haviva M; Hortsch, Michael

2018-02-14

Over the last 20 years, virtual microscopy has become the predominant modus of teaching the structural organization of cells, tissues, and organs, replacing the use of optical microscopes and glass slides in a traditional histology or pathology laboratory setting. Although virtual microscopy image files can easily be duplicated, creating them requires not only quality histological glass slides but also an expensive whole slide microscopic scanner and massive data storage devices. These resources are not available to all educators and researchers, especially at new institutions in developing countries. This leaves many schools without access to virtual microscopy resources. The Virtual Microscopy Database (VMD) is a new resource established to address this problem. It is a virtual image file-sharing website that allows researchers and educators easy access to a large repository of virtual histology and pathology image files. With the support from the American Association of Anatomists (Bethesda, MD) and MBF Bioscience Inc. (Williston, VT), registration and use of the VMD are currently free of charge. However, the VMD site is restricted to faculty and staff of research and educational institutions. Virtual Microscopy Database users can upload their own collection of virtual slide files, as well as view and download image files for their own non-profit educational and research purposes that have been deposited by other VMD clients. Anat Sci Educ. © 2018 American Association of Anatomists. © 2018 American Association of Anatomists.

Fourier transform infrared spectroscopy microscopic imaging classification based on spatial-spectral features

Science.gov (United States)

Liu, Lian; Yang, Xiukun; Zhong, Mingliang; Liu, Yao; Jing, Xiaojun; Yang, Qin

2018-04-01

The discrete fractional Brownian incremental random (DFBIR) field is used to describe the irregular, random, and highly complex shapes of natural objects such as coastlines and biological tissues, for which traditional Euclidean geometry cannot be used. In this paper, an anisotropic variable window (AVW) directional operator based on the DFBIR field model is proposed for extracting spatial characteristics of Fourier transform infrared spectroscopy (FTIR) microscopic imaging. Probabilistic principal component analysis first extracts spectral features, and then the spatial features of the proposed AVW directional operator are combined with the former to construct a spatial-spectral structure, which increases feature-related information and helps a support vector machine classifier to obtain more efficient distribution-related information. Compared to Haralick’s grey-level co-occurrence matrix, Gabor filters, and local binary patterns (e.g. uniform LBPs, rotation-invariant LBPs, uniform rotation-invariant LBPs), experiments on three FTIR spectroscopy microscopic imaging datasets show that the proposed AVW directional operator is more advantageous in terms of classification accuracy, particularly for low-dimensional spaces of spatial characteristics.

Femtosecond photoelectron point projection microscope

International Nuclear Information System (INIS)

Quinonez, Erik; Handali, Jonathan; Barwick, Brett

2013-01-01

By utilizing a nanometer ultrafast electron source in a point projection microscope we demonstrate that images of nanoparticles with spatial resolutions of the order of 100 nanometers can be obtained. The duration of the emission process of the photoemitted electrons used to make images is shown to be of the order of 100 fs using an autocorrelation technique. The compact geometry of this photoelectron point projection microscope does not preclude its use as a simple ultrafast electron microscope, and we use simple analytic models to estimate temporal resolutions that can be expected when using it as a pump-probe ultrafast electron microscope. These models show a significant increase in temporal resolution when comparing to ultrafast electron microscopes based on conventional designs. We also model the microscopes spectroscopic abilities to capture ultrafast phenomena such as the photon induced near field effect

Virtual pinhole confocal microscope

Energy Technology Data Exchange (ETDEWEB)

George, J.S.; Rector, D.M.; Ranken, D.M. [Los Alamos National Lab., NM (United States). Biophysics Group; Peterson, B. [SciLearn Inc. (United States); Kesteron, J. [VayTech Inc. (United States)

1999-06-01

Scanned confocal microscopes enhance imaging capabilities, providing improved contrast and image resolution in 3-D, but existing systems have significant technical shortcomings and are expensive. Researchers at Los Alamos National Laboratory have developed a novel approach--virtual pinhole confocal microscopy--that uses state of the art illumination, detection, and data processing technologies to produce an imager with a number of advantages: reduced cost, faster imaging, improved efficiency and sensitivity, improved reliability and much greater flexibility. Work at Los Alamos demonstrated proof of principle; prototype hardware and software have been used to demonstrate technical feasibility of several implementation strategies. The system uses high performance illumination, patterned in time and space. The authors have built functional confocal imagers using video display technologies (LCD or DLP) and novel scanner based on a micro-lens array. They have developed a prototype system for high performance data acquisition and processing, designed to support realtime confocal imaging. They have developed algorithms to reconstruct confocal images from a time series of spatially sub-sampled images; software development remains an area of active development. These advances allow the collection of high quality confocal images (in fluorescence, reflectance and transmission modes) with equipment that can inexpensively retrofit to existing microscopes. Planned future extensions to these technologies will significantly enhance capabilities for microscopic imaging in a variety of applications, including confocal endoscopy, and confocal spectral imaging.

Automatic segmentation of Leishmania parasite in microscopic images using a modified CV level set method

Science.gov (United States)

Farahi, Maria; Rabbani, Hossein; Talebi, Ardeshir; Sarrafzadeh, Omid; Ensafi, Shahab

2015-12-01

Visceral Leishmaniasis is a parasitic disease that affects liver, spleen and bone marrow. According to World Health Organization report, definitive diagnosis is possible just by direct observation of the Leishman body in the microscopic image taken from bone marrow samples. We utilize morphological and CV level set method to segment Leishman bodies in digital color microscopic images captured from bone marrow samples. Linear contrast stretching method is used for image enhancement and morphological method is applied to determine the parasite regions and wipe up unwanted objects. Modified global and local CV level set methods are proposed for segmentation and a shape based stopping factor is used to hasten the algorithm. Manual segmentation is considered as ground truth to evaluate the proposed method. This method is tested on 28 samples and achieved 10.90% mean of segmentation error for global model and 9.76% for local model.

Imaging of soft and hard materials using a Boersch phase plate in a transmission electron microscope

Energy Technology Data Exchange (ETDEWEB)

Alloyeau, D., E-mail: alloyeau.damien@gmail.com [National Center for Electron Microscopy, Lawrence Berkeley National Laboratory, One Cyclotron Road, MS/72, Berkeley, CA 94720 (United States); Hsieh, W.K. [National Center for Electron Microscopy, Lawrence Berkeley National Laboratory, One Cyclotron Road, MS/72, Berkeley, CA 94720 (United States); Anderson, E.H.; Hilken, L. [Center for X-ray Optics, Lawrence Berkeley National Laboratory, Berkeley CA 94720 (United States); Benner, G. [Carl Zeiss NTS GmbH, Oberkochen 73447 (Germany); Meng, X. [Electrical Engineering and Computer Sciences, UC Berkeley, Berkeley, CA 94720-1770 (United States); Chen, F.R. [Department of Engineering and System Science, National Tsing Hua University, Hsinchu, Taiwan (China); Kisielowski, C. [National Center for Electron Microscopy, Lawrence Berkeley National Laboratory, One Cyclotron Road, MS/72, Berkeley, CA 94720 (United States)

2010-04-15

Using two levels of electron beam lithography, vapor phase deposition techniques, and FIB etching, we have fabricated an electrostatic Boersch phase plate for contrast enhancement of weak phase objects in a transmission electron microscope. The phase plate has suitable dimensions for the imaging of small biological samples without compromising the high-resolution capabilities of the microscope. A micro-structured electrode allows for phase tuning of the unscattered electron beam, which enables the recording of contrast enhanced in-focus images and in-line holograms. We have demonstrated experimentally that our phase plate improves the contrast of carbon nanotubes while maintaining high-resolution imaging performance, which is demonstrated for the case of an AlGaAs heterostructure. The development opens a new way to study interfaces between soft and hard materials.

Automatic detection of the hippocampal region associated with Alzheimer's disease from microscopic images of mice brain

Science.gov (United States)

Albaidhani, Tahseen; Hawkes, Cheryl; Jassim, Sabah; Al-Assam, Hisham

2016-05-01

The hippocampus is the region of the brain that is primarily associated with memory and spatial navigation. It is one of the first brain regions to be damaged when a person suffers from Alzheimer's disease. Recent research in this field has focussed on the assessment of damage to different blood vessels within the hippocampal region from a high throughput brain microscopic images. The ultimate aim of our research is the creation of an automatic system to count and classify different blood vessels such as capillaries, veins, and arteries in the hippocampus region. This work should provide biologists with efficient and accurate tools in their investigation of the causes of Alzheimer's disease. Locating the boundary of the Region of Interest in the hippocampus from microscopic images of mice brain is the first essential stage towards developing such a system. This task benefits from the variation in colour channels and texture between the two sides of the hippocampus and the boundary region. Accordingly, the developed initial step of our research to locating the hippocampus edge uses a colour-based segmentation of the brain image followed by Hough transforms on the colour channel that isolate the hippocampus region. The output is then used to split the brain image into two sides of the detected section of the boundary: the inside region and the outside region. Experimental results on a sufficiently number of microscopic images demonstrate the effectiveness of the developed solution.

A frameless stereotaxic operating microscope for neurosurgery

International Nuclear Information System (INIS)

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

1989-01-01

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

A frameless stereotaxic operating microscope for neurosurgery.

Science.gov (United States)

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

1989-06-01

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

Scanning tunneling microscopic images and scanning tunneling spectra for coupled rectangular quantum corrals

International Nuclear Information System (INIS)

Mitsuoka, Shigenori; Tamura, Akira

2011-01-01

Assuming that an electron confined by double δ-function barriers lies in a quasi-stationary state, we derived eigenstates and eigenenergies of the electron. Such an electron has a complex eigenenergy, and the imaginary part naturally leads to the lifetime of the electron associated with tunneling through barriers. We applied this point of view to the electron confined in a rectangular quantum corral (QC) on a noble metal surface, and obtained scanning tunneling microscopic images and a scanning tunneling spectrum consistent with experimental ones. We investigated the electron states confined in coupled QCs and obtained the coupled states constructed with bonding and anti-bonding states. Using those energy levels and wavefunctions we specified scanning tunneling microscope (STM) images and scanning tunneling spectra (STS) for the doubly and triply coupled QCs. In addition we pointed out the feature of resonant electron states associated with the same QCs at both ends of the triply coupled QCs.

Fermi surface contours obtained from scanning tunneling microscope images around surface point defects

International Nuclear Information System (INIS)

Khotkevych-Sanina, N V; Kolesnichenko, Yu A; Van Ruitenbeek, J M

2013-01-01

We present a theoretical analysis of the standing wave patterns in scanning tunneling microscope (STM) images, which occur around surface point defects. We consider arbitrary dispersion relations for the surface states and calculate the conductance for a system containing a small-size tunnel contact and a surface impurity. We find rigorous theoretical relations between the interference patterns in the real-space STM images, their Fourier transforms and the Fermi contours of two-dimensional electrons. We propose a new method for reconstructing Fermi contours of surface electron states, directly from the real-space STM images around isolated surface defects. (paper)

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

Science.gov (United States)

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.

Nanosecond Time-Resolved Microscopic Gate-Modulation Imaging of Polycrystalline Organic Thin-Film Transistors

Science.gov (United States)

Matsuoka, Satoshi; Tsutsumi, Jun'ya; Matsui, Hiroyuki; Kamata, Toshihide; Hasegawa, Tatsuo

2018-02-01

We develop a time-resolved microscopic gate-modulation (μ GM ) imaging technique to investigate the temporal evolution of the channel current and accumulated charges in polycrystalline pentacene thin-film transistors (TFTs). A time resolution of as high as 50 ns is achieved by using a fast image-intensifier system that could amplify a series of instantaneous optical microscopic images acquired at various time intervals after the stepped gate bias is switched on. The differential images obtained by subtracting the gate-off image allows us to acquire a series of temporal μ GM images that clearly show the gradual propagation of both channel charges and leaked gate fields within the polycrystalline channel layers. The frontal positions for the propagations of both channel charges and leaked gate fields coincide at all the time intervals, demonstrating that the layered gate dielectric capacitors are successively transversely charged up along the direction of current propagation. The initial μ GM images also indicate that the electric field effect is originally concentrated around a limited area with a width of a few micrometers bordering the channel-electrode interface, and that the field intensity reaches a maximum after 200 ns and then decays. The time required for charge propagation over the whole channel region with a length of 100 μ m is estimated at about 900 ns, which is consistent with the measured field-effect mobility and the temporal-response model for organic TFTs. The effect of grain boundaries can be also visualized by comparison of the μ GM images for the transient and the steady states, which confirms that the potential barriers at the grain boundaries cause the transient shift in the accumulated charges or the transient accumulation of additional charges around the grain boundaries.

High-resolution electron microscope image analysis approach for superconductor YBa2Cu3O7-x

International Nuclear Information System (INIS)

Xu, J.; Lu, F.; Jia, C.; Hua, Z.

1991-01-01

In this paper, an HREM (High-resolution electron microscope) image analysis approach has been developed. The image filtering, segmentation and particles extraction based on gray-scale mathematical morphological operations, are performed on the original HREM image. The final image is a pseudocolor image, with the background removed, relatively uniform brightness, filtered slanting elongation, regular shape for every kind of particle, and particle boundaries that no longer touch each other so that the superconducting material structure can be shown clearly

eSIP: A Novel Solution-Based Sectioned Image Property Approach for Microscope Calibration.

Directory of Open Access Journals (Sweden)

Malte Butzlaff

Full Text Available Fluorescence confocal microscopy represents one of the central tools in modern sciences. Correspondingly, a growing amount of research relies on the development of novel microscopic methods. During the last decade numerous microscopic approaches were developed for the investigation of various scientific questions. Thereby, the former qualitative imaging methods became replaced by advanced quantitative methods to gain more and more information from a given sample. However, modern microscope systems being as complex as they are, require very precise and appropriate calibration routines, in particular when quantitative measurements should be compared over longer time scales or between different setups. Multispectral beads with sub-resolution size are often used to describe the point spread function and thus the optical properties of the microscope. More recently, a fluorescent layer was utilized to describe the axial profile for each pixel, which allows a spatially resolved characterization. However, fabrication of a thin fluorescent layer with matching refractive index is technically not solved yet. Therefore, we propose a novel type of calibration concept for sectioned image property (SIP measurements which is based on fluorescent solution and makes the calibration concept available for a broader number of users. Compared to the previous approach, additional information can be obtained by application of this extended SIP chart approach, including penetration depth, detected number of photons, and illumination profile shape. Furthermore, due to the fit of the complete profile, our method is less susceptible to noise. Generally, the extended SIP approach represents a simple and highly reproducible method, allowing setup independent calibration and alignment procedures, which is mandatory for advanced quantitative microscopy.

SUPERVISED AUTOMATIC HISTOGRAM CLUSTERING AND WATERSHED SEGMENTATION. APPLICATION TO MICROSCOPIC MEDICAL COLOR IMAGES

Directory of Open Access Journals (Sweden)

Olivier Lezoray

2011-05-01

Full Text Available In this paper, an approach to the segmentation of microscopic color images is addressed, and applied to medical images. The approach combines a clustering method and a region growing method. Each color plane is segmented independently relying on a watershed based clustering of the plane histogram. The marginal segmentation maps intersect in a label concordance map. The latter map is simplified based on the assumption that the color planes are correlated. This produces a simplified label concordance map containing labeled and unlabeled pixels. The formers are used as an image of seeds for a color watershed. This fast and robust segmentation scheme is applied to several types of medical images.

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

Science.gov (United States)

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.

Science.gov (United States)

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.

Handy Microscopic Close-Range Videogrammetry

Science.gov (United States)

Esmaeili, F.; Ebadi, H.

2017-09-01

The modeling of small-scale objects is used in different applications such as medicine, industry, and cultural heritage. The capability of modeling small-scale objects using imaging with the help of hand USB digital microscopes and use of videogrammetry techniques has been implemented and evaluated in this paper. Use of this equipment and convergent imaging of the environment for modeling, provides an appropriate set of images for generation of three-dimensional models. The results of the measurements made with the help of a microscope micrometer calibration ruler have demonstrated that self-calibration of a hand camera-microscope set can help obtain a three-dimensional detail extraction precision of about 0.1 millimeters on small-scale environments.

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

Science.gov (United States)

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

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

Science.gov (United States)

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

2015-07-01

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

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

Science.gov (United States)

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

2015-07-01

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

Intensity-based segmentation and visualization of cells in 3D microscopic images using the GPU

Science.gov (United States)

Kang, Mi-Sun; Lee, Jeong-Eom; Jeon, Woong-ki; Choi, Heung-Kook; Kim, Myoung-Hee

2013-02-01

3D microscopy images contain abundant astronomical data, rendering 3D microscopy image processing time-consuming and laborious on a central processing unit (CPU). To solve these problems, many people crop a region of interest (ROI) of the input image to a small size. Although this reduces cost and time, there are drawbacks at the image processing level, e.g., the selected ROI strongly depends on the user and there is a loss in original image information. To mitigate these problems, we developed a 3D microscopy image processing tool on a graphics processing unit (GPU). Our tool provides efficient and various automatic thresholding methods to achieve intensity-based segmentation of 3D microscopy images. Users can select the algorithm to be applied. Further, the image processing tool provides visualization of segmented volume data and can set the scale, transportation, etc. using a keyboard and mouse. However, the 3D objects visualized fast still need to be analyzed to obtain information for biologists. To analyze 3D microscopic images, we need quantitative data of the images. Therefore, we label the segmented 3D objects within all 3D microscopic images and obtain quantitative information on each labeled object. This information can use the classification feature. A user can select the object to be analyzed. Our tool allows the selected object to be displayed on a new window, and hence, more details of the object can be observed. Finally, we validate the effectiveness of our tool by comparing the CPU and GPU processing times by matching the specification and configuration.

Pancam and microscopic imager observations of dust on the Spirit Rovers

DEFF Research Database (Denmark)

Vaughan....[], Alicia F.; Johnson, Jeffrey R.; Walter, Goetz

2010-01-01

This work describes dust deposits on the Spirit Rover over 2000 sols through examination of Pancam and Microscopic Imager observations of specific locations on the rover body, including portions of the solar array, Pancam and Mini-TES calibration targets, and the magnets. This data set reveals...... the three "cleaning events" experienced by Spirit to date, the spectral properties of dust, and the tendency of dust particles to form aggregates 100 um and larger...

Microscope on Mars

Science.gov (United States)

2004-01-01

This image taken at Meridiani Planum, Mars by the panoramic camera on the Mars Exploration Rover Opportunity shows the rover's microscopic imager (circular device in center), located on its instrument deployment device, or 'arm.' The image was acquired on the ninth martian day or sol of the rover's mission.

Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging.

Science.gov (United States)

Zhao, Qiaole; Schelen, Ben; Schouten, Raymond; van den Oever, Rein; Leenen, René; van Kuijk, Harry; Peters, Inge; Polderdijk, Frank; Bosiers, Jan; Raspe, Marcel; Jalink, Kees; Geert Sander de Jong, Jan; van Geest, Bert; Stoop, Karel; Young, Ian Ted

2012-12-01

We have built an all-solid-state camera that is directly modulated at the pixel level for frequency-domain fluorescence lifetime imaging microscopy (FLIM) measurements. This novel camera eliminates the need for an image intensifier through the use of an application-specific charge coupled device design in a frequency-domain FLIM system. The first stage of evaluation for the camera has been carried out. Camera characteristics such as noise distribution, dark current influence, camera gain, sampling density, sensitivity, linearity of photometric response, and optical transfer function have been studied through experiments. We are able to do lifetime measurement using our modulated, electron-multiplied fluorescence lifetime imaging microscope (MEM-FLIM) camera for various objects, e.g., fluorescein solution, fixed green fluorescent protein (GFP) cells, and GFP-actin stained live cells. A detailed comparison of a conventional microchannel plate (MCP)-based FLIM system and the MEM-FLIM system is presented. The MEM-FLIM camera shows higher resolution and a better image quality. The MEM-FLIM camera provides a new opportunity for performing frequency-domain FLIM.

Field-portable pixel super-resolution colour microscope.

Directory of Open Access Journals (Sweden)

Alon Greenbaum

Full Text Available 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.

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

Science.gov (United States)

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

Reconstruction of an Non-Monochromatically Illuminated Object Imaged through an Electron Microscope with a Fluctuating Electromagnetic Field

NARCIS (Netherlands)

Hoenders, B.J.

1975-01-01

It is shown that a weak phase object imaged by an electron microscope within the presence of instabilities of the lense currents and the acceleration voltage, fluctuating electromagnetic field, can be reconstructed from the intensity distribution in the image plane. Perfectly incoherent illumination

Color Laser Microscope

Science.gov (United States)

Awamura, D.; Ode, T.; Yonezawa, M.

1987-04-01

A color laser microscope utilizing a new color laser imaging system has been developed for the visual inspection of semiconductors. The light source, produced by three lasers (Red; He-Ne, Green; Ar, Blue; He-Cd), is deflected horizontally by an AOD (Acoustic Optical Deflector) and vertically by a vibration mirror. The laser beam is focused in a small spot which is scanned over the sample at high speed. The light reflected back from the sample is reformed to contain linear information by returning to the original vibration mirror. The linear light is guided to the CCD image sensor where it is converted into a video signal. Individual CCD image sensors are used for each of the three R, G, or B color image signals. The confocal optical system with its laser light source yields a color TV monitor image with no flaring and a much sharper resolution than that of the conventional optical microscope. The AOD makes possible a high speed laser scan and a NTSC or PAL TV video signal is produced in real time without any video memory. Since the light source is composed of R, G, and B laser beams, color separation superior to that of white light illumination is achieved. Because of the photometric linearity of the image detector, the R, G, and B outputs of the system are most suitably used for hue analysis. The CCD linear image sensors in the optical system produce no geometrical distortion, and good color registration is available principally. The output signal can be used for high accuracy line width measuring. The many features of the color laser microscope make it ideally suited for the visual inspection of semiconductor processing. A number of these systems have already been installed in such a capacity. The Color Laser Microscope can also be a very useful tool for the fields of material engineering and biotechnology.

Reducing charging effects in scanning electron microscope images by Rayleigh contrast stretching method (RCS).

Science.gov (United States)

Wan Ismail, W Z; Sim, K S; Tso, C P; Ting, H Y

2011-01-01

To reduce undesirable charging effects in scanning electron microscope images, Rayleigh contrast stretching is developed and employed. First, re-scaling is performed on the input image histograms with Rayleigh algorithm. Then, contrast stretching or contrast adjustment is implemented to improve the images while reducing the contrast charging artifacts. This technique has been compared to some existing histogram equalization (HE) extension techniques: recursive sub-image HE, contrast stretching dynamic HE, multipeak HE and recursive mean separate HE. Other post processing methods, such as wavelet approach, spatial filtering, and exponential contrast stretching, are compared as well. Overall, the proposed method produces better image compensation in reducing charging artifacts. Copyright © 2011 Wiley Periodicals, Inc.

A method for automatic grain segmentation of multi-angle cross-polarized microscopic images of sandstone

Science.gov (United States)

Jiang, Feng; Gu, Qing; Hao, Huizhen; Li, Na; Wang, Bingqian; Hu, Xiumian

2018-06-01

Automatic grain segmentation of sandstone is to partition mineral grains into separate regions in the thin section, which is the first step for computer aided mineral identification and sandstone classification. The sandstone microscopic images contain a large number of mixed mineral grains where differences among adjacent grains, i.e., quartz, feldspar and lithic grains, are usually ambiguous, which make grain segmentation difficult. In this paper, we take advantage of multi-angle cross-polarized microscopic images and propose a method for grain segmentation with high accuracy. The method consists of two stages, in the first stage, we enhance the SLIC (Simple Linear Iterative Clustering) algorithm, named MSLIC, to make use of multi-angle images and segment the images as boundary adherent superpixels. In the second stage, we propose the region merging technique which combines the coarse merging and fine merging algorithms. The coarse merging merges the adjacent superpixels with less evident boundaries, and the fine merging merges the ambiguous superpixels using the spatial enhanced fuzzy clustering. Experiments are designed on 9 sets of multi-angle cross-polarized images taken from the three major types of sandstones. The results demonstrate both the effectiveness and potential of the proposed method, comparing to the available segmentation methods.

Neurite density imaging versus imaging of microscopic anisotropy in diffusion MRI: A model comparison using spherical tensor encoding.

Science.gov (United States)

Lampinen, Björn; Szczepankiewicz, Filip; Mårtensson, Johan; van Westen, Danielle; Sundgren, Pia C; Nilsson, Markus

2017-02-15

In diffusion MRI (dMRI), microscopic diffusion anisotropy can be obscured by orientation dispersion. Separation of these properties is of high importance, since it could allow dMRI to non-invasively probe elongated structures such as neurites (axons and dendrites). However, conventional dMRI, based on single diffusion encoding (SDE), entangles microscopic anisotropy and orientation dispersion with intra-voxel variance in isotropic diffusivity. SDE-based methods for estimating microscopic anisotropy, such as the neurite orientation dispersion and density imaging (NODDI) method, must thus rely on model assumptions to disentangle these features. An alternative approach is to directly quantify microscopic anisotropy by the use of variable shape of the b-tensor. Along those lines, we here present the 'constrained diffusional variance decomposition' (CODIVIDE) method, which jointly analyzes data acquired with diffusion encoding applied in a single direction at a time (linear tensor encoding, LTE) and in all directions (spherical tensor encoding, STE). We then contrast the two approaches by comparing neurite density estimated using NODDI with microscopic anisotropy estimated using CODIVIDE. Data were acquired in healthy volunteers and in glioma patients. NODDI and CODIVIDE differed the most in gray matter and in gliomas, where NODDI detected a neurite fraction higher than expected from the level of microscopic diffusion anisotropy found with CODIVIDE. The discrepancies could be explained by the NODDI tortuosity assumption, which enforces a connection between the neurite density and the mean diffusivity of tissue. Our results suggest that this assumption is invalid, which leads to a NODDI neurite density that is inconsistent between LTE and STE data. Using simulations, we demonstrate that the NODDI assumptions result in parameter bias that precludes the use of NODDI to map neurite density. With CODIVIDE, we found high levels of microscopic anisotropy in white matter

High-speed atomic force microscope imaging: Adaptive multiloop mode

Science.gov (United States)

Ren, Juan; Zou, Qingze; Li, Bo; Lin, Zhiqun

2014-07-01

In this paper, an imaging mode (called the adaptive multiloop mode) of atomic force microscope (AFM) is proposed to substantially increase the speed of tapping mode (TM) imaging while preserving the advantages of TM imaging over contact mode (CM) imaging. Due to its superior image quality and less sample disturbances over CM imaging, particularly for soft materials such as polymers, TM imaging is currently the most widely used imaging technique. The speed of TM imaging, however, is substantially (over an order of magnitude) lower than that of CM imaging, becoming the major bottleneck of this technique. Increasing the speed of TM imaging is challenging as a stable probe tapping on the sample surface must be maintained to preserve the image quality, whereas the probe tapping is rather sensitive to the sample topography variation. As a result, the increase of imaging speed can quickly lead to loss of the probe-sample contact and/or annihilation of the probe tapping, resulting in image distortion and/or sample deformation. The proposed adaptive multiloop mode (AMLM) imaging overcomes these limitations of TM imaging through the following three efforts integrated together: First, it is proposed to account for the variation of the TM deflection when quantifying the sample topography; second, an inner-outer feedback control loop to regulate the TM deflection is added on top of the tapping-feedback control loop to improve the sample topography tracking; and, third, an online iterative feedforward controller is augmented to the whole control system to further enhance the topography tracking, where the next-line sample topography is predicted and utilized to reduce the tracking error. The added feedback regulation of the TM deflection ensures the probe-sample interaction force remains near the minimum for maintaining a stable probe-sample interaction. The proposed AMLM imaging is tested and demonstrated by imaging a poly(tert-butyl acrylate) sample in experiments. The

Infrared up-conversion microscope

DEFF Research Database (Denmark)

2014-01-01

There is presented an up-conversion infrared microscope (110) arranged for imaging an associated object (130), wherein the up-conversion infrared microscope (110) comprises a non-linear crystal (120) arranged for up-conversion of infrared electromagnetic radiation, and wherein an objective optical...

Neural Network for Nanoscience Scanning Electron Microscope Image Recognition.

Science.gov (United States)

Modarres, Mohammad Hadi; Aversa, Rossella; Cozzini, Stefano; Ciancio, Regina; Leto, Angelo; Brandino, Giuseppe Piero

2017-10-16

In this paper we applied transfer learning techniques for image recognition, automatic categorization, and labeling of nanoscience images obtained by scanning electron microscope (SEM). Roughly 20,000 SEM images were manually classified into 10 categories to form a labeled training set, which can be used as a reference set for future applications of deep learning enhanced algorithms in the nanoscience domain. The categories chosen spanned the range of 0-Dimensional (0D) objects such as particles, 1D nanowires and fibres, 2D films and coated surfaces, and 3D patterned surfaces such as pillars. The training set was used to retrain on the SEM dataset and to compare many convolutional neural network models (Inception-v3, Inception-v4, ResNet). We obtained compatible results by performing a feature extraction of the different models on the same dataset. We performed additional analysis of the classifier on a second test set to further investigate the results both on particular cases and from a statistical point of view. Our algorithm was able to successfully classify around 90% of a test dataset consisting of SEM images, while reduced accuracy was found in the case of images at the boundary between two categories or containing elements of multiple categories. In these cases, the image classification did not identify a predominant category with a high score. We used the statistical outcomes from testing to deploy a semi-automatic workflow able to classify and label images generated by the SEM. Finally, a separate training was performed to determine the volume fraction of coherently aligned nanowires in SEM images. The results were compared with what was obtained using the Local Gradient Orientation method. This example demonstrates the versatility and the potential of transfer learning to address specific tasks of interest in nanoscience applications.

Imaging optical scattering of butterfly wing scales with a microscope.

Science.gov (United States)

Fu, Jinxin; Yoon, Beom-Jin; Park, Jung Ok; Srinivasarao, Mohan

2017-08-06

A new optical method is proposed to investigate the reflectance of structurally coloured objects, such as Morpho butterfly wing scales and cholesteric liquid crystals. Using a reflected-light microscope and a digital single-lens reflex (DSLR) camera, we have successfully measured the two-dimensional reflection pattern of individual wing scales of Morpho butterflies. We demonstrate that this method enables us to measure the bidirectional reflectance distribution function (BRDF). The scattering image observed in the back focal plane of the objective is projected onto the camera sensor by inserting a Bertrand lens in the optical path of the microscope. With monochromatic light illumination, we quantify the angle-dependent reflectance spectra from the wing scales of Morpho rhetenor by retrieving the raw signal from the digital camera sensor. We also demonstrate that the polarization-dependent reflection of individual wing scales is readily observed using this method, using the individual wing scales of Morpho cypris . In an effort to show the generality of the method, we used a chiral nematic fluid to illustrate the angle-dependent reflectance as seen by this method.

A preliminary investigation: the impact of microscopic condenser on depth of field in cytogenetic imaging

Science.gov (United States)

Ren, Liqiang; Qiu, Yuchen; Li, Zheng; Li, Yuhua; Zheng, Bin; Li, Shibo; Chen, Wei R.; Liu, Hong

2013-02-01

As one of the important components of optical microscopes, the condenser has a considerable impact on system performance, especially on the depth of field (DOF). DOF is a critical technical feature in cytogenetic imaging that may affect the efficiency and accuracy of clinical diagnosis. The purpose of this study is to investigate the influence of microscopic condenser on DOF using a prototype of transmitted optical microscope, based on objective and subjective evaluations. After the description of the relationship between condenser and objective lens and the theoretical analysis of the condenser impact on system numerical aperture and DOF, a standard resolution pattern and several cytogenetic samples are adopted to assess the condenser impact on DOF, respectively. The experimental results of these objective and subjective evaluations are in agreement with the theoretical analysis and show that, under the specific intermediate range of condenser numerical aperture ( NAcond ), the DOF value decreases with the increase of NAcond . Although the above qualitative results are obtained under the experimental conditions with a specific prototype system, the methods presented in this preliminary investigation could offer useful guidelines for optimizing operational parameters in cytogenetic imaging.

Nucleus and cytoplasm segmentation in microscopic images using K-means clustering and region growing.

Science.gov (United States)

Sarrafzadeh, Omid; Dehnavi, Alireza Mehri

2015-01-01

Segmentation of leukocytes acts as the foundation for all automated image-based hematological disease recognition systems. Most of the time, hematologists are interested in evaluation of white blood cells only. Digital image processing techniques can help them in their analysis and diagnosis. The main objective of this paper is to detect leukocytes from a blood smear microscopic image and segment them into their two dominant elements, nucleus and cytoplasm. The segmentation is conducted using two stages of applying K-means clustering. First, the nuclei are segmented using K-means clustering. Then, a proposed method based on region growing is applied to separate the connected nuclei. Next, the nuclei are subtracted from the original image. Finally, the cytoplasm is segmented using the second stage of K-means clustering. The results indicate that the proposed method is able to extract the nucleus and cytoplasm regions accurately and works well even though there is no significant contrast between the components in the image. In this paper, a method based on K-means clustering and region growing is proposed in order to detect leukocytes from a blood smear microscopic image and segment its components, the nucleus and the cytoplasm. As region growing step of the algorithm relies on the information of edges, it will not able to separate the connected nuclei more accurately in poor edges and it requires at least a weak edge to exist between the nuclei. The nucleus and cytoplasm segments of a leukocyte can be used for feature extraction and classification which leads to automated leukemia detection.

Designing a large field-of-view two-photon microscope using optical invariant analysis.

Science.gov (United States)

Bumstead, Jonathan R; Park, Jasmine J; Rosen, Isaac A; Kraft, Andrew W; Wright, Patrick W; Reisman, Matthew D; Côté, Daniel C; Culver, Joseph P

2018-04-01

Conventional two-photon microscopy (TPM) is capable of imaging neural dynamics with subcellular resolution, but it is limited to a field-of-view (FOV) diameter [Formula: see text]. Although there has been recent progress in extending the FOV in TPM, a principled design approach for developing large FOV TPM (LF-TPM) with off-the-shelf components has yet to be established. Therefore, we present a design strategy that depends on analyzing the optical invariant of commercially available objectives, relay lenses, mirror scanners, and emission collection systems in isolation. Components are then selected to maximize the space-bandwidth product of the integrated microscope. In comparison with other LF-TPM systems, our strategy simplifies the sequence of design decisions and is applicable to extending the FOV in any microscope with an optical relay. The microscope we constructed with this design approach can image [Formula: see text] lateral and [Formula: see text] axial resolution over a 7-mm diameter FOV, which is a 100-fold increase in FOV compared with conventional TPM. As a demonstration of the potential that LF-TPM has on understanding the microarchitecture of the mouse brain across interhemispheric regions, we performed in vivo imaging of both the cerebral vasculature and microglia cell bodies over the mouse cortex.

Automated Image Analysis of Lung Branching Morphogenesis from Microscopic Images of Fetal Rat Explants

Science.gov (United States)

Rodrigues, Pedro L.; Rodrigues, Nuno F.; Duque, Duarte; Granja, Sara; Correia-Pinto, Jorge; Vilaça, João L.

2014-01-01

Background. Regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. This work presents a new methodology to accurately quantify the epithelial, outer contour, and peripheral airway buds of lung explants during cellular development from microscopic images. Methods. The outer contour was defined using an adaptive and multiscale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelium was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds was counted as the skeleton branched ends from a skeletonized image of the lung inner epithelia. Results. The time for lung branching morphometric analysis was reduced in 98% in contrast to the manual method. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Nonsignificant differences were found between the automatic and manual results in all culture days. Conclusions. The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lighting characteristics and allowing a reliable comparison between different researchers. PMID:25250057

Automated Image Analysis of Lung Branching Morphogenesis from Microscopic Images of Fetal Rat Explants

Directory of Open Access Journals (Sweden)

Pedro L. Rodrigues

2014-01-01

Full Text Available Background. Regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. This work presents a new methodology to accurately quantify the epithelial, outer contour, and peripheral airway buds of lung explants during cellular development from microscopic images. Methods. The outer contour was defined using an adaptive and multiscale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelium was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds was counted as the skeleton branched ends from a skeletonized image of the lung inner epithelia. Results. The time for lung branching morphometric analysis was reduced in 98% in contrast to the manual method. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Nonsignificant differences were found between the automatic and manual results in all culture days. Conclusions. The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lighting characteristics and allowing a reliable comparison between different researchers.

Microscope-Integrated Intraoperative Ultrahigh-Speed Swept-Source Optical Coherence Tomography for Widefield Retinal and Anterior Segment Imaging.

Science.gov (United States)

Lu, Chen D; Waheed, Nadia K; Witkin, Andre; Baumal, Caroline R; Liu, Jonathan J; Potsaid, Benjamin; Joseph, Anthony; Jayaraman, Vijaysekhar; Cable, Alex; Chan, Kinpui; Duker, Jay S; Fujimoto, James G

2018-02-01

To demonstrate the feasibility of retinal and anterior segment intraoperative widefield imaging using an ultrahigh-speed, swept-source optical coherence tomography (SS-OCT) surgical microscope attachment. A prototype post-objective SS-OCT using a 1,050-nm wavelength, 400 kHz A-scan rate, vertical cavity surface-emitting laser (VCSEL) light source was integrated to a commercial ophthalmic surgical microscope after the objective. Each widefield OCT data set was acquired in 3 seconds (1,000 × 1,000 A-scans, 12 × 12 mm 2 for retina and 10 × 10 mm 2 for anterior segment). Intraoperative SS-OCT was performed in 20 eyes of 20 patients. In six of seven membrane peels and five of seven rhegmatogenous retinal detachment repair surgeries, widefield retinal imaging enabled evaluation pre- and postoperatively. In all seven cataract cases, anterior imaging evaluated the integrity of the posterior lens capsule. Ultrahigh-speed SS-OCT enables widefield intraoperative viewing in the posterior and anterior eye. Widefield imaging visualizes ocular structures and pathology without requiring OCT realignment. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:94-102.]. Copyright 2018, SLACK Incorporated.

Scanning electron microscope cathodoluminescence imaging of subgrain boundaries, twins and planar deformation features in quartz

Science.gov (United States)

Hamers, M. F.; Pennock, G. M.; Drury, M. R.

2017-04-01

The study of deformation features has been of great importance to determine deformation mechanisms in quartz. Relevant microstructures in both growth and deformation processes include dislocations, subgrains, subgrain boundaries, Brazil and Dauphiné twins and planar deformation features (PDFs). Dislocations and twin boundaries are most commonly imaged using a transmission electron microscope (TEM), because these cannot directly be observed using light microscopy, in contrast to PDFs. Here, we show that red-filtered cathodoluminescence imaging in a scanning electron microscope (SEM) is a useful method to visualise subgrain boundaries, Brazil and Dauphiné twin boundaries. Because standard petrographic thin sections can be studied in the SEM, the observed structures can be directly and easily correlated to light microscopy studies. In contrast to TEM preparation methods, SEM techniques are non-destructive to the area of interest on a petrographic thin section.

Science applications of a multispectral microscopic imager for the astrobiological exploration of Mars.

Science.gov (United States)

Núñez, Jorge I; Farmer, Jack D; Sellar, R Glenn; Swayze, Gregg A; Blaney, Diana L

2014-02-01

Future astrobiological missions to Mars are likely to emphasize the use of rovers with in situ petrologic capabilities for selecting the best samples at a site for in situ analysis with onboard lab instruments or for caching for potential return to Earth. Such observations are central to an understanding of the potential for past habitable conditions at a site and for identifying samples most likely to harbor fossil biosignatures. The Multispectral Microscopic Imager (MMI) provides multispectral reflectance images of geological samples at the microscale, where each image pixel is composed of a visible/shortwave infrared spectrum ranging from 0.46 to 1.73 μm. This spectral range enables the discrimination of a wide variety of rock-forming minerals, especially Fe-bearing phases, and the detection of hydrated minerals. The MMI advances beyond the capabilities of current microimagers on Mars by extending the spectral range into the infrared and increasing the number of spectral bands. The design employs multispectral light-emitting diodes and an uncooled indium gallium arsenide focal plane array to achieve a very low mass and high reliability. To better understand and demonstrate the capabilities of the MMI for future surface missions to Mars, we analyzed samples from Mars-relevant analog environments with the MMI. Results indicate that the MMI images faithfully resolve the fine-scale microtextural features of samples and provide important information to help constrain mineral composition. The use of spectral endmember mapping reveals the distribution of Fe-bearing minerals (including silicates and oxides) with high fidelity, along with the presence of hydrated minerals. MMI-based petrogenetic interpretations compare favorably with laboratory-based analyses, revealing the value of the MMI for future in situ rover-mediated astrobiological exploration of Mars. Mars-Microscopic imager-Multispectral imaging-Spectroscopy-Habitability-Arm instrument.

The influence of the microscope lamp filament colour temperature on the process of digital images of histological slides acquisition standardization.

Science.gov (United States)

Korzynska, Anna; Roszkowiak, Lukasz; Pijanowska, Dorota; Kozlowski, Wojciech; Markiewicz, Tomasz

2014-01-01

The aim of this study is to compare the digital images of the tissue biopsy captured with optical microscope using bright field technique under various light conditions. The range of colour's variation in immunohistochemically stained with 3,3'-Diaminobenzidine and Haematoxylin tissue samples is immense and coming from various sources. One of them is inadequate setting of camera's white balance to microscope's light colour temperature. Although this type of error can be easily handled during the stage of image acquisition, it can be eliminated with use of colour adjustment algorithms. The examination of the dependence of colour variation from microscope's light temperature and settings of the camera is done as an introductory research to the process of automatic colour standardization. Six fields of view with empty space among the tissue samples have been selected for analysis. Each field of view has been acquired 225 times with various microscope light temperature and camera white balance settings. The fourteen randomly chosen images have been corrected and compared, with the reference image, by the following methods: Mean Square Error, Structural SIMilarity and visual assessment of viewer. For two types of backgrounds and two types of objects, the statistical image descriptors: range, median, mean and its standard deviation of chromaticity on a and b channels from CIELab colour space, and luminance L, and local colour variability for objects' specific area have been calculated. The results have been averaged for 6 images acquired in the same light conditions and camera settings for each sample. The analysis of the results leads to the following conclusions: (1) the images collected with white balance setting adjusted to light colour temperature clusters in certain area of chromatic space, (2) the process of white balance correction for images collected with white balance camera settings not matched to the light temperature moves image descriptors into proper

Ultrabright planar optodes for luminescence life-time based microscopic imaging of O₂ dynamics in biofilms

DEFF Research Database (Denmark)

Staal, Marc Jaap; Borisov, S M; Rickelt, L F

2011-01-01

New transparent optodes for life-time based microscopic imaging of O2 were developed by spin-coating a µm-thin layer of a highly luminescent cyclometalated iridium(III) coumarin complex in polystyrene onto glass cover slips. Compared to similar thin-film O2 optodes based on a ruthenium(II) polypy......New transparent optodes for life-time based microscopic imaging of O2 were developed by spin-coating a µm-thin layer of a highly luminescent cyclometalated iridium(III) coumarin complex in polystyrene onto glass cover slips. Compared to similar thin-film O2 optodes based on a ruthenium...

Image processing of muscle striations below the resolution limit of the light microscope

International Nuclear Information System (INIS)

Burns, D.H.; Holdren, D.N.; Periasamy, A.; Everts, W.C.; Pollack, G.H.

1985-01-01

We describe the use of digital deconvolution in the study of muscle striations below the resolution imposed by the optical diffraction of our video light microscope. To use deconvolution procedures on muscle images, the transfer function of the optical system is first characterized. This is accomplished by imaging a step object and fitting the image with the combination of a first order Bessel and Guassian function using a non-linear least squares approach. Due to the ill-conditioned nature of deconvolution, however, ambiguity in the reconstruction is sometimes found. To allow better estimation of the true object, separate deconvolution approaches are used and the reconstructions compared. In this manner, the fine structure of muscle striations is determined

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

Science.gov (United States)

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

Science.gov (United States)

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.

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

Science.gov (United States)

Stemmer, A

1995-04-01

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

Capturing and displaying microscopic images used in medical diagnostics and forensic science using 4K video resolution - an application in higher education.

Science.gov (United States)

Maier, Hans; de Heer, Gert; Ortac, Ajda; Kuijten, Jan

2015-11-01

To analyze, interpret and evaluate microscopic images, used in medical diagnostics and forensic science, video images for educational purposes were made with a very high resolution of 4096 × 2160 pixels (4K), which is four times as many pixels as High-Definition Video (1920 × 1080 pixels). The unprecedented high resolution makes it possible to see details that remain invisible to any other video format. The images of the specimens (blood cells, tissue sections, hair, fibre, etc.) are recorded using a 4K video camera which is attached to a light microscope. After processing, this resulted in very sharp and highly detailed images. This material was then used in education for classroom discussion. Spoken explanation by experts in the field of medical diagnostics and forensic science was also added to the high-resolution video images to make it suitable for self-study. © 2015 The Authors. Journal of Microscopy published by John Wiley & Sons Ltd on behalf of Royal Microscopical Society.

Scanning Electron Microscope Calibration Using a Multi-Image Non-Linear Minimization Process

Science.gov (United States)

Cui, Le; Marchand, Éric

2015-04-01

A scanning electron microscope (SEM) calibrating approach based on non-linear minimization procedure is presented in this article. A part of this article has been published in IEEE International Conference on Robotics and Automation (ICRA), 2014. . Both the intrinsic parameters and the extrinsic parameters estimations are achieved simultaneously by minimizing the registration error. The proposed approach considers multi-images of a multi-scale calibration pattern view from different positions and orientations. Since the projection geometry of the scanning electron microscope is different from that of a classical optical sensor, the perspective projection model and the parallel projection model are considered and compared with distortion models. Experiments are realized by varying the position and the orientation of a multi-scale chessboard calibration pattern from 300× to 10,000×. The experimental results show the efficiency and the accuracy of this approach.

Macro-SICM: A Scanning Ion Conductance Microscope for Large-Range Imaging.

Science.gov (United States)

Schierbaum, Nicolas; Hack, Martin; Betz, Oliver; Schäffer, Tilman E

2018-04-17

The scanning ion conductance microscope (SICM) is a versatile, high-resolution imaging technique that uses an electrolyte-filled nanopipet as a probe. Its noncontact imaging principle makes the SICM uniquely suited for the investigation of soft and delicate surface structures in a liquid environment. The SICM has found an ever-increasing number of applications in chemistry, physics, and biology. However, a drawback of conventional SICMs is their relatively small scan range (typically 100 μm × 100 μm in the lateral and 10 μm in the vertical direction). We have developed a Macro-SICM with an exceedingly large scan range of 25 mm × 25 mm in the lateral and 0.25 mm in the vertical direction. We demonstrate the high versatility of the Macro-SICM by imaging at different length scales: from centimeters (fingerprint, coin) to millimeters (bovine tongue tissue, insect wing) to micrometers (cellular extensions). We applied the Macro-SICM to the study of collective cell migration in epithelial wound healing.

First test model of the optical microscope which images the whole vertical particle tracks without any depth scanning

International Nuclear Information System (INIS)

Soroko, L.M.

2001-01-01

The first test model of the optical microscope which produces the in focus image of the whole vertical particle track without depth scanning is described. The in focus image of the object consisting of the linear array of the point-like elements was obtained. A comparison with primary out of focus image of such an object has been made

Imaging of Norway spruce early somatic embryos with the ESEM, Cryo-SEM and laser scanning microscope.

Science.gov (United States)

Neděla, Vilém; Hřib, Jiří; Havel, Ladislav; Hudec, Jiří; Runštuk, Jiří

2016-05-01

This article describes the surface structure of Norway spruce early somatic embryos (ESEs) as a typical culture with asynchronous development. The microstructure of extracellular matrix covering ESEs were observed using the environmental scanning electron microscope as a primary tool and using the scanning electron microscope with cryo attachment and laser electron microscope as a complementary tool allowing our results to be proven independently. The fresh samples were observed in conditions of the air environment of the environmental scanning electron microscope (ESEM) with the pressure from 550Pa to 690Pa and the low temperature of the sample from -18°C to -22°C. The samples were studied using two different types of detector to allow studying either the thin surface structure or material composition. The scanning electron microscope with cryo attachment was used for imaging frozen extracellular matrix microstructure with higher resolution. The combination of both electron microscopy methods was suitable for observation of "native" plant samples, allowing correct evaluation of our results, free of error and artifacts. Copyright © 2016 Elsevier Ltd. All rights reserved.

Design of a transmission electron positron microscope

International Nuclear Information System (INIS)

Doyama, Masao; Inoue, M.; Kogure, Y.; Hayashi, Y.; Yoshii, T.; Kurihara, T.; Tsuno, K.

2003-01-01

This paper reports the plans and design of positron-electron microscopes being built at KEK (High Energy Accelerator Research Organization), Tsukuba, Japan. A used electron microscope is altered. The kinetic energies of positrons produced by accelerators or by nuclear decays are not a unique value but show a spread over in a wide range. Positron beam is guided to a transmission electron microscope (JEM100SX). Positrons are moderated by a tungsten foil, are accelerated and are focused on a nickel sheet. The monochromatic focused beam is injected into an electron microscope. The focusing and aberration of positrons are the same as electrons in a magnetic system which are used in commercial electron microscopes. Imaging plates are used to record positron images for the transmission electron microscope. (author)

A high-resolution multimode digital microscope system.

Science.gov (United States)

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

2013-01-01

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

Compact plane illumination plugin device to enable light sheet fluorescence imaging of multi-cellular organisms on an inverted wide-field microscope

OpenAIRE

Guan, Zeyi; Lee, Juhyun; Jiang, Hao; Dong, Siyan; Jen, Nelson; Hsiai, Tzung; Ho, Chih-Ming; Fei, Peng

2015-01-01

We developed a compact plane illumination plugin (PIP) device which enabled plane illumination and light sheet fluorescence imaging on a conventional inverted microscope. The PIP device allowed the integration of microscope with tunable laser sheet profile, fast image acquisition, and 3-D scanning. The device is both compact, measuring approximately 15 by 5 by 5 cm, and cost-effective, since we employed consumer electronics and an inexpensive device molding method. We demonstrated that PIP pr...

Weight preserving image registration for monitoring disease progression in lung CT.

Science.gov (United States)

Gorbunova, Vladlena; Lol, Pechin; Ashraf, Haseem; Dirksen, Asger; Nielsen, Mads; de Bruijne, Marleen

2008-01-01

We present a new image registration based method for monitoring regional disease progression in longitudinal image studies of lung disease. A free-form image registration technique is used to match a baseline 3D CT lung scan onto a following scan. Areas with lower intensity in the following scan compared with intensities in the deformed baseline image indicate local loss of lung tissue that is associated with progression of emphysema. To account for differences in lung intensity owing to differences in the inspiration level in the two scans rather than disease progression, we propose to adjust the density of lung tissue with respect to local expansion or compression such that the total weight of the lungs is preserved during deformation. Our method provides a good estimation of regional destruction of lung tissue for subjects with a significant difference in inspiration level between CT scans and may result in a more sensitive measure of disease progression than standard quantitative CT measures.

A widefield fluorescence microscope with a linear image sensor for image cytometry of biospecimens: Considerations for image quality optimization

Energy Technology Data Exchange (ETDEWEB)

Hutcheson, Joshua A.; Majid, Aneeka A.; Powless, Amy J.; Muldoon, Timothy J., E-mail: tmuldoon@uark.edu [Department of Biomedical Engineering, University of Arkansas, 120 Engineering Hall, Fayetteville, Arkansas 72701 (United States)

2015-09-15

Linear image sensors have been widely used in numerous research and industry applications to provide continuous imaging of moving objects. Here, we present a widefield fluorescence microscope with a linear image sensor used to image translating objects for image cytometry. First, a calibration curve was characterized for a custom microfluidic chamber over a span of volumetric pump rates. Image data were also acquired using 15 μm fluorescent polystyrene spheres on a slide with a motorized translation stage in order to match linear translation speed with line exposure periods to preserve the image aspect ratio. Aspect ratios were then calculated after imaging to ensure quality control of image data. Fluorescent beads were imaged in suspension flowing through the microfluidics chamber being pumped by a mechanical syringe pump at 16 μl min{sup −1} with a line exposure period of 150 μs. The line period was selected to acquire images of fluorescent beads with a 40 dB signal-to-background ratio. A motorized translation stage was then used to transport conventional glass slides of stained cellular biospecimens. Whole blood collected from healthy volunteers was stained with 0.02% (w/v) proflavine hemisulfate was imaged to highlight leukocyte morphology with a 1.56 mm × 1.28 mm field of view (1540 ms total acquisition time). Oral squamous cells were also collected from healthy volunteers and stained with 0.01% (w/v) proflavine hemisulfate to demonstrate quantifiable subcellular features and an average nuclear to cytoplasmic ratio of 0.03 (n = 75), with a resolution of 0.31 μm pixels{sup −1}.

Wolter x-ray microscope calibration

International Nuclear Information System (INIS)

Gerassimenko, M.

1986-06-01

A 22 x Wolter microscope was calibrated after several months of operation in the Lawrence Livermore National laboratory (LLNL) Inertial Confinement Fusion program. Placing a point x-ray source at the microscope focus, I recorded the image plane spectrum, as well as the direct spectrum, and from the ratio of these two spectra derived an accurate estimate of the microscope solid angle in the 1 to 4 keV range. The solid angle was also calculated using the microscope geometry and composition. Comparison of this calculated value with the solid angle that was actually measured suggests contamination of the microscope surface

Wolter x-ray microscope calibration

International Nuclear Information System (INIS)

Gerassimenko, M.

1986-01-01

A 22 x Wolter microscope was calibrated after several months of operation in the Lawrence Livermore National Laboratory (LLNL) Inertial Confinement Fusion program. Placing a point x-ray source at the microscope focus, I recorded the image plane spectrum, as well as the direct spectrum, and from the ratio of these two spectra derived an accurate estimate of the microscope solid angle in the 1-4 keV range. The solid angle was also calculated using the microscope geometry and composition. Comparison of this calculated value with the solid angle that was actually measured suggests contamination of the microscope surface

Recent progress in the microscopic description of small and large amplitude collective motion

Energy Technology Data Exchange (ETDEWEB)

Lacroix, D., E-mail: lacroix@ipno.in2p3.fr; Tanimura, Y. [Institut de Physique Nucléaire, IN2P3-CNRS, Université Paris-Sud, F-91406 Orsay Cedex (France); Ayik, S. [Physics Department, Tennessee Technological University, Cookeville, Tennessee 38505 (United States); Scamps, G. [Department of Physics, Tohoku University, Sendai 980-8578 (Japan); Simenel, C. [Department of Nuclear Physics, Research School of Physics and Engineering, Australian National University, Canberra, Australian Capital Territory 2601 (Australia); Yilmaz, B. [Physics Department, Faculty of Sciences, Ankara University, 06100, Ankara (Turkey)

2015-10-15

Dynamical mean-field theory has recently attracted much interests to provide a unified framework for the description of many aspects of nuclear dynamics [1, 2, 3, 4, 5] (for recent reviews see [6, 7]). In particular, the inclusion of pairing correlation has opened new perspectives [8, 9, 10, 11, 12]. A summary of recent applications including giant resonances and transfer reactions will be made in this talk [13, 14, 15, 16]. While new progresses have been made with the use of sophisticated effective interactions and the development of symmetry unrestricted applications, mean-field dynamics suffer from the poor treatment of quantum fluctuations in collective space. As a consequence, these theories are successful in describing average properties of many different experimental observations but generally fail to account realistically for the width of experimental distribution. The increase of predictive power of dynamical mean-field theory is facing the difficulty of going beyond the independent particle or quasi-particle picture. Nevertheless, in the last decade, novel methods have been proposed to prepare the next generation of microscopic mean-field codes able to account for both average properties and fluctuations around the average. A review of recent progresses in this direction as well as recent applications to heavy-ion collisions will be given [17, 18].

Haemozoin Detection in Mouse Liver Histology Using Simple Polarized Light Microscope

Directory of Open Access Journals (Sweden)

DWI RAMADHANI

2014-03-01

Full Text Available The presence of malarial pigment (haemozoin due to Plasmodium infection is a common histopathological effect in mouse liver. Previous research showed that by using a polarized light microscope, researchers were better able to detect haemozoin in mouse liver histology section. Thus, the aim of this research was to compare the haemozoin area observed by a conventional vs. simple polarized light microscope by using image processing analysis. A total of 40 images produced from both conventional light microscope and simple polarized light microscope were collected. All images were analyzed using ImageJ 1.47 software to measure the haemozoin areas. Our results showed that non birefringent haemozoin and birefringent haemozoin area was significantly different. This was because when using conventional light microscope the brown area that contained images of non birefringent haemozoin images also contained Kupffer cells which appeared as the same brown color as haemozoin. In contrast, haemozoin gave bright effect and can be easily differentiated with Kupffer cells in the birefringent haemozoin images. This study concluded that haemozoin detection in mouse liver histology using a simple polarized light microscope was more accurate compared to that of conventional light microscope.

Image transfer with spatial coherence for aberration corrected transmission electron microscopes

International Nuclear Information System (INIS)

Hosokawa, Fumio; Sawada, Hidetaka; Shinkawa, Takao; Sannomiya, Takumi

2016-01-01

The formula of spatial coherence involving an aberration up to six-fold astigmatism is derived for aberration-corrected transmission electron microscopy. Transfer functions for linear imaging are calculated using the newly derived formula with several residual aberrations. Depending on the symmetry and origin of an aberration, the calculated transfer function shows characteristic symmetries. The aberrations that originate from the field’s components, having uniformity along the z direction, namely, the n-fold astigmatism, show rotational symmetric damping of the coherence. The aberrations that originate from the field’s derivatives with respect to z, such as coma, star, and three lobe, show non-rotational symmetric damping. It is confirmed that the odd-symmetric wave aberrations have influences on the attenuation of an image via spatial coherence. Examples of image simulations of haemoglobin and Si [211] are shown by using the spatial coherence for an aberration-corrected electron microscope. - Highlights: • The formula of partial coherence for aberration corrected TEM is derived. • Transfer functions are calculated with several residual aberrations. • The calculated transfer function shows the characteristic damping. • The odd-symmetric wave aberrations can cause the attenuation of image via coherence. • The examples of aberration corrected TEM image simulations are shown.

Image transfer with spatial coherence for aberration corrected transmission electron microscopes

Energy Technology Data Exchange (ETDEWEB)

Hosokawa, Fumio, E-mail: hosokawa@bio-net.co.jp [BioNet Ltd., 2-3-28 Nishikityo, Tachikwa, Tokyo (Japan); Tokyo Institute of Technology, 4259 Nagatsuta, Midoriku, Yokohama 226-8503 (Japan); Sawada, Hidetaka [JEOL (UK) Ltd., JEOL House, Silver Court, Watchmead, Welwyn Garden City, Herts AL7 1LT (United Kingdom); Shinkawa, Takao [BioNet Ltd., 2-3-28 Nishikityo, Tachikwa, Tokyo (Japan); Sannomiya, Takumi [Tokyo Institute of Technology, 4259 Nagatsuta, Midoriku, Yokohama 226-8503 (Japan)

2016-08-15

The formula of spatial coherence involving an aberration up to six-fold astigmatism is derived for aberration-corrected transmission electron microscopy. Transfer functions for linear imaging are calculated using the newly derived formula with several residual aberrations. Depending on the symmetry and origin of an aberration, the calculated transfer function shows characteristic symmetries. The aberrations that originate from the field’s components, having uniformity along the z direction, namely, the n-fold astigmatism, show rotational symmetric damping of the coherence. The aberrations that originate from the field’s derivatives with respect to z, such as coma, star, and three lobe, show non-rotational symmetric damping. It is confirmed that the odd-symmetric wave aberrations have influences on the attenuation of an image via spatial coherence. Examples of image simulations of haemoglobin and Si [211] are shown by using the spatial coherence for an aberration-corrected electron microscope. - Highlights: • The formula of partial coherence for aberration corrected TEM is derived. • Transfer functions are calculated with several residual aberrations. • The calculated transfer function shows the characteristic damping. • The odd-symmetric wave aberrations can cause the attenuation of image via coherence. • The examples of aberration corrected TEM image simulations are shown.

Simultaneous topography imaging and broadband nanomechanical mapping on atomic force microscope

Science.gov (United States)

Li, Tianwei; Zou, Qingze

2017-12-01

In this paper, an approach is proposed to achieve simultaneous imaging and broadband nanomechanical mapping of soft materials in air by using an atomic force microscope. Simultaneous imaging and nanomechanical mapping are needed, for example, to correlate the morphological and mechanical evolutions of the sample during dynamic phenomena such as the cell endocytosis process. Current techniques for nanomechanical mapping, however, are only capable of capturing static elasticity of the material, or the material viscoelasticity in a narrow frequency band around the resonant frequency(ies) of the cantilever used, not competent for broadband nanomechanical mapping, nor acquiring topography image of the sample simultaneously. These limitations are addressed in this work by enabling the augmentation of an excitation force stimuli of rich frequency spectrum for nanomechanical mapping in the imaging process. Kalman-filtering technique is exploited to decouple and split the mixed signals for imaging and mapping, respectively. Then the sample indentation generated is quantified online via a system-inversion method, and the effects of the indentation generated and the topography tracking error on the topography quantification are taken into account. Moreover, a data-driven feedforward-feedback control is utilized to track the sample topography. The proposed approach is illustrated through experimental implementation on a polydimethylsiloxane sample with a pre-fabricated pattern.

Red Blood Cell Count Automation Using Microscopic Hyperspectral Imaging Technology.

Science.gov (United States)

Li, Qingli; Zhou, Mei; Liu, Hongying; Wang, Yiting; Guo, Fangmin

2015-12-01

Red blood cell counts have been proven to be one of the most frequently performed blood tests and are valuable for early diagnosis of some diseases. This paper describes an automated red blood cell counting method based on microscopic hyperspectral imaging technology. Unlike the light microscopy-based red blood count methods, a combined spatial and spectral algorithm is proposed to identify red blood cells by integrating active contour models and automated two-dimensional k-means with spectral angle mapper algorithm. Experimental results show that the proposed algorithm has better performance than spatial based algorithm because the new algorithm can jointly use the spatial and spectral information of blood cells.

Chemical imaging of cotton fibers using an infrared microscope and a focal-plane array detector

Science.gov (United States)

In this presentation, the chemical imaging of cotton fibers with an infrared microscope and a Focal-Plane Array (FPA) detector will be discussed. Infrared spectroscopy can provide us with information on the structure and quality of cotton fibers. In addition, FPA detectors allow for simultaneous spe...

Compact plane illumination plugin device to enable light sheet fluorescence imaging of multi-cellular organisms on an inverted wide-field microscope.

Science.gov (United States)

Guan, Zeyi; Lee, Juhyun; Jiang, Hao; Dong, Siyan; Jen, Nelson; Hsiai, Tzung; Ho, Chih-Ming; Fei, Peng

2016-01-01

We developed a compact plane illumination plugin (PIP) device which enabled plane illumination and light sheet fluorescence imaging on a conventional inverted microscope. The PIP device allowed the integration of microscope with tunable laser sheet profile, fast image acquisition, and 3-D scanning. The device is both compact, measuring approximately 15 by 5 by 5 cm, and cost-effective, since we employed consumer electronics and an inexpensive device molding method. We demonstrated that PIP provided significant contrast and resolution enhancement to conventional microscopy through imaging different multi-cellular fluorescent structures, including 3-D branched cells in vitro and live zebrafish embryos. Imaging with the integration of PIP greatly reduced out-of-focus contamination and generated sharper contrast in acquired 2-D plane images when compared with the stand-alone inverted microscope. As a result, the dynamic fluid domain of the beating zebrafish heart was clearly segmented and the functional monitoring of the heart was achieved. Furthermore, the enhanced axial resolution established by thin plane illumination of PIP enabled the 3-D reconstruction of the branched cellular structures, which leads to the improvement on the functionality of the wide field microscopy.

Microscopic and macroscopic models for the onset and progression of Alzheimer's disease

Science.gov (United States)

Bertsch, Michiel; Franchi, Bruno; Carla Tesi, Maria; Tosin, Andrea

2017-10-01

In the first part of this paper we review a mathematical model for the onset and progression of Alzheimer’s disease (AD) that was developed in subsequent steps over several years. The model is meant to describe the evolution of AD in vivo. In Achdou et al (2013 J. Math. Biol. 67 1369-92) we treated the problem at a microscopic scale, where the typical length scale is a multiple of the size of the soma of a single neuron. Subsequently, in Bertsch et al (2017 Math. Med. Biol. 34 193-214) we concentrated on the macroscopic scale, where brain neurons are regarded as a continuous medium, structured by their degree of malfunctioning. In the second part of the paper we consider the relation between the microscopic and the macroscopic models. In particular we show under which assumptions the kinetic transport equation, which in the macroscopic model governs the evolution of the probability measure for the degree of malfunctioning of neurons, can be derived from a particle-based setting. The models are based on aggregation and diffusion equations for β-Amyloid (Aβ from now on), a protein fragment that healthy brains regularly produce and eliminate. In case of dementia Aβ monomers are no longer properly washed out and begin to coalesce forming eventually plaques. Two different mechanisms are assumed to be relevant for the temporal evolution of the disease: (i) diffusion and agglomeration of soluble polymers of amyloid, produced by damaged neurons; (ii) neuron-to-neuron prion-like transmission. In the microscopic model we consider mechanism (i), modelling it by a system of Smoluchowski equations for the amyloid concentration (describing the agglomeration phenomenon), with the addition of a diffusion term as well as of a source term on the neuronal membrane. At the macroscopic level instead we model processes (i) and (ii) by a system of Smoluchowski equations for the amyloid concentration, coupled to a kinetic-type transport equation for the distribution function of the

Robotic autopositioning of the operating microscope.

Science.gov (United States)

Oppenlander, Mark E; Chowdhry, Shakeel A; Merkl, Brandon; Hattendorf, Guido M; Nakaji, Peter; Spetzler, Robert F

2014-06-01

Use of the operating microscope has become pervasive since its introduction to the neurosurgical world. Neuronavigation fused with the operating microscope has allowed accurate correlation of the focal point of the microscope and its location on the downloaded imaging study. However, the robotic ability of the Pentero microscope has not been utilized to orient the angle of the microscope or to change its focal length to hone in on a predefined target. To report a novel technology that allows automatic positioning of the operating microscope onto a set target and utilization of a planned trajectory, either determined with the StealthStation S7 by using preoperative imaging or intraoperatively with the microscope. By utilizing the current motorized capabilities of the Zeiss OPMI Pentero microscope, a robotic autopositioning feature was developed in collaboration with Surgical Technologies, Medtronic, Inc. (StealthStation S7). The system is currently being tested at the Barrow Neurological Institute. Three options were developed for automatically positioning the microscope: AutoLock Current Point, Align Parallel to Plan, and Point to Plan Target. These options allow the microscope to pivot around the lesion, hover in a set plane parallel to the determined trajectory, or rotate and point to a set target point, respectively. Integration of automatic microscope positioning into the operative workflow has potential to increase operative efficacy and safety. This technology is best suited for precise trajectories and entry points into deep-seated lesions.

Evaluation of an X-ray-excited optical microscope for chemical imaging of metal and other surfaces.

Science.gov (United States)

Sabbe, Pieter-Jan; Dowsett, Mark; Hand, Matthew; Grayburn, Rosie; Thompson, Paul; Bras, Wim; Adriaens, Annemie

2014-12-02

The application of a modular system for the nondestructive chemical imaging of metal and other surfaces is described using heritage metals as an example. The custom-built X-ray-excited optical luminescence (XEOL) microscope, XEOM 1, images the chemical state and short-range atomic order of the top 200 nm of both amorphous and crystalline surfaces. A broad X-ray beam is used to illuminate large areas (up to 4 mm(2)) of the sample, and the resulting XEOL emission is collected simultaneously for each pixel by a charge-coupled device sensor to form an image. The input X-ray energy is incremented across a range typical for the X-ray absorption near-edge structure (XANES) and an image collected for each increment. The use of large-footprint beams combined with parallel detection allows the power density to be kept low and facilitates complete nondestructive XANES mapping on a reasonable time scale. In this study the microscope was evaluated by imaging copper surfaces with well-defined patterns of different corrosion products (cuprite Cu2O and nantokite CuCl). The images obtained show chemical contrast, and filtering the XEOL light allowed different corrosion products to be imaged separately. Absorption spectra extracted from software-selected regions of interest exhibit characteristic XANES fingerprints for the compounds present. Moreover, when the X-ray absorption edge positions were extracted from each spectrum, an oxidation state map of the sample could be compiled. The results show that this method allows one to obtain nondestructive and noninvasive information at the micrometer scale while using full-field imaging.

as-PSOCT: Volumetric microscopic imaging of human brain architecture and connectivity.

Science.gov (United States)

Wang, Hui; Magnain, Caroline; Wang, Ruopeng; Dubb, Jay; Varjabedian, Ani; Tirrell, Lee S; Stevens, Allison; Augustinack, Jean C; Konukoglu, Ender; Aganj, Iman; Frosch, Matthew P; Schmahmann, Jeremy D; Fischl, Bruce; Boas, David A

2018-01-15

Polarization sensitive optical coherence tomography (PSOCT) with serial sectioning has enabled the investigation of 3D structures in mouse and human brain tissue samples. By using intrinsic optical properties of back-scattering and birefringence, PSOCT reliably images cytoarchitecture, myeloarchitecture and fiber orientations. In this study, we developed a fully automatic serial sectioning polarization sensitive optical coherence tomography (as-PSOCT) system to enable volumetric reconstruction of human brain samples with unprecedented sample size and resolution. The 3.5 μm in-plane resolution and 50 μm through-plane voxel size allow inspection of cortical layers that are a single-cell in width, as well as small crossing fibers. We show the abilities of as-PSOCT in quantifying layer thicknesses of the cerebellar cortex and creating microscopic tractography of intricate fiber networks in the subcortical nuclei and internal capsule regions, all based on volumetric reconstructions. as-PSOCT provides a viable tool for studying quantitative cytoarchitecture and myeloarchitecture and mapping connectivity with microscopic resolution in the human brain. Copyright © 2017 Elsevier Inc. All rights reserved.

High-resolution imaging of ultracold fermions in microscopically tailored optical potentials

International Nuclear Information System (INIS)

Zimmermann, B; Mueller, T; Meineke, J; Esslinger, T; Moritz, H

2011-01-01

We report on the local probing and preparation of an ultracold Fermi gas on the length scale of one micrometer, i.e. of the order of the Fermi wavelength. The essential tool of our experimental setup is a pair of identical, high-resolution microscope objectives. One of the microscope objectives allows local imaging of the trapped Fermi gas of 6 Li atoms with a maximum resolution of 660 nm, while the other enables the generation of arbitrary optical dipole potentials on the same length scale. Employing a two-dimensional (2D) acousto-optical deflector, we demonstrate the formation of several trapping geometries, including a tightly focused single optical dipole trap, a 4x4 site 2D optical lattice and an 8 site ring lattice configuration. Furthermore, we show the ability to load and detect a small number of atoms in these trapping potentials. A site separation down to one micrometer in combination with the low mass of 6 Li results in tunneling rates that are sufficiently large for the implementation of Hubbard models with the designed geometries.

Study on mechanical properties and damage behaviors of Kevlar fiber reinforced epoxy composites by digital image correlation technique under optical microscope

Science.gov (United States)

Gao, Xiang; Shao, Wenquan; Ji, Hongwei

2010-10-01

Kevlar fiber-reinforced epoxy (KFRE) composites are widely used in the fields of aerospace, weapon, shipping, and civil industry, due to their outstanding capabilities. In this paper, mechanical properties and damage behaviors of KFRE laminate (02/902) were tested and studied under tension condition. To precisely measure the tensile mechanical properties of the material and investigate its micro-scale damage evolution, a micro-image measuring system with in-situ tensile device was designed. The measuring system, by which the in-situ tensile test can be carried out and surface morphology evolution of the tensile specimen can be visually monitored and recorded during the process of loading, includes an ultra-long working distance zoom microscope and a in-situ tensile loading device. In this study, a digital image correlation method (DICM) was used to calculate the deformation of the tensile specimen under different load levels according to the temporal series images captured by an optical microscope and CCD camera. Then, the elastic modulus and Poisson's ratio of the KFRE was obtained accordingly. The damage progresses of the KFRE laminates were analyzed. Experimental results indicated that: (1) the KFRE laminate (02/902) is almost elastic, its failure mode is brittle tensile fracture.(2) Mechanical properties parameters of the material are as follows: elastic modulus is 14- 16GPa, and tensile ultimate stress is 450-480 Mpa respectively. (3) The damage evolution of the material is that cracks appear in epoxy matrix firstly, then, with the increasing of the tensile loading, matrix cracks add up and extend along a 45° angle direction with tensile load. Furthermore, decohesion between matrix and fibers as well as delamination occurs. Eventually, fibers break and the material is damaged.

In vivo microscopic imaging of the bronchial mucosa using an endo-cytoscopy system.

Science.gov (United States)

Shibuya, Kiyoshi; Fujiwara, Taiki; Yasufuku, Kazuhiro; Alaa, Mohamed; Chiyo, Masako; Nakajima, Takahiro; Hoshino, Hidehisa; Hiroshima, Kenzo; Nakatani, Yukio; Yoshino, Ichiro

2011-05-01

We investigated the capabilities of an endo-cytoscopy system (ECS) that enables microscopic imaging of the tracheobronchial tree during bronchoscopy, including normal bronchial epithelium, dysplastic mucosa and squamous cell carcinoma. The newly developed ECS has a 3.2 mm diameter that can be passed through the 4.2 mm working channel of a mother endoscope for insertion of the ECS. It has a high magnification of 570× on a 17 in. video monitor. Twenty-two patients (7 squamous cell carcinoma, 11 squamous dysplasia and 4 after PDT therapies) were underwent white light, NBI light and AFI bronchoscopy. Both abnormal areas of interest and normal bronchial mucosa were stained with 0.5% methylene blue and examined with ECS at high magnification (570×). Histological examinations using haematoxylin and eosin staining were made of biopsied specimens. Analyzed ECS images were compared with the corresponding histological examinations. In normal bronchial mucosa, ciliated columnar epithelial cells were visible. In bronchial squamous dysplasia, superficial cells with abundant cytoplasm were arranged regularly. In squamous cell carcinoma, large, polymorphic tumor cells showed increased cellular densities with irregular stratified patterns. These ECS images corresponded well with the light-microscopic examination of conventional histology. ECS was useful for the discrimination between normal bronchial epithelial cells and dysplastic cells or malignant cells during bronchoscopy in real time. This novel technology has an excellent potential to provide in vivo diagnosis during bronchoscopic examinations. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

Nanoscale imaging of the growth and division of bacterial cells on planar substrates with the atomic force microscope

Energy Technology Data Exchange (ETDEWEB)

Van Der Hofstadt, M. [Institut de Bioenginyeria de Catalunya (IBEC), C/ Baldiri i Reixac 11-15, 08028 Barcelona (Spain); Hüttener, M.; Juárez, A. [Institut de Bioenginyeria de Catalunya (IBEC), C/ Baldiri i Reixac 11-15, 08028 Barcelona (Spain); Departament de Microbiologia, Universitat de Barcelona, Avinguda Diagonal 645, 08028 Barcelona (Spain); Gomila, G., E-mail: ggomila@ibecbarcelona.eu [Institut de Bioenginyeria de Catalunya (IBEC), C/ Baldiri i Reixac 11-15, 08028 Barcelona (Spain); Departament d' Electronica, Universitat de Barcelona, C/ Marti i Franqués 1, 08028 Barcelona (Spain)

2015-07-15

With the use of the atomic force microscope (AFM), the Nanomicrobiology field has advanced drastically. Due to the complexity of imaging living bacterial processes in their natural growing environments, improvements have come to a standstill. Here we show the in situ nanoscale imaging of the growth and division of single bacterial cells on planar substrates with the atomic force microscope. To achieve this, we minimized the lateral shear forces responsible for the detachment of weakly adsorbed bacteria on planar substrates with the use of the so called dynamic jumping mode with very soft cantilever probes. With this approach, gentle imaging conditions can be maintained for long periods of time, enabling the continuous imaging of the bacterial cell growth and division, even on planar substrates. Present results offer the possibility to observe living processes of untrapped bacteria weakly attached to planar substrates. - Highlights: • Gelatine coatings used to weakly attach bacterial cells onto planar substrates. • Use of the dynamic jumping mode as a non-perturbing bacterial imaging mode. • Nanoscale resolution imaging of unperturbed single living bacterial cells. • Growth and division of single bacteria cells on planar substrates observed.

A quadruple-scanning-probe force microscope for electrical property measurements of microscopic materials

International Nuclear Information System (INIS)

Higuchi, Seiji; Kubo, Osamu; Kuramochi, Hiromi; Aono, Masakazu; Nakayama, Tomonobu

2011-01-01

Four-terminal electrical measurement is realized on a microscopic structure in air, without a lithographic process, using a home-built quadruple-scanning-probe force microscope (QSPFM). The QSPFM has four probes whose positions are individually controlled by obtaining images of a sample in the manner of atomic force microscopy (AFM), and uses the probes as contacting electrodes for electrical measurements. A specially arranged tuning fork probe (TFP) is used as a self-detection force sensor to operate each probe in a frequency modulation AFM mode, resulting in simultaneous imaging of the same microscopic feature on an insulator using the four TFPs. Four-terminal electrical measurement is then demonstrated in air by placing each probe electrode in contact with a graphene flake exfoliated on a silicon dioxide film, and the sheet resistance of the flake is measured by the van der Pauw method. The present work shows that the QSPFM has the potential to measure the intrinsic electrical properties of a wide range of microscopic materials in situ without electrode fabrication.

Trace Element Mapping of a Biological Specimen by a Full-Field X-ray Fluorescence Imaging Microscope with a Wolter Mirror

International Nuclear Information System (INIS)

Hoshino, Masato; Yamada, Norimitsu; Ishino, Toyoaki; Namiki, Takashi; Watanabe, Norio; Aoki, Sadao

2007-01-01

A full-field X-ray fluorescence imaging microscope with a Wolter mirror was applied to the element mapping of alfalfa seeds. The X-ray fluorescence microscope was built at the Photon Factory BL3C2 (KEK). X-ray fluorescence images of several growing stages of the alfalfa seeds were obtained. X-ray fluorescence energy spectra were measured with either a solid state detector or a CCD photon counting method. The element distributions of iron and zinc which were included in the seeds were obtained using a photon counting method

Ordering of diagnostic information in encoded medical images. Accuracy progression

Science.gov (United States)

Przelaskowski, A.; Jóźwiak, R.; Krzyżewski, T.; Wróblewska, A.

2008-03-01

A concept of diagnostic accuracy progression for embedded coding of medical images was presented. Implementation of JPEG2000 encoder with a modified PCRD optimization algorithm was realized and initially verified as a tool for accurate medical image streaming. Mean square error as a distortion measure was replaced by other numerical measures to revise quality progression according to diagnostic importance of successively encoded image information. A faster increment of image diagnostic importance during reconstruction of initial packets of code stream was reached. Modified Jasper code was initially tested on a set of mammograms containing clusters of microcalcifications and malignant masses, and other radiograms. Teleradiologic applications were considered as the first area of interests.

Aiming of Kirkpatrick-Baez microscope based on auxiliary optical system

International Nuclear Information System (INIS)

Huang Shengling; Mu Baozhong; Yi Shengzhen; Wang Xin; Wang Zhanshan; Ding Yongkun; Miao Wenyong; Dong Jianjun

2009-01-01

An auxiliary optical system has been designed, which can provide precise positioning for aiming Kirkpatrick-Baez (KB) microscope object location. An 8 keV X-ray imaging system by KB microscope with periodic multilayer films has been designed. The field of view and depth of field in the resolution of 5 μm are got, and then the corresponding point and depth of field in diagnostic experiments are calculated. Based on the object-image relations and precision of the KB microscope, an auxiliary visible light imaging system is designed and X-ray imaging experiments are performed, which can achieve equivalent aiming between the visible imaging system and the KB microscope. The results show that ±20 μm vertical axis plane and ±300 μm axial accuracy are achieved through the auxiliary optical path, which can meet the object point positioning requirements of the KB microscope. (authors)

Environmental TEM in an Aberration Corrected Microscope

DEFF Research Database (Denmark)

Hansen, Thomas Willum; Wagner, Jakob Birkedal

‐resolution imaging. A gaseous atmosphere in the pole‐piece gap of the objective lens of the microscope alters both the incoming electron wave prior to interaction with the sample and the outgoing wave below the sample. Whereas conventional TEM samples are usually thin (below 10‐20 nm), the gas in the environmental...... the microscope column. The effects of gas on the electron wave in the objective lens are not well understood and needs further attention. Imaging samples with a simple geometry, such as gold particles on a flat graphene substrate and analyzing the variations in contrast, provides a means for understanding...... results from imaging in various elemental as well as di‐molecular gases and their effect on imaging and spectroscopy in the environmental transmission electron microscope....

A fourth order PDE based fuzzy c- means approach for segmentation of microscopic biopsy images in presence of Poisson noise for cancer detection.

Science.gov (United States)

Kumar, Rajesh; Srivastava, Subodh; Srivastava, Rajeev

2017-07-01

For cancer detection from microscopic biopsy images, image segmentation step used for segmentation of cells and nuclei play an important role. Accuracy of segmentation approach dominate the final results. Also the microscopic biopsy images have intrinsic Poisson noise and if it is present in the image the segmentation results may not be accurate. The objective is to propose an efficient fuzzy c-means based segmentation approach which can also handle the noise present in the image during the segmentation process itself i.e. noise removal and segmentation is combined in one step. To address the above issues, in this paper a fourth order partial differential equation (FPDE) based nonlinear filter adapted to Poisson noise with fuzzy c-means segmentation method is proposed. This approach is capable of effectively handling the segmentation problem of blocky artifacts while achieving good tradeoff between Poisson noise removals and edge preservation of the microscopic biopsy images during segmentation process for cancer detection from cells. The proposed approach is tested on breast cancer microscopic biopsy data set with region of interest (ROI) segmented ground truth images. The microscopic biopsy data set contains 31 benign and 27 malignant images of size 896 × 768. The region of interest selected ground truth of all 58 images are also available for this data set. Finally, the result obtained from proposed approach is compared with the results of popular segmentation algorithms; fuzzy c-means, color k-means, texture based segmentation, and total variation fuzzy c-means approaches. The experimental results shows that proposed approach is providing better results in terms of various performance measures such as Jaccard coefficient, dice index, Tanimoto coefficient, area under curve, accuracy, true positive rate, true negative rate, false positive rate, false negative rate, random index, global consistency error, and variance of information as compared to other

Chip-scale fluorescence microscope based on a silo-filter complementary metal-oxide semiconductor image sensor.

Science.gov (United States)

Ah Lee, Seung; Ou, Xiaoze; Lee, J Eugene; Yang, Changhuei

2013-06-01

We demonstrate a silo-filter (SF) complementary metal-oxide semiconductor (CMOS) image sensor for a chip-scale fluorescence microscope. The extruded pixel design with metal walls between neighboring pixels guides fluorescence emission through the thick absorptive filter to the photodiode of a pixel. Our prototype device achieves 13 μm resolution over a wide field of view (4.8 mm × 4.4 mm). We demonstrate bright-field and fluorescence longitudinal imaging of living cells in a compact, low-cost configuration.

Cancer cell imaging by stable wet near-field scanning optical microscope with resonance tracking method

International Nuclear Information System (INIS)

Park, Kyoung-Duck; Park, Doo-Jae; Jeong, Mun-Seok; Choi, Geun-Chang; Lee, Seung-Gol; Byeon, Clare-Chisu; Choi, Soo-Bong

2014-01-01

We report on a successful topographical and optical imaging of various cancer cells in liquid and in air by using a stable wet near-field scanning optical microscope that utilizes a resonance tracking method. We observed a clear dehydration which gives rise to a decrease in the cell volume down to 51%. In addition, a micro-ball lens effect due to the round-shaped young cancer cells was observed from near-field imaging, where the refractive index of young cancer cells was deduced.

Cancer cell imaging by stable wet near-field scanning optical microscope with resonance tracking method

Energy Technology Data Exchange (ETDEWEB)

Park, Kyoung-Duck [Sungkyunkwan University, Suwon (Korea, Republic of); Inha University, Incheon (Korea, Republic of); Park, Doo-Jae; Jeong, Mun-Seok [Sungkyunkwan University, Suwon (Korea, Republic of); Choi, Geun-Chang [Seoul National University, Seoul (Korea, Republic of); Lee, Seung-Gol [Inha University, Incheon (Korea, Republic of); Byeon, Clare-Chisu [Kyungpook National University, Daegu (Korea, Republic of); Choi, Soo-Bong [Incheon National University, Incheon (Korea, Republic of)

2014-05-15

We report on a successful topographical and optical imaging of various cancer cells in liquid and in air by using a stable wet near-field scanning optical microscope that utilizes a resonance tracking method. We observed a clear dehydration which gives rise to a decrease in the cell volume down to 51%. In addition, a micro-ball lens effect due to the round-shaped young cancer cells was observed from near-field imaging, where the refractive index of young cancer cells was deduced.

Miniaturized integration of a fluorescence microscope

Science.gov (United States)

Ghosh, Kunal K.; Burns, Laurie D.; Cocker, Eric D.; Nimmerjahn, Axel; Ziv, Yaniv; Gamal, Abbas El; Schnitzer, Mark J.

2013-01-01

The light microscope is traditionally an instrument of substantial size and expense. Its miniaturized integration would enable many new applications based on mass-producible, tiny microscopes. Key prospective usages include brain imaging in behaving animals towards relating cellular dynamics to animal behavior. Here we introduce a miniature (1.9 g) integrated fluorescence microscope made from mass-producible parts, including semiconductor light source and sensor. This device enables high-speed cellular-level imaging across ∼0.5 mm2 areas in active mice. This capability allowed concurrent tracking of Ca2+ spiking in >200 Purkinje neurons across nine cerebellar microzones. During mouse locomotion, individual microzones exhibited large-scale, synchronized Ca2+ spiking. This is a mesoscopic neural dynamic missed by prior techniques for studying the brain at other length scales. Overall, the integrated microscope is a potentially transformative technology that permits distribution to many animals and enables diverse usages, such as portable diagnostics or microscope arrays for large-scale screens. PMID:21909102

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

Science.gov (United States)

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

2014-06-01

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

MIDAS - an atomic force microscope for in-situ imaging of cometary dust particles

International Nuclear Information System (INIS)

Fehringer, H.M.; Ruedenauer, F.G.; Steiger, W.

1997-02-01

Comets are interesting bodies, since they are considered to consist of matter remaining in essentially unchanged chemistry from the presolar nebula. Investigation of cometary matter therefore permits to draw conclusion s with respect to the composition of presolar matter. The atomic force microscope MIDAS will be the first instrument to analyze, within ESA's ROSETTA-mission priestine cometary matter in the form of dust particles emitted by comet WIRTANEN during its perihelion in 2013. Within this project, a dust model has been developed, permitting estimation of dust collection times required for statistically significant imaging of cometary particles. The dynamics of dust collection has been developed and experimental dust collection surfaces have been produced making use of modem nanostructuring techniques. Mechanical properties of 3-dimensional piezo-control elements, which are an essential part of the MIDAS microscope, have been determined. (author)

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

Energy Technology Data Exchange (ETDEWEB)

Baumbach, S., E-mail: baumbach@rheinahrcampus.de; Wilhein, T. [Institute for X-Optics, University of Applied Sciences Koblenz, RheinAhrCampus Remagen, Joseph-Rovan-Allee 2, D-53424 Remagen (Germany); Kanngießer, B.; Malzer, W. [Institute for Optics and Atomic Physics, Technical University of Berlin, Hardenbergstrasse 36, D-10623 Berlin (Germany); Stiel, H. [Max-Born-Institute, Max-Born-Strasse 2A, D-12489 Berlin (Germany)

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 detector limited spatial resolution of 210 nm is measured on x-ray images of Siemens star test patterns.

MRT letter: Guided filtering of image focus volume for 3D shape recovery of microscopic objects.

Science.gov (United States)

Mahmood, Muhammad Tariq

2014-12-01

In this letter, a shape from focus (SFF) method is proposed that utilizes the guided image filtering to enhance the image focus volume efficiently. First, image focus volume is computed using a conventional focus measure. Then each layer of image focus volume is filtered using guided filtering. In this work, the all-in-focus image, which can be obtained from the initial focus volume, is used as guidance image. Finally, improved depth map is obtained from the filtered image focus volume by maximizing the focus measure along the optical axis. The proposed SFF method is efficient and provides better depth maps. The improved performance is highlighted by conducting several experiments using image sequences of simulated and real microscopic objects. The comparative analysis demonstrates the effectiveness of the proposed SFF method. © 2014 Wiley Periodicals, Inc.

A mini-microscope for in situ monitoring of cells.

Science.gov (United States)

Kim, Sang Bok; Koo, Kyo-in; Bae, Hojae; Dokmeci, Mehmet R; Hamilton, Geraldine A; Bahinski, Anthony; Kim, Sun Min; Ingber, Donald E; Khademhosseini, Ali

2012-10-21

A mini-microscope was developed for in situ monitoring of cells by modifying off-the-shelf components of a commercial webcam. The mini-microscope consists of a CMOS imaging module, a small plastic lens and a white LED illumination source. The CMOS imaging module was connected to a laptop computer through a USB port for image acquisition and analysis. Due to its compact size, 8 × 10 × 9 cm, the present microscope is portable and can easily fit inside a conventional incubator, and enables real-time monitoring of cellular behaviour. Moreover, the mini-microscope can be used for imaging cells in conventional cell culture flasks, such as Petri dishes and multi-well plates. To demonstrate the operation of the mini-microscope, we monitored the cellular migration of mouse 3T3 fibroblasts in a scratch assay in medium containing three different concentrations of fetal bovine serum (5, 10, and 20%) and demonstrated differential responses depending on serum levels. In addition, we seeded embryonic stem cells inside poly(ethylene glycol) microwells and monitored the formation of stem cell aggregates in real time using the mini-microscope. Furthermore, we also combined a lab-on-a-chip microfluidic device for microdroplet generation and analysis with the mini-microscope and observed the formation of droplets under different flow conditions. Given its cost effectiveness, robust imaging and portability, the presented platform may be useful for a range of applications for real-time cellular imaging using lab-on-a-chip devices at low cost.

Scanless nonlinear optical microscope for image reconstruction and space-time correlation analysis

Science.gov (United States)

Ceffa, N. G.; Radaelli, F.; Pozzi, P.; Collini, M.; Sironi, L.; D'alfonso, L.; Chirico, G.

2017-06-01

Optical Microscopy has been applied to life science from its birth and reached widespread application due to its major advantages: limited perturbation of the biological tissue and the easy accessibility of the light sources. However, as the spatial and time resolution requirements and the time stability of the microscopes increase, researchers are struggling against some of its limitations: limited transparency and the refractivity of the living tissue to light and the field perturbations induced by the path in the tissue. We have developed a compact stand-alone, completely scan-less, optical setup that allows to acquire non-linear excitation images and to measure the sample dynamics simultaneously on an ensemble of arbitrary chosen regions of interests. The image is obtained by shining a square array of spots on the sample obtained by a spatial light modulator and by shifting it (10 ms refresh time) on the sample. The final image is computed from the superposition of (100-1000) images. Filtering procedures can be applied to the raw images of the excitation array before building the image. We discuss results that show how this setup can be used for the correction of wave front aberrations induced by turbid samples (such as living tissues) and for the computation of space-time cross-correlations in complex networks.

Novel low-dose imaging technique for characterizing atomic structures through scanning transmission electron microscope

Science.gov (United States)

Su, Chia-Ping; Syu, Wei-Jhe; Hsiao, Chien-Nan; Lai, Ping-Shan; Chen, Chien-Chun

2017-08-01

To investigate dislocations or heterostructures across interfaces is now of great interest to condensed matter and materials scientists. With the advances in aberration-corrected electron optics, the scanning transmission electron microscope has demonstrated its excellent capability of characterizing atomic structures within nanomaterials, and well-resolved atomic-resolution images can be obtained through long-exposure data acquisition. However, the sample drifting, carbon contamination, and radiation damage hinder further analysis, such as deriving three-dimensional (3D) structures from a series of images. In this study, a method for obtaining atomic-resolution images with significantly reduced exposure time was developed, using which an original high-resolution image with approximately one tenth the electron dose can be obtained by combining a fast-scan high-magnification image and a slow-scan low-magnification image. The feasibility of obtaining 3D atomic structures using the proposed approach was demonstrated through multislice simulation. Finally, the feasibility and accuracy of image restoration were experimentally verified. This general method cannot only apply to electron microscopy but also benefit to image radiation-sensitive materials using various light sources.

21 CFR 864.3600 - Microscopes and accessories.

Science.gov (United States)

2010-04-01

... enlarge images of specimens, preparations, and cultures for medical purposes. Variations of microscopes... light. (3) Inverted stage microscopes, which permit examination of tissue cultures or other biological... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Microscopes and accessories. 864.3600 Section 864...

Atomic force microscope featuring an integrated optical microscope

NARCIS (Netherlands)

Putman, C.A.J.; Putman, Constant A.J.; de Grooth, B.G.; van Hulst, N.F.; Greve, Jan

1992-01-01

The atomic force microscope (AFM) is used to image the surface of both conductors and nonconductors. Biological specimens constitute a large group of nonconductors. A disadvantage of most AFM's is the fact that relatively large areas of the sample surface have to be scanned to pinpoint a biological

[Quantitative data analysis for live imaging of bone.

Science.gov (United States)

Seno, Shigeto

Bone tissue is a hard tissue, it was difficult to observe the interior of the bone tissue alive. With the progress of microscopic technology and fluorescent probe technology in recent years, it becomes possible to observe various activities of various cells forming bone society. On the other hand, the quantitative increase in data and the diversification and complexity of the images makes it difficult to perform quantitative analysis by visual inspection. It has been expected to develop a methodology for processing microscopic images and data analysis. In this article, we introduce the research field of bioimage informatics which is the boundary area of biology and information science, and then outline the basic image processing technology for quantitative analysis of live imaging data of bone.

Science Applications of a Multispectral Microscopic Imager for the Astrobiological Exploration of Mars

Science.gov (United States)

Farmer, Jack D.; Sellar, R. Glenn; Swayze, Gregg A.; Blaney, Diana L.

2014-01-01

Abstract Future astrobiological missions to Mars are likely to emphasize the use of rovers with in situ petrologic capabilities for selecting the best samples at a site for in situ analysis with onboard lab instruments or for caching for potential return to Earth. Such observations are central to an understanding of the potential for past habitable conditions at a site and for identifying samples most likely to harbor fossil biosignatures. The Multispectral Microscopic Imager (MMI) provides multispectral reflectance images of geological samples at the microscale, where each image pixel is composed of a visible/shortwave infrared spectrum ranging from 0.46 to 1.73 μm. This spectral range enables the discrimination of a wide variety of rock-forming minerals, especially Fe-bearing phases, and the detection of hydrated minerals. The MMI advances beyond the capabilities of current microimagers on Mars by extending the spectral range into the infrared and increasing the number of spectral bands. The design employs multispectral light-emitting diodes and an uncooled indium gallium arsenide focal plane array to achieve a very low mass and high reliability. To better understand and demonstrate the capabilities of the MMI for future surface missions to Mars, we analyzed samples from Mars-relevant analog environments with the MMI. Results indicate that the MMI images faithfully resolve the fine-scale microtextural features of samples and provide important information to help constrain mineral composition. The use of spectral endmember mapping reveals the distribution of Fe-bearing minerals (including silicates and oxides) with high fidelity, along with the presence of hydrated minerals. MMI-based petrogenetic interpretations compare favorably with laboratory-based analyses, revealing the value of the MMI for future in situ rover-mediated astrobiological exploration of Mars. Key Words: Mars—Microscopic imager—Multispectral imaging�

Foldscope: origami-based paper microscope.

Directory of Open Access Journals (Sweden)

James S Cybulski

Full Text Available Here we describe an ultra-low-cost origami-based approach for large-scale manufacturing of microscopes, specifically demonstrating brightfield, darkfield, and fluorescence microscopes. Merging principles of optical design with origami enables high-volume fabrication of microscopes from 2D media. Flexure mechanisms created via folding enable a flat compact design. Structural loops in folded paper provide kinematic constraints as a means for passive self-alignment. This light, rugged instrument can survive harsh field conditions while providing a diversity of imaging capabilities, thus serving wide-ranging applications for cost-effective, portable microscopes in science and education.

Magnetic resonance dacryocystography: comparison between conventional surface coils and microscopic coils

International Nuclear Information System (INIS)

Abreu Junior, Luiz de; Wolosker, Angela Maria Borri; Borri, Maria Lucia; Galvao Filho, Mario de Melo; Hartmann, Luiz Guilherme de Carvalho; D'Ippolito, Giuseppe; Castro, Claudio Campi de

2008-01-01

Objective: Magnetic resonance imaging has been utilized in the evaluation of the lacrimal apparatus with some advantages over conventional dacryocystography. The present study was aimed at acquiring high resolution images utilizing microscopic coils for evaluating typical structures of the lacrimal apparatus as compared with the findings observed with conventional surface coils. Materials and methods: Five asymptomatic volunteers with no history of epiphora were submitted to high-field magnetic resonance imaging with microscopic and conventional surface coils, and STIR sequence after instillation of saline solution. The definition of normal anatomic structures of lacrimal apparatuses was compared utilizing conventional and microscopic surface coils. Based on a consensual scoring system, the mean values for each structure were calculated by two observers. Results: In 90% of cases, higher scores were attributed to images acquired with the microscopic coil. On average, a 1.17 point increase was observed in the scoring of anatomic structures imaged with the microscopic coil. Additionally, a subjective improvement was observed in the signal-to-noise ratio with the microscopic coil. Conclusion: Magnetic resonance dacryocystography with microscopic coils is the appropriate method for evaluating the lacrimal apparatus, providing images with better quality as compared with those acquired with conventional surface coils. (author)

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

Science.gov (United States)

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.

Pore sub-features reproducibility in direct microscopic and Livescan images--their reliability in personal identification.

Science.gov (United States)

Gupta, Abhishek; Sutton, Raul

2010-07-01

Third level features have been reported to have equal discriminatory power as second level details in establishing personal identification. Pore area, as an extended set third level sub-feature, has been studied by minimizing possible factors that could affect pore size. The reproducibility of pore surface area has been studied using direct microscopic and 500 ppi Livescan images. Direct microscopic pore area measurements indicated that the day on which the pore area was measured had a significant impact on the measured pore area. Pore area measurement was shown to be difficult to estimate in 500 ppi Livescan measurements owing to lack of resolution. It is not possible to reliably use pore area as an identifying feature in fingerprint examination.

A pragmatic guide to multiphoton microscope design

Science.gov (United States)

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

2016-01-01

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

Comparison of Electron Imaging Modes for Dimensional Measurements in the Scanning Electron Microscope.

Science.gov (United States)

Postek, Michael T; Vladár, András E; Villarrubia, John S; Muto, Atsushi

2016-08-01

Dimensional measurements from secondary electron (SE) images were compared with those from backscattered electron (BSE) and low-loss electron (LLE) images. With the commonly used 50% threshold criterion, the lines consistently appeared larger in the SE images. As the images were acquired simultaneously by an instrument with the capability to operate detectors for both signals at the same time, the differences cannot be explained by the assumption that contamination or drift between images affected the SE, BSE, or LLE images differently. Simulations with JMONSEL, an electron microscope simulator, indicate that the nanometer-scale differences observed on this sample can be explained by the different convolution effects of a beam with finite size on signals with different symmetry (the SE signal's characteristic peak versus the BSE or LLE signal's characteristic step). This effect is too small to explain the >100 nm discrepancies that were observed in earlier work on different samples. Additional modeling indicates that those discrepancies can be explained by the much larger sidewall angles of the earlier samples, coupled with the different response of SE versus BSE/LLE profiles to such wall angles.

Anisotropic contrast optical microscope.

Science.gov (United States)

Peev, D; Hofmann, T; Kananizadeh, N; Beeram, S; Rodriguez, E; Wimer, S; Rodenhausen, K B; Herzinger, C M; Kasputis, T; Pfaunmiller, E; Nguyen, A; Korlacki, R; Pannier, A; Li, Y; Schubert, E; Hage, D; Schubert, M

2016-11-01

An optical microscope is described that reveals contrast in the Mueller matrix images of a thin, transparent, or semi-transparent specimen located within an anisotropic object plane (anisotropic filter). The specimen changes the anisotropy of the filter and thereby produces contrast within the Mueller matrix images. Here we use an anisotropic filter composed of a semi-transparent, nanostructured thin film with sub-wavelength thickness placed within the object plane. The sample is illuminated as in common optical microscopy but the light is modulated in its polarization using combinations of linear polarizers and phase plate (compensator) to control and analyze the state of polarization. Direct generalized ellipsometry data analysis approaches permit extraction of fundamental Mueller matrix object plane images dispensing with the need of Fourier expansion methods. Generalized ellipsometry model approaches are used for quantitative image analyses. These images are obtained from sets of multiple images obtained under various polarizer, analyzer, and compensator settings. Up to 16 independent Mueller matrix images can be obtained, while our current setup is limited to 11 images normalized by the unpolarized intensity. We demonstrate the anisotropic contrast optical microscope by measuring lithographically defined micro-patterned anisotropic filters, and we quantify the adsorption of an organic self-assembled monolayer film onto the anisotropic filter. Comparison with an isotropic glass slide demonstrates the image enhancement obtained by our method over microscopy without the use of an anisotropic filter. In our current instrument, we estimate the limit of detection for organic volumetric mass within the object plane of ≈49 fg within ≈7 × 7 μm 2 object surface area. Compared to a quartz crystal microbalance with dissipation instrumentation, where contemporary limits require a total load of ≈500 pg for detection, the instrumentation demonstrated here improves

An optical super-microscope for far-field, real-time imaging beyond the diffraction limit.

Science.gov (United States)

Wong, Alex M H; Eleftheriades, George V

2013-01-01

Optical microscopy suffers from a fundamental resolution limitation arising from the diffractive nature of light. While current solutions to sub-diffraction optical microscopy involve combinations of near-field, non-linear and fine scanning operations, we hereby propose and demonstrate the optical super-microscope (OSM) - a superoscillation-based linear imaging system with far-field working and observation distances - which can image an object in real-time and with sub-diffraction resolution. With our proof-of-principle prototype we report a point spread function with a spot size clearly reduced from the diffraction limit, and demonstrate corresponding improvements in two-point resolution experiments. Harnessing a new understanding of superoscillations, based on antenna array theory, our OSM achieves far-field, sub-diffraction optical imaging of an object without the need for fine scanning, data post-processing or object pre-treatment. Hence the OSM can be used in a wide variety of imaging applications beyond the diffraction limit, including real-time imaging of moving objects.

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

Science.gov (United States)

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.

A high-energy x-ray microscope for inertial confinement fusion

International Nuclear Information System (INIS)

Marshall, F.J.; Bennett, G.R.

1999-01-01

We have developed a microscope capable of imaging x-ray emission from inertial confinement fusion targets in the range of 7 - 9 keV. Imaging is accomplished with a Kirkpatrick-Baez type, four-image microscope coated with a WB 4 C multilayer having a 2d period of 140 Angstrom. This microscope design (a standard used on the University of Rochester close-quote s OMEGA laser system) is capable of 5 μm resolution over a region large enough to image an imploded target (∼400 μm). This design is capable of being extended to ∼40 keV if state-of-the-art, short-spacing, multilayer coatings are used (∼25 Angstrom), and has been configured to obtain 3 μm resolution with the appropriate choice of mirror size. As such, this type of microscope could serve as a platform for multiframe, hard x-ray imaging on the National Ignition Facility. Characterization of the microscope and laboratory measurements of the energy response made with a cw x-ray source will be shown. copyright 1999 American Institute of Physics

Real-time progressive hyperspectral image processing endmember finding and anomaly detection

CERN Document Server

Chang, Chein-I

2016-01-01

The book covers the most crucial parts of real-time hyperspectral image processing: causality and real-time capability. Recently, two new concepts of real time hyperspectral image processing, Progressive Hyperspectral Imaging (PHSI) and Recursive Hyperspectral Imaging (RHSI). Both of these can be used to design algorithms and also form an integral part of real time hyperpsectral image processing. This book focuses on progressive nature in algorithms on their real-time and causal processing implementation in two major applications, endmember finding and anomaly detection, both of which are fundamental tasks in hyperspectral imaging but generally not encountered in multispectral imaging. This book is written to particularly address PHSI in real time processing, while a book, Recursive Hyperspectral Sample and Band Processing: Algorithm Architecture and Implementation (Springer 2016) can be considered as its companion book. Includes preliminary background which is essential to those who work in hyperspectral ima...

Science 101: How Does an Electron Microscope Work?

Science.gov (United States)

Robertson, Bill

2013-01-01

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

Thimble microscope system

Science.gov (United States)

Kamal, Tahseen; Rubinstein, Jaden; Watkins, Rachel; Cen, Zijian; Kong, Gary; Lee, W. M.

2016-12-01

Wearable computing devices, e.g. Google Glass, Smart watch, embodies the new human design frontier, where technology interfaces seamlessly with human gestures. During examination of any subject in the field (clinic, surgery, agriculture, field survey, water collection), our sensory peripherals (touch and vision) often go hand-in-hand. The sensitivity and maneuverability of the human fingers are guided with tight distribution of biological nerve cells, which perform fine motor manipulation over a range of complex surfaces that is often out of sight. Our sight (or naked vision), on the other hand, is generally restricted to line of sight that is ill-suited to view around corner. Hence, conventional imaging methods are often resort to complex light guide designs (periscope, endoscopes etc) to navigate over obstructed surfaces. Using modular design strategies, we constructed a prototype miniature microscope system that is incorporated onto a wearable fixture (thimble). This unique platform allows users to maneuver around a sample and take high resolution microscopic images. In this paper, we provide an exposition of methods to achieve a thimble microscopy; microscope lens fabrication, thimble design, integration of miniature camera and liquid crystal display.

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

Science.gov (United States)

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

2018-01-01

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

Segmentation of White Blood Cells From Microscopic Images Using a Novel Combination of K-Means Clustering and Modified Watershed Algorithm.

Science.gov (United States)

Ghane, Narjes; Vard, Alireza; Talebi, Ardeshir; Nematollahy, Pardis

2017-01-01

Recognition of white blood cells (WBCs) is the first step to diagnose some particular diseases such as acquired immune deficiency syndrome, leukemia, and other blood-related diseases that are usually done by pathologists using an optical microscope. This process is time-consuming, extremely tedious, and expensive and needs experienced experts in this field. Thus, a computer-aided diagnosis system that assists pathologists in the diagnostic process can be so effective. Segmentation of WBCs is usually a first step in developing a computer-aided diagnosis system. The main purpose of this paper is to segment WBCs from microscopic images. For this purpose, we present a novel combination of thresholding, k-means clustering, and modified watershed algorithms in three stages including (1) segmentation of WBCs from a microscopic image, (2) extraction of nuclei from cell's image, and (3) separation of overlapping cells and nuclei. The evaluation results of the proposed method show that similarity measures, precision, and sensitivity respectively were 92.07, 96.07, and 94.30% for nucleus segmentation and 92.93, 97.41, and 93.78% for cell segmentation. In addition, statistical analysis presents high similarity between manual segmentation and the results obtained by the proposed method.

Cellscope Aquatic: a Lab Quality, Portable Cellphone-Based Microscope for On-Site Collection of Algae Images

Science.gov (United States)

Steinberg, S. J.; Howard, M. D.

2016-02-01

Collecting algae samples from the field presents issues of specimen damage or degradation caused by preservation methods, handling and transport to laboratory facilities for identification. Traditionally, in-field collection of high quality microscopic images has not been possible due to the size, weight and fragility of high quality instruments and training of field staff in species identification. Scientists at the Southern California Coastal Water Research Project (SCCWRP) in collaboration with the Fletcher Lab, University of California Berkeley, Department of Bioengineering, tested and translated Fletcher's original medical CellScope for use in environmental monitoring applications. Field tests conducted by SCCWRP in 2014 led to modifications of the clinical CellScope to one better suited to in-field microscopic imaging for aquatic organisms. SCCWRP subsequently developed a custom cell-phone application to acquire microscopic imagery using the "CellScope Aquatic "in combination with other cell-phone derived field data (e.g. GPS location, date, time and other field observations). Data and imagery collected in-field may be transmitted in real-time to a web-based data system for tele-taxonomy evaluation and assessment by experts in the office. These hardware and software tools was tested in field in a variety of conditions and settings by multiple algae experts during the spring and summer of 2015 to further test and refine the CellScope Aquatic platform. The CellScope Aquatic provides an easy-to-use, affordable, lightweight, professional quality, data collection platform for environmental monitoring. Our ongoing efforts will focus on development of real-time expert systems for data analysis and image processing, to provide onsite feedback to field scientists.

High-speed multiframe dynamic transmission electron microscope image acquisition system with arbitrary timing

Science.gov (United States)

Reed, Bryan W.; DeHope, William J.; Huete, Glenn; LaGrange, Thomas B.; Shuttlesworth, Richard M.

2015-10-20

An electron microscope is disclosed which has a laser-driven photocathode and an arbitrary waveform generator (AWG) laser system ("laser"). The laser produces a train of temporally-shaped laser pulses of a predefined pulse duration and waveform, and directs the laser pulses to the laser-driven photocathode to produce a train of electron pulses. An image sensor is used along with a deflector subsystem. The deflector subsystem is arranged downstream of the target but upstream of the image sensor, and has two pairs of plates arranged perpendicular to one another. A control system controls the laser and a plurality of switching components synchronized with the laser, to independently control excitation of each one of the deflector plates. This allows each electron pulse to be directed to a different portion of the image sensor, as well as to be provided with an independently set duration and independently set inter-pulse spacings.

Tapping mode imaging and measurements with an inverted atomic force microscope.

Science.gov (United States)

Chan, Sandra S F; Green, John-Bruce D

2006-07-18

This report demonstrates the successful use of the inverted atomic force microscope (i-AFM) for tapping mode AFM imaging of cantilever-supported samples. i-AFM is a mode of AFM operation in which a sample supported on a tipless cantilever is imaged by one of many tips in a microfabricated tip array. Tapping mode is an intermittent contact mode whereby the cantilever is oscillated at or near its resonance frequency, and the amplitude and/or phase are used to image the sample. In the process of demonstrating that tapping mode images could be obtained in the i-AFM design, it was observed that the amplitude of the cantilever oscillation decreased markedly as the cantilever and tip array were approached. The source of this damping of the cantilever oscillations was identified to be the well-known "squeeze film damping", and the extent of damping was a direct consequence of the relatively shorter tip heights for the tip arrays, as compared to those of commercially available tapping mode cantilevers with integrated tips. The functional form for the distance dependence of the damping coefficient is in excellent agreement with previously published models for squeeze film damping, and the values for the fitting parameters make physical sense. Although the severe damping reduces the cantilever free amplitude substantially, we found that we were still able to access the low-amplitude regime of oscillation necessary for attractive tapping mode imaging of fragile molecules.

Compact, Light-weight and Cost-effective Microscope based on Lensless Incoherent Holography for Telemedicine Applications

Science.gov (United States)

Mudanyali, Onur; Tseng, Derek; Oh, Chulwoo; Isikman, Serhan O.; Sencan, Ikbal; Bishara, Waheb; Oztoprak, Cetin; Seo, Sungkyu; Khademhosseini, Bahar; Ozcan, Aydogan

2010-01-01

Despite the rapid progress in optical imaging, most of the advanced microscopy modalities still require complex and costly set-ups that unfortunately limit their use beyond well equipped laboratories. In the meantime, microscopy in resource-limited settings has requirements significantly different from those encountered in advanced laboratories, and such imaging devices should be cost-effective, compact, light-weight and appropriately accurate and simple to be usable by minimally trained personnel. Furthermore, these portable microscopes should ideally be digitally integrated as part of a telemedicine network that connects various mobile health-care providers to a central laboratory or hospital. Toward this end, here we demonstrate a lensless on-chip microscope weighing ~46 grams with dimensions smaller than 4.2cm × 4.2cm × 5.8cm that achieves sub-cellular resolution over a large field of view of ~24 mm2. This compact and light-weight microscope is based on digital in-line holography and does not need any lenses, bulky optical/mechanical components or coherent sources such as lasers. Instead, it utilizes a simple light-emitting-diode (LED) and a compact opto-electronic sensor-array to record lensless holograms of the objects, which then permits rapid digital reconstruction of regular transmission or differential interference contrast (DIC) images of the objects. Because this lensless incoherent holographic microscope has orders-of-magnitude improved light collection efficiency and is very robust to mechanical misalignments it may offer a cost-effective tool especially for telemedicine applications involving various global health problems in resource limited settings. PMID:20401422

A mini-microscope for in situ monitoring of cells†‡

Science.gov (United States)

Kim, Sang Bok; Koo, Kyo-in; Bae, Hojae; Dokmeci, Mehmet R.; Hamilton, Geraldine A.; Bahinski, Anthony; Kim, Sun Min; Ingber, Donald E.

2013-01-01

A mini-microscope was developed for in situ monitoring of cells by modifying off-the-shelf components of a commercial webcam. The mini-microscope consists of a CMOS imaging module, a small plastic lens and a white LED illumination source. The CMOS imaging module was connected to a laptop computer through a USB port for image acquisition and analysis. Due to its compact size, 8 × 10 × 9 cm, the present microscope is portable and can easily fit inside a conventional incubator, and enables real-time monitoring of cellular behaviour. Moreover, the mini-microscope can be used for imaging cells in conventional cell culture flasks, such as Petri dishes and multi-well plates. To demonstrate the operation of the mini-microscope, we monitored the cellular migration of mouse 3T3 fibroblasts in a scratch assay in medium containing three different concentrations of fetal bovine serum (5, 10, and 20%) and demonstrated differential responses depending on serum levels. In addition, we seeded embryonic stem cells inside poly(ethylene glycol) microwells and monitored the formation of stem cell aggregates in real time using the mini-microscope. Furthermore, we also combined a lab-on-a-chip microfluidic device for microdroplet generation and analysis with the mini-microscope and observed the formation of droplets under different flow conditions. Given its cost effectiveness, robust imaging and portability, the presented platform may be useful for a range of applications for real-time cellular imaging using lab-on-a-chip devices at low cost. PMID:22911426

Mobile microscope complex GIB-1

International Nuclear Information System (INIS)

Belyakov, A.V.; Gorbachev, A.N.

2002-01-01

To study microstructure in operating pipelines of power units a mobile microscope system is developed and successfully used. The system includes a portable microscope, a monitor, power supply and a portable computer. The monitor is used for surveying images from a video camera mounted on the microscope. The magnification on visual examination constitutes x 100 and x 500. Diameters of pipelines examined should not be less than 130 mm. Surface preparation for microstructural studies includes routine mechanical rough grinding and polishing with subsequent etching [ru

Designs for a quantum electron microscope.

Science.gov (United States)

Kruit, P; Hobbs, R G; Kim, C-S; Yang, Y; Manfrinato, V R; Hammer, J; Thomas, S; Weber, P; Klopfer, B; Kohstall, C; Juffmann, T; Kasevich, M A; Hommelhoff, P; Berggren, K K

2016-05-01

One of the astounding consequences of quantum mechanics is that it allows the detection of a target using an incident probe, with only a low probability of interaction of the probe and the target. This 'quantum weirdness' could be applied in the field of electron microscopy to generate images of beam-sensitive specimens with substantially reduced damage to the specimen. A reduction of beam-induced damage to specimens is especially of great importance if it can enable imaging of biological specimens with atomic resolution. Following a recent suggestion that interaction-free measurements are possible with electrons, we now analyze the difficulties of actually building an atomic resolution interaction-free electron microscope, or "quantum electron microscope". A quantum electron microscope would require a number of unique components not found in conventional transmission electron microscopes. These components include a coherent electron beam-splitter or two-state-coupler, and a resonator structure to allow each electron to interrogate the specimen multiple times, thus supporting high success probabilities for interaction-free detection of the specimen. Different system designs are presented here, which are based on four different choices of two-state-couplers: a thin crystal, a grating mirror, a standing light wave and an electro-dynamical pseudopotential. Challenges for the detailed electron optical design are identified as future directions for development. While it is concluded that it should be possible to build an atomic resolution quantum electron microscope, we have also identified a number of hurdles to the development of such a microscope and further theoretical investigations that will be required to enable a complete interpretation of the images produced by such a microscope. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Atmospheric scanning electron microscope for correlative microscopy.

Science.gov (United States)

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

2012-01-01

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

Operation of a scanning near field optical microscope in reflection in combination with a scanning force microscope

NARCIS (Netherlands)

van Hulst, N.F.; Moers, M.H.P.; Moers, M.H.P.; Noordman, O.F.J.; Noordman, O.F.J.; Faulkner, T.; Segerink, Franciscus B.; van der Werf, Kees; de Grooth, B.G.; Bölger, B.; Bölger, B.

1992-01-01

Images obtained with a scanning near field optical microscope (SNOM) operating in reflection are presented. We have obtained the first results with a SiN tip as optical probe. The instrument is simultaneously operated as a scanning force microscope (SFM). Moreover, the instrument incorporates an

Chromosome structure investigated with the atomic force microscope

NARCIS (Netherlands)

de Grooth, B.G.; Putman, C.A.J.; Putman, Constant A.; van der Werf, Kees; van Hulst, N.F.; van Oort, G.; van Oort, Geeske; Greve, Jan; Manne, Srinivas

1992-01-01

We have developed an atomic force microscope (AFM) with an integrated optical microscope. The optical microscope consists of an inverted epi-illumination system that yields images in reflection or fluorescence of the sample. With this system it is possible to quickly locate an object of interest. A

X-ray analysis of a single aerosol particle with combination of scanning electron microscope and synchrotron radiation X-ray microscope

International Nuclear Information System (INIS)

Toyoda, Masatoshi; Kaibuchi, Kazuki; Nagasono, Mitsuru; Terada, Yasuko; Tanabe, Teruo; Hayakawa, Shinjiro; Kawai, Jun

2004-01-01

We developed a microscope by a combination of synchrotron radiation X-ray fluorescence (SR-XRF) microscope and scanning electron microscope (SEM) with an energy dispersive X-ray spectrometer (EDX). SR-XRF is appropriate to detect trace and micro amount of elements and sensitive to heavy elements in an analyte but it cannot observe the real time image. SEM-EDX can observe the secondary electron image of a single particle in real time and is appropriate to detect lighter elements. This combination microscope can ensure the identification of the XRF spectrum to the SEM image without transferring the sample. For aerosol analysis, it is important to analyze each particle. The present method makes feasible to analyze not only the average elemental composition as the total particles but also elemental composition of each particle, which is dependent on the particle shape and size. The microscope was applied to an individual aerosol particle study. The X-ray spectra were different among the particles, but also different between SR-XRF and SEM-EDX for the same particle, due to the difference in fluorescence yields between X-ray excitation and electron excitation

Dark-field imaging based on post-processed electron backscatter diffraction patterns of bulk crystalline materials in a scanning electron microscope.

Science.gov (United States)

Brodusch, Nicolas; Demers, Hendrix; Gauvin, Raynald

2015-01-01

Dark-field (DF) images were acquired in the scanning electron microscope with an offline procedure based on electron backscatter diffraction (EBSD) patterns (EBSPs). These EBSD-DF images were generated by selecting a particular reflection on the electron backscatter diffraction pattern and by reporting the intensity of one or several pixels around this point at each pixel of the EBSD-DF image. Unlike previous studies, the diffraction information of the sample is the basis of the final image contrast with a pixel scale resolution at the EBSP providing DF imaging in the scanning electron microscope. The offline facility of this technique permits the selection of any diffraction condition available in the diffraction pattern and displaying the corresponding image. The high number of diffraction-based images available allows a better monitoring of deformation structures compared to electron channeling contrast imaging (ECCI) which is generally limited to a few images of the same area. This technique was applied to steel and iron specimens and showed its high capability in describing more rigorously the deformation structures around micro-hardness indents. Due to the offline relation between the reference EBSP and the EBSD-DF images, this new technique will undoubtedly greatly improve our knowledge of deformation mechanism and help to improve our understanding of the ECCI contrast mechanisms. Copyright © 2014 Elsevier B.V. All rights reserved.

Use of fundus autofluorescence images to predict geographic atrophy progression.

Science.gov (United States)

Bearelly, Srilaxmi; Khanifar, Aziz A; Lederer, David E; Lee, Jane J; Ghodasra, Jason H; Stinnett, Sandra S; Cousins, Scott W

2011-01-01

Fundus autofluorescence imaging has been shown to be helpful in predicting progression of geographic atrophy (GA) secondary to age-related macular degeneration. We assess the ability of fundus autofluorescence imaging to predict rate of GA progression using a simple categorical scheme. Subjects with GA secondary to age-related macular degeneration with fundus autofluorescence imaging acquired at least 12 months apart were included. Rim area focal hyperautofluorescence was defined as percentage of the 500-μm-wide margin bordering the GA that contained increased autofluorescence. Rim area focal hyperautofluorescence on baseline fundus autofluorescence images was assessed and categorized depending on the extent of rim area focal hyperautofluorescence (Category 1: ≤33%; Category 2: between 33 and 67%; Category 3: ≥67%). Total GA areas at baseline and follow-up were measured to calculate change in GA progression. Forty-five eyes of 45 subjects were included; average duration of follow-up was 18.5 months. Median growth rates differed among categories of baseline rim area focal hyperautofluorescence (P = 0.01 among Categories 1, 2, and 3; P = 0.008 for Category 1 compared with Category 3, Jonckheere-Terpstra test). A simple categorical scheme that stratifies the amount of increased autofluorescence in the 500-μm margin bordering GA may be used to differentiate faster and slower progressors.

Single DNA imaging and length quantification through a mobile phone microscope

Science.gov (United States)

Wei, Qingshan; Luo, Wei; Chiang, Samuel; Kappel, Tara; Mejia, Crystal; Tseng, Derek; Chan, Raymond Yan L.; Yan, Eddie; Qi, Hangfei; Shabbir, Faizan; Ozkan, Haydar; Feng, Steve; Ozcan, Aydogan

2016-03-01

The development of sensitive optical microscopy methods for the detection of single DNA molecules has become an active research area which cultivates various promising applications including point-of-care (POC) genetic testing and diagnostics. Direct visualization of individual DNA molecules usually relies on sophisticated optical microscopes that are mostly available in well-equipped laboratories. For POC DNA testing/detection, there is an increasing need for the development of new single DNA imaging and sensing methods that are field-portable, cost-effective, and accessible for diagnostic applications in resource-limited or field-settings. For this aim, we developed a mobile-phone integrated fluorescence microscopy platform that allows imaging and sizing of single DNA molecules that are stretched on a chip. This handheld device contains an opto-mechanical attachment integrated onto a smartphone camera module, which creates a high signal-to-noise ratio dark-field imaging condition by using an oblique illumination/excitation configuration. Using this device, we demonstrated imaging of individual linearly stretched λ DNA molecules (48 kilobase-pair, kbp) over 2 mm2 field-of-view. We further developed a robust computational algorithm and a smartphone app that allowed the users to quickly quantify the length of each DNA fragment imaged using this mobile interface. The cellphone based device was tested by five different DNA samples (5, 10, 20, 40, and 48 kbp), and a sizing accuracy of <1 kbp was demonstrated for DNA strands longer than 10 kbp. This mobile DNA imaging and sizing platform can be very useful for various diagnostic applications including the detection of disease-specific genes and quantification of copy-number-variations at POC settings.

Textures of the soils and rocks at Gusev crater from Spirit's Microscopic Imager

DEFF Research Database (Denmark)

Herkenhoff, K.E.; Squyres, S.W.; Arvidson, R.

2004-01-01

The Microscopic Imager on the Spirit rover analyzed the textures of the soil and rocks at Gusev crater on Mars at a resolution of 100 micrometers. Weakly bound agglomerates of dust are present in the soil near the Columbia Memorial Station. Some of the brushed or abraded rock surfaces show igneous...... textures and evidence for alteration rinds, coatings, and veins consistent with secondary mineralization. The rock textures are consistent with a volcanic origin and subsequent alteration and/or weathering by impact events, wind, and possibly water....

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

Science.gov (United States)

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.

Quantitative measurements with x-ray microscopes in laser-fusion experiments

International Nuclear Information System (INIS)

Marshall, F.J.; Su, Q.

1995-01-01

X-ray imaging of laser-fusion target implosions has been performed on the University of Rochester's OMEGA laser system by means of grazing-incidence optical imaging with Kirkpatrick--Baez (KB) microscopes. High spatial resolution imaging (∼5 μm) of hard x-ray emission (up to ∼7 keV) has been achieved. New grazing-incidence optics are currently being fabricated for the OMEGA Upgrade experimental laser-fusion facility. Projected performance indicates that resolution may be increased to ∼2 μm at the center of the field of view and sensitivity extended to ∼8 keV. Uses of KB microscopes on the OMEGA Upgrade will include hard x-ray imaging, grating-dispersed imaged spectroscopy, and framed imaging. A novel technique for monochromatic imaging with KB microscopes has also been demonstrated enabling images of target emission in a narrow energy band (10 to 20 eV) to be obtained

A high-resolution mini-microscope system for wireless real-time monitoring.

Science.gov (United States)

Wang, Zongjie; Boddeda, Akash; Parker, Benjamin; Samanipour, Roya; Ghosh, Sanjoy; Menard, Frederic; Kim, Keekyoung

2017-09-04

Compact, cost-effective and high-performance microscope that enables the real-time imaging of cells and lab-on-a-chip devices is highly demanded for cell biology and biomedical engineering. This paper aims to present the design and application of an inexpensive wireless mini-microscope with resolution up to 2592 × 1944 pixels and speed up to 90 fps. The mini-microscope system was built on a commercial embedded system (Raspberry Pi). We modified a camera module and adopted an inverse dual lens system to obtain the clear field of view and appropriate magnification for tens of micrometer objects. The system was capable of capturing time-lapse images and transferring image data wirelessly. The entire system can be operated wirelessly and cordlessly in a conventional cell culturing incubator. The developed mini-microscope was used to monitor the attachment and proliferation of NIH-3T3 and HEK 293 cells inside an incubator for 50 hours. In addition, the mini-microscope was used to monitor a droplet generation process in a microfluidic device. The high-quality images captured by the mini-microscope enabled us an automated analysis of experimental parameters. The successful applications prove the great potential of the developed mini-microscope for monitoring various biological samples and microfluidic devices. This paper presents the design of a high resolution mini-microscope system that enables the wireless real-time imaging of cells inside the incubator. This system has been verified to be a useful tool to obtain high-quality images and videos for the automated quantitative analysis of biological samples and lab-on-a-chip devices in the long term.

Polarized differential-phase laser scanning microscope

International Nuclear Information System (INIS)

Chou Chien; Lyu, C.-W.; Peng, L.-C.

2001-01-01

A polarized differential-phase laser scanning microscope, which combines a polarized optical heterodyne Mach-Zehnder interferometer and a differential amplifier to scan the topographic image of a surface, is proposed. In the experiment the differential amplifier, which acts as a PM-AM converter, in the experiment, converting phase modulation (PM) into amplitude modulation (AM). Then a novel, to our knowledge, phase demodulator was proposed and implemented for the differential-phase laser scanning microscope. An optical grating (1800 lp/mm) was imaged. The lateral and the depth resolutions of the imaging system were 0.5 μm and 1 nm, respectively. The detection accuracy, which was limited by the reflectivity variation of the test surface, is discussed

Imaging sequential dehydrogenation of methanol on Cu(110) with a scanning tunneling microscope.

Science.gov (United States)

Kitaguchi, Y; Shiotari, A; Okuyama, H; Hatta, S; Aruga, T

2011-05-07

Adsorption of methanol and its dehydrogenation on Cu(110) were studied by using a scanning tunneling microscope (STM). Upon adsorption at 12 K, methanol preferentially forms clusters on the surface. The STM could induce dehydrogenation of methanol sequentially to methoxy and formaldehyde. This enabled us to study the binding structures of these products in a single-molecule limit. Methoxy was imaged as a pair of protrusion and depression along the [001] direction. This feature is fully consistent with the previous result that it adsorbs on the short-bridge site with the C-O axis tilted along the [001] direction. The axis was induced to flip back and forth by vibrational excitations with the STM. Two configurations were observed for formaldehyde, whose structures were proposed based on their characteristic images and motions.

Simple and versatile modifications allowing time gated spectral acquisition, imaging and lifetime profiling on conventional wide-field microscopes

International Nuclear Information System (INIS)

Pal, Robert; Beeby, Andrew

2014-01-01

An inverted microscope has been adapted to allow time-gated imaging and spectroscopy to be carried out on samples containing responsive lanthanide probes. The adaptation employs readily available components, including a pulsed light source, time-gated camera, spectrometer and photon counting detector, allowing imaging, emission spectroscopy and lifetime measurements. Each component is controlled by a suite of software written in LabVIEW and is powered via conventional USB ports. (technical note)

New deconvolution method for microscopic images based on the continuous Gaussian radial basis function interpolation model.

Science.gov (United States)

Chen, Zhaoxue; Chen, Hao

2014-01-01

A deconvolution method based on the Gaussian radial basis function (GRBF) interpolation is proposed. Both the original image and Gaussian point spread function are expressed as the same continuous GRBF model, thus image degradation is simplified as convolution of two continuous Gaussian functions, and image deconvolution is converted to calculate the weighted coefficients of two-dimensional control points. Compared with Wiener filter and Lucy-Richardson algorithm, the GRBF method has an obvious advantage in the quality of restored images. In order to overcome such a defect of long-time computing, the method of graphic processing unit multithreading or increasing space interval of control points is adopted, respectively, to speed up the implementation of GRBF method. The experiments show that based on the continuous GRBF model, the image deconvolution can be efficiently implemented by the method, which also has a considerable reference value for the study of three-dimensional microscopic image deconvolution.

Line-scanning tomographic optical microscope with isotropic transfer function

International Nuclear Information System (INIS)

Gajdátsy, Gábor; Dudás, László; Erdélyi, Miklós; Szabó, Gábor

2010-01-01

An imaging method and optical system, referred to as a line-scanning tomographic optical microscope (LSTOM) using a combination of line-scanning technique and CT reconstruction principle, is proposed and studied theoretically and experimentally. In our implementation a narrow focus line is scanned over the sample and the reflected light is measured in a confocal arrangement. One such scan is equivalent to a transverse projection in tomography. Repeating the scanning procedure in several directions, a number of transverse projections are recorded from which the image can be obtained using conventional CT reconstruction algorithms. The resolution of the image is independent of the spatial dimensions and structure of the applied detector; furthermore, the transfer function of the system is isotropic. The imaging performance of the implemented confocal LSTOM was compared with a point-scanning confocal microscope, based on recorded images. These images demonstrate that the resolution of the confocal LSTOM exceeds (by 15%) the resolution limit of a point-scanning confocal microscope

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

Science.gov (United States)

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

2014-01-01

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

Reconstructing 3D profiles of flux distribution in array of unshunted Josephson junctions from 2D scanning SQUID microscope images

International Nuclear Information System (INIS)

Nascimento, F.M.; Sergeenkov, S.; Araujo-Moreira, F.M.

2012-01-01

By using a specially designed algorithm (based on utilizing the so-called Hierarchical Data Format), we report on successful reconstruction of 3D profiles of local flux distribution within artificially prepared arrays of unshunted Nb-AlO x -Nb Josephson junctions from 2D surface images obtained via the scanning SQUID microscope. The analysis of the obtained results suggest that for large sweep areas, the local flux distribution significantly deviates from the conventional picture and exhibits a more complicated avalanche-type behavior with a prominent dendritic structure. -- Highlights: ► The penetration of external magnetic field into an array of Nb-AlO x -Nb Josephson junctions is studied. ► Using Scanning SQUID Microscope, 2D images of local flux distribution within array are obtained. ► Using specially designed pattern recognition algorithm, 3D flux profiles are reconstructed from 2D images.

Examination of Scanning Electron Microscope and Computed Tomography Images of PICA

Science.gov (United States)

Lawson, John W.; Stackpoole, Margaret M.; Shklover, Valery

2010-01-01

Micrographs of PICA (Phenolic Impregnated Carbon Ablator) taken using a Scanning Electron Microscope (SEM) and 3D images taken with a Computed Tomography (CT) system are examined. PICA is a carbon fiber based composite (Fiberform ) with a phenolic polymer matrix. The micrographs are taken at different surface depths and at different magnifications in a sample after arc jet testing and show different levels of oxidative removal of the charred matrix (Figs 1 though 13). CT scans, courtesy of Xradia, Inc. of Concord CA, were captured for samples of virgin PICA, charred PICA and raw Fiberform (Fig. 14). We use these images to calculate the thermal conductivity (TC) of these materials using correlation function (CF) methods. CF methods give a mathematical description of how one material is embedded in another and is thus ideally suited for modeling composites like PICA. We will evaluate how the TC of the materials changes as a function of surface depth. This work is in collaboration with ETH-Zurich, which has expertise in high temperature materials and TC modeling (including CF methods).

Algorithm for detection of overlapped red blood cells in microscopic images of blood smears

OpenAIRE

Romero-Rondón, Miguel Fabián; Sanabria-Rosas, Laura Melissa; Bautista-Rozo, Lola Xiomara; Mendoza-Castellanos, Alfonso

2016-01-01

The hemogram is one of the most requested medical tests as it presents details about the three cell series in the blood: red series, white series and platelet series. To make some diagnostics, the specialist must undertake the test manually, observing the blood cells under the microscope, which implies a great physical effort. In order to facilitate this work, different digital image processing techniques to detect and classify red blood cells have been proposed. However, a common problem is ...

Color capable sub-pixel resolving optofluidic microscope and its application to blood cell imaging for malaria diagnosis.

Directory of Open Access Journals (Sweden)

Seung Ah Lee

Full Text Available Miniaturization of imaging systems can significantly benefit clinical diagnosis in challenging environments, where access to physicians and good equipment can be limited. Sub-pixel resolving optofluidic microscope (SROFM offers high-resolution imaging in the form of an on-chip device, with the combination of microfluidics and inexpensive CMOS image sensors. In this work, we report on the implementation of color SROFM prototypes with a demonstrated optical resolution of 0.66 µm at their highest acuity. We applied the prototypes to perform color imaging of red blood cells (RBCs infected with Plasmodium falciparum, a particularly harmful type of malaria parasites and one of the major causes of death in the developing world.

A fluorescence scanning electron microscope

International Nuclear Information System (INIS)

Kanemaru, Takaaki; Hirata, Kazuho; Takasu, Shin-ichi; Isobe, Shin-ichiro; Mizuki, Keiji; Mataka, Shuntaro; Nakamura, Kei-ichiro

2009-01-01

Fluorescence techniques are widely used in biological research to examine molecular localization, while electron microscopy can provide unique ultrastructural information. To date, correlative images from both fluorescence and electron microscopy have been obtained separately using two different instruments, i.e. a fluorescence microscope (FM) and an electron microscope (EM). In the current study, a scanning electron microscope (SEM) (JEOL JXA8600 M) was combined with a fluorescence digital camera microscope unit and this hybrid instrument was named a fluorescence SEM (FL-SEM). In the labeling of FL-SEM samples, both Fluolid, which is an organic EL dye, and Alexa Fluor, were employed. We successfully demonstrated that the FL-SEM is a simple and practical tool for correlative fluorescence and electron microscopy.

Sub-wavelength imaging by depolarization in a reflection near-field optical microscope using an uncoated fiber probe

DEFF Research Database (Denmark)

Madsen, Steen; Bozhevolnyi, Sergey I.; Hvam, Jørn Märcher

1998-01-01

We present a reflection scanning near-field optical microscope utilizing counter-directional light propagation in an uncoated fiber probe, cross-polarized detection and shear-force feedback. Topographical and near-field optical imaging with a scanning speed of up to 10 mu m/s and a lateral...... resolution better than 40 nm are demonstrated with a latex projection test sample. Determination of the optical resolution as well as correlation between topographical and near-field optical images are discussed. (C) 1998 Elsevier Science B.V....

First Sample Delivery to Mars Microscope

Science.gov (United States)

2008-01-01

The Robotic Arm on NASA's Phoenix Mars Lander has just delivered the first sample of dug-up soil to the spacecraft's microscope station in this image taken by the Surface Stereo Imager during the mission's Sol 17 (June 12), or 17th Martian day after landing. The scoop is positioned above the box containing key parts of Phoenix's Microscopy, Electrochemistry and Conductivity Analyzer, or MECA, instrument suite. It has sprinkled a small amount of soil into a notch in the MECA box where the microscope's sample wheel is exposed. The wheel turns to present sample particles on various substrates to the Optical Microscope for viewing. The scoop is about 8.5 centimeters (3.3 inches) wide. The top of the MECA box is 20 centimeters (7.9 inches) wide. This image has been lightened to make details more visible. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

Progress in Molecular Imaging in Endoscopy and Endomicroscopy for Cancer Imaging

Directory of Open Access Journals (Sweden)

Supang Khondee

2013-01-01

Full Text Available Imaging is an essential tool for effective cancer management. Endoscopes are important medical instruments for performing in vivo imaging in hollow organs. Early detection of cancer can be achieved with surveillance using endoscopy, and has been shown to reduce mortality and to improve outcomes. Recently, great advancements have been made in endoscopic instruments, including new developments in optical designs, light sources, optical fibers, miniature scanners, and multimodal systems, allowing for improved resolution, greater tissue penetration, and multispectral imaging. In addition, progress has been made in the development of highly-specific optical probes, allowing for improved specificity for molecular targets. Integration of these new endoscopic instruments with molecular probes provides a unique opportunity for significantly improving patient outcomes and has potential to further improve early detection, image guided therapy, targeted therapy, and personalized medicine. This work summarizes current and evolving endoscopic technologies, and provides an overview of various promising optical molecular probes.

High-speed multi-frame dynamic transmission electron microscope image acquisition system with arbitrary timing

Science.gov (United States)

Reed, Bryan W.; DeHope, William J.; Huete, Glenn; LaGrange, Thomas B.; Shuttlesworth, Richard M.

2016-02-23

An electron microscope is disclosed which has a laser-driven photocathode and an arbitrary waveform generator (AWG) laser system ("laser"). The laser produces a train of temporally-shaped laser pulses each being of a programmable pulse duration, and directs the laser pulses to the laser-driven photocathode to produce a train of electron pulses. An image sensor is used along with a deflector subsystem. The deflector subsystem is arranged downstream of the target but upstream of the image sensor, and has a plurality of plates. A control system having a digital sequencer controls the laser and a plurality of switching components, synchronized with the laser, to independently control excitation of each one of the deflector plates. This allows each electron pulse to be directed to a different portion of the image sensor, as well as to enable programmable pulse durations and programmable inter-pulse spacings.

A high resolution ion microscope for cold atoms

International Nuclear Information System (INIS)

Stecker, Markus; Schefzyk, Hannah; Fortágh, József; Günther, Andreas

2017-01-01

We report on an ion-optical system that serves as a microscope for ultracold ground state and Rydberg atoms. The system is designed to achieve a magnification of up to 1000 and a spatial resolution in the 100 nm range, thereby surpassing many standard imaging techniques for cold atoms. The microscope consists of four electrostatic lenses and a microchannel plate in conjunction with a delay line detector in order to achieve single particle sensitivity with high temporal and spatial resolution. We describe the design process of the microscope including ion-optical simulations of the imaging system and characterize aberrations and the resolution limit. Furthermore, we present the experimental realization of the microscope in a cold atom setup and investigate its performance by patterned ionization with a structure size down to 2.7 μ m. The microscope meets the requirements for studying various many-body effects, ranging from correlations in cold quantum gases up to Rydberg molecule formation. (paper)

A Method for Improving the Progressive Image Coding Algorithms

Directory of Open Access Journals (Sweden)

Ovidiu COSMA

2014-12-01

Full Text Available This article presents a method for increasing the performance of the progressive coding algorithms for the subbands of images, by representing the coefficients with a code that reduces the truncation error.

Microscopic and macroscopic models for the onset and progression of Alzheimer's disease

International Nuclear Information System (INIS)

Bertsch, Michiel; Franchi, Bruno; Tesi, Maria Carla; Tosin, Andrea

2017-01-01

In the first part of this paper we review a mathematical model for the onset and progression of Alzheimer’s disease (AD) that was developed in subsequent steps over several years. The model is meant to describe the evolution of AD in vivo . In Achdou et al (2013 J. Math. Biol . 67 1369–92) we treated the problem at a microscopic scale, where the typical length scale is a multiple of the size of the soma of a single neuron. Subsequently, in Bertsch et al (2017 Math. Med. Biol . 34 193–214) we concentrated on the macroscopic scale, where brain neurons are regarded as a continuous medium, structured by their degree of malfunctioning. In the second part of the paper we consider the relation between the microscopic and the macroscopic models. In particular we show under which assumptions the kinetic transport equation, which in the macroscopic model governs the evolution of the probability measure for the degree of malfunctioning of neurons, can be derived from a particle-based setting. The models are based on aggregation and diffusion equations for β -Amyloid (A β from now on), a protein fragment that healthy brains regularly produce and eliminate. In case of dementia A β monomers are no longer properly washed out and begin to coalesce forming eventually plaques. Two different mechanisms are assumed to be relevant for the temporal evolution of the disease: (i) diffusion and agglomeration of soluble polymers of amyloid, produced by damaged neurons; (ii) neuron-to-neuron prion-like transmission. In the microscopic model we consider mechanism (i), modelling it by a system of Smoluchowski equations for the amyloid concentration (describing the agglomeration phenomenon), with the addition of a diffusion term as well as of a source term on the neuronal membrane. At the macroscopic level instead we model processes (i) and (ii) by a system of Smoluchowski equations for the amyloid concentration, coupled to a kinetic-type transport equation for the distribution

Progression Analysis and Stage Discovery in Continuous Physiological Processes Using Image Computing

Directory of Open Access Journals (Sweden)

Ferrucci Luigi

2010-01-01

Full Text Available We propose an image computing-based method for quantitative analysis of continuous physiological processes that can be sensed by medical imaging and demonstrate its application to the analysis of morphological alterations of the bone structure, which correlate with the progression of osteoarthritis (OA. The purpose of the analysis is to quantitatively estimate OA progression in a fashion that can assist in understanding the pathophysiology of the disease. Ultimately, the texture analysis will be able to provide an alternative OA scoring method, which can potentially reflect the progression of the disease in a more direct fashion compared to the existing clinically utilized classification schemes based on radiology. This method can be useful not just for studying the nature of OA, but also for developing and testing the effect of drugs and treatments. While in this paper we demonstrate the application of the method to osteoarthritis, its generality makes it suitable for the analysis of other progressive clinical conditions that can be diagnosed and prognosed by using medical imaging.

Atomic imaging using secondary electrons in a scanning transmission electron microscope: experimental observations and possible mechanisms.

Science.gov (United States)

Inada, H; Su, D; Egerton, R F; Konno, M; Wu, L; Ciston, J; Wall, J; Zhu, Y

2011-06-01

We report detailed investigation of high-resolution imaging using secondary electrons (SE) with a sub-nanometer probe in an aberration-corrected transmission electron microscope, Hitachi HD2700C. This instrument also allows us to acquire the corresponding annular dark-field (ADF) images both simultaneously and separately. We demonstrate that atomic SE imaging is achievable for a wide range of elements, from uranium to carbon. Using the ADF images as a reference, we studied the SE image intensity and contrast as functions of applied bias, atomic number, crystal tilt, and thickness to shed light on the origin of the unexpected ultrahigh resolution in SE imaging. We have also demonstrated that the SE signal is sensitive to the terminating species at a crystal surface. A possible mechanism for atomic-scale SE imaging is proposed. The ability to image both the surface and bulk of a sample at atomic-scale is unprecedented, and can have important applications in the field of electron microscopy and materials characterization. Copyright © 2010 Elsevier B.V. All rights reserved.

A new self-made digital slide scanner and microscope for imaging and quantification of fluorescent microspheres

DEFF Research Database (Denmark)

Henning, William; Bjerglund Andersen, Julie; Højgaard, Liselotte

2015-01-01

Objective: A low-cost microscope slide scanner was constructed for the purpose of digital imaging of newborn piglet brain tissue and to quantify fluorescent microspheres in tissue. Methods: Using a standard digital single-lens reflex (DSLR) camera, fluorescent imaging of newborn piglet brain tissue......-field imaging was tested by adding light diffuser film. Results: Cost of the slide scanner was a fraction of the cost of a commercial slide scanner. The slide scanner was able to image a large number of tissue slides in a semiautomatic manner and provided a large field of view (FOV) of 101 mm2 combined...... and an automatic algorithm to detect fluorescent microspheres in tissue was developed and validated and showed an acceptable difference to “gold standard” manual counting. The slide scanner can be regarded as a low-cost alternative for researchers when digital slide imaging and quantification of fluorescent...

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

Science.gov (United States)

Pawłowska, Monika; Legutko, Diana; Stefaniuk, Marzena

2017-01-01

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

z calibration of the atomic force microscope by means of a pyramidal tip

DEFF Research Database (Denmark)

Jensen, Flemming

1993-01-01

A new method for imaging the probe tip of an atomic force microscope cantilever by the atomic force microscope itself (self-imaging) is presented. The self-imaging is accomplished by scanning the probe tip across a sharper tip on the surface. By using a pyramidal probe tip with a very well......-defined aspect ratio, this technique provides an excellent z-calibration standard for the atomic force microscope....

Design and analysis of a cross-type structured-illumination confocal microscope for high speed and high resolution

International Nuclear Information System (INIS)

Kim, Young-Duk; Ahn, MyoungKi; Kim, Taejoong; Gweon, DaeGab; Yoo, Hongki

2012-01-01

There have been many studies about a super resolution microscope for many years. A super resolution microscope can detect the physical phenomena or morphology of a biological sample more precisely than conventional microscopes. The structured-illumination microscope (SIM) is one of the technologies that demonstrate super resolution. However, the conventional SIM requires more time to obtain one resolution-enhanced image than other super resolution microscopes. More specifically, the conventional SIM uses three images with a 120° phase difference for each direction and three different directions are image-processed to make one resolution enhancement by increasing the optical transfer function in three directions. In this paper, we present a novel cross structured-illumination confocal microscope (CSICM) that takes the advantage of the technology of both SIM and the confocal microscope. The CSICM uses only two directions with three phase difference images, for a total of six images. By reducing the number of images that must be obtained, the total image acquisition time and image reconstruction time in obtaining the final output images can be decreased, and the confocal microscope provides axial information of the sample automatically. We demonstrate our method of cross illumination and evaluate the performance of the CSICM and compare it to the conventional SIM and the confocal microscope. (paper)

Automatic Recognition of Acute Myelogenous Leukemia in Blood Microscopic Images Using K-means Clustering and Support Vector Machine.

Science.gov (United States)

Kazemi, Fatemeh; Najafabadi, Tooraj Abbasian; Araabi, Babak Nadjar

2016-01-01

Acute myelogenous leukemia (AML) is a subtype of acute leukemia, which is characterized by the accumulation of myeloid blasts in the bone marrow. Careful microscopic examination of stained blood smear or bone marrow aspirate is still the most significant diagnostic methodology for initial AML screening and considered as the first step toward diagnosis. It is time-consuming and due to the elusive nature of the signs and symptoms of AML; wrong diagnosis may occur by pathologists. Therefore, the need for automation of leukemia detection has arisen. In this paper, an automatic technique for identification and detection of AML and its prevalent subtypes, i.e., M2-M5 is presented. At first, microscopic images are acquired from blood smears of patients with AML and normal cases. After applying image preprocessing, color segmentation strategy is applied for segmenting white blood cells from other blood components and then discriminative features, i.e., irregularity, nucleus-cytoplasm ratio, Hausdorff dimension, shape, color, and texture features are extracted from the entire nucleus in the whole images containing multiple nuclei. Images are classified to cancerous and noncancerous images by binary support vector machine (SVM) classifier with 10-fold cross validation technique. Classifier performance is evaluated by three parameters, i.e., sensitivity, specificity, and accuracy. Cancerous images are also classified into their prevalent subtypes by multi-SVM classifier. The results show that the proposed algorithm has achieved an acceptable performance for diagnosis of AML and its common subtypes. Therefore, it can be used as an assistant diagnostic tool for pathologists.

Molecular Imaging of Apoptosis: From Micro to Macro

Science.gov (United States)

Zeng, Wenbin; Wang, Xiaobo; Xu, Pengfei; Liu, Gang; Eden, Henry S.; Chen, Xiaoyuan

2015-01-01

Apoptosis, or programmed cell death, is involved in numerous human conditions including neurodegenerative diseases, ischemic damage, autoimmune disorders and many types of cancer, and is often confused with other types of cell death. Therefore strategies that enable visualized detection of apoptosis would be of enormous benefit in the clinic for diagnosis, patient management, and development of new therapies. In recent years, improved understanding of the apoptotic machinery and progress in imaging modalities have provided opportunities for researchers to formulate microscopic and macroscopic imaging strategies based on well-defined molecular markers and/or physiological features. Correspondingly, a large collection of apoptosis imaging probes and approaches have been documented in preclinical and clinical studies. In this review, we mainly discuss microscopic imaging assays and macroscopic imaging probes, ranging in complexity from simple attachments of reporter moieties to proteins that interact with apoptotic biomarkers, to rationally designed probes that target biochemical changes. Their clinical translation will also be our focus. PMID:25825597

Immobilization method of yeast cells for intermittent contact mode imaging using the atomic force microscope

International Nuclear Information System (INIS)

De, Tathagata; Chettoor, Antony M.; Agarwal, Pranav; Salapaka, Murti V.; Nettikadan, Saju

2010-01-01

The atomic force microscope (AFM) is widely used for studying the surface morphology and growth of live cells. There are relatively fewer reports on the AFM imaging of yeast cells (Kasas and Ikai, 1995), (Gad and Ikai, 1995). Yeasts have thick and mechanically strong cell walls and are therefore difficult to attach to a solid substrate. In this report, a new immobilization technique for the height mode imaging of living yeast cells in solid media using AFM is presented. The proposed technique allows the cell surface to be almost completely exposed to the environment and studied using AFM. Apart from the new immobilization protocol, for the first time, height mode imaging of live yeast cell surface in intermittent contact mode is presented in this report. Stable and reproducible imaging over a 10-h time span is observed. A significant improvement in operational stability will facilitate the investigation of growth patterns and surface patterns of yeast cells.

Electron microscope studies. Progress report, 1 July 1964--1 June 1992

Energy Technology Data Exchange (ETDEWEB)

Crewe, A.V.; Kapp, O.H.

1992-07-01

This is a report covering the research performed in the Crewe laboratory between 1964 and 1992. Because of limitations of space we have provided relatively brief summaries of the major research directions of the facility during these years. A complete bibliography has been included and we have referenced groups of pertinent publications at the beginning of each section. This report summarizes our efforts to develop better electron microscopes and chronicles many of the experimental programs, in materials science and biology, that acted both as a stimulus to better microscope design and also as a testing ground for many instrumental innovations.

Speckle-free and halo-free low coherent Mach-Zehnder quantitative-phase-imaging module as a replacement of objective lens in conventional inverted microscopes

Science.gov (United States)

Yamauchi, Toyohiko; Yamada, Hidenao; Matsui, Hisayuki; Yasuhiko, Osamu; Ueda, Yukio

2018-02-01

We developed a compact Mach-Zehnder interferometer module to be used as a replacement of the objective lens in a conventional inverted microscope (Nikon, TS100-F) in order to make them quantitative phase microscopes. The module has a 90-degree-flipped U-shape; the dimensions of the module are 160 mm by 120 mm by 40 mm and the weight is 380 grams. The Mach-Zehnder interferometer equipped with the separate reference and sample arms was implemented in this U-shaped housing and the path-length difference between the two arms was manually adjustable. The sample under test was put on the stage of the microscope and a sample light went through it. Both arms had identical achromatic lenses for image formation and the lateral positions of them were also manually adjustable. Therefore, temporally and spatially low coherent illumination was applicable because the users were able to balance precisely the path length of the two arms and to overlap the two wavefronts. In the experiment, spectrally filtered LED light for illumination (wavelength = 633 nm and bandwidth = 3 nm) was input to the interferometer module via a 50 micrometer core optical fiber. We have successfully captured full-field interference images by a camera put on the trinocular tube of the microscope and constructed quantitative phase images of the cultured cells by means of the quarter-wavelength phase shifting algorithm. The resultant quantitative phase images were speckle-free and halo-free due to spectrally and spatially low coherent illumination.

The compound microscope: optical tube length or parfocalization?

International Nuclear Information System (INIS)

Simon, J M; Comastri, S A

2005-01-01

In various well-known textbooks for undergraduate students of physics, the compound microscope is described as having a standardized 'optical tube length'. On the other hand, in order to fulfil the parfocalization condition required by the human visual system to understand the relation between what is viewed with and without the microscope, the distance between the object and its image through the objective must remain constant as objectives are interchanged. In this paper, we show that these two requirements are not compatible in microscopes containing a revolver with various objectives and that the 'optical tube length' (which differs from the mechanical tube length) cannot be standardized. Moreover, we consider the Deutsche Industrie Norm (DIN) and the Japanese Industry Standards (JIS) norms employed in the microscope industry for standardization of the object-to-intermediate image distance, the parfocal distance and the mechanical tube length

A dual-mode mobile phone microscope using the onboard camera flash and ambient light.

Science.gov (United States)

Orth, A; Wilson, E R; Thompson, J G; Gibson, B C

2018-02-19

Mobile phone microscopes are a natural platform for point-of-care imaging, but current solutions require an externally powered illumination source, thereby adding bulk and cost. We present a mobile phone microscope that uses the internal flash or sunlight as the illumination source, thereby reducing complexity whilst maintaining functionality and performance. The microscope is capable of both brightfield and darkfield imaging modes, enabling microscopic visualisation of samples ranging from plant to mammalian cells. We describe the microscope design principles, assembly process, and demonstrate its imaging capabilities through the visualisation of unlabelled cell nuclei to observing the motility of cattle sperm and zooplankton.

Intra-operative application of optical coherence tomography with an operating microscope.

Science.gov (United States)

Just, T; Lankenau, E; Hüttmann, G; Pau, H W

2009-09-01

To introduce the use of optical coherence tomography with an operating microscope for intra-operative evaluation of the human larynx. A specially equipped operating microscope with integrated spectral domain optical coherence tomography apparatus was used during microlaryngoscopy. Technical improvements in optical coherence tomography equipment (e.g. pilot beam, variable focal distance, improved image quality and integration into an operating microscope) have enabled greater sensitivity and imaging speed and a non-contact approach. Spectral domain optical coherence tomography now enables a better correlation between optical coherence tomography images and histological findings. With this new technology, the precision of biopsy can be improved during microlaryngoscopy. Use of this new optical coherence tomography technology, integrated into an operating microscope, enables the surgeon to define the biopsy site location and resection plane precisely, while the optical zoom of the operating microscope can be used over the complete range.

High-speed imaging upgrade for a standard sample scanning atomic force microscope using small cantilevers

Energy Technology Data Exchange (ETDEWEB)

Adams, Jonathan D.; Nievergelt, Adrian; Erickson, Blake W.; Yang, Chen; Dukic, Maja; Fantner, Georg E., E-mail: georg.fantner@epfl.ch [Ecole Polytechnique Fédérale de Lausanne, Lausanne (Switzerland)

2014-09-15

We present an atomic force microscope (AFM) head for optical beam deflection on small cantilevers. Our AFM head is designed to be small in size, easily integrated into a commercial AFM system, and has a modular architecture facilitating exchange of the optical and electronic assemblies. We present two different designs for both the optical beam deflection and the electronic readout systems, and evaluate their performance. Using small cantilevers with our AFM head on an otherwise unmodified commercial AFM system, we are able to take tapping mode images approximately 5–10 times faster compared to the same AFM system using large cantilevers. By using additional scanner turnaround resonance compensation and a controller designed for high-speed AFM imaging, we show tapping mode imaging of lipid bilayers at line scan rates of 100–500 Hz for scan areas of several micrometers in size.

A new computerized moving stage for optical microscopes

Science.gov (United States)

Hatiboglu, Can Ulas; Akin, Serhat

2004-06-01

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

Region of interest and windowing-based progressive medical image delivery using JPEG2000

Science.gov (United States)

Nagaraj, Nithin; Mukhopadhyay, Sudipta; Wheeler, Frederick W.; Avila, Ricardo S.

2003-05-01

An important telemedicine application is the perusal of CT scans (digital format) from a central server housed in a healthcare enterprise across a bandwidth constrained network by radiologists situated at remote locations for medical diagnostic purposes. It is generally expected that a viewing station respond to an image request by displaying the image within 1-2 seconds. Owing to limited bandwidth, it may not be possible to deliver the complete image in such a short period of time with traditional techniques. In this paper, we investigate progressive image delivery solutions by using JPEG 2000. An estimate of the time taken in different network bandwidths is performed to compare their relative merits. We further make use of the fact that most medical images are 12-16 bits, but would ultimately be converted to an 8-bit image via windowing for display on the monitor. We propose a windowing progressive RoI technique to exploit this and investigate JPEG 2000 RoI based compression after applying a favorite or a default window setting on the original image. Subsequent requests for different RoIs and window settings would then be processed at the server. For the windowing progressive RoI mode, we report a 50% reduction in transmission time.

Analysis of photon-scanning tunneling microscope images of inhomogeneous samples: Determination of the local refractive index of channel waveguides

International Nuclear Information System (INIS)

Bourillot, E.; Fornel, F. de.; Goudonnet, J.P.

1995-01-01

Channel waveguides are imaged by a photon-scanning tunneling microscope (PSTM). The polarization of the light and its orientation with respect to the guide aids are shown to be very important parameters in the analysis of the images of such samples. We simulated image formation for the plane of incidence parallel to the axis of the guide. Our theoretical results are qualitatively in agreement with our measurements. These results show the ability of the PSTM to give information about the local refractive-index variations of a sample. 21 refs., 14 figs

Non-scanning x-ray fluorescence microscope: application to real time micro-imaging

International Nuclear Information System (INIS)

Sakurai, K.; Eba, H.

2000-01-01

So far, x-ray fluorescence (XRF) micro-imaging has been performed by a 2D positional scan of a sample against a collimated beam. Obtaining information on specific elements in a nondestructive manner is an attractive prospect for many scientific applications. Furthermore, a synchrotron micro-beam can enhance the spatial resolution down to 0.1 μm. However, the total measuring time becomes quite long (a few hours to a half day), since one needs a number of scanning points in order to obtain a high-quality image. It is possible to obtain an x-ray image with 1 M pixels and with 20 μm resolution in a very short time of 20 sec - 3 min using a non-scanning XRF microscope, which is based on completely different concept. In the present report, we discuss the application of this technique to real time micro-imaging. The experiments were carried out at BL-4A, Photon Factory, Tsukuba, Japan. We employed a grazing-incidence arrangement to make primary x-rays illuminate the whole sample surface. We adopted parallel-beam optics and extremely-close-geometry in order to detect x-ray fluorescence with a CCD camera. The selective-excitation capability of tunable monochromatic synchrotron radiation is a feasible method for distinguishing the elements of interest. One can obtain an image of each element by differentiating the images obtained above and below the absorption edges of interest. The growth of metallic dendrites from a solution dropped on a substrate was studied successfully. Several different growth patterns, corresponding to concentration and other conditions for diffusion, were observed as x-ray images. Since the present technique requires only 40 sec for each shot, it is possible to record a growing process through repeated exposures like a movie. The authors would like to thank Prof. A. Iida (Photon Factory) for his valuable comments. (author)

Confocal laser microscopic imaging of conspicuous facial pores in vivo: relation between the appearance and the internal structure of skin.

Science.gov (United States)

Sugata, Keiichi; Nishijima, Takafumi; Kitahara, Takashi; Takema, Yoshinori

2008-05-01

Conspicuous facial pores are one of the more serious esthetic defects of most concern to women. Previous microscopic observations of the skin surface around conspicuous pores have discovered large hollows and uneven skin tone. In this study, the observation area was extended from the skin surface to deeper skin to find the characteristic features of conspicuous pores in a wider spectrum. First, a magnified surface image of the cheek skin was obtained using a video microscope. Second, replicas were collected from the same area. Third, the horizontal cross-sectioned images of the epidermis and papillary dermis in different depths were non-invasively obtained using in vivo confocal laser scanning microscopy. These images were compared with each other to find a correlation between features of the skin surface and those of deeper layers. In cross-sectioned images of conspicuous pores, a strongly undulated epidermal-dermal junction was commonly observed around a pore's opening. Areas with this feature correlated well to the areas with larger hollows and an uneven skin tone. Our results indicate that there is a positive correlation between the incidence of the characteristic feature at the epidermal-dermal junction and the visual appearance of a pore.

Imaging the p-n junction in a gallium nitride nanowire with a scanning microwave microscope

Energy Technology Data Exchange (ETDEWEB)

Imtiaz, Atif [Physical Measurement Laboratory, National Institute of Standards and Technology, Boulder, Colorado 80305 (United States); Department of Electrical, Computer, and Energy Engineering, University of Colorado, Boulder, Colorado 80309 (United States); Wallis, Thomas M.; Brubaker, Matt D.; Blanchard, Paul T.; Bertness, Kris A.; Sanford, Norman A.; Kabos, Pavel, E-mail: kabos@boulder.nist.gov [Physical Measurement Laboratory, National Institute of Standards and Technology, Boulder, Colorado 80305 (United States); Weber, Joel C. [Physical Measurement Laboratory, National Institute of Standards and Technology, Boulder, Colorado 80305 (United States); Department of Mechanical Engineering, University of Colorado, Boulder, Colorado 80309 (United States); Coakley, Kevin J. [Information Technology Laboratory, National Institute of Standards and Technology, Boulder, Colorado 80305 (United States)

2014-06-30

We used a broadband, atomic-force-microscope-based, scanning microwave microscope (SMM) to probe the axial dependence of the charge depletion in a p-n junction within a gallium nitride nanowire (NW). SMM enables the visualization of the p-n junction location without the need to make patterned electrical contacts to the NW. Spatially resolved measurements of S{sub 11}{sup ′}, which is the derivative of the RF reflection coefficient S{sub 11} with respect to voltage, varied strongly when probing axially along the NW and across the p-n junction. The axial variation in S{sub 11}{sup ′}  effectively mapped the asymmetric depletion arising from the doping concentrations on either side of the junction. Furthermore, variation of the probe tip voltage altered the apparent extent of features associated with the p-n junction in S{sub 11}{sup ′} images.

Design and implementation of a cost-effective microscope for fabrication and imaging

International Nuclear Information System (INIS)

Trout, G; Basu, S

2009-01-01

The use of lasers and optical systems for advanced research and demonstrative purposes has traditionally been cost-prohibitive for many researchers. In this note, we present the design and optimization of a low-cost microscopy setup capable of imaging, fabrication or photopolymerization via multiphoton excitation of a photoactivator and the study of processes such as diffusion using fluorescence recovery after photobleaching (FRAP). The setup features a continuous wave (CW) Ar-ion laser, a pulsed Nd 3+ :YAG laser, an inverted microscope with a CCD camera and appropriate optics. The setup is cost-effective and puts a once-expensive setup within reach of more researchers interested in micron- and sub-micron-scale processes. (technical design note)

Analysis of Zebrafish Kidney Development with Time-lapse Imaging Using a Dissecting Microscope Equipped for Optical Sectioning.

Science.gov (United States)

Perner, Birgit; Schnerwitzki, Danny; Graf, Michael; Englert, Christoph

2016-04-07

In order to understand organogenesis, the spatial and temporal alterations that occur during development of tissues need to be recorded. The method described here allows time-lapse analysis of normal and impaired kidney development in zebrafish embryos by using a fluorescence dissecting microscope equipped for structured illumination and z-stack acquisition. To visualize nephrogenesis, transgenic zebrafish (Tg(wt1b:GFP)) with fluorescently labeled kidney structures were used. Renal defects were triggered by injection of an antisense morpholino oligonucleotide against the Wilms tumor gene wt1a, a factor known to be crucial for kidney development. The advantage of the experimental setup is the combination of a zoom microscope with simple strategies for re-adjusting movements in x, y or z direction without additional equipment. To circumvent focal drift that is induced by temperature variations and mechanical vibrations, an autofocus strategy was applied instead of utilizing a usually required environmental chamber. In order to re-adjust the positional changes due to a xy-drift, imaging chambers with imprinted relocation grids were employed. In comparison to more complex setups for time-lapse recording with optical sectioning such as confocal laser scanning or light sheet microscopes, a zoom microscope is easy to handle. Besides, it offers dissecting microscope-specific benefits such as high depth of field and an extended working distance. The method to study organogenesis presented here can also be used with fluorescence stereo microscopes not capable of optical sectioning. Although limited for high-throughput, this technique offers an alternative to more complex equipment that is normally used for time-lapse recording of developing tissues and organ dynamics.

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

Science.gov (United States)

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.

Computer aided solution for segmenting the neuron line in hippocampal microscope images

Science.gov (United States)

Albaidhani, Tahseen; Jassim, Sabah; Al-Assam, Hisham

2017-05-01

The brain Hippocampus component is known to be responsible for memory and spatial navigation. Its functionality depends on the status of different blood vessels within the Hippocampus and is severely impaired by Alzheimer's disease as a result blockage of increasing number of blood vessels by accumulation of amyloid-beta (Aβ) protein. Accurate counting of blood vessels within the Hippocampus of mice brain, from microscopic images, is an active research area for the understanding of Alzheimer's disease. Here, we report our work on automatic detection of the Region of Interest, i.e. the region in which blood vessels are located. This area typically falls between the hippocampus edge and the line of neurons within the Hippocampus. This paper proposes a new method to detect and exclude the neuron line to improve the accuracy of blood vessel counting because some neurons on it might lead to false positive cases as they look like blood vessels. Our proposed solution is based on using trainable segmentation approach with morphological operations, taking into account variation in colour, intensity values, and image texture. Experiments on a sufficient number of microscopy images of mouse brain demonstrate the effectiveness of the developed solution in preparation for blood vessels counting.

Generic distortion model for metrology under optical microscopes

Science.gov (United States)

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

2018-04-01

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

Designs for a quantum electron microscope

Energy Technology Data Exchange (ETDEWEB)

Kruit, P., E-mail: p.kruit@tudelft.nl [Department of Imaging Physics, Delft University of Technology, Lorentzweg 1, 2628CJ Delft (Netherlands); Hobbs, R.G.; Kim, C-S.; Yang, Y.; Manfrinato, V.R. [Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States); Hammer, J.; Thomas, S.; Weber, P. [Department of Physics, Friedrich Alexander University Erlangen-Nürnberg (FAU), Staudtstrasse 1, d-91058 Erlangen (Germany); Klopfer, B.; Kohstall, C.; Juffmann, T.; Kasevich, M.A. [Department of Physics, Stanford University, Stanford, California 94305 (United States); Hommelhoff, P. [Department of Physics, Friedrich Alexander University Erlangen-Nürnberg (FAU), Staudtstrasse 1, d-91058 Erlangen (Germany); Berggren, K.K. [Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States)

2016-05-15

One of the astounding consequences of quantum mechanics is that it allows the detection of a target using an incident probe, with only a low probability of interaction of the probe and the target. This ‘quantum weirdness’ could be applied in the field of electron microscopy to generate images of beam-sensitive specimens with substantially reduced damage to the specimen. A reduction of beam-induced damage to specimens is especially of great importance if it can enable imaging of biological specimens with atomic resolution. Following a recent suggestion that interaction-free measurements are possible with electrons, we now analyze the difficulties of actually building an atomic resolution interaction-free electron microscope, or “quantum electron microscope”. A quantum electron microscope would require a number of unique components not found in conventional transmission electron microscopes. These components include a coherent electron beam-splitter or two-state-coupler, and a resonator structure to allow each electron to interrogate the specimen multiple times, thus supporting high success probabilities for interaction-free detection of the specimen. Different system designs are presented here, which are based on four different choices of two-state-couplers: a thin crystal, a grating mirror, a standing light wave and an electro-dynamical pseudopotential. Challenges for the detailed electron optical design are identified as future directions for development. While it is concluded that it should be possible to build an atomic resolution quantum electron microscope, we have also identified a number of hurdles to the development of such a microscope and further theoretical investigations that will be required to enable a complete interpretation of the images produced by such a microscope. - Highlights: • Quantum electron microscopy has the potential of reducing radiation damage. • QEM requires a fraction of the electron wave to pass through the sample

Designs for a quantum electron microscope

International Nuclear Information System (INIS)

Kruit, P.; Hobbs, R.G.; Kim, C-S.; Yang, Y.; Manfrinato, V.R.; Hammer, J.; Thomas, S.; Weber, P.; Klopfer, B.; Kohstall, C.; Juffmann, T.; Kasevich, M.A.; Hommelhoff, P.; Berggren, K.K.

2016-01-01

One of the astounding consequences of quantum mechanics is that it allows the detection of a target using an incident probe, with only a low probability of interaction of the probe and the target. This ‘quantum weirdness’ could be applied in the field of electron microscopy to generate images of beam-sensitive specimens with substantially reduced damage to the specimen. A reduction of beam-induced damage to specimens is especially of great importance if it can enable imaging of biological specimens with atomic resolution. Following a recent suggestion that interaction-free measurements are possible with electrons, we now analyze the difficulties of actually building an atomic resolution interaction-free electron microscope, or “quantum electron microscope”. A quantum electron microscope would require a number of unique components not found in conventional transmission electron microscopes. These components include a coherent electron beam-splitter or two-state-coupler, and a resonator structure to allow each electron to interrogate the specimen multiple times, thus supporting high success probabilities for interaction-free detection of the specimen. Different system designs are presented here, which are based on four different choices of two-state-couplers: a thin crystal, a grating mirror, a standing light wave and an electro-dynamical pseudopotential. Challenges for the detailed electron optical design are identified as future directions for development. While it is concluded that it should be possible to build an atomic resolution quantum electron microscope, we have also identified a number of hurdles to the development of such a microscope and further theoretical investigations that will be required to enable a complete interpretation of the images produced by such a microscope. - Highlights: • Quantum electron microscopy has the potential of reducing radiation damage. • QEM requires a fraction of the electron wave to pass through the sample

Progress in microscopic direct reaction modeling of nucleon induced reactions

Energy Technology Data Exchange (ETDEWEB)

Dupuis, M.; Bauge, E.; Hilaire, S.; Lechaftois, F.; Peru, S.; Pillet, N.; Robin, C. [CEA, DAM, DIF, Arpajon (France)

2015-12-15

A microscopic nuclear reaction model is applied to neutron elastic and direct inelastic scatterings, and pre-equilibrium reaction. The JLM folding model is used with nuclear structure information calculated within the quasi-particle random phase approximation implemented with the Gogny D1S interaction. The folding model for direct inelastic scattering is extended to include rearrangement corrections stemming from both isoscalar and isovector density variations occurring during a transition. The quality of the predicted (n,n), (n,n{sup '}), (n,xn) and (n,n{sup '}γ) cross sections, as well as the generality of the present microscopic approach, shows that it is a powerful tool that can help improving nuclear reactions data quality. Short- and long-term perspectives are drawn to extend the present approach to more systems, to include missing reactions mechanisms, and to consistently treat both structure and reaction problems. (orig.)

The research progress of dual-modality probes for molecular imaging

International Nuclear Information System (INIS)

Cao Feng; Chen Yue

2010-01-01

Various imaging modalities have been exploited to investigate the anatomic or functional dissemination of tissues in the body. However, no single imaging modality allows overall structural, functional, and molecular information as each imaging modality has its own unique strengths and weaknesses. The combination of two imaging modalities that investigates the strengths of different methods might offer the prospect of improved diagnostic abilities. As more and more dual-modality imaging system have become clinically adopted, significant progress has been made toward the creation of dual-modality imaging probes, which can be used as novel tools for future multimodality systems. These all-in-one probes take full advantage of two different imaging modalities and could provide comprehensive information for clinical diagnostics. This review discusses the advantages and challenges in developing dual-modality imaging probes. (authors)

Microscope and method of use

Science.gov (United States)

Bongianni, Wayne L.

1984-01-01

A method and apparatus for electronically focusing and electronically scanning microscopic specimens are given. In the invention, visual images of even moving, living, opaque specimens can be acoustically obtained and viewed with virtually no time needed for processing (i.e., real time processing is used). And planar samples are not required. The specimens (if planar) need not be moved during scanning, although it will be desirable and possible to move or rotate nonplanar specimens (e.g., laser fusion targets) against the lens of the apparatus. No coupling fluid is needed, so specimens need not be wetted. A phase acoustic microscope is also made from the basic microscope components together with electronic mixers.

Adaptive striping watershed segmentation method for processing microscopic images of overlapping irregular-shaped and multicentre particles.

Science.gov (United States)

Xiao, X; Bai, B; Xu, N; Wu, K

2015-04-01

Oversegmentation is a major drawback of the morphological watershed algorithm. Here, we study and reveal that the oversegmentation is not only because of the irregular shapes of the particle images, which people are familiar with, but also because of some particles, such as ellipses, with more than one centre. A new parameter, the striping level, is introduced and the criterion for striping parameter is built to help find the right markers prior to segmentation. An adaptive striping watershed algorithm is established by applying a procedure, called the marker searching algorithm, to find the markers, which can effectively suppress the oversegmentation. The effectiveness of the proposed method is validated by analysing some typical particle images including the images of gold nanorod ensembles. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

A novel optical microscope for imaging large embryos and tissue volumes with sub-cellular resolution throughout.

Science.gov (United States)

McConnell, Gail; Trägårdh, Johanna; Amor, Rumelo; Dempster, John; Reid, Es; Amos, William Bradshaw

2016-09-23

Current optical microscope objectives of low magnification have low numerical aperture and therefore have too little depth resolution and discrimination to perform well in confocal and nonlinear microscopy. This is a serious limitation in important areas, including the phenotypic screening of human genes in transgenic mice by study of embryos undergoing advanced organogenesis. We have built an optical lens system for 3D imaging of objects up to 6 mm wide and 3 mm thick with depth resolution of only a few microns instead of the tens of microns currently attained, allowing sub-cellular detail to be resolved throughout the volume. We present this lens, called the Mesolens, with performance data and images from biological specimens including confocal images of whole fixed and intact fluorescently-stained 12.5-day old mouse embryos.

Multisite two-photon imaging of neurons on multielectrode arrays

Science.gov (United States)

Potter, Steve M.; Lukina, Natalia; Longmuir, Kenneth J.; Wu, Yan

2001-04-01

We wish to understand how neural systems store, recall, and process information. We are using cultured networks of cortical neurons grown on microelectrode arrays as a model system for studying the emergent properties of ensembles of living neurons. We have developed a 2-way communication interface between the cultured network and a computer- generated animal, the Neurally Controlled Animat. Neural activity is used to control the behavior of the Animat, and 2- photon time-lapse imaging is carried out in order to observe the morphological changes that might underlie changes in neural processing. The 2-photon microscope is ideal for repeated imaging over hours or days, with submicron resolution and little photodamage. We have designed a computer-controlled microscope stage that allows imaging several locations in sequence, in order to collect more image data. For the latest progress, see: http://www.caltech.edu/~pinelab/PotterGroup.htm.

Eye-orbit NMR imaging: works in progress

International Nuclear Information System (INIS)

Cabanis, E.A.; Iba-Zizen, M.T.; Tourbah, A.; Stievenart, J.L.; Nguyen, T.H.; Trocme, P.; Mottier, N.; Thibierge, M.; Haut, J.; Hamard, H.

1995-01-01

Progress in brain and eyes magnetic resonance imagery follows improvements in data processing. Most of the 45000 examinations performed since 1984 concerns the head (90%) and a third of them concerns the optic system (from the cornea to the calcarine fissure). The majority of the explorations has been done with a SIGNA GE device (1.5 T, 4 and 5x versions). Image treatment is performed on Advantage Windows stations. Proton spectroscopy is becoming a major tool in myelin evaluation. The spatial resolution of bi-dimensional descriptive anatomy has increased (see for instance the ciliary body and the axonal group of the optical nerve). Problem remains in the fusion of colour or grey scale X-ray scanning images with magnetic resonance images for surface or transparency tri-dimensional analysis of regional anatomy. Dynamic ( 4 D ) anatomy of cone muscles is favourable to the comprehension of theoretical data or precise clinical situations. (J.S.)

Many-beam effects in electron microscope images of lattice defects

International Nuclear Information System (INIS)

Izui, Kazuhiko; Nishida, Takahiko; Furuno, Shigemi; Otsu, Hitoshi

1974-01-01

Multi-beam effects in electron microscopic images were investigated. A computation program was developed on the basis of a matrix theory of the multi-beam effects. The matrix theory for a perfect crystal and an imperfect crystal is described, and expression for absorption coefficient is presented. The amplitude of electron wave penetrating through lattice defects is expressed by using scattering matrices which correspond to crystal slices. Calculation of extinction distance was performed, and compared with experimental results. In case of systematic reflection, the difference between two beams and from four to eight beams approximation was small, while a large effect was seen in case of accidental reflection. The intensity profile of bend contour was calculated for silicon and copper-aluminum alloy. Distance between submaxima becomes short with increase of thickness. The change in stacking fault fringes with diffraction condition was investigated. Samples were copper-aluminum alloy. Systematic behavior of the fringes was obtained, and the calculated results reproduced experimental ones. (Kato, T.)

Full-field x-ray nano-imaging at SSRF

Science.gov (United States)

Deng, Biao; Ren, Yuqi; Wang, Yudan; Du, Guohao; Xie, Honglan; Xiao, Tiqiao

2013-09-01

Full field X-ray nano-imaging focusing on material science is under developing at SSRF. A dedicated full field X-ray nano-imaging beamline based on bending magnet will be built in the SSRF phase-II project. The beamline aims at the 3D imaging of the nano-scale inner structures. The photon energy range is of 5-14keV. The design goals with the field of view (FOV) of 20μm and a spatial resolution of 20nm are proposed at 8 keV, taking a Fresnel zone plate (FZP) with outermost zone width of 25 nm. Futhermore, an X-ray nano-imaging microscope is under developing at the SSRF BL13W beamline, in which a larger FOV will be emphasized. This microscope is based on a beam shaper and a zone plate using both absorption contrast and Zernike phase contrast, with the optimized energy set to 10keV. The detailed design and the progress of the project will be introduced.

A full-field transmission x-ray microscope for time-resolved imaging of magnetic nanostructures

Energy Technology Data Exchange (ETDEWEB)

Ewald, J.; Nisius, T.; Abbati, G.; Baumbach, S.; Overbuschmann, J.; Wilhein, T. [Institute for X-Optics (IXO), Hochschule Koblenz, Joseph-Rovan-Allee 2, 53424 Remagen (Germany); Wessels, P.; Wieland, M.; Drescher, M. [The Hamburg Centre for Ultrafast Imaging (CUI), University of Hamburg, Luruper Chaussee 149, 22761 Hamburg (Germany); Institut für Experimentalphysik, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg (Germany); Vogel, A. [Institut für Angewandte Physik, University of Hamburg, Jungiusstraße 11, 20355 Hamburg (Germany); Viefhaus, J. [Deutsches Elektronen-Synchrotron (DESY), Notkestraße 85, 22607 Hamburg (Germany); Meier, G. [The Hamburg Centre for Ultrafast Imaging (CUI), University of Hamburg, Luruper Chaussee 149, 22761 Hamburg (Germany); Max Planck Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, 22761 Hamburg (Germany)

2016-01-28

Sub-nanosecond magnetization dynamics of small permalloy (Ni{sub 80}Fe{sub 20}) elements has been investigated with a new full-field transmission microscope at the soft X-ray beamline P04 of the high brilliance synchrotron radiation source PETRA III. The soft X-ray microscope generates a flat-top illumination field of 20 μm diameter using a grating condenser. A tilted nanostructured magnetic sample can be excited by a picosecond electric current pulse via a coplanar waveguide. The transmitted light of the sample plane is directly imaged by a micro zone plate with < 65 nm resolution onto a 2D gateable X-ray detector to select one particular bunch in the storage ring that probes the time evolution of the dynamic information successively via XMCD spectromicroscopy in a pump-probe scheme. In the experiments it was possible to generate a homogeneously magnetized state in patterned magnetic layers by a strong magnetic Oersted field pulse of 200 ps duration and directly observe the recovery to the initial flux-closure vortex patterns.

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

Science.gov (United States)

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.

Progressive image denoising through hybrid graph Laplacian regularization: a unified framework.

Science.gov (United States)

Liu, Xianming; Zhai, Deming; Zhao, Debin; Zhai, Guangtao; Gao, Wen

2014-04-01

Recovering images from corrupted observations is necessary for many real-world applications. In this paper, we propose a unified framework to perform progressive image recovery based on hybrid graph Laplacian regularized regression. We first construct a multiscale representation of the target image by Laplacian pyramid, then progressively recover the degraded image in the scale space from coarse to fine so that the sharp edges and texture can be eventually recovered. On one hand, within each scale, a graph Laplacian regularization model represented by implicit kernel is learned, which simultaneously minimizes the least square error on the measured samples and preserves the geometrical structure of the image data space. In this procedure, the intrinsic manifold structure is explicitly considered using both measured and unmeasured samples, and the nonlocal self-similarity property is utilized as a fruitful resource for abstracting a priori knowledge of the images. On the other hand, between two successive scales, the proposed model is extended to a projected high-dimensional feature space through explicit kernel mapping to describe the interscale correlation, in which the local structure regularity is learned and propagated from coarser to finer scales. In this way, the proposed algorithm gradually recovers more and more image details and edges, which could not been recovered in previous scale. We test our algorithm on one typical image recovery task: impulse noise removal. Experimental results on benchmark test images demonstrate that the proposed method achieves better performance than state-of-the-art algorithms.

Commissioning and modification of the low temperature scanning polarization microscope (TTSPM) and imaging of the local magnetic flux density distribution in superconducting niobium samples

International Nuclear Information System (INIS)

Gruenzweig, Matthias Sebastian Peter

2014-01-01

The dissertation is separated into two different parts, which will be presented in the following. Part I of the dissertation is about the commissioning and the modification of the ''low-temperature scanning polarization microscope'' which was designed in a previous dissertation of Stefan Guenon [1]. A scanning polarization microscope has certain advantages compared to conventional polarization microscopes. With a scanning polarization microscope it is easily possible to achieve a high illumination intensity, which is important to realize a high signal-to-noise ratio. Moreover, the confocal design of the scanning polarization microscope improves the resolution of the microscope by a factor of 1.4. Normally, it is not necessary to post-process the images by means of differential frame method to eliminate the contrast of non-magnetic origin. In contrast to conventional polarization microscopes the low-temperature scanning polarization microscope is able to image electronic transport properties via beam-induced voltage variation in addition to the magneto-optical effects. In this dissertation, it was possible to demonstrate the performance capability of the scanning polarization microscope at room temperature as well as at low temperatures. The investigation of the polar Kerr-effect has been carried out with a BaFe 12 O 19 -test sample whereas the measurements of the longitudinal Kerr-effect have been carried out with an in-plane magnetized acceleration sensor. Furthermore, an independent room temperature construction for out-of-plane measurements in a magnetic field up to 1 Tesla has been designed and implemented within the framework of a diploma thesis, supervised by the author of this dissertation. Using this construction, it was possible to gain experimental results regarding the interlayer exchange coupling between iron-terbium alloys (Fe 1-x Tb x ) and cobalt-platinum multilayers (vertical stroke Co/Pt vertical stroke n ). Indeed, it has been

On the resolution of the electron microscopic radioautography

International Nuclear Information System (INIS)

Uchida, Kazuko; Daimon, Tateo; Kawai, Kazuhiro

1981-01-01

The aim of electron microscopic radioautography is to reveal the exact localization of certain substances at the macromolecular level. In order to attain this object the establishment of a fine grain development method is indispensable. Some of latent images are formed at the contact surface between the polyhedral halide silver grain and the section surface, where the impact of #betta# particles come directly from the section involved, and since it is in contact with the section it remains in place even after development and gelatin removal. This latent image finally becomes a developed silver grain in the electron microscope radioautogram. Although the limit of resolution in electron microscopic radioautography is supposed to be the diameter of halide silver grains in emulsion, it may be improved by considering the fact that the contact area between the halide silver grain and the section surface is the minimum unit of resolution. The minimum resolution of electron microscopic radioautography was determined histologically to be about 100A. (author)

Single-pulse CARS based multimodal nonlinear optical microscope for bioimaging.

Science.gov (United States)

Kumar, Sunil; Kamali, Tschackad; Levitte, Jonathan M; Katz, Ori; Hermann, Boris; Werkmeister, Rene; Považay, Boris; Drexler, Wolfgang; Unterhuber, Angelika; Silberberg, Yaron

2015-05-18

Noninvasive label-free imaging of biological systems raises demand not only for high-speed three-dimensional prescreening of morphology over a wide-field of view but also it seeks to extract the microscopic functional and molecular details within. Capitalizing on the unique advantages brought out by different nonlinear optical effects, a multimodal nonlinear optical microscope can be a powerful tool for bioimaging. Bringing together the intensity-dependent contrast mechanisms via second harmonic generation, third harmonic generation and four-wave mixing for structural-sensitive imaging, and single-beam/single-pulse coherent anti-Stokes Raman scattering technique for chemical sensitive imaging in the finger-print region, we have developed a simple and nearly alignment-free multimodal nonlinear optical microscope that is based on a single wide-band Ti:Sapphire femtosecond pulse laser source. Successful imaging tests have been realized on two exemplary biological samples, a canine femur bone and collagen fibrils harvested from a rat tail. Since the ultra-broad band-width femtosecond laser is a suitable source for performing high-resolution optical coherence tomography, a wide-field optical coherence tomography arm can be easily incorporated into the presented multimodal microscope making it a versatile optical imaging tool for noninvasive label-free bioimaging.

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

Science.gov (United States)

Patel, Shreyas J.

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

Mapping microscopic order in plant and mammalian cells and tissues: novel differential polarization attachment for new generation confocal microscopes (DP-LSM)

Science.gov (United States)

Steinbach, G.; Pawlak, K.; Pomozi, I.; Tóth, E. A.; Molnár, A.; Matkó, J.; Garab, G.

2014-03-01

Elucidation of the molecular architecture of complex, highly organized molecular macro-assemblies is an important, basic task for biology. Differential polarization (DP) measurements, such as linear (LD) and circular dichroism (CD) or the anisotropy of the fluorescence emission (r), which can be carried out in a dichrograph or spectrofluorimeter, respectively, carry unique, spatially averaged information about the molecular organization of the sample. For inhomogeneous samples—e.g. cells and tissues—measurements on macroscopic scale are not satisfactory, and in some cases not feasible, thus microscopic techniques must be applied. The microscopic DP-imaging technique, when based on confocal laser scanning microscope (LSM), allows the pixel by pixel mapping of anisotropy of a sample in 2D and 3D. The first DP-LSM configuration, which, in fluorescence mode, allowed confocal imaging of different DP quantities in real-time, without interfering with the ‘conventional’ imaging, was built on a Zeiss LSM410. It was demonstrated to be capable of determining non-confocally the linear birefringence (LB) or LD of a sample and, confocally, its FDLD (fluorescence detected LD), the degree of polarization (P) and the anisotropy of the fluorescence emission (r), following polarized and non-polarized excitation, respectively (Steinbach et al 2009 Acta Histochem.111 316-25). This DP-LSM configuration, however, cannot simply be adopted to new generation microscopes with considerably more compact structures. As shown here, for an Olympus FV500, we designed an easy-to-install DP attachment to determine LB, LD, FDLD and r, in new-generation confocal microscopes, which, in principle, can be complemented with a P-imaging unit, but specifically to the brand and type of LSM.

Mapping microscopic order in plant and mammalian cells and tissues: novel differential polarization attachment for new generation confocal microscopes (DP-LSM)

International Nuclear Information System (INIS)

Steinbach, G; Pawlak, K; Garab, G; Pomozi, I; Tóth, E A; Molnár, A; Matkó, J

2014-01-01

Elucidation of the molecular architecture of complex, highly organized molecular macro-assemblies is an important, basic task for biology. Differential polarization (DP) measurements, such as linear (LD) and circular dichroism (CD) or the anisotropy of the fluorescence emission (r), which can be carried out in a dichrograph or spectrofluorimeter, respectively, carry unique, spatially averaged information about the molecular organization of the sample. For inhomogeneous samples—e.g. cells and tissues—measurements on macroscopic scale are not satisfactory, and in some cases not feasible, thus microscopic techniques must be applied. The microscopic DP-imaging technique, when based on confocal laser scanning microscope (LSM), allows the pixel by pixel mapping of anisotropy of a sample in 2D and 3D. The first DP-LSM configuration, which, in fluorescence mode, allowed confocal imaging of different DP quantities in real-time, without interfering with the ‘conventional’ imaging, was built on a Zeiss LSM410. It was demonstrated to be capable of determining non-confocally the linear birefringence (LB) or LD of a sample and, confocally, its FDLD (fluorescence detected LD), the degree of polarization (P) and the anisotropy of the fluorescence emission (r), following polarized and non-polarized excitation, respectively (Steinbach et al 2009 Acta Histochem.111 316–25). This DP-LSM configuration, however, cannot simply be adopted to new generation microscopes with considerably more compact structures. As shown here, for an Olympus FV500, we designed an easy-to-install DP attachment to determine LB, LD, FDLD and r, in new-generation confocal microscopes, which, in principle, can be complemented with a P-imaging unit, but specifically to the brand and type of LSM. (paper)

Development of a fluorescent microscope combined with a real-time autoradiography system

International Nuclear Information System (INIS)

Rai, Hiroki; Kanno, Satomi; Hayashi, Yoshitake; Nihei, Naoto; Nakanishi, Tomoko M.

2008-01-01

For combination with microscope, we developed real-time autoradiography system for micro-scale analysis with adjustment of the CsI(Ti) scintillator thickness for higher resolution and applying tapered fiber optic plate for magnification of autoradiograph image. We combined real-time autoradiography system with an inverted fluorescent microscope so that an autoradiograph image as well as fluorescent image, bright-field image can be acquired at the same time. In the case of observation of sliced soybean stalk traced 45 CaCl, the fluorescent and bright-field image was acquired which magnified to 50 times, the autoradiograph image of 45 Ca distribution in the tissue was acquired in almost same scale. The new microscopic system which can acquire autoradiograph image of labeled signals (low molecular weight) is expected to develop the signal transduction study and gene expression, combined with fluorescent protein techniques such as GFP etc. (author)

Distinction between amorphous and healed planar deformation features in shocked quartz using composite color scanning electron microscope cathodoluminescence (SEM-CL) imaging

Science.gov (United States)

Hamers, Maartje F.; Pennock, Gill M.; Herwegh, Marco; Drury, Martyn R.

2016-10-01

Planar deformation features (PDFs) in quartz are one of the most reliable and most widely used forms of evidence for hypervelocity impact. PDFs can be identified in scanning electron microscope cathodoluminescence (SEM-CL) images, but not all PDFs show the same CL behavior: there are nonluminescent and red luminescent PDFs. This study aims to explain the origin of the different CL emissions in PDFs. Focused ion beam (FIB) thin foils were prepared of specific sample locations selected in composite color SEM-CL images and were analyzed in a transmission electron microscope (TEM). The FIB preparation technique allowed a direct, often one-to-one correlation between the CL images and the defect structure observed in TEM. This correlation shows that composite color SEM-CL imaging allows distinction between amorphous PDFs on one hand and healed PDFs and basal Brazil twins on the other: nonluminescent PDFs are amorphous, while healed PDFs and basal Brazil twins are red luminescent, with a dominant emission peak at 650 nm. We suggest that the red luminescence is the result of preferential beam damage along dislocations, fluid inclusions, and twin boundaries. Furthermore, a high-pressure phase (possibly stishovite) in PDFs can be detected in color SEM-CL images by its blue luminescence.

Nuclear medicine and imaging research. Progress report, January 1, 1981-December 31, 1981

International Nuclear Information System (INIS)

Beck, R.N.; Cooper, M.C.

1981-09-01

The Progress Report for the period January 1, 1981-December 31, 1981 of the Franklin Memorial Research Institute discusses instrumentation and quantitative methods of evaluation in nuclear medicine and imaging research. Imaging systems and image evaluation are discussed in four projects: Radiation Detector Studies, Dual Purpose Scanner for Thyroid Imaging, Instrumentation for Image Processing and Enhancement, and Energy-Coded Processing in Nuclear Medicine

Automatic quantification of mitochondrial fragmentation from two-photon microscope images of mouse brain tissue.

Science.gov (United States)

Lihavainen, E; Kislin, M; Toptunov, D; Khiroug, L; Ribeiro, A S

2015-12-01

The morphology of mitochondria can inform about their functional state and, thus, about cell vitality. For example, fragmentation of the mitochondrial network is associated with many diseases. Recent advances in neuronal imaging have enabled the observation of mitochondria in live brains for long periods of time, enabling the study of their dynamics in animal models of diseases. To aid these studies, we developed an automatic method, based on supervised learning, for quantifying the degree of mitochondrial fragmentation in tissue images acquired via two-photon microscopy from transgenic mice, which exclusively express Enhanced cyan fluorescent protein (ECFP) under Thy1 promoter, targeted to the mitochondrial matrix in subpopulations of neurons. We tested the method on images prior to and after cardiac arrest, and found it to be sensitive to significant changes in mitochondrial morphology because of the arrest. We conclude that the method is useful in detecting morphological abnormalities in mitochondria and, likely, in other subcellular structures as well. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Capturing and displaying microscopic images used in medical diagnostics and forensic science using 4K video resolution – an application in higher education

NARCIS (Netherlands)

Jan Kuijten; Ajda Ortac; Hans Maier; Gert de Heer

2015-01-01

To analyze, interpret and evaluate microscopic images, used in medical diagnostics and forensic science, video images for educational purposes were made with a very high resolution of 4096 × 2160 pixels (4K), which is four times as many pixels as High-Definition Video (1920 × 1080 pixels).

A multi-modal stereo microscope based on a spatial light modulator.

Science.gov (United States)

Lee, M P; Gibson, G M; Bowman, R; Bernet, S; Ritsch-Marte, M; Phillips, D B; Padgett, M J

2013-07-15

Spatial Light Modulators (SLMs) can emulate the classic microscopy techniques, including differential interference (DIC) contrast and (spiral) phase contrast. Their programmability entails the benefit of flexibility or the option to multiplex images, for single-shot quantitative imaging or for simultaneous multi-plane imaging (depth-of-field multiplexing). We report the development of a microscope sharing many of the previously demonstrated capabilities, within a holographic implementation of a stereo microscope. Furthermore, we use the SLM to combine stereo microscopy with a refocusing filter and with a darkfield filter. The instrument is built around a custom inverted microscope and equipped with an SLM which gives various imaging modes laterally displaced on the same camera chip. In addition, there is a wide angle camera for visualisation of a larger region of the sample.

Preparation of theoretical scanning tunneling microscope images of adsorbed molecules: a theoretical study of benzene on the Cu(110) surface

International Nuclear Information System (INIS)

Shapter, J.G.; Rogers, B.L.; Ford, M.J.

2003-01-01

Full text: Since its development in 1982, the Scanning Tunneling Microscope (STM) has developed into a powerful tool for the study of surfaces and adsorbates. However, the utility of the technique can be further enhanced through the development of techniques for generating theoretical STM images. This is particularly true when studying molecules adsorbed on a substrate, as the results are often interpreted superficially due to an inadequate understanding of the orbital overlap probed in the experiment. A method of preparing theoretical scanning tunneling microscope (STM) images using comparatively inexpensive desktop computers and the commercially available CRYSTAL98 package is presented through a study of benzene adsorbed on the Cu(110) surface. Density Functional Theory (DFT) and Hartree-Fock (HF) methods are used to model clean Cu(110) slabs of various thicknesses and to simulate the adsorption of benzene onto these slabs. Eight possible orientations of benzene on the Cu(110) surface are proposed, and the optimum orientation according to the calculations is presented. Theoretical STM images of the Cu(110) surface and benzene adsorbed on the Cu(110) surface are compared with experimental STM images of the system from a published study. Significant differences are observed and are examined in detail

First application experiments with the Stockholm compact soft x-ray microscope

International Nuclear Information System (INIS)

Bertilson, M; Hofsten, O von; Lindblom, M; Holmberg, A; Takman, P; Vogt, U; Hertz, H; Thieme, J

2009-01-01

Most soft x-ray microscopes operating in the water window (λ = 2.3 - 4.4 nm) rely on synchrotron radiation sources. In the future we believe scientists will use soft x-ray microscopes as one imaging tool among others in their own laboratory. For this purpose we have developed a full field soft x-ray microscope with a laser-plasma source compact enough to fit on an optical table. In this contribution we describe the current status of this microscope now featuring stable operation at λ = 3.37 nm or λ 2.48 nm. In-house fabricated single element zone plates offering the possibility to perform phase contrast imaging have been implemented. We also report on the first application experiments for compact soft x-ray microscopy, including results from studies of clay minerals and colloids existing in nature and results from phase optics experiments. Planned upgrades of the microscope include increasing the source brightness, implementing more efficient condenser optics, and installing a cryo sample stage for tomography. These improvements will open up for further applications, especially in the field of biological imaging.

On-line transmission electron microscopic image analysis of chromatin texture for differentiation of thyroid gland tumors.

Science.gov (United States)

Kriete, A; Schäffer, R; Harms, H; Aus, H M

1987-06-01

Nuclei of the cells from the thyroid gland were analyzed in a transmission electron microscope by direct TV scanning and on-line image processing. The method uses the advantages of a visual-perception model to detect structures in noisy and low-contrast images. The features analyzed include area, a form factor and texture parameters from the second derivative stage. Three tumor-free thyroid tissues, three follicular adenomas, three follicular carcinomas and three papillary carcinomas were studied. The computer-aided cytophotometric method showed that the most significant differences were the statistics of the chromatin texture features of homogeneity and regularity. These findings document the possibility of an automated differentiation of tumors at the ultrastructural level.

[Remote Slit Lamp Microscope Consultation System Based on Web].

Science.gov (United States)

Chen, Junfa; Zhuo, Yong; Liu, Zuguo; Chen, Yanping

2015-11-01

To realize the remote operation of the slit lamp microscope for department of ophthalmology consultation, and visual display the real-time status of remote slit lamp microscope, a remote slit lamp microscope consultation system based on B/S structure is designed and implemented. Through framing the slit lamp microscope on the website system, the realtime acquisition and transmission of remote control and image data is realized. The three dimensional model of the slit lamp microscope is established and rendered on the web by using WebGL technology. The practical application results can well show the real-time interactive of the remote consultation system.

Combined reflection and transmission microscope for telemedicine applications in field settings.

Science.gov (United States)

Biener, Gabriel; Greenbaum, Alon; Isikman, Serhan O; Lee, Kelvin; Tseng, Derek; Ozcan, Aydogan

2011-08-21

We demonstrate a field-portable upright and inverted microscope that can image specimens in both reflection and transmission modes. This compact and cost-effective dual-mode microscope weighs only ∼135 grams (image of the specimen onto an opto-electronic sensor-array that is positioned above the beam-splitter cube. In addition to this, the illumination beam is also partially transmitted through the same specimen, which then casts lensfree in-line holograms of the same objects onto a second opto-electronic sensor-array that is positioned underneath the beam-splitter cube. By rapid digital reconstruction of the acquired lensfree holograms, transmission images (both phase and amplitude) of the same specimen are also created. We tested the performance of this field-portable microscope by imaging various micro-particles, blood smears as well as a histopathology slide corresponding to skin tissue. Being compact, light-weight and cost-effective, this combined reflection and transmission microscope might especially be useful for telemedicine applications in resource limited settings. This journal is © The Royal Society of Chemistry 2011

Optical design of Kirkpatrick-Baez microscope for ICF

International Nuclear Information System (INIS)

Mu Baozhong; Yi Shengzhen; Huang Shengling; Wang Zhanshan

2008-01-01

A new flux-resolution optical design method of Kirkpatrick-Baez microscope (KB microscope) is proposed. In X-ray imaging diagnostics of inertial confinement fusion(ICF), spatial resolution and flux are always the key parameters. While the traditional optical design of KB microscope is to correct on-axis spherical aberration and astigmatic aberration, flux-resolution method is based on lateral aberration of full field and astigmatic aberration. Thus the spatial resolution related to field dimension and light flux can be estimated. By the expressions of spatial resolution and actual limits in ICF, rules of how to set original structure and optical design flow are summarized. An instance is presented and it shows that the design has met the original targets and overcome the shortcomings of image characterization in compressed core by traditional spherical aberration correction. (authors)

Optical Computed-Tomographic Microscope for Three-Dimensional Quantitative Histology