A Simplified, Low-Cost Method for Polarized Light Microscopy

PubMed Central

Maude, Richard J.; Buapetch, Wanchana; Silamut, Kamolrat

2009-01-01

Malaria pigment is an intracellular inclusion body that appears in blood and tissue specimens on microscopic examination and can help in establishing the diagnosis of malaria. In simple light microscopy, it can be difficult to discern from cellular background and artifacts. It has long been known that if polarized light microscopy is used, malaria pigment can be much easier to distinguish. However, this technique is rarely used because of the need for a relatively costly polarization microscope. We describe a simple and economical technique to convert any standard light microscope suitable for examination of malaria films into a polarization microscope. PMID:19861611

Dual-Channel Endoscopic Indocyanine Green Fluorescence Angiography for Clipping of Cerebral Aneurysms.

PubMed

Cho, Won-Sang; Kim, Jeong Eun; Kang, Hyun-Seung; Ha, Eun Jin; Jung, Minwoong; Lee, Choonghee; Shin, Il Hyung; Kang, Uk

2017-04-01

Neuroendoscopy is useful for assessing status of perforators, parent arteries, and aneurysms beyond the straight line of microscopic view during aneurysm clipping. We aimed to evaluate the clinical usefulness of our endoscopic indocyanine green angiography (ICGA) system, which can simultaneously display visible light and indocyanine green fluorescent images. Surgical clipping of 16 unruptured aneurysms in 10 patients was performed via the keyhole approach. Using our endoscopic ICGA and commercial microscopic ICGA systems, we prospectively compared 10 targeted cerebral aneurysms at the posterior communicating (n = 4) and anterior choroidal (n = 6) arteries. Microscopic ICGA and endoscopic ICGA were feasible during surgery. Microscopic ICGA displayed 50% of branch orifices, 100% of branch trunks, and 20% of exact clip positions, whereas endoscopic ICGA showed 100% of these. Based on endoscopic ICGA findings such as incomplete clipping and compromise of parent arteries or branches, clips were repositioned in 2 cases, and additional clips were applied in 2 cases. Complete occlusion and residual neck states were achieved in 6 and 4 aneurysms after surgery. There were no neurologic deficits within 3 months after surgery except for frontalis palsy and anosmia in each patient. The endoscopic ICGA system with dual imaging of visible light and indocyanine green fluorescence was very useful for assessing geometry of aneurysms and surrounding vessels before clipping and for evaluating completeness of clip position after clipping. Copyright © 2017 Elsevier Inc. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

2010-02-01

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

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

PubMed

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

2014-10-06

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

Implementation of cost-effective diffuse light source mechanism to reduce specular reflection and halo effects for resistor-image processing

NASA Astrophysics Data System (ADS)

Chen, Yung-Sheng; Wang, Jeng-Yau

2015-09-01

Light source plays a significant role to acquire a qualified image from objects for facilitating the image processing and pattern recognition. For objects possessing specular surface, the phenomena of reflection and halo appearing in the acquired image will increase the difficulty of information processing. Such a situation may be improved by the assistance of valuable diffuse light source. Consider reading resistor via computer vision, due to the resistor's specular reflective surface it will face with a severe non-uniform luminous intensity on image yielding a higher error rate in recognition without a well-controlled light source. A measurement system including mainly a digital microscope embedded in a replaceable diffuse cover, a ring-type LED embedded onto a small pad carrying a resistor for evaluation, and Arduino microcontrollers connected with PC, is presented in this paper. Several replaceable cost-effective diffuse covers made by paper bowl, cup and box inside pasted with white paper are presented for reducing specular reflection and halo effects and compared with a commercial diffuse some. The ring-type LED can be flexibly configured to be a full or partial lighting based on the application. For each self-made diffuse cover, a set of resistors with 4 or 5 color bands are captured via digital microscope for experiments. The signal-to-noise ratio from the segmented resistor-image is used for performance evaluation. The detected principal axis of resistor body is used for the partial LED configuration to further improve the lighting condition. Experimental results confirm that the proposed mechanism can not only evaluate the cost-effective diffuse light source but also be extended as an automatic recognition system for resistor reading.

Microscope use in clinical veterinary practice and potential implications for veterinary school curricula.

PubMed

Stewart, Sherry M; Dowers, Kristy L; Cerda, Jacey R; Schoenfeld-Tacher, Regina M; Kogan, Lori R

2014-01-01

Microscopy (skill of using a microscope) and the concepts of cytology (study of cells) and histology (study of tissues) are most often taught in professional veterinary medicine programs through the traditional method of glass slides and light microscopes. Several limiting factors in veterinary training programs are encouraging educators to explore innovative options for teaching microscopy skills and the concepts of cytology and histology. An anonymous online survey was administered through the Colorado Veterinary Medical Association to Colorado veterinarians working in private practice. It was designed to assess their current usage of microscopes for cytological and histological evaluation of specimens and their perceptions of microscope use in their veterinary education. The first part of the survey was answered by 183 veterinarians, with 104 indicating they had an onsite diagnostic lab. Analysis pertaining to the use of the microscope in practice and in veterinary programs was conducted on this subset. Most respondents felt the amount of time spent in the curriculum using a microscope was just right for basic microscope use and using the microscope for viewing and learning about normal and abnormal histological sections and clinical cytology. Participants felt more emphasis could be placed on clinical and diagnostic cytology. Study results suggest that practicing veterinarians frequently use microscopes for a wide variety of cytological diagnostics. However, only two respondents indicated they prepared samples for histological evaluation. Veterinary schools should consider these results against the backdrop of pressure to implement innovative teaching techniques to meet the changing needs of the profession.

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

NASA Astrophysics Data System (ADS)

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

2018-04-01

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

Development of Low-Cost Inverted Microscope to Detect Early Growth of Mycobacterium tuberculosis in MODS Culture

PubMed Central

Zimic, Mirko; Velazco, Abner; Comina, Germán; Coronel, Jorge; Fuentes, Patricia; Luna, Carmen G.; Sheen, Patricia; Gilman, Robert H.; Moore, David A. J.

2010-01-01

Background The microscopic observation drug susceptibility (MODS) assay for rapid, low-cost detection of tuberculosis and multidrug resistant tuberculosis depends upon visualization of the characteristic cording colonies of Mycobacterium tuberculosis in liquid media. This has conventionally required an inverted light microscope in order to inspect the MODS culture plates from below. Few tuberculosis laboratories have this item and the capital cost of $5,000 for a high-end microscope could be a significant obstacle to MODS roll-out. Methodology We hypothesized that the precise definition provided by costly high-specification inverted light microscopes might not be necessary for pattern recognition. Significance In this work we describe the development of a low-cost artesenal inverted microscope that can operate in both a standard or digital mode to effectively replace the expensive commercial inverted light microscope, and an integrated system that could permit a local and remote diagnosis of tuberculosis. PMID:20351778

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

NASA Technical Reports Server (NTRS)

Wade, Lawrence A. (Inventor)

2010-01-01

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

Electron microscopic evaluation of a gold glaucoma micro shunt after explantation.

PubMed

Berk, Thomas A; Tam, Diamond Y; Werner, Liliana; Mamalis, Nick; Ahmed, Iqbal Ike K

2015-03-01

We present a case of an explanted gold glaucoma micro shunt (GMS Plus) and the subsequent light and electron microscopic analyses. The shunt was implanted in a patient with medically refractive glaucoma. The intraocular pressure (IOP) was stable at 12 mm Hg 6 months postoperatively but spiked to 26 mm Hg 6 months later; membranous growth was visible on the implant gonioscopically. A second gold micro shunt was placed 2 years after the first. The IOP was 7 mm Hg 1 week postoperatively but increased to 23 mm Hg 3 weeks later; similar membranous growth was visible on this implant. One of the shunts was explanted, and light and scanning electron microscopic analyses revealed encapsulation around the shunt exterior and connective tissue invasion of the microstructure. This represents the first electron microscopic analysis of an explanted gold glaucoma micro shunt and the first unequivocal images of the fibrotic pseudo-capsule traversing its microchannels and fenestrations. Dr. Ahmed is a consultant to and has received research grants from Solx, Inc. No other author has a financial or proprietary interest in any material or method mentioned. Copyright © 2015 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

Evaluation of a Mobile Phone-Based Microscope for Screening of Schistosoma haematobium Infection in Rural Ghana.

PubMed

Bogoch, Isaac I; Koydemir, Hatice C; Tseng, Derek; Ephraim, Richard K D; Duah, Evans; Tee, Joseph; Andrews, Jason R; Ozcan, Aydogan

2017-06-01

AbstractSchistosomiasis affects over 170 million people in Africa. Here we compare a novel, low-cost mobile phone microscope to a conventional light microscope for the label-free diagnosis of Schistosoma haematobium infections in a rural Ghanaian school setting. We tested the performance of our handheld microscope using 60 slides that were randomly chosen from an ongoing epidemiologic study in school-aged children. The mobile phone microscope had a sensitivity of 72.1% (95% confidence interval [CI]: 56.1-84.2), specificity of 100% (95% CI: 75.9-100), positive predictive value of 100% (95% CI: 86.3-100), and a negative predictive value of 57.1% (95% CI: 37.4-75.0). With its modest sensitivity and high specificity, this handheld and cost-effective mobile phone-based microscope is a stepping-stone toward developing a powerful tool in clinical and public health settings where there is limited access to conventional laboratory diagnostic support.

A light field microscope imaging spectrometer based on the microlens array

NASA Astrophysics Data System (ADS)

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

2017-10-01

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

Coherent imaging with incoherent light in digital holographic microscopy

NASA Astrophysics Data System (ADS)

Chmelik, Radim

2012-01-01

Digital holographic microscope (DHM) allows for imaging with a quantitative phase contrast. In this way it becomes an important instrument, a completely non-invasive tool for a contrast intravital observation of living cells and a cell drymass density distribution measurement. A serious drawback of current DHMs is highly coherent illumination which makes the lateral resolution worse and impairs the image quality by a coherence noise and a parasitic interference. An uncompromising solution to this problem can be found in the Leith concept of incoherent holography. An off-axis hologram can be formed with arbitrary degree of light coherence in systems equipped with an achromatic interferometer and thus the resolution and the image quality typical for an incoherent-light wide-field microscopy can be achieved. In addition, advanced imaging modes based on limited coherence can be utilized. The typical example is a coherence-gating effect which provides a finite axial resolution and makes DHM image similar to that of a confocal microscope. These possibilities were described theoretically using the formalism of three-dimensional coherent transfer functions and proved experimentally by the coherence-controlled holographic microscope which is DHM based on the Leith achromatic interferometer. Quantitative-phase-contrast imaging is demonstrated with incoherent light by the living cancer cells observation and their motility evaluation. The coherence-gating effect was proved by imaging of model samples through a scattering layer and living cells inside an opalescent medium.

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

PubMed

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

2015-10-01

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

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

PubMed

Keller, Ole

2005-08-01

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

Enhancing the performance of the light field microscope using wavefront coding

PubMed Central

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

2014-01-01

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

Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development

PubMed Central

Bertrand, Vincent; Lenne, Pierre-François

2014-01-01

Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of a light sheet based microscope, which is composed of an upright microscope and a small set of opto-mechanical elements for the generation of the light sheet. The protocol describes how to build, align the microscope and characterize the light sheet. In addition, it details how to implement the method for in toto imaging of C. elegans embryos using a simple observation chamber. The method allows the capture of 3D two-colors time-lapse movies over few hours of development. This should ease the tracking of cell shape, cell divisions and tagged proteins over long periods of time. PMID:24836407

Light-driven liquid microlenses

NASA Astrophysics Data System (ADS)

Angelini, A.; Pirani, F.; Frascella, F.; Ricciardi, S.; Descrovi, E.

2017-02-01

We propose a liquid polymeric compound based on photo-responsive azo-polymers to be used as light-activated optical element with tunable and reversible functionalities. The interaction of a laser beam locally modifies the liquid density thus producing a refractive index gradient. The laser induced refractive index profiles are observed along the optical axis of the microscope to evaluate the total phase shift induced and along the orthogonal direction to provide the axial distribution of the refractive index variation. The focusing and imaging properties of the liquid lenses as functions of the light intensity are illustrated.

Application of automatic image analysis for morphometric studies of peroxisomes stained cytochemically for catalase. II. Light-microscopic application.

PubMed

Beier, K; Fahimi, H D

1987-01-01

The feasibility of the application of a television-based image analyzer, the Texture Analysis System (TAS, Leitz Wetzlar, FRG) in conjunction with a light microscope for morphometric studies of hepatic peroxisomes has been investigated. Rat liver peroxisomes were stained with the alkaline-DAB method for localization of catalase and semithin (0.25 and 1 micron) sections of plastic-embedded material were examined under an oil immersion objective. The TAS detected the peroxisomal profiles selectively and determined their morphometric parameters automatically. The same parameters were obtained also by morphometric analysis of electron micrographs from the same material. The volume density of peroxisomes determined by TAS in semithin sections of normal liver, after correction for section thickness, is quite close to the corresponding value obtained by morphometry of electron micrographs. The difference is approximately 20%. In animals treated with the hypolipidemic drug bezafibrate, which causes proliferation of peroxisomes, TAS detected readily the increase in volume density of peroxisomes in semithin sections. In comparison with electron microscopy, however, the light-microscopic approach seems to underestimate the proliferation. The lower resolution of the light microscope and overlapping of neighbouring particles in relatively thick sections used for light-microscopic analysis may account for the differences. The present study has demonstrated the usefulness of automatic image analysis in conjunction with selective cytochemical staining of peroxisomes for morphometry of this organelle in rat liver. The light-microscopic approach is not only faster but is also extremely economical by obviating the use of an electron microscope.

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

Wang, Yimeng; Zhang, Xinping, E-mail: Zhangxinping@bjut.edu.cn; Zhang, Jian

We investigate stretching-induced microscopic deformations spatially distributed in a flexible plate of polydimethylsiloxane (PDMS) and their applications in the broadening of the output spectrum of a femtosecond optical parametric oscillator. The hologram of the stretched PDMS plate was used to evaluate indirectly the microscopic deformations. The experimental results show that these deformations exhibit weak scattering and diffraction of light and induce negligible cavity loss, ensuring practical applications of the PDMS plate as an intracavity device for lasers. In combination with the thickness reduction of the PDMS plate through stretching, the distributed deformations enable smooth tuning of the output spectrum.

LC-lens array with light field algorithm for 3D biomedical applications

NASA Astrophysics Data System (ADS)

Huang, Yi-Pai; Hsieh, Po-Yuan; Hassanfiroozi, Amir; Martinez, Manuel; Javidi, Bahram; Chu, Chao-Yu; Hsuan, Yun; Chu, Wen-Chun

2016-03-01

In this paper, liquid crystal lens (LC-lens) array was utilized in 3D bio-medical applications including 3D endoscope and light field microscope. Comparing with conventional plastic lens array, which was usually placed in 3D endoscope or light field microscope system to record image disparity, our LC-lens array has higher flexibility of electrically changing its focal length. By using LC-lens array, the working distance and image quality of 3D endoscope and microscope could be enhanced. Furthermore, the 2D/3D switching ability could be achieved if we turn off/on the electrical power on LClens array. In 3D endoscope case, a hexagonal micro LC-lens array with 350um diameter was placed at the front end of a 1mm diameter endoscope. With applying electric field on LC-lens array, the 3D specimen would be recorded as from seven micro-cameras with different disparity. We could calculate 3D construction of specimen with those micro images. In the other hand, if we turn off the electric field on LC-lens array, the conventional high resolution 2D endoscope image would be recorded. In light field microscope case, the LC-lens array was placed in front of the CMOS sensor. The main purpose of LC-lens array is to extend the refocusing distance of light field microscope, which is usually very narrow in focused light field microscope system, by montaging many light field images sequentially focusing on different depth. With adjusting focal length of LC-lens array from 2.4mm to 2.9mm, the refocusing distance was extended from 1mm to 11.3mm. Moreover, we could use a LC wedge to electrically shift the optics axis and increase the resolution of light field.

Identification and quantitative evaluation of the fiber structure in the pathological tissue using Mueller matrix microscope

NASA Astrophysics Data System (ADS)

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

2017-02-01

Fiber structure changes in the various pathological processes, such as the increase of fibrosis in liver diseases, the derangement of fiber in cervical cancer and so on. Currently, clinical pathologic diagnosis is regarded as the golden criterion, but different doctors with discrepancy in knowledge and experience may obtain different conclusions. Up to a point, quantitative evaluation of the fiber structure in the pathological tissue can be of great service to quantitative diagnosis. Mueller matrix measurement is capable of probing comprehensive microstructural information of samples and different wavelength of lights can provide more information. In this paper, we use a Mueller matrix microscope with light sources in six different wavelength. We use unstained, dewaxing liver tissue slices in four stages and the pathological biopsy of the filtration channels from rabbit eyes as samples. We apply the Mueller matrix polar decomposition (MMPD) parameter δ which corresponds to retardance to liver slices. The mean value of abnormal region get bigger when the level of fibrosis get higher and light in short wavelength is more sensitive to the microstructure of fiber. On the other hand, we use the Mueller matrix transformation (MMT) parameter Φ which is associated to the angel of fast axis in the analysis of the slices of the filtration channels from rabbit eyes. The value of kurtosis and the value of skewness shows big difference between new born region and normal region and can reveal the arrangement of fiber. These results indicate that the Mueller matrix microscope has great potential in auxiliary diagnosis.

Ultrastructure of cholinergic neurons in the laterodorsal tegmental nucleus of the rat: interaction with catecholamine fibers.

PubMed

Kubota, Y; Leung, E; Vincent, S R

1992-01-01

The ultrastructure of choline acetyltransferase (ChAT)-immunoreactive neurons in the laterodorsal tegmental nucleus (TLD) of the rat was investigated by immunohistochemical techniques. The immunoreactive neurons were medium to large in size, with a few elongated dendrites, contained well-developed cytoplasm, and a nucleus with deep infoldings. They received many nonimmunoreactive, mostly asymmetric synaptic inputs on their soma and dendrites. ChAT-immunoreactive, usually myelinated, axons were occasionally seen in TLD. Only one immunoreactive axon terminal was observed within TLD, and it made synaptic contact with a nonimmunoreactive neuronal perikaryon. The synaptic interactions between ChAT-immunoreactive neurons and tyrosine hydroxylase (TH)-immunoreactive fibers in the TLD were investigated with a double immunohistochemical staining method. ChAT-immunoreactivity detected with a beta-galactosidase method was light blue-green in the light microscope and formed dot-like electron dense particles at the electron microscopic level. TH-immunoreactivity, visualized with a nickel-enhanced immunoperoxidase method, was dark blue-black in the light microscope and diffusely opaque in the electron microscope. Therefore, the difference between these two kinds of immunoreactivity could be quite easily distinguished at both light and electron microscopic levels. In the light microscope, TH-positive fibers were often closely apposed to ChAT-immunoreactive cell bodies and dendrites in TLD. In the electron microscope, the cell soma and proximal dendrites of ChAT-immunoreactive neurons received synaptic contacts from TH-immunoreactive axon terminals. These results provide a morphological basis for catecholaminergic regulation of the cholinergic reticular system.

Microscopic observation of magnetic bacteria in the magnetic field of a rotating permanent magnet.

PubMed

Smid, Pieter; Shcherbakov, Valeriy; Petersen, Nikolai

2015-09-01

Magnetotactic bacteria are ubiquitous and can be found in both freshwater and marine environments. Due to intracellular chains of magnetic single domain particles, they behave like swimming compass needles. In external magnetic fields like the Earth's magnetic field, a torque is acting on the chain. This will cause the bacterium to be rotated and aligned with the external field. The swimming direction of magnetotactic bacteria can be controlled with external magnetic fields, which makes it convenient to study them under a light microscope. Usually, a special set of coils arranged around a light microscope is used to control the swimming magnetotactic bacteria. Here, we present a simple mechanical system with a permanent magnet, which produces a rotating magnetic field of nearly constant amplitude in the focal plane of a light microscope. The device is placed beside the light microscope and easily adaptable to almost any microscope and thus convenient for field experiments. To describe the trajectories qualitatively, a theoretical model of the trajectories is presented. This device can be used to control the swimming direction of magnetotactic bacteria and also for studying their magnetic and hydrodynamic properties.

Choice and maintenance of equipment for electron crystallography.

PubMed

Mills, Deryck J; Vonck, Janet

2013-01-01

The choice of equipment for an electron crystallography laboratory will ultimately be determined by the available budget; nevertheless, the ideal lab will have two electron microscopes: a dedicated 300 kV cryo-EM with a field emission gun and a smaller LaB(6) machine for screening. The high-end machine should be equipped with photographic film or a very large CCD or CMOS camera for 2D crystal data collection; the screening microscope needs a mid-size CCD for rapid evaluation of crystal samples. The microscope room installations should provide adequate space and a special environment that puts no restrictions on the collection of high-resolution data. Equipment for specimen preparation includes a carbon coater, glow discharge unit, light microscope, plunge freezer, and liquid nitrogen containers and storage dewars. When photographic film is to be used, additional requirements are a film desiccator, dark room, optical diffractometer, and a film scanner. Having the electron microscopes and ancillary equipment well maintained and always in optimum condition facilitates the production of high-quality data.

Scanning Miniature Microscopes without Lenses

NASA Technical Reports Server (NTRS)

Wang, Yu

2009-01-01

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

Integrative advances for OCT-guided ophthalmic surgery and intraoperative OCT: microscope integration, surgical instrumentation, and heads-up display surgeon feedback.

PubMed

Ehlers, Justis P; Srivastava, Sunil K; Feiler, Daniel; Noonan, Amanda I; Rollins, Andrew M; Tao, Yuankai K

2014-01-01

To demonstrate key integrative advances in microscope-integrated intraoperative optical coherence tomography (iOCT) technology that will facilitate adoption and utilization during ophthalmic surgery. We developed a second-generation prototype microscope-integrated iOCT system that interfaces directly with a standard ophthalmic surgical microscope. Novel features for improved design and functionality included improved profile and ergonomics, as well as a tunable lens system for optimized image quality and heads-up display (HUD) system for surgeon feedback. Novel material testing was performed for potential suitability for OCT-compatible instrumentation based on light scattering and transmission characteristics. Prototype surgical instruments were developed based on material testing and tested using the microscope-integrated iOCT system. Several surgical maneuvers were performed and imaged, and surgical motion visualization was evaluated with a unique scanning and image processing protocol. High-resolution images were successfully obtained with the microscope-integrated iOCT system with HUD feedback. Six semi-transparent materials were characterized to determine their attenuation coefficients and scatter density with an 830 nm OCT light source. Based on these optical properties, polycarbonate was selected as a material substrate for prototype instrument construction. A surgical pick, retinal forceps, and corneal needle were constructed with semi-transparent materials. Excellent visualization of both the underlying tissues and surgical instrument were achieved on OCT cross-section. Using model eyes, various surgical maneuvers were visualized, including membrane peeling, vessel manipulation, cannulation of the subretinal space, subretinal intraocular foreign body removal, and corneal penetration. Significant iterative improvements in integrative technology related to iOCT and ophthalmic surgery are demonstrated.

Optimizing the performance of dual-axis confocal microscopes via Monte-Carlo scattering simulations and diffraction theory.

PubMed

Chen, Ye; Liu, Jonathan T C

2013-06-01

Dual-axis confocal (DAC) microscopy has been found to exhibit superior rejection of out-of-focus and multiply scattered background light compared to conventional single-axis confocal microscopy. DAC microscopes rely on the use of separated illumination and collection beam paths that focus and intersect at a single focal volume (voxel) within tissue. While it is generally recognized that the resolution and contrast of a DAC microscope depends on both the crossing angle of the DAC beams, 2θ, and the focusing numerical aperture of the individual beams, α, a detailed study to investigate these dependencies has not been performed. Contrast and resolution are considered as two main criteria to assess the performance of a point-scanned DAC microscope (DAC-PS) and a line-scanned DAC microscope (DAC-LS) as a function of θ and α. The contrast and resolution of these designs are evaluated by Monte-Carlo scattering simulations and diffraction theory calculations, respectively. These results can be used for guiding the optimal designs of DAC-PS and DAC-LS microscopes.

Ultrastructural characterization of pulmonary neoplasms. II. The role of electron microscopy in characterization of uncommon epithelial pulmonary neoplasms, metastatic neoplasms to and from lung, and other tumors, including mesenchymal neoplasms.

PubMed

Herrera, G A; Alexander, C B; Jones, J M

1985-01-01

Ultrastructural analysis through better resolution adds significant information to the evaluation and classification of primary pulmonary neoplasms. Light microscopy is limited in the evaluation of lung neoplasms. In some cases the light microscopic appearance may be entirely misleading, whereas in others it is inconclusive. Immunocytochemistry provides information on cytoplasmic differentiation of various tumors and hence more data on their corresponding phenotypes. The data from immunocytochemistry without corresponding objective electron microscopic evaluation may be very difficult to interpret. Correlation of historical, gross, light, electron microscopic, and immunocytochemical data is essential for a final accurate diagnosis (fig. 20). Fine needle aspiration of pulmonary neoplasms is becoming very fashionable and a diagnosis, including type of neoplasm, is expected on the basis of examination of a limited number of cells which further emphasizes the importance of ultrastructural characterization in helping to establish an accurate diagnosis [63-69]. The current classification of pulmonary neoplasms may need to be modified in the near future to incorporate the newly created data [70-72]. At the present time, there appears to be, at least, a need for a 'double standard', as Sobin [73] has suggested, which would permit the evaluation of the biologic significance of the ultrastructural and immunocytochemical findings (as applied to classification of neoplasms) in an effort to derive meaningful clinicopathologic correlations. Figure 20 emphasizes the additive role which should be played by the various diagnostic modalities to enable a morphologic assessment which would be an accurate predictor of biologic behavior. With an accurate assessment of biologic behavior, a more appropriate and rational approach for therapy is possible. There is also an important role for ultrastructural analysis in metastatic pleural and pulmonary neoplasms, primarily adenocarcinomas, as well as in the differential diagnosis of pulmonary neoplasms versus other tumors that may be similar in histological appearance. The role of ultrastructure in mesenchymal neoplasms is also crucial in defining specific neoplastic cell populations and in some cases in the differentiation from other non-mesenchymal tumors. It seems that routine electron microscopic examination of pulmonary neoplasms provides additional information that may be of great value in the management of patients and in understanding the differentiation, and perhaps histogenesis, of pulmonary neoplasms.(ABSTRACT TRUNCATED AT 400 WORDS)

Very low risk of light-induced retinal damage during Boston keratoprosthesis surgery: a rabbit study.

PubMed

Salvador-Culla, Borja; Behlau, Irmgard; Sayegh, Rony R; Stacy, Rebecca C; Dohlman, Claes H; Delori, François

2014-02-01

The aim of this study was to assess the possibility of light damage to the retina by a surgical microscope during implantation of a Boston Keratoprosthesis (B-KPro) in rabbits. The retinal irradiance from a Zeiss OPMI Lumera S7 operating microscope was measured at the working distance (16.5 cm). Light transmittance through an isolated B-KPro was measured. A B-KPro was implanted into 1 eye of 12 rabbits with the optic covered during the procedure. The operated eyes were then continuously exposed to a fixed light intensity under the microscope for 1 hour. Fluorescein angiography was carried out on days 2 and 9 postsurgery, after which the animals were euthanized. Further, we compared the potential of these retinal exposures to well-accepted light safety guidelines applicable to humans. Light transmittance of B-KPro revealed a blockage of short wavelengths (<390 nm) and of long wavelengths (1660-1750 nm) of light. In addition, the surgical microscope filtered a part of the blue, ultraviolet, and infrared wavelengths. Neither fluorescein angiography nor a histological examination showed any morphological retinal changes in our rabbits. Moreover, the retinal exposures were well below the safety limits. Modern surgical microscopes have filters incorporated in them that block the most damaging wavelengths of light. The B-KPro is made of 100% poly(methyl methacrylate), which makes it in itself a blocker of short wavelengths of light. No damage could be demonstrated in the animal study, and the retinal exposures were well below the safety limits. Together, these results suggest that light exposures during B-KPro surgery present a low risk of photochemical damage to the retina.

Proceedings of the Workshop on Nondestructive Evaluation (NDE) of Titanium Alloys (2nd) Held on 2-4 February 1982 at the Naval Research Laboratory, Underwater Sound Reference Detachment, Orlando, Florida.

DTIC Science & Technology

1984-07-01

phase. The second step, swabbing with a 1 ml HF, 2 ml HN0 3 , 97 ml water solution, removed the stain, leaving a light field of alpha phase material in...microscope (SEM), the beta phase appeared as finely dispersed light lines in a dark - field of alpha phase material. 4.2 Ultrasonic Measurements The...appropriate couplants were used in this research. Aerotech couplant • (a light oil) was used for the wave velocity measurements. A 2 mm thick elastomer

Comparison of two methods of visual magnification for removal of adhesive flash during bracket placement using two types of orthodontic bonding agents

PubMed Central

Alencar, Estefania Queiroga de Santana e; Nobrega, Maria de Lourdes Martins; Dametto, Fabio Roberto; dos Santos, Patrícia Bittencourt Dutra; Pinheiro, Fabio Henrique de Sá Leitão

2016-01-01

ABSTRACT Objective: This study aimed to evaluate the effectiveness of two methods of visual magnification (operating microscope and light head magnifying glass) for removal of composite flash around orthodontic metal brackets. Material and Methods: Brackets were bonded in the center of the clinical crown of sixty well-preserved human premolars. Half of the sample was bonded with conventional Transbond XT (3M Unitek TM, USA), whereas the other half was bonded with Transbond TM Plus Color Change (3M Unitek TM, USA). For each type of composite, the choice of method to remove the flash was determined by randomly distributing the teeth into the following subgroups: A (removal by naked eye, n = 10), B (removal with the aid of light head magnifying glass, under 4x magnification, n = 10), and C (removal with the aid of an operating microscope, under 40x magnification, n = 10). Brackets were debonded and teeth taken to a scanning electron microscope (SS-x-550, Shimadzu, Japan) for visualization of their buccal surface. Quantification of composite flash was performed with Image Pro Plus software, and values were compared by Kruskal-Wallis test and Dunn’s post-hoc test at 5% significance level. Results: Removal of pigmented orthodontic adhesive with the aid of light head magnifying glass proved, in general, to be advantageous in comparison to all other methods. Conclusion: There was no advantage in using Transbond TM Plus Color Change alone. Further studies are necessary to draw a more definitive conclusion in regards to the benefits of using an operating microscope. PMID:28125139

Development of SEM/STEM-WDX for highly sensitive detection of light elements

NASA Astrophysics Data System (ADS)

Anan, Y.; Koguchi, M.; Kimura, T.; Sekiguchi, T.

2018-02-01

In this study, to detect the light element lithium (Li) and to detect low dosed Boron (B) in the local area at nm order, we developed an analytical electron microscope equipped with an improved serial (S)-type WDX (wavelength dispersive X-ray spectroscopy) system. In detail, to detect Li, we developed a high-conductivity multi-capillary X-ray (MCX) lens, and a diffractor with a lattice spacing (d) of 15 nm, and with a spacing variation (δ d) of 0.8 nm. Moreover, to detect low dosed light element B, we designed a high-conductivity MCX lens based on the soft X-ray reflectivity in the capillary and calculation. We developed a large-solid-angle MCX lens whose conductivity of the characteristic X-rays of B became 20 times higher than that of an MCX lens with a 30-mm focal length. Our developed analytical electron microscope was applied to a LiAl specimen and a low B-doped Si substrate specimen, and the performance of this analytical electron microscope was evaluated. As a results, this analytical electron microscope could detect the characteristic X-rays of Li with a minimum mass fraction (MMF) of 8.4 atomic % (at. %). The energy resolution was 1 eV at 55 eV. From the results of measuring the line profile of B for the unpatterned B-implantation area on a B-doped Si substrate specimen, the measured line profile data were in good agreement with secondary ion mass spectrometry data up to a depth of 100 nm with a B concentration of 0.05 at. %.

Infrared microscope inspection apparatus

DOEpatents

Forman, S.E.; Caunt, J.W.

1985-02-26

Apparatus and system for inspecting infrared transparents, such as an array of photovoltaic modules containing silicon solar cells, includes an infrared microscope, at least three sources of infrared light placed around and having their axes intersect the center of the object field and means for sending the reflected light through the microscope. The apparatus is adapted to be mounted on an X-Y translator positioned adjacent the object surface. 4 figs.

Infrared microscope inspection apparatus

DOEpatents

Forman, Steven E.; Caunt, James W.

1985-02-26

Apparatus and system for inspecting infrared transparents, such as an array of photovoltaic modules containing silicon solar cells, includes an infrared microscope, at least three sources of infrared light placed around and having their axes intersect the center of the object field and means for sending the reflected light through the microscope. The apparatus is adapted to be mounted on an X-Y translator positioned adjacent the object surface.

Light propagation and interaction observed with electrons.

PubMed

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

2016-01-01

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

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

PubMed

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

2011-08-01

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

Volumetric bioimaging based on light field microscopy with temporal focusing illumination

NASA Astrophysics Data System (ADS)

Hsu, Feng-Chun; Sie, Yong Da; Lai, Feng-Jie; Chen, Shean-Jen

2018-02-01

Light field technique at a single shot can get the whole volume image of observed sample. Therefore, the original frame rate of the optical system can be taken as the volumetric image rate. For dynamically imaging whole micron-scale biosample, a light field microscope with temporal focusing illumination has been developed. In the light field microscope, the f-number of the microlens array (MLA) is adopted to match that of the objective; hence, the subimages via adjacent lenslets do not overlay each other. A three-dimensional (3D) deconvolution algorithm is utilized to deblur the out-of-focusing part. Conventional light field microscopy (LFM) illuminates whole volume sample even noninteresting parts; nevertheless, whole volume excitation causes even more damage on bio-sample and also increase the background noise from the out of range. Therefore, temporal focusing is integrated into the light field microscope for selecting the illumination volume. Herein, a slit on the back focal plane of the objective is utilized to control the axial excitation confinement for selecting the illumination volume. As a result, the developed light field microscope with the temporal focusing multiphoton illumination (TFMPI) can reconstruct 3D images within the selected volume, and the lateral resolution approaches to the theoretical value. Furthermore, the 3D Brownian motion of two-micron fluorescent beads is observed as the criterion of dynamic sample. With superior signal-to-noise ratio and less damage to tissue, the microscope is potential to provide volumetric imaging for vivo sample.

Hyperspectral stimulated emission depletion microscopy and methods of use thereof

DOEpatents

Timlin, Jerilyn A; Aaron, Jesse S

2014-04-01

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

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

PubMed

Stemmer, A

1995-04-01

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

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

PubMed

Paddock, S W

1994-05-01

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

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

NASA Astrophysics Data System (ADS)

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

2010-02-01

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

Preparation of polymeric Janus particles by directional UV-induced reactions.

PubMed

Liu, Lianying; Ren, Mingwei; Yang, Wantai

2009-09-15

Polymeric Janus particles are obtained by UV-induced selective surface grafting polymerizations and coupling reactions, in virtue of the light-absorption of photoreactive materials such as the immobilized photoinitiator and spread photoinitiator solution on the surfaces exposed to UV light and the sheltering of densely arrayed immovable particles from light. Varying the monomers or macromolecules applied in photografting polymerization or coupling reaction, and choosing diverse polymeric particles of various size, bicolor and amphiphilic Janus particles could be successfully achieved. Observations by fluorescence microscope, scanning electron microscope ,and transmission electron microscope confirmed the asymmetrical morphology of the resultant Janus particles.

21 CFR 864.3600 - Microscopes and accessories.

Code of Federal Regulations, 2010 CFR

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...

Femtosecond resolution of soft mode dynamics in structural phase transitions

NASA Technical Reports Server (NTRS)

Dougherty, Thomas P.; Wiederrecht, Gary P.; Nelson, Keith A.; Garrett, Mark H.; Jensen, Hans P.; Warde, Cardinal

1992-01-01

The microscopic pathway along which ions or molecules in a crystal move during structural phase transition can often be described in terms of a collective vibrational mode of the lattice. In many cases, this mode, called a 'soft' phonon mode because of its characteristically low frequency near the phase transition temperature, is difficult to characterize through conventional frequency-domain spectroscopies such as light or neutron scattering. A femtosecond time-domain analog of light-scattering spectroscopy called impulsive stimulated Raman scattering (ISRS) has been used to examine the soft modes of two perovskite ferroelectric crystals. The low-frequency lattice dynamics of KNbO3 and BaTiO3 are clarified in a manner that permits critical evaluation of microscopic models for their ferroelectric transitions. The results illustrate the advantages of ISRS over conventional Raman spectroscopy of low-frequency, heavily damped soft modes.

Ectopic decidua and metastatic squamous carcinoma: presentation in a single pelvic lymph node.

PubMed

Cobb, C J

1988-06-01

The presence of ectopic decidua in pelvic lymph nodes from patients with squamous carcinoma of the cervix makes evaluation for metastatic disease difficult due to the light microscopic similarity between decidua and sheets of squamous epithelial cells. A patient is present in whom decidualized endometriosis was intimately associated with metastatic moderately differentiate squamous carcinoma in a single pelvic lymph node. This phenomenon afforded an excellent opportunity to study the unique morphologic features that distinguish these two entities. A prior report of this kind was not found. In the absence of obvious squamous differentiation (i.e., intercellular bridges, dyskeratosis, and keratin "pearl" formation), as is frequently the case with squamous carcinoma of the cervix, the light microscopic features that are most useful in distinguishing squamous carcinoma from decidua include the presence of well-defined nests of cohesive cells, nuclear hyperchromasia, and cellular pleomorphism.

Plasma cell quantification in bone marrow by computer-assisted image analysis.

PubMed

Went, P; Mayer, S; Oberholzer, M; Dirnhofer, S

2006-09-01

Minor and major criteria for the diagnosis of multiple meloma according to the definition of the WHO classification include different categories of the bone marrow plasma cell count: a shift from the 10-30% group to the > 30% group equals a shift from a minor to a major criterium, while the < 10% group does not contribute to the diagnosis. Plasma cell fraction in the bone marrow is therefore critical for the classification and optimal clinical management of patients with plasma cell dyscrasias. The aim of this study was (i) to establish a digital image analysis system able to quantify bone marrow plasma cells and (ii) to evaluate two quantification techniques in bone marrow trephines i.e. computer-assisted digital image analysis and conventional light-microscopic evaluation. The results were compared regarding inter-observer variation of the obtained results. Eighty-seven patients, 28 with multiple myeloma, 29 with monoclonal gammopathy of undetermined significance, and 30 with reactive plasmocytosis were included in the study. Plasma cells in H&E- and CD138-stained slides were quantified by two investigators using light-microscopic estimation and computer-assisted digital analysis. The sets of results were correlated with rank correlation coefficients. Patients were categorized according to WHO criteria addressing the plasma cell content of the bone marrow (group 1: 0-10%, group 2: 11-30%, group 3: > 30%), and the results compared by kappa statistics. The degree of agreement in CD138-stained slides was higher for results obtained using the computer-assisted image analysis system compared to light microscopic evaluation (corr.coeff. = 0.782), as was seen in the intra- (corr.coeff. = 0.960) and inter-individual results correlations (corr.coeff. = 0.899). Inter-observer agreement for categorized results (SM/PW: kappa 0.833) was in a high range. Computer-assisted image analysis demonstrated a higher reproducibility of bone marrow plasma cell quantification. This might be of critical importance for diagnosis, clinical management and prognostics when plasma cell numbers are low, which makes exact quantifications difficult.

Virtual microscopes in podiatric medical education.

PubMed

Becker, John H

2006-01-01

In many medical schools, microscopes are being replaced as teaching tools by computers with software that emulates the use of a light microscope. This article chronicles the adoption of "virtual microscopes" by a podiatric medical school and presents the results of educational research on the effectiveness of this adoption in a histology course. If the trend toward virtual microscopy in education continues, many 21st-century physicians will not be trained to operate a light microscope. The replacement of old technologies by new is discussed. The fundamental question is whether all podiatric physicians should be trained in the use of a particular tool or only those who are likely to use it in their own practice.

Integrative Advances for OCT-Guided Ophthalmic Surgery and Intraoperative OCT: Microscope Integration, Surgical Instrumentation, and Heads-Up Display Surgeon Feedback

PubMed Central

Ehlers, Justis P.; Srivastava, Sunil K.; Feiler, Daniel; Noonan, Amanda I.; Rollins, Andrew M.; Tao, Yuankai K.

2014-01-01

Purpose To demonstrate key integrative advances in microscope-integrated intraoperative optical coherence tomography (iOCT) technology that will facilitate adoption and utilization during ophthalmic surgery. Methods We developed a second-generation prototype microscope-integrated iOCT system that interfaces directly with a standard ophthalmic surgical microscope. Novel features for improved design and functionality included improved profile and ergonomics, as well as a tunable lens system for optimized image quality and heads-up display (HUD) system for surgeon feedback. Novel material testing was performed for potential suitability for OCT-compatible instrumentation based on light scattering and transmission characteristics. Prototype surgical instruments were developed based on material testing and tested using the microscope-integrated iOCT system. Several surgical maneuvers were performed and imaged, and surgical motion visualization was evaluated with a unique scanning and image processing protocol. Results High-resolution images were successfully obtained with the microscope-integrated iOCT system with HUD feedback. Six semi-transparent materials were characterized to determine their attenuation coefficients and scatter density with an 830 nm OCT light source. Based on these optical properties, polycarbonate was selected as a material substrate for prototype instrument construction. A surgical pick, retinal forceps, and corneal needle were constructed with semi-transparent materials. Excellent visualization of both the underlying tissues and surgical instrument were achieved on OCT cross-section. Using model eyes, various surgical maneuvers were visualized, including membrane peeling, vessel manipulation, cannulation of the subretinal space, subretinal intraocular foreign body removal, and corneal penetration. Conclusions Significant iterative improvements in integrative technology related to iOCT and ophthalmic surgery are demonstrated. PMID:25141340

Evaluation of a new soft tipped injector for the implantation of foldable intraocular lenses.

PubMed

Kleinmann, Guy; Apple, David J

2007-08-01

To evaluate the R-INJ-04 soft-tipped injector, a new injector with an integral round nozzle manufactured by Rayner Intraocular Lenses, England. 16 Rayner C-flex intraocular lenses (IOLs; Rayner Intraocular lenses, England) ranging between +10 and +30 D (2 for each power) were tested. An ophthalmic viscoelastic device (Healon, AMO, Santa Ana, California, USA) was applied to the injectors. The IOLs were loaded according to the company injector's instructions for use and were injected into a Petri dish. After the injection, all the IOLs and nozzles were evaluated by gross (macroscopic) and microscopic analyses and photographed under a light microscope. One lens of each power and the cartridge used for the implantation were then sent for further analysis by scanning electron microscopy (SEM). The rest of the IOLs were tested for power and modulation transfer function (MTF). All the injections were successful. No damage to the IOLs or to the injectors was found by gross examination, light microscopy and SEM. No deposits were found on the IOL optical surfaces or haptics. Power and MTF analysis showed a close match with the original measurements. Our results suggest that the R-INJ-04 soft-tipped injector is safe for the implantation of the C-flex IOL with power range from 10 to 30 D. No structural damage to the IOLs or to the injectors was found, and the lens power and light transmission properties were not damaged in any way by the injection process.

Spectral confocal reflection microscopy using a white light source

NASA Astrophysics Data System (ADS)

Booth, M.; Juškaitis, R.; Wilson, T.

2008-08-01

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

A portable fluorescence microscopic imaging system for cholecystectomy

NASA Astrophysics Data System (ADS)

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

2016-03-01

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

Surface plasmon resonance microscopy: achieving a quantitative optical response

PubMed Central

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

2016-01-01

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

Evaluation of physical changes in wood during colonization by Ceriporiopsis submervispora

Treesearch

Chris Hunt; Alex Wiedenhoeft; Eric Horn; Carl Houtman

2001-01-01

Mechanical, chemical, and light microscopic methods were used to observe wood cell wall changes during colonization by Ceriporiopsis submervispora. Maximum crushing load, dynamic mechanical analysis (DMA), and quantitative Simon’s staining were found to be the most useful methods for tracking biopulping action. MOE and loss modulus trended downward within 2 days of...

Proximal tubulopathies associated with monoclonal light chains: the spectrum of clinicopathologic manifestations and molecular pathogenesis.

PubMed

Herrera, Guillermo A

2014-10-01

Lesions associated with monoclonal light and heavy chains display a variety of glomerular, tubular interstitial, and vascular manifestations. While some of the entities are well recognized, including light and heavy chain deposition diseases, AL (light chain) and AH (heavy chain) amyloidosis, and light chain ("myeloma") cast nephropathy, other lesions centered on proximal tubules are much less accurately identified, properly diagnosed, and adequately understood in terms of pathogenesis and molecular mechanisms involved. These proximal tubule-centered lesions are typically associated with monoclonal light chains and have not been reported in patients with circulating monoclonal heavy chains. To determine the incidence of proximal tubulopathies in a series of patients with monoclonal light chain-related renal lesions and characterize them with an emphasis on clinical correlations and elucidation of molecular mechanisms involved in their pathogenesis. A study of 5410 renal biopsies with careful evaluation of light microscopic, immunofluorescence, and electron microscopic findings was conducted to identify these monoclonal light/heavy chain-related lesions. In selected cases, ultrastructural immunolabeling was performed to better illustrate and understand molecular mechanisms involved or to resolve specific diagnostic difficulties. In all, 2.5% of the biopsies were diagnosed as demonstrating renal pathology associated with monoclonal light or heavy chains. Of these, approximately 46% were classified as proximal tubule-centered lesions, also referred to as monoclonal light chain-associated proximal tubulopathies. These proximal tubulopathies were divided into 4 groups defined by characteristic immunomorphologic manifestations associated with specific clinical settings. These are important lesions whose recognition in the different clinical settings is extremely important for patients' clinical management, therapeutic purposes, and prognosis. These entities have been segregated into 4 distinct variants, conceptualized morphologically and clinically. Specific mechanisms involved in their pathogenesis are proposed.

7 CFR 353.9 - Standards for accreditation of non-government facilities to perform laboratory seed health...

Code of Federal Regulations, 2013 CFR

2013-01-01

... seed requires a stereo microscope. Visual examination of tissue requires a compound light microscope... equipment; fluorescent microscopes; plate readers; spectrophotometers; and the appropriate assay materials...

7 CFR 353.9 - Standards for accreditation of non-government facilities to perform laboratory seed health...

Code of Federal Regulations, 2011 CFR

2011-01-01

... seed requires a stereo microscope. Visual examination of tissue requires a compound light microscope... equipment; fluorescent microscopes; plate readers; spectrophotometers; and the appropriate assay materials...

7 CFR 353.9 - Standards for accreditation of non-government facilities to perform laboratory seed health...

Code of Federal Regulations, 2010 CFR

2010-01-01

... seed requires a stereo microscope. Visual examination of tissue requires a compound light microscope... equipment; fluorescent microscopes; plate readers; spectrophotometers; and the appropriate assay materials...

7 CFR 353.9 - Standards for accreditation of non-government facilities to perform laboratory seed health...

Code of Federal Regulations, 2014 CFR

2014-01-01

... seed requires a stereo microscope. Visual examination of tissue requires a compound light microscope... equipment; fluorescent microscopes; plate readers; spectrophotometers; and the appropriate assay materials...

Microscopic theory of light-induced deformation in amorphous side-chain azobenzene polymers.

PubMed

Toshchevikov, V; Saphiannikova, M; Heinrich, G

2009-04-16

We propose a microscopic theory of light-induced deformation of side-chain azobenzene polymers taking into account the internal structure of polymer chains. Our theory is based on the fact that interaction of chromophores with the polarized light leads to the orientation anisotropy of azobenzene macromolecules which is accompanied by the appearance of mechanical stress. It is the first microscopic theory which provides the value of the light-induced stress larger than the yield stress. This result explains a possibility for the inscription of surface relief gratings in glassy side-chain azobenzene polymers. For some chemical architectures, elongation of a sample demonstrates a nonmonotonic behavior with the light intensity and can change its sign (a stretched sample starts to be uniaxially compressed), in agreement with experiments. Using a viscoplastic approach, we show that the irreversible strain of a sample, which remains after the light is switched off, decreases with increasing temperature and can disappear at certain temperature below the glass transition temperature. This theoretical prediction is also confirmed by recent experiments.

Rapid identification of Salmonella serotypes through hyperspectral microscopy with different lighting sources

USDA-ARS?s Scientific Manuscript database

Hyperspectral microscope imaging (HMI) has the potential to classify foodborne pathogenic bacteria at cell level by combining microscope images with a spectrophotometer. In this study, the spectra generated from HMIs of five live Salmonella serovars from two light sources, metal halide (MH) and tun...

Use of a white light supercontinuum laser for confocal interference-reflection microscopy

PubMed Central

Chiu, L-D; Su, L; Reichelt, S; Amos, WB

2012-01-01

Shortly after its development, the white light supercontinuum laser was applied to confocal scanning microscopy as a more versatile substitute for the multiple monochromatic lasers normally used for the excitation of fluorescence. This light source is now available coupled to commercial confocal fluorescence microscopes. We have evaluated a supercontinuum laser as a source for a different purpose: confocal interferometric imaging of living cells and artificial models by interference reflection. We used light in the range 460–700 nm where this source provides a reasonably flat spectrum, and obtained images free from fringe artefacts caused by the longer coherence length of conventional lasers. We have also obtained images of cytoskeletal detail that is difficult to see with a monochromatic laser. PMID:22432542

Integrated optical motor.

PubMed

Kelemen, Lóránd; Valkai, Sándor; Ormos, Pál

2006-04-20

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

Light and electron microscopic observation of regenerated fungiform taste buds in patients with recovered taste function after severing chorda tympani nerve.

PubMed

Saito, Takehisa; Ito, Tetsufumi; Narita, Norihiko; Yamada, Takechiyo; Manabe, Yasuhiro

2011-11-01

The aim of this study was to evaluate the mean number of regenerated fungiform taste buds per papilla and perform light and electron microscopic observation of taste buds in patients with recovered taste function after severing the chorda tympani nerve during middle ear surgery. We performed a biopsy on the fungiform papillae (FP) in the midlateral region of the dorsal surface of the tongue from 5 control volunteers (33 total FP) and from 7 and 5 patients with and without taste recovery (34 and 29 FP, respectively) 3 years 6 months to 18 years after surgery. The specimens were observed by light and transmission electron microscopy. The taste function was evaluated by electrogustometry. The mean number of taste buds in the FP of patients with completely recovered taste function was significantly smaller (1.9 +/- 1.4 per papilla; p < 0.01) than that of the control subjects (3.8 +/- 2.2 per papilla). By transmission electron microscopy, 4 distinct types of cell (type I, II, III, and basal cells) were identified in the regenerated taste buds. Nerve fibers and nerve terminals were also found in the taste buds. It was clarified that taste buds containing taste cells and nerve endings do regenerate in the FP of patients with recovered taste function.

Titania modified activated carbon prepared from sugarcane bagasse: adsorption and photocatalytic degradation of methylene blue under visible light irradiation.

PubMed

El-Salamony, R A; Amdeha, E; Ghoneim, S A; Badawy, N A; Salem, K M; Al-Sabagh, A M

2017-12-01

Activated carbon (AC), prepared from sugarcane bagasse waste through a low-temperature chemical carbonization treatment, was used as a support for nano-TiO 2 . TiO 2 supported on AC (xTiO 2 -AC) catalysts (x = 10, 20, 50, and 70 wt.%) were prepared through a mechano-mixing method. The photocatalysts were characterized by Raman, X-ray diffraction analysis, FTIR, S BET , field emission scanning electron microscope, and optical technique. The adsorption and photo-activity of the prepared catalysts (xTiO 2 -AC) were evaluated using methylene blue (MB) dye. The photocatalytic degradation of MB was evaluated under UVC irradiation and visible light. The degradation percentage of the 100 ppm MB at neutral pH using 20TiO 2 -AC reaches 96 and 91 after 180 min under visible light and UV irradiation, respectively. In other words, these catalysts are more active under visible light than under UV light irradiation, opening the possibility of using solar light for this application.

Aqueous carrier waveguide in a flow cytometer

DOEpatents

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

1995-01-01

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

Miniature self-contained vacuum compatible electronic imaging microscope

DOEpatents

Naulleau, Patrick P.; Batson, Phillip J.; Denham, Paul E.; Jones, Michael S.

2001-01-01

A vacuum compatible CCD-based microscopic camera with an integrated illuminator. The camera can provide video or still feed from the microscope contained within a vacuum chamber. Activation of an optional integral illuminator can provide light to illuminate the microscope subject. The microscope camera comprises a housing with a objective port, modified objective, beam-splitter, CCD camera, and LED illuminator.

Design considerations of a real-time clinical confocal microscope

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1991-06-01

A real-time clinical confocal light microscope provides the ophthalmologist with a new tool for the observation of the cornea and the ocular lens. In addition, the ciliary body, the iris, and the sclera can be observed. The real-time light microscopic images have high contrast and resolution. The transverse resolution is about one half micron and the range resolution is one micron. The following observations were made with visible light: corneal epithelial cells, wing cells, basal cells, Bowman's membrane, nerve fibers, basal lamina, fibroblast nuclei, Descemet's membrane, endothelial cells. Observation of the in situ ocular lens showed lens capsule, lens epithelium, lens fibrils, the interior of lens fibrils. The applications of the confocal microscope include: eye banking, laser refractive surgery, observation of wound healing, observation of the iris, the sciera, the ciliary body, the ocular lens, and the intraocular lens. Digital image processing can produce three-dimensional reconstructions of the cornea and the ocular lens.

Proper alignment of the microscope.

PubMed

Rottenfusser, Rudi

2013-01-01

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

Faster and less phototoxic 3D fluorescence microscopy using a versatile compressed sensing scheme

PubMed Central

Woringer, Maxime; Darzacq, Xavier; Zimmer, Christophe

2017-01-01

Three-dimensional fluorescence microscopy based on Nyquist sampling of focal planes faces harsh trade-offs between acquisition time, light exposure, and signal-to-noise. We propose a 3D compressed sensing approach that uses temporal modulation of the excitation intensity during axial stage sweeping and can be adapted to fluorescence microscopes without hardware modification. We describe implementations on a lattice light sheet microscope and an epifluorescence microscope, and show that images of beads and biological samples can be reconstructed with a 5-10 fold reduction of light exposure and acquisition time. Our scheme opens a new door towards faster and less damaging 3D fluorescence microscopy. PMID:28788909

Miniaturized integration of a fluorescence microscope

PubMed Central

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

Miniaturized integration of a fluorescence microscope.

PubMed

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

2011-09-11

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 for relating cellular dynamics to animal behavior. Here we introduce a miniature (1.9 g) integrated fluorescence microscope made from mass-producible parts, including a semiconductor light source and sensor. This device enables high-speed cellular 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.

Light, electron microscopic and immunohistochemical study of the effect of low-dose aspirin during the proestrus phase on rat endometrium in the preimplantation period.

PubMed

Ateş, Utku; Baka, Meral; Turgut, Mehmet; Uyanikgil, Yiğit; Ulker, Sibel; Yilmaz, Ozlem; Tavmergen, Erol; Yurtseven, Mine

2007-04-01

To evaluate structural alterations in rat endometrium at preimplantation following treatment with aspirin beginning from proestrus by light microscopy, electron microscopy and immunohistochemical techniques. Twenty rats were divided into control (n = 10) and experimental (n = 10) groups. Experimental rats were treated with low-dose aspirin daily (2 mg/kg/day) during estrus, beginning from the proestrus phase, mated at end of cycle and treated with aspirin. Untreated pregnant rats were the control group. Rats in both groups were sacrificed at the 84th pregnancy hour; the uterus was rapidly removed and dissected free of surrounding adipose tissue. Uteri specimens from nonpregnant rats were transferred into fixative solution and processed for light, electron microscopic and immunohistochemical study. Light and electron microscopy of endometrium from control rats conformed to mid-diestrus phase; endometrial histology of the aspirin-treated group conformed to late diestrus phase. The endometrial layer was significantly thicker in the aspirin-treated group compared to the untreated control group (p <0.001). No significant difference was found in vessel number between groups. Staining with alphaV integrin was more dense in the aspirin-treated group. Based on histologic findings, we suggest low-dose aspirin has positive effects on preparing endometrium before implantation.

A computer-assisted microscopic analysis of bone tissue developed inside a polyactive polymer implanted into an equine articular surface.

PubMed

Albert, Réka; Vásárhelyi, Gábor; Bodó, Gábor; Kenyeres, Annamária; Wolf, Ervin; Papp, Tamás; Terdik, Tünde; Módis, László; Felszeghy, Szabolcs

2012-09-01

One of the most promising applications for the restoration of small or moderately sized focal articular lesions is mosaicplasty (MP). Although recurrent hemarthrosis is a rare complication after MP, recently, various strategies have been designed to find an effective filling material to prevent postoperative bleeding from the donor site. The porous biodegradable polymer Polyactive (PA; a polyethylene glycol terephthalate - polybutylene terephthalate copolymer) represents a promising solution in this respect. A histological evaluation of the longterm PA-filled donor sites obtained from 10 experimental horses was performed. In this study, attention was primarily focused on the bone tissue developed in the plug. A computer-assisted image analysis and quantitative polarized light microscopic measurements of decalcified, longitudinally sectioned, dimethylmethylene blue (DMMB)- and picrosirius red (PS) stained sections revealed that the coverage area of the bone trabecules in the PA-filled donor tunnels was substantially (25%) enlarged compared to the neighboring cancellous bone. For this quantification, identical ROIs (regions of interest) were used and compared. The birefringence retardation values were also measured with a polarized light microscope using monochromatic light. Identical retardation values could be recorded from the bone trabeculae developed in the PA and in the neighboring bone, which indicates that the collagen orientation pattern does not differ significantly among these bone trabecules. Based on our new data, we speculate that PA promotes bone formation, and some of the currently identified degradation products of PA may enhance osteo-conduction and osteoinduction inside the donor canal.

A numerical analysis of the Born approximation for image formation modeling of differential interference contrast microscopy for human embryos

NASA Astrophysics Data System (ADS)

Trattner, Sigal; Feigin, Micha; Greenspan, Hayit; Sochen, Nir

2008-03-01

The differential interference contrast (DIC) microscope is commonly used for the visualization of live biological specimens. It enables the view of the transparent specimens while preserving their viability, being a non-invasive modality. Fertility clinics often use the DIC microscope for evaluation of human embryos quality. Towards quantification and reconstruction of the visualized specimens, an image formation model for DIC imaging is sought and the interaction of light waves with biological matter is examined. In many image formation models the light-matter interaction is expressed via the first Born approximation. The validity region of this approximation is defined in a theoretical bound which limits its use to very small specimens with low dielectric contrast. In this work the Born approximation is investigated via the Helmholtz equation, which describes the interaction between the specimen and light. A solution on the lens field is derived using the Gaussian Legendre quadrature formulation. This numerical scheme is considered both accurate and efficient and has shortened significantly the computation time as compared to integration methods that required a great amount of sampling for satisfying the Whittaker - Shannon sampling theorem. By comparing the numerical results with the theoretical values it is shown that the theoretical bound is not directly relevant to microscopic imaging and is far too limiting. The numerical exhaustive experiments show that the Born approximation is inappropriate for modeling the visualization of thick human embryos.

Sample holder for axial rotation of specimens in 3D microscopy.

PubMed

Bruns, T; Schickinger, S; Schneckenburger, H

2015-10-01

In common light microscopy, observation of samples is only possible from one perspective. However, especially for larger three-dimensional specimens observation from different views is desirable. Therefore, we are presenting a sample holder permitting rotation of the specimen around an axis perpendicular to the light path of the microscope. Thus, images can be put into a defined multidimensional context, enabling reliable three-dimensional reconstructions. The device can be easily adapted to a great variety of common light microscopes and is suitable for various applications in science, education and industry, where the observation of three-dimensional specimens is essential. Fluorescence z-projection images of copepods and ixodidae ticks at different rotation angles obtained by confocal laser scanning microscopy and light sheet fluorescence microscopy are reported as representative results. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Improvement of malaria diagnostic system based on acridine orange staining.

PubMed

Kimura, Masatsugu; Teramoto, Isao; Chan, Chim W; Idris, Zulkarnain Md; Kongere, James; Kagaya, Wataru; Kawamoto, Fumihiko; Asada, Ryoko; Isozumi, Rie; Kaneko, Akira

2018-02-07

Rapid diagnosis of malaria using acridine orange (AO) staining and a light microscope with a halogen lamp and interference filter was deployed in some malaria-endemic countries. However, it has not been widely adopted because: (1) the lamp was weak as an excitation light and the set-up did not work well under unstable power supply; and, (2) the staining of samples was frequently inconsistent. The halogen lamp was replaced by a low-cost, blue light-emitting diode (LED) lamp. Using a reformulated AO solution, the staining protocol was revised to make use of a concentration gradient instead of uniform staining. To evaluate this new AO diagnostic system, a pilot field study was conducted in the Lake Victoria basin in Kenya. Without staining failure, malaria infection status of about 100 samples was determined on-site per one microscopist per day, using the improved AO diagnostic system. The improved AO diagnosis had both higher overall sensitivity (46.1 vs 38.9%: p = 0.08) and specificity (99.0 vs 96.3%) than the Giemsa method (N = 1018), using PCR diagnosis as the standard. Consistent AO staining of thin blood films and rapid evaluation of malaria parasitaemia with the revised protocol produced superior results relative to the Giemsa method. This AO diagnostic system can be set up easily at low cost using an ordinary light microscope. It may supplement rapid diagnostic tests currently used in clinical settings in malaria-endemic countries, and may be considered as an inexpensive tool for case surveillance in malaria-eliminating countries.

Auricular burns associated with operating microscope use during otologic surgery.

PubMed

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

2014-02-01

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

A light and scanning electron microscopic evaluation of electro-discharge-compacted porous titanium implants in rabbit tibia.

PubMed

Drummond, J F; Dominici, J T; Sammon, P J; Okazaki, K; Geissler, R; Lifland, M I; Anderson, S A; Renshaw, W

1995-01-01

This study used light and scanning electron microscopic (SEM) histomorphometric methods to quantitate the rate of osseointegration of totally porous titanium alloy (Ti-6Al-4V) implants prepared by a novel fabrication technique--electrodischarge compaction (EDC). EDC was used to fuse 150-250-micrometer spherical titanium alloy beads into 4 X 6 mm cylindrical implants through application of a 300-microsecond pulse of high-voltage/high-current density. Two sterilized implants were surgically placed into each tibia of 20 New Zealand white rabbits and left in situ for periods corresponding to 2, 4, 8, 12, and 24 weeks. At each time point, 4 rabbits were humanely killed, and the implants with surrounding bone were removed, fixed, and sectioned for light and SEM studies. The degree of osseointegration was quantitated by means of a True Grid Digitizing Pad and Jandel Scan Version 3.9 software on an IBM PS/2 computer. The total pore area occupied by bone was divided by the total pore area available for bone ingrowth, and a Bone Ingrowth Factor (BIF) was calculated as a percent. The light microscopic results showed BIFs of 4% at week 2, 47% at week 4, 62% at week 8, 84% at week 12, and greater than 90% at week 24. The SEM results showed BIFs of 5% at week 2, 34% at week 4, 69% at week 8, 75% at week 12, and in excess of 90% at week 24. The results of this study show that EDC implants are biocompatible and support rapid osseointegration in the rabbit tibia and suggest that, after additional studies, they may be suitable for use as dental implants in humans.

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

PubMed

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

2013-01-01

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

Remote microscopy and volumetric imaging on the surface of icy satellites

NASA Astrophysics Data System (ADS)

Soto, Alejandro; Nowicki, Keith; Howett, Carly; Feldkhun, Daniel; Retherford, Kurt D.

2017-10-01

With NASA PIDDP support we have applied recent advancements in Fourier-domain microscopy to develop an instrument capable of microscopic imaging from meter-scale distances for use on a planetary lander on the surface of an icy satellite or other planetary bodies. Without moving parts, our instrument projects dynamic patterns of laser light onto a distant target using a lightweight large-aperture reflector, which then collects the light scattered or fluoresced by the target on a fast photon-bucket detector. Using Fourier Transform based techniques, we reconstruct an image from the detected light. The remote microscope has been demonstrated to produce 2D images with better than 15 micron lateral resolution for targets at a distance of 5 meters and is capable of linearly proportionally higher resolution at shorter distances. The remote microscope is also capable of providing three-dimensional (3D) microscopic imaging capabilities, allowing future surface scientists to explore the morphology of microscopic features in surface ices, for example. The instrument enables microscopic in-situ imaging during day or night without the use of a robotic arm, greatly facilitating the surface operations for a lander or rover while expanding the area of investigation near a landing site for improved science targeting. We are developing this remote microscope for in-situ planetary exploration as a collaboration between the Southwest Research Institute, LambdaMetrics, and the University of Colorado.

The impact of loupes and microscopes on vision in endodontics.

PubMed

Perrin, P; Neuhaus, K W; Lussi, A

2014-05-01

To report on an intraradicular visual test in a simulated clinical setting under different optical conditions. Miniaturized visual tests with E-optotypes (bar distance from 0.01 to 0.05 mm) were fixed inside the root canal system of an extracted maxillary molar at different locations: at the orifice, a depth of 5 mm and the apex. The tooth was mounted in a phantom head for a simulated clinical setting. Unaided vision was compared with Galilean loupes (2.5× magnification) with integrated light source and an operating microscope (6× magnification). The influence of the dentists' age within two groups was evaluated: <40 years (n = 9) and ≥40 years (n = 15). Some younger dentists were able to identify the E-optotypes at the orifice, but otherwise, natural vision did not reveal any measurable result. With Galilean loupes, the younger dentists <40 years could see a 0.05 mm structure at the root canal orifice, in contrast to the older group ≥40 years. Only the microscope allowed the observation of structures inside the root canal, independent of age. Unaided vision and Galilean loupes with an integrated light source could not provide any measurable vision inside the root canal, but younger dentists <40 years could detect with Galilean loupes a canal orifice corresponding to the tip of the smallest endodontic instruments. Dentists over 40 years of age were dependent on the microscope to inspect the root canal system. © 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Aqueous carrier waveguide in a flow cytometer

DOEpatents

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

1995-12-12

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

Confocal multispot microscope for fast and deep imaging in semicleared tissues

NASA Astrophysics Data System (ADS)

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.

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

PubMed

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

2007-05-01

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

Volumetric Light-field Encryption at the Microscopic Scale

PubMed Central

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

2017-01-01

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

Volumetric Light-field Encryption at the Microscopic Scale

NASA Astrophysics Data System (ADS)

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

2017-01-01

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

Applications of Blue Light-curing Acrylic Resin to Forensic Sample Preparation and Microtomy.

PubMed

Groves, Ethan; Palenik, Christopher S

2016-03-01

This study discusses the results of an evaluation of a one-part blue light-curing acrylic resin for embedding trace evidence prior to the preparation of thin sections with a microtome. Through a comparison to several epoxy resins, the physical properties relevant to both trace evidence examination and analytical microscopy in general, including as viscosity, clarity, color, hardness, and cure speed, were explored. Finally, thin sections from paint samples embedded in this acrylic resin were evaluated to determine if, through smearing or impregnation, the resin contributed to the infrared spectra. The results of this study show that blue light-curing acrylic resins provide the desired properties of an embedding medium, generate high-quality thin sections, and can significantly simplify the preparation of paint chips, fibers and a multitude of other types of microscopic samples in the forensic trace evidence laboratory. © 2015 American Academy of Forensic Sciences.

EFFECTS OF CHEMICAL PROCESSING AND OXIDE ETHYLENE STERILIZATION ON CORTICAL AND CANCELLOUS RAT BONE: A LIGHT AND ELECTRON SCANNING MICROSCOPY STUDY

PubMed Central

Castiglia, Marcello Teixeira; da Silva, Juliano Voltarelli F.; Frezarim Thomazini, José Armendir; Volpon, José Batista

2015-01-01

To evaluate, under microscopic examination, the structural changes displayed by the trabecular and cortical bones after being processed chemically and sterilized by ethylene oxide. Methods: Samples of cancellous and cortical bones obtained from young female albinus rats (Wistar) were assigned to four groups according to the type of treatment: Group I- drying; Group II- drying and ethylene oxide sterilization; III- chemical treatment; IV- chemical treatment and ethylene oxide sterilization. Half of this material was analyzed under ordinary light microscope and the other half using scanning electron microscopy. Results: In all the samples, regardless the group, there was good preservation of the general morphology. For samples submitted to the chemical processing there was better preservation of the cellular content, whereas there was amalgamation of the fibres when ethylene oxide was used. Conclusion: Treatment with ethylene oxide caused amalgamation of the fibers, possibly because of heating and the chemical treatment contributed to a better cellular preservation of the osseous structure. PMID:26998450

Bi-directional transmission of molecular information by photon or electron beams passing in the close vicinity of specific molecules, and its clinical and basic research applications: 1) Diagnosis of humans or animal patients without any direct contact; 2) Light microscopic and electron microscopic localization of neuro-transmitters, heavy metals, Oncogen C-fos (AB2), etc. of intracellular fine structures of normal and abnormal single cells using light or electro-microscopic indirect Bi-Digital O-Ring Test.

PubMed

Omura, Y; Losco, M; Omura, A K; Takeshige, C; Hisamitsu, T; Nakajima, H; Soejima, K; Yamamoto, S; Ishikawa, H; Kagoshima, T

1992-01-01

In 1985, Omura, Y. discovered that, when specific molecules were placed anywhere in the close vicinity of the path of a light beam (laser), their molecular information, as well as information on electrical & magnetic fields, is transmitted bi-directionally along the path of this light beam. Namely, this information is transmitted in the direction the light beam is projected and towards the direction from which the light beam is coming. This finding was applied to the following clinical and basic research: 1) In the past, using indirect Bi-Digital O-Ring Test, human or animal patients were diagnosed through an intermediate third person holding a good electrical conducting probe, the tip of which was touching the part of the patient to be examined. However, in order to diagnose the patient in isolation from a distance, or a dangerous or unmanagable unanesthesized animal, such as a lion or tiger, the author succeeded in making a diagnosis by replacing the metal conducting probe with a soft laser beam which is held by the one hand of the third person whose index finger is placed in close vicinity of the laser beam generated by a battery-powered penlight-type solid state laser generator. Thus, diagnosis within visible distance, without direct patient contact, became a reality. 2) Using a projection light microscope, by giving indirect Bi-Digital O-Ring Test while contacting with a fine electro-conductive probe on the magnified fine structure of normal and abnormal cells, various normal and abnormal intracellular substances were localized through a third person holding a pure reference control substance with the same hand that is holding the probe as an intermediary for the indirect Bi-Digital O-Ring Test. Instead of the photon beam in a light microscope, the author found that, using an electron beam passing through the close vicinity of specific molecules of specimens in an electron microscope, the molecular information is transmitted to the magnified fluorescent screen, and an indirect Bi-Digital O-Ring Test could be performed through a projected penlight-type solid state soft laser beam on the magnified intracellular structure through an observation glass window. Using the magnified fine structure of the cells, by either a light projection microscopic field or electron microscope, in various cancer cells of both humans and animals, Oncogen C-fos (AB2) and mercury were found inside of the nucleus. Integrin alpha 5 beta 1 was found on cell membranes and nuclear cell membranes of cancer cells. Acetylcholine was not found anywhere within cancer cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Injection moulding of plastic parts with laser textured surfaces with optical applications

NASA Astrophysics Data System (ADS)

Pina-Estany, J.; García-Granada, A. A.; Corull-Massana, E.

2018-05-01

The purpose of this work is to manufacture micro and nanotextured surfaces on plastic injection moulds with the aim of replicating them and obtaining plastic parts with optical applications. Different patterns are manufactured with nanosecond and femtosecond lasers in order to obtain three different optical applications: (i) homogeneous light diffusion (ii) 1D light directionality and (iii) 2D light directionality. Induction heating is used in the injections in order to improve the textures degree of replication. The steel mould and the plastic parts are analyzed with a confocal/focus variation microscope and with a surface roughness tester. A mock-up and a luminance camera are used to evaluate the homogeneity and luminance of the homogeneous light diffusion application in comparison with the current industrial solutions.

Malaria diagnosis under field conditions in the Venezuelan Amazon.

PubMed

Metzger, W G; Vivas-Martínez, S; Rodriguez, I; Gonçalves, J; Bongard, E; Fanello, C I; Vivas, L; Magris, M

2008-01-01

To improve practical, accurate diagnosis of malaria in the Amazon rainforest of Venezuela, two rapid diagnostic tests (RDT) (OptiMAL-IT) and FalciVax) and a laboratory light microscope, used in the field with a battery-operated head lamp as an external light source, were evaluated against the standard laboratory microscope procedure for malaria detection. One hundred and thirty-six Yanomami patients were studied for the presence of malaria parasites. Thirty-three patients (24%) were positive for malaria (Plasmodium falciparum, P. vivax, P. malariae). Twenty-one (64%) of the positive patients had <100 parasites/microl. Both RDTs showed poor sensitivity (24.2% for OptiMAL-IT) and 36.4% for FalciVax) but good specificity (99% both for OptiMAL-IT) and FalciVax). Field and laboratory microscopy showed sensitivities of 94% and 91%, respectively. The kappa coefficient was 0.90, indicating a high agreement between field and laboratory microscopy. We conclude that (i) adequate slide reading cannot be substituted by either of the two RDTs in the Venezuelan Amazon and (ii) the use of a light source such as that described above makes slide reading more feasible than hitherto in remote areas without electricity.

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

PubMed

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

2015-12-01

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

Comparison of LED and Conventional Fluorescence Microscopy for Detection of Acid Fast Bacilli in a Low-Incidence Setting

PubMed Central

Minion, Jessica; Pai, Madhukar; Ramsay, Andrew; Menzies, Dick; Greenaway, Christina

2011-01-01

Introduction Light emitting diode fluorescence microscopes have many practical advantages over conventional mercury vapour fluorescence microscopes, which would make them the preferred choice for laboratories in both low- and high-resource settings, provided performance is equivalent. Methods In a nested case-control study, we compared diagnostic accuracy and time required to read slides with the Zeiss PrimoStar iLED, LW Scientific Lumin, and a conventional fluorescence microscope (Leica DMLS). Mycobacterial culture was used as the reference standard, and subgroup analysis by specimen source and organism isolated were performed. Results There was no difference in sensitivity or specificity between the three microscopes, and agreement was high for all comparisons and subgroups. The Lumin and the conventional fluorescence microscope were equivalent with respect to time required to read smears, but the Zeiss iLED was significantly time saving compared to both. Conclusions Light emitting diode microscopy should be considered by all tuberculosis diagnostic laboratories, including those in high income countries, as a replacement for conventional fluorescence microscopes. Our findings provide support to the recent World Health Organization policy recommending that conventional fluorescence microscopy be replaced by light emitting diode microscopy using auramine staining in all settings where fluorescence microscopy is currently used. PMID:21811622

Accessible microscopy workstation for students and scientists with mobility impairments.

PubMed

Duerstock, Bradley S

2006-01-01

An integrated accessible microscopy workstation was designed and developed to allow persons with mobility impairments to control all aspects of light microscopy with minimal human assistance. This system, named AccessScope, is capable of performing brightfield and fluorescence microscopy, image analysis, and tissue morphometry requisite for undergraduate science courses to graduate-level research. An accessible microscope is necessary for students and scientists with mobility impairments to be able to use a microscope independently to better understand microscopical imaging concepts and cell biology. This knowledge is not always apparent by simply viewing a catalog of histological images. The ability to operate a microscope independently eliminates the need to hire an assistant or rely on a classmate and permits one to take practical laboratory examinations by oneself. Independent microscope handling is also crucial for graduate students and scientists with disabilities to perform scientific research. By making a personal computer as the user interface for controlling AccessScope functions, different upper limb mobility impairments could be accommodated by using various computer input devices and assistive technology software. Participants with a range of upper limb mobility impairments evaluated the prototype microscopy workstation. They were able to control all microscopy functions including loading different slides without assistance.

Effects of radiation upon the light-sensing elements of the retina as characterized by scanning electron microscopy

NASA Technical Reports Server (NTRS)

Malachowski, M. J.; Tobias, C. A.; Leith, J. T.

1977-01-01

A model system using Necturus maculosus, the common mudpuppy, was established for evaluating effects of radiation upon the light-sensing elements of the retina. Accelerated heavy ions of helium and neon from the Berkeley Bevalac were used. A number of criteria were chosen to characterize radiation damage by observing morphological changes with the scanning electron microscope. The studies indicated retina sensitivity to high-LET (neon) particles at radiation levels below 10 rads (7 particles per visual element) whereas no significant effects were seen from fast helium ions below 50 rads.

Transition of a dental histology course from light to virtual microscopy.

PubMed

Weaker, Frank J; Herbert, Damon C

2009-10-01

The transition of the dental histology course at the University of Texas Health Science Center at San Antonio Dental School was completed gradually over a five-year period. A pilot project was initially conducted to study the feasibility of integrating virtual microscopy into a traditional light microscopic lecture and laboratory course. Because of the difficulty of procuring quality calcified and decalcified sections of teeth, slides from the student loan collection in the oral histology block of the course were outsourced for conversion to digital images and placed on DVDs along with a slide viewer. The slide viewer mimicked the light microscope, allowing horizontal and vertical movement and changing of magnification, and, in addition, a feature to capture static images. In a survey, students rated the ease of use of the software, quality of the images, maneuverability of the images, and questions regarding use of the software, effective use of laboratory, and faculty time. Because of the positive support from the students, our entire student loan collection of 153 glass slides was subsequently converted to virtual images and distributed on an Apricorn pocket external hard drive. Students were asked to assess the virtual microscope over a four-year period. As a result of the surveys, light microscopes have been totally eliminated, and microscope exams have been replaced with project slide examinations. In the future, we plan to expand our virtual slides and incorporate computer testing.

Images created in a model eye during simulated cataract surgery can be the basis for images perceived by patients during cataract surgery

PubMed Central

Inoue, M; Uchida, A; Shinoda, K; Taira, Y; Noda, T; Ohnuma, K; Bissen-Miyajima, H; Hirakata, A

2014-01-01

Purpose To evaluate the images created in a model eye during simulated cataract surgery. Patients and methods This study was conducted as a laboratory investigation and interventional case series. An artificial opaque lens, a clear intraocular lens (IOL), or an irrigation/aspiration (I/A) tip was inserted into the ‘anterior chamber' of a model eye with the frosted posterior surface corresponding to the retina. Video images were recorded of the posterior surface of the model eye from the rear during simulated cataract surgery. The video clips were shown to 20 patients before cataract surgery, and the similarity of their visual perceptions to these images was evaluated postoperatively. Results The images of the moving lens fragments and I/A tip and the insertion of the IOL were seen from the rear. The image through the opaque lens and the IOL without moving objects was the light of the surgical microscope from the rear. However, when the microscope light was turned off after IOL insertion, the images of the microscope and operating room were observed by the room illumination from the rear. Seventy percent of the patients answered that the visual perceptions of moving lens fragments were similar to the video clips and 55% reported similarity with the IOL insertion. Eighty percent of the patients recommended that patients watch the video clip before their scheduled cataract surgery. Conclusions The patients' visual perceptions during cataract surgery can be reproduced in the model eye. Watching the video images preoperatively may help relax the patients during surgery. PMID:24788007

Compact, light-weight and cost-effective microscope based on lensless incoherent holography for telemedicine applications.

PubMed

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

2010-06-07

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 approximately 46 grams with dimensions smaller than 4.2 cm x 4.2 cm x 5.8 cm that achieves sub-cellular resolution over a large field of view of approximately 24 mm(2). 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.

The optics of microscope image formation.

PubMed

Wolf, David E

2013-01-01

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

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

PubMed

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.

Imaging arrangement and microscope

DOEpatents

Pertsinidis, Alexandros; Chu, Steven

2015-12-15

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

In situ flat embedding of monolayers and cell relocation in the acrylic resin LR white for comparative light and electron microscopy studies.

PubMed

Steiner, M; Schöfer, C; Mosgoeller, W

1994-12-01

A simple and reliable method has been developed for the in situ LR White embedding of cell monolayers grown on glass cover-slips. Combined with cytochemical or immunological procedures, this technique allows light and/or electron microscopy investigations of a large number of cells in the same horizontal plane within a relatively short period of time. It can be applied to cells grown on microgrid finder cover-slips which allows a distinct site of even an individual cell of a monolayer to be studied at first at the light microscope level and subsequently at the electron microscope level. Hence, it is also suitable for controlling manipulation of single cells, followed by their serial sectioning after relocation in the electron microscope.

CW laser use in biomedical research and practice

NASA Astrophysics Data System (ADS)

Matthopoulos, D. P.

2003-04-01

The communication of humans with their surrouding is achieved through their senses and the related organs. Visual communication using the eyes is made possible because the various sources of light, natural i.e. the sun or the lightning, or artificial such as Lasers, emit electromagnetic radiation which is either reflected or scattered by surfaces. This radiation received by eyes is processed in the brain where the images of the environment are developed. The luminous processing can be either macro- or microscopic. The macroscopic processing is the result of light coming from the sun or from wide range lamps, while the microscopic results from light coming from wide range lamps, mercury lamps, lasers or electron beam. The microscopic processing is the subject we are dealing with in this presentation.

Evaluation of sperm motility with CASA-Mot: which factors may influence our measurements?

PubMed

Yeste, Marc; Bonet, Sergi; Rodríguez-Gil, Joan E; Rivera Del Álamo, Maria M

2018-03-14

Computer-aided sperm analysis (CASA) is now routinely used in IVF clinics, animal breeding centres and research laboratories. Although CASA provides a more objective way to evaluate sperm parameters, a significant number of factors can affect these measurements. This paper classifies these factors into four categories: (1) sample and slide (e.g. preincubation time, type of specimen and type of chamber slide); (2) microscope (e.g. light source and microscope stage); (3) hardware and software, including the settings of each system; and (4) user-related factors. We review the effects of the different factors in each category on the measurements made and emphasise the need to take measures to standardise evaluations. The take-home message of the present article is that there are several commercial and useful CASA systems, and all are appropriate for routine analysis. Non-commercial systems may also be good choices when the user needs to adapt the device to specific experimental conditions. In both cases (commercial and non-commercial), it is important that standard protocols are put in place for evaluation, as well as methods to validate the system.

Effect of phosphate treatment of Acid-etched implants on mineral apposition rates near implants in a dog model.

PubMed

Foley, Christine Hyon; Kerns, David G; Hallmon, William W; Rivera-Hidalgo, Francisco; Nelson, Carl J; Spears, Robert; Dechow, Paul C; Opperman, Lynne A

2010-01-01

This study evaluated the effects of phosphate coating of acid-etched titanium on the mineral apposition rate (MAR) and new bone-to-implant contact (BIC) in a canine model. Titanium implants (2.2 3 4 mm) with acid-etched surfaces that were electrolytically phosphated or not were placed in 48 mandibular sites in six foxhounds. Tetracycline and calcein dyes were administered 1 week after implant placement and 1 week before sacrifice. At 12 weeks after implant placement, the animals were sacrificed. MAR and BIC were evaluated using fluorescence microscopy. Light microscopic and histologic evaluations were performed on undecalcified sections. Microscopic evaluation showed the presence of healthy osteoblasts lining bone surfaces near implants. Similar BIC was observed in phosphated and nonphosphated titanium implant sites. MAR was significantly higher around the nonphosphated titanium implant surfaces than around the phosphated titanium samples. No significant differences were found between dogs or implant sites. Acid-etched implants showed significantly higher MARs compared to acid-etched, phosphate-coated implants. Int J Maxillofac Implants 2010;25:278-286.

Visualization of Neutrophil Extracellular Traps and Fibrin Meshwork in Human Fibrinopurulent Inflammatory Lesions: III. Correlative Light and Electron Microscopic Study

PubMed Central

Onouchi, Takanori; Shiogama, Kazuya; Mizutani, Yasuyoshi; Takaki, Takashi; Tsutsumi, Yutaka

2016-01-01

Neutrophil extracellular traps (NETs) released from dead neutrophils at the site of inflammation represent webs of neutrophilic DNA stretches dotted with granule-derived antimicrobial proteins, including lactoferrin, and play important roles in innate immunity against microbial infection. We have shown the coexistence of NETs and fibrin meshwork in varied fibrinopurulent inflammatory lesions at both light and electron microscopic levels. In the present study, correlative light and electron microscopy (CLEM) employing confocal laser scanning microscopy and scanning electron microscopy was performed to bridge light and electron microscopic images of NETs and fibrin fibrils in formalin-fixed, paraffin-embedded, autopsied lung sections of legionnaire’s pneumonia. Lactoferrin immunoreactivity and 4'-6-diamidino-2-phenylindole (DAPI) reactivity were used as markers of NETs, and fibrin was probed by fibrinogen gamma chain. Of note is that NETs light microscopically represented as lactoferrin and DAPI-colocalized dots, 2.5 μm in diameter. CLEM gave super-resolution images of NETs and fibrin fibrils: “Dotted” NETs were ultrastructurally composed of fine filaments and masses of 58 nm-sized globular materials. A fibrin fibril consisted of clusters of smooth-surfaced filaments. NETs filaments (26 nm in diameter) were significantly thinner than fibrin filaments (295 nm in diameter). Of note is that CLEM was applicable to formalin-fixed, paraffin-embedded sections of autopsy material. PMID:27917008

Williams configures the LMM

NASA Image and Video Library

2016-04-18

ISS047e066551 (04/18/2016) --- NASA astronaut Jeff Williams configures the station’s Light Microscopy Module (LMM), a modified commercial, highly flexible, state-of-the-art light imaging microscope facility that provides researchers with powerful diagnostic hardware and software. The LMM enables novel research of microscopic phenomena in microgravity, with the capability of remotely acquiring and downloading digital images and videos across many levels of magnification.

Interactive effects of melatonin, exercise and diabetes on liver glycogen levels.

PubMed

Bicer, Mursel; Akil, Mustafa; Avunduk, Mustafa Cihat; Kilic, Mehmet; Mogulkoc, Rasim; Baltaci, Abdulkerim Kasim

2011-01-01

This study aimed to examine the effects of melatonin supplementation on liver glycogen levels in rats with streptozotocin- induced diabetes and subjected to acute swimming exercise. Eighty Sprague-Dawley type adult male rats were divided into eight groups: Group 1, general control; Group 2, melatonin-supplemented control; Group 3, melatonin-supplemented diabetes; Group 4, swimming control; Group 5, melatonin-supplemented swimming; Group 6, melatonin-supplemented diabetic swimming; Group 7, diabetic swimming; Group 8, diabetic control. Melatonin was supplemented at a dose of 3 mg/kg/day intraperitoneally for four weeks. Liver tissue samples were collected and evaluated using a Nikon Eclipse E400 light microscope. All images obtained from the light microscope were transferred to PC medium and evaluated using Clemex PE 3.5 image analysis software. The lowest liver glycogen levels in the study were found in group 4. Liver glycogen levels in groups 3, 6, 7 and 8 (the diabetic groups) were higher than group 4, but lower than those in groups 1 and 2. The lowest liver glycogen levels were obtained in groups 1 and 2. The study indicates that melatonin supplementation maintains the liver glycogen levels that decrease in acute swimming exercise, while induced diabetes prevents this maintenance effect in rats.

Studying aerosol light scattering based on aspect ratio distribution observed by fluorescence microscope.

PubMed

Li, Li; Zheng, Xu; Li, Zhengqiang; Li, Zhanhua; Dubovik, Oleg; Chen, Xingfeng; Wendisch, Manfred

2017-08-07

Particle shape is crucial to the properties of light scattered by atmospheric aerosol particles. A method of fluorescence microscopy direct observation was introduced to determine the aspect ratio distribution of aerosol particles. The result is comparable with that of the electron microscopic analysis. The measured aspect ratio distribution has been successfully applied in modeling light scattering and further in simulation of polarization measurements of the sun/sky radiometer. These efforts are expected to improve shape retrieval from skylight polarization by using directly measured aspect ratio distribution.

Light Microscopy Microscope Experiment

NASA Image and Video Library

2016-02-04

Ground testing for the first confocal Light Microscopy Microscope (LMM) Experiment. Procter and Gamble is working with NASA Glenn scientists to prepare for a study that examines product stabilizers in a microgravity environment. The particles in the tube glow orange because they have been fluorescently tagged with a dye that reacts to green laser lights to allow construction of a 3D image point by point. The experiment, which will be sent to the ISS later this year, will help P&G develop improved product stabilizers to extend shelf life and develop more environmentally friendly packaging.

The Potential Protective Effects of 2-aminoethyl Diphenylborinate against Inner Ear Acoustic Trauma: Experimental Study Using Transmission and Scanning Electron Microscopy.

PubMed

Kaymakçı, Mustafa; Acar, Mustafa; Burukoglu, Dilek; Kutlu, Hatice Mehtap; Shojaolsadati, Paria; Cingi, Cemal; Bayar Muluk, Nuray

2015-04-01

In this prospective experimental study, we investigated the preventive effects of 2-aminoethyl diphenylborinate (2-APB) in rats exposed to acoustic trauma (AT). Light microscopic, transmission electron microscopic (TEM), and scanning electron microscopic (SEM) examinations were performed. Eighteen healthy Wistar albino rats were divided into the following three groups: groups 1 (control), 2 (AT), and 3 (AT+APB). The rats in groups 2 and 3 were exposed to AT; in group 3 rats, 2-APB at 2 mg/kg was also administered, initially transperitoneally, after 10 min. During the light microscopic, TEM, and SEM examinations, the structures of the cochlear hair cells, stereocilia, and Deiter's cells were normal in the control group. In the AT group, the organ of Corti and proximate structures were damaged according to the light microscopic examination. During the TEM examination, intense cellular damage and stereocilia loss were detected, while during the SEM examination, extensive damage and stereocilia loss were observed. Decreased damage with preserved cochlear structure was detected during the light microscopic examination in the AT+APB group than in the AT group. During the TEM and SEM examinations, although stereocilia loss occurred in the AT+APB group, near-normal cell, cilia, and tectorial membrane structures were also observed in the AT+APB group compared with the AT group. 2-APB may have protective effects against AT damage of the cochlea. The main mechanism underlying this effect is the inhibition of the vasoconstriction of the cochlear spiral modiolar artery, thereby improving cochlear blood flow. We conclude that 2-APB may also be effective if used immediately following AT.

Laser speckle contrast imaging using light field microscope approach

NASA Astrophysics Data System (ADS)

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.

A multi-modal stereo microscope based on a spatial light modulator.

PubMed

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.

Lensfree microscopy on a cellphone

PubMed Central

Tseng, Derek; Mudanyali, Onur; Oztoprak, Cetin; Isikman, Serhan O.; Sencan, Ikbal; Yaglidere, Oguzhan; Ozcan, Aydogan

2010-01-01

We demonstrate lensfree digital microscopy on a cellphone. This compact and light-weight holographic microscope installed on a cellphone does not utilize any lenses, lasers or other bulky optical components and it may offer a cost-effective tool for telemedicine applications to address various global health challenges. Weighing ~38 grams (<1.4 ounces), this lensfree imaging platform can be mechanically attached to the camera unit of a cellphone where the samples are loaded from the side, and are vertically illuminated by a simple light-emitting diode (LED). This incoherent LED light is then scattered from each micro-object to coherently interfere with the background light, creating the lensfree hologram of each object on the detector array of the cellphone. These holographic signatures captured by the cellphone permit reconstruction of microscopic images of the objects through rapid digital processing. We report the performance of this lensfree cellphone microscope by imaging various sized micro-particles, as well as red blood cells, white blood cells, platelets and a waterborne parasite (Giardia lamblia). PMID:20445943

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

PubMed

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

2009-09-15

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

Digital photography for the light microscope: results with a gated, video-rate CCD camera and NIH-image software.

PubMed

Shaw, S L; Salmon, E D; Quatrano, R S

1995-12-01

In this report, we describe a relatively inexpensive method for acquiring, storing and processing light microscope images that combines the advantages of video technology with the powerful medium now termed digital photography. Digital photography refers to the recording of images as digital files that are stored, manipulated and displayed using a computer. This report details the use of a gated video-rate charge-coupled device (CCD) camera and a frame grabber board for capturing 256 gray-level digital images from the light microscope. This camera gives high-resolution bright-field, phase contrast and differential interference contrast (DIC) images but, also, with gated on-chip integration, has the capability to record low-light level fluorescent images. The basic components of the digital photography system are described, and examples are presented of fluorescence and bright-field micrographs. Digital processing of images to remove noise, to enhance contrast and to prepare figures for printing is discussed.

Fluorescence microscope (Cyscope) for malaria diagnosis in pregnant women in Medani Hospital, Sudan.

PubMed

Hassan, Saad El-Din H; Haggaz, Abd Elrahium D; Mohammed-Elhassan, Ehab B; Malik, Elfatih M; Adam, Ishag

2011-09-24

Accuracy of diagnosis is the core for malaria control. Although microscopy is the gold standard in malaria diagnosis, its reliability is largely dependent on user skill. We compared performance of Cyscope fluorescence microscope with the Giemsa stained light microscopy for the diagnosis of malaria among pregnant women at Medani Hospital in Central Sudan. The area is characterized by unstable malaria transmission. Socio-demographic characteristics and obstetrics history were gathered using pre-tested questionnaires. Blood samples were collected from febrile pregnant women who were referred as malaria case following initial diagnosis by general microscopist. During the study period 128 febrile pregnant women presented at the hospital. Among them, Plasmodium falciparum malaria was detected in 82 (64.1%) and 80 (62.5%) by the Giemsa-stained light microscopy and the Cyscope fluorescence microscope, respectively. The sensitivity of the Cyscope fluorescence microscope was 97.6% (95% CI: 92.2%-99.6%). Out of 46 which were negative by Giemsa-stained light microscopy, 5 were positive by the Cyscope fluorescence microscope. This is translated in specificity of 89.1% (95% CI: 77.5%-95.9%). The positive and negative predictive value of Cyscope fluorescence microscope was 94.1% (95% CI: 87.4% -97.8%) and 95.3% (95% CI: 85.4% - 99.2%), respectively. This study has shown that Cyscope fluorescence microscope is a reliable diagnostic, sensitive and specific in diagnosing P. falciparum malaria among pregnant women in this setting. Further studies are needed to determine effectiveness in diagnosing other Plasmodium species and to compare it with other diagnostic tools e.g. rapid diagnostic tests and PCR.

Staining-free malaria diagnostics by multispectral and multimodality light-emitting-diode microscopy

NASA Astrophysics Data System (ADS)

Merdasa, Aboma; Brydegaard, Mikkel; Svanberg, Sune; Zoueu, Jeremie T.

2013-03-01

We report an accurate optical differentiation technique between healthy and malaria-infected erythrocytes by quasi-simultaneous measurements of transmittance, reflectance, and scattering properties of unstained blood smears using a multispectral and multimode light-emitting diode microscope. We propose a technique for automated imaging, identification, and counting of malaria-infected erythrocytes for real-time and cost-effective parasitaemia diagnosis as an effective alternative to the manual screening of stained blood smears, now considered to be the gold standard in malaria diagnosis. We evaluate the performance of our algorithm against manual estimations of an expert and show a spectrally resolved increased scattering from malaria-infected blood cells.

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

Fläschner, G.; Ruschmeier, K.; Schwarz, A., E-mail: aschwarz@physnet.uni-hamburg.de

The sensitivity of atomic force microscopes is fundamentally limited by the cantilever temperature, which can be, in principle, determined by measuring its thermal spectrum and applying the equipartition theorem. However, the mechanical response can be affected by the light field inside the cavity of a Fabry-Perot interferometer due to light absorption, radiation pressure, photothermal forces, and laser noise. By evaluating the optomechanical Hamiltonian, we are able to explain the peculiar distance dependence of the mechanical quality factor as well as the appearance of thermal spectra with symmetrical Lorentzian as well as asymmetrical Fano line shapes. Our results can be appliedmore » to any type of mechanical oscillator in an interferometer-based detection system.« less

Optical depth localization of nitrogen-vacancy centers in diamond with nanometer accuracy.

PubMed

Häußler, Andreas J; Heller, Pascal; McGuinness, Liam P; Naydenov, Boris; Jelezko, Fedor

2014-12-01

Precise positioning of nitrogen-vacancy (NV) centers is crucial for their application in sensing and quantum information. Here we present a new purely optical technique enabling determination of the NV position with nanometer resolution. We use a confocal microscope to determine the position of individual emitters along the optical axis. Using two separate detection channels, it is possible to simultaneously measure reflected light from the diamond surface and fluorescent light from the NV center and statistically evaluate both signals. An accuracy of 2.6 nm for shallow NV centers was achieved and is consistent with other techniques for depth determination.

Optimal resolution in Fresnel incoherent correlation holographic fluorescence microscopy

PubMed Central

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

2011-01-01

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

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

PubMed

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

2016-01-01

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

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

PubMed Central

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

2016-01-01

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

The HVAC Challenges of Upgrading an Old Lab for High-end Light Microscopes

PubMed Central

Richard, R.; Martone, P.; Callahan, L.M.

2014-01-01

The University of Rochester Medical Center forms the centerpiece of the University of Rochester's health research, teaching, patient care, and community outreach missions. Within this large facility of over 5 million square feet, demolition and remodeling of existing spaces is a constant activity. With more than $145 million in federal research funding, lab space is frequently repurposed and renovated to support this work. The URMC Medical Center Facilities Organization supporting small to medium space renovations is constantly challenged and constrained by the existing mechanical infrastructure and budgets to deliver a renovated space that functions within the equipment environmental parameters. One recent project, sponsored by the URMC Shared Resources Laboratory, demonstrates these points. The URMC Light Microscopy Shared Resource Laboratory requested renovation of a 121 sq. ft. room in a 40 year old building which would enable placement of a laser capture microdissection microscope and a Pascal 5 laser scanning confocal microscope with the instruments separated by a blackout curtain. This poster discusses the engineering approach implemented to bring an older lab into the environmental specifications needed for the proper operation of the high-end light microscopes.

From Animaculum to single molecules: 300 years of the light microscope.

PubMed

Wollman, Adam J M; Nudd, Richard; Hedlund, Erik G; Leake, Mark C

2015-04-01

Although not laying claim to being the inventor of the light microscope, Antonj van Leeuwenhoek (1632-1723) was arguably the first person to bring this new technological wonder of the age properly to the attention of natural scientists interested in the study of living things (people we might now term 'biologists'). He was a Dutch draper with no formal scientific training. From using magnifying glasses to observe threads in cloth, he went on to develop over 500 simple single lens microscopes (Baker & Leeuwenhoek 1739 Phil. Trans. 41, 503-519. (doi:10.1098/rstl.1739.0085)) which he used to observe many different biological samples. He communicated his finding to the Royal Society in a series of letters (Leeuwenhoek 1800 The select works of Antony Van Leeuwenhoek, containing his microscopical discoveries in many of the works of nature, vol. 1) including the one republished in this edition of Open Biology. Our review here begins with the work of van Leeuwenhoek before summarizing the key developments over the last ca 300 years, which has seen the light microscope evolve from a simple single lens device of van Leeuwenhoek's day into an instrument capable of observing the dynamics of single biological molecules inside living cells, and to tracking every cell nucleus in the development of whole embryos and plants.

Acute unclassified leukemia: A clinicopathologic study with diagnostic implications of electron microscopy.

PubMed

Youness, E; Trujillo, J M; Ahearn, M J; McCredie, K B; Cork, A

1980-01-01

By rigid cytological and cytochemical criteria, the diagnosis of acute and undifferentiated leukemia was established in 22 patients. According to defined criteria, the leukemic cells could not be classified by conventional light microscopic techniques employed in the study of hematopoietic tissue. Cytochemical studies including peroxidase, periodic acid schiff (PAS) and nonspecific esterase (alpha napthyl butyrate-reacting esterase) stains were done on fresh bone marrow samples, and the percentage of positive leukemia cells for each of these stains was determined on 200 cells. In this series of leukemias, cytochemistry at the light microscope level did not contribute to further classification. Subsequent electron microscopic examination of bone marrow samples from these patients confirmed the immaturity and nuclear/cytoplasmic asynchrony of the leukemic cells. Several in vivo neoplastic markers, such as nuclear blebs, increased nuclear bodies, and cytoplasmic fibrillar bundles could be demonstrated in these cells. Fourteen cases from this series exhibited peroxidase-positive developmental granule formation at the ultrastructural level and were reclassified as acute granulocyte leukemia (AGL). One case was reclassified as lymphoma (poor differentiated type), one case was diagnosed as acute monocytic leukemia (AmonoL), and six cases remained in the undifferentiated category (AUL). Clinical and laboratory features, response to treatment, and survival data were evaluated for these patients. This study demonstrated that electron microscopy is useful in the cytological diagnosis of human leukemia.

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

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

Scanning optical microscope with long working distance objective

DOEpatents

Cloutier, Sylvain G.

2010-10-19

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

Acquisition of a High Resolution Field Emission Scanning Electron Microscope for the Analysis of Returned Samples

NASA Technical Reports Server (NTRS)

Nittler, Larry R.

2003-01-01

This grant furnished funds to purchase a state-of-the-art scanning electron microscope (SEM) to support our analytical facilities for extraterrestrial samples. After evaluating several instruments, we purchased a JEOL 6500F thermal field emission SEM with the following analytical accessories: EDAX energy-dispersive x-ray analysis system with fully automated control of instrument and sample stage; EDAX LEXS wavelength-dispersive x-ray spectrometer for high sensitivity light-element analysis; EDAX/TSL electron backscatter diffraction (EBSD) system with software for phase identification and crystal orientation mapping; Robinson backscatter electron detector; and an in situ micro-manipulator (Kleindiek). The total price was $550,000 (with $150,000 of the purchase supported by Carnegie institution matching funds). The microscope was delivered in October 2002, and most of the analytical accessories were installed by January 2003. With the exception of the wavelength spectrometer (which has been undergoing design changes) everything is working well and the SEM is in routine use in our laboratory.

Microstructure Evolution and Mechanical Properties of Mg-14%Li-1%Al Alloy During the High-Pressure Torsion

NASA Astrophysics Data System (ADS)

Tian, Chenguang; Lu, Huimin; Zhao, Liyuan

The super-light LA141 (Mg-14%Li-1%Al) alloy was produced and processed by high-pressure torsion (HPT) under the imposed pressure of 3 GPa and different shear strains γ through 3, 6, 9 and 12 turns at room temperature (RT). The microstructure evolution of the alloy during the HPT treatment was investigated by transmission electron microscope (TEM) and optical microscope (OM). It turned out that the grains were substantially refined, and the optical microscope revealed that the grains of HPT processed samples at the edge of the disc were finer by comparison with the ones near the center of the disc. Later, Vickers indentation analysis was used to evaluate the micro-hardness of deformed samples, and tension test was employed to obtain the strength and elongation at room temperature. The results indicated that the micro-hardness and tensile strength had increased to a certain extent, and the elongation had been significantly improved.

TiO2 nanosheets decorated with B4C nanoparticles as photocatalysts for solar fuel production under visible light irradiation

NASA Astrophysics Data System (ADS)

Zhang, Xiaojie; Yang, Jipeng; Cai, Tiancong; Zuo, Guoqiang; Tang, Changqing

2018-06-01

Boron carbide (B4C) nanoparticles-decorated anatase titanium dioxide (TiO2) nanosheets photocatalysts were synthesized by a hydrothermal method in the presence of hydrofluoric acid and characterized by field emission scanning electron microscope, high-resolution transmission electron microscope, UV-vis diffuse reflectance spectra, photoluminescence spectra, etc. With metallic Pt nanoparticles as a co-catalyst, the as-synthesized B4C/TiO2 composites were evaluated using photocatalytic CO2 or H2O reduction to solar fuels such as methane and hydrogen. Under either simulated sunlight or visible light irradiation, coupling p-type B4C with n-type anatase TiO2 significantly improved the photocatalytic performance. Both photoluminescence and transient photocurrent measurements indicated that the interfacial coupling effect between B4C and anatase TiO2 could significantly promote photo-excited charges separations. On the basis of measurements and literatures, a possible mechanism of excited charges transfer at the B4C-anatase TiO2 heterojunction interface during irradiation was deduced.

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

NASA Astrophysics Data System (ADS)

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

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

Transmission electron microscope sample holder with optical features

DOEpatents

Milas, Mirko [Port Jefferson, NY; Zhu, Yimei [Stony Brook, NY; Rameau, Jonathan David [Coram, NY

2012-03-27

A sample holder for holding a sample to be observed for research purposes, particularly in a transmission electron microscope (TEM), generally includes an external alignment part for directing a light beam in a predetermined beam direction, a sample holder body in optical communication with the external alignment part and a sample support member disposed at a distal end of the sample holder body opposite the external alignment part for holding a sample to be analyzed. The sample holder body defines an internal conduit for the light beam and the sample support member includes a light beam positioner for directing the light beam between the sample holder body and the sample held by the sample support member.

Microcircuit testing and fabrication, using scanning electron microscopes

NASA Technical Reports Server (NTRS)

Nicolas, D. P.

1975-01-01

Scanning electron microscopes are used to determine both user-induced damages and manufacturing defects subtle enough to be missed by conventional light microscopy. Method offers greater depth of field and increased working distances.

ultraLM and miniLM: Locator tools for smart tracking of fluorescent cells in correlative light and electron microscopy.

PubMed

Brama, Elisabeth; Peddie, Christopher J; Wilkes, Gary; Gu, Yan; Collinson, Lucy M; Jones, Martin L

2016-12-13

In-resin fluorescence (IRF) protocols preserve fluorescent proteins in resin-embedded cells and tissues for correlative light and electron microscopy, aiding interpretation of macromolecular function within the complex cellular landscape. Dual-contrast IRF samples can be imaged in separate fluorescence and electron microscopes, or in dual-modality integrated microscopes for high resolution correlation of fluorophore to organelle. IRF samples also offer a unique opportunity to automate correlative imaging workflows. Here we present two new locator tools for finding and following fluorescent cells in IRF blocks, enabling future automation of correlative imaging. The ultraLM is a fluorescence microscope that integrates with an ultramicrotome, which enables 'smart collection' of ultrathin sections containing fluorescent cells or tissues for subsequent transmission electron microscopy or array tomography. The miniLM is a fluorescence microscope that integrates with serial block face scanning electron microscopes, which enables 'smart tracking' of fluorescent structures during automated serial electron image acquisition from large cell and tissue volumes.

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

PubMed

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.

Thin laser light sheet microscope for microbial oceanography

NASA Astrophysics Data System (ADS)

Fuchs, Eran; Jaffe, Jules S.; Long, Richard A.; Azam, Farooq

2002-01-01

Despite a growing need, oceanographers are limited by existing technological constrains and are unable to observe aquatic microbes in their natural setting. In order to provide a simple and easy to implement solution for such studies, a new Thin Light Sheet Microscope (TLSM) has been developed. The TLSM utilizes a well-defined sheet of laser light, which has a narrow (23 micron) axial dimension over a 1 mm x 1 mm field of view. This light sheet is positioned precisely within the depth of field of the microscope’s objective lens. The technique thus utilizes conventional microscope optics but replaces the illumination system. The advantages of the TLSM are two-fold: First, it concentrates light only where excitation is needed, thus maximizing the efficiency of the illumination source. Secondly, the TLSM maximizes image sharpness while at the same time minimizing the level of background noise. Particles that are not located within the objective's depth of field are not illuminated and therefore do not contribute to an out-of-focus image. Images from a prototype system that used SYBR Green I fluorescence stain in order to localize single bacteria are reported. The bacteria were in a relatively large and undisturbed volume of 4ml, which contained natural seawater. The TLSM can be used for fresh water studies of bacteria with no modification. The microscope permits the observation of interactions at the microscale and has potential to yield insights into how microbes structure pelagic ecosystems.

Scintigraphic evaluation of osteoblastic activity in extraction sockets treated with platelet-rich fibrin.

PubMed

Gürbüzer, Bahadir; Pikdöken, Levent; Tunali, Mustafa; Urhan, Muammer; Küçükodaci, Zafer; Ercan, Feriha

2010-05-01

To evaluate the effect of platelet-rich fibrin (PRF) on the early bone healing process with bone scintigraphy based on technetium-99m methylene diphosphonate uptake in third molar extraction sockets. Fourteen patients with bilaterally soft tissue impacted third mandibular molars were included in the study. The right and left impacted third molars were surgically extracted in the same session. PRF was randomly administered into one of the extraction sockets, whereas the contralateral sockets were left without treatment. Four weeks after surgery, scintigrams were obtained to evaluate scintigraphic differences between PRF-treated and non-PRF-treated sockets. After completion of the clinical study, PRF samples were evaluated by light and scanning electron microscopy. The average increase in technetium-99m methylene diphosphonate uptake as an indication of enhanced bone healing did not differ significantly between PRF-treated and non-PRF-treated sockets 4 weeks postoperatively (P > .05). Abundant fibrin and inflammatory cells were observed by light microscopic examination of PRF samples. Scanning electron microscopic analysis of PRF revealed the existence of platelet aggregates in a fibrin network and crystalline particles on the outer surface of PRF. PRF might not lead to enhanced bone healing in soft tissue impacted mandibular third molar extraction sockets 4 weeks after surgery. PRF exhibits the potential characteristics of an autologous fibrin matrix. However, whether the presence of crystal-like particles on the outer surface of PRF alters bone healing should be investigated further. Copyright 2010 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

Photodermatitis and photokeratoconjunctivitis in a ball python (Python regius) and a blue-tongue skink (Tiliqua spp.).

PubMed

Gardiner, David W; Baines, Frances M; Pandher, Karamjeet

2009-12-01

A male ball python (Python regius) and a female blue tongue skink (Tiliqua spp.) of unknown age were evaluated for anorexia, lethargy, excessive shedding, corneal opacity (python), and weight loss (skink) of approximately three weeks' duration. These animals represented the worst affected animals from a private herpetarium where many animals exhibited similar signs. At necropsy, the python had bilateral corneal opacity and scattered moderate dysecdysis. The skink had mild dysecdysis, poor body condition, moderate intestinal nematodiasis, and mild liver atrophy. Microscopic evaluation revealed epidermal erosion and ulceration, with severe epidermal basal cell degeneration and necrosis, and superficial dermatitis (python and skink). Severe bilateral ulcerative keratoconjunctivitis with bacterial colonization was noted in the ball python. Microscopic findings within the skin and eyes were suggestive of ultraviolet (UV) radiation damage or of photodermatitis and photokeratoconjunctivitis. Removal of the recently installed new lamps from the terrariums of the surviving reptiles resulted in resolution of clinical signs. Evaluation of a sample lamp of the type associated with these cases revealed an extremely high UV output, including very-short-wavelength UVB, neither found in natural sunlight nor emitted by several other UVB lamps unassociated with photokeratoconjunctivitis. Exposure to high-intensity and/or inappropriate wavelengths of UV radiation may be associated with significant morbidity, and even mortality, in reptiles. Veterinarians who are presented with reptiles with ocular and/or cutaneous disease of unapparent cause should fully evaluate the specifics of the vivarium light sources. Further research is needed to determine the characteristics of appropriate and of toxic UV light for reptiles kept in captivity.

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

PubMed

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 chromatic space but simultaneously the value of luminance changes. So the process of the image unification in a sense of colour fidelity can be solved in separate introductory stage before the automatic image analysis.

Hybrid of two-photon microscopy and optical multimodality imaging for multi-scale imaging of small animals

NASA Astrophysics Data System (ADS)

Li, Tianmeng; Hui, Hui; Ma, He; Yang, Xin; Tian, Jie

2018-02-01

Non-invasive imaging technologies, such as magnetic resonance imaging (MRI) and optical multimodality imaging methods, are commonly used for diagnosing and supervising the development of inflammatory bowel disease (IBD). These in vivo imaging methods can provide morphology changes information of IBD in macro-scale. However, it is difficult to investigate the intestinal wall in molecular and cellular level. State-of-art light-sheet and two-photon microscopy have the ability to acquire the changes for IBD in micro-scale. The aim of this work is to evaluate the size of the enterocoel and the thickness of colon wall using both MRI for in vivo imaging, and light-sheet and two-photon microscope for in vitro imaging. C57BL/6 mice were received 3.5% Dextran sodium sulfate (DSS) in the drinking water for 5 days to build IBD model. Mice were imaged with MRI on days 0, 6 to observe colitis progression. After MRI imaging, the mice were sacrificed to take colons for tissue clearing. Then, light-sheet and two-photon microscopies are used for in vitro imaging of the cleared samples. The experimental group showed symptoms of bloody stools, sluggishness and weight loss. It showed that the colon wall was thicker while the enterocoel was narrower compare to control group. The more details are observed using light-sheet and two-photon microscope. It is demonstrated that hybrid of MRI in macro-scale and light-sheet and two-photon microscopy in micro-scale imaging is feasible for colon inflammation diagnosing and supervising.

Dual-labeling with 5-aminolevulinic acid and fluorescein for fluorescence-guided resection of high-grade gliomas: technical note.

PubMed

Suero Molina, Eric; Wölfer, Johannes; Ewelt, Christian; Ehrhardt, André; Brokinkel, Benjamin; Stummer, Walter

2018-02-01

OBJECTIVE Fluorescence guidance with 5-aminolevulinic acid (5-ALA) helps improve resections of malignant gliomas. However, one limitation is the low intensity of blue light for background illumination. Fluorescein has recently been reintroduced into neurosurgery, and novel microscope systems are available for visualizing this fluorochrome, which highlights all perfused tissues but has limited selectivity for tumor detection. Here, the authors investigate a combination of both fluorochromes: 5-ALA for distinguishing tumor and fluorescein for providing tissue fluorescence of adjacent brain tissue. METHODS The authors evaluated 6 patients who harbored cerebral lesions suggestive of high-grade glioma. Patients received 5-ALA (20 mg/kg) orally 4 hours before induction of anesthesia. Low-dose fluorescein (3 mg/kg intravenous) was injected immediately after anesthesia induction. Pentero microscopes (equipped either with Yellow 560 or Blue 400 filters) were used to visualize fluorescence. To simultaneously visualize both fluorochromes, the Yellow 560 module was combined with external blue light illumination (D-light C System). RESULTS Fluorescein-induced fluorescence created a useful background for protoporphyrin IX (PPIX) fluorescence, which appeared orange to red, surrounded by greenly fluorescent normal brain and edematous tissue. Green brain-tissue fluorescence was helpful in augmenting background. Levels of blue illumination that were too strong obscured PPIX fluorescence. Unspecific extravasation of fluorescein was noted at resection margins, which did not interfere with PPIX fluorescence detection. CONCLUSIONS Dual labeling with both PPIX and fluorescein fluorescence is feasible and gives superior background information during fluorescence-guided resections. The authors believe that this technique carries potential as a next step in fluorescence-guided resections if it is completely integrated into the surgical microscope.

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

PubMed

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.

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

PubMed Central

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

2018-01-01

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

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

PubMed

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

2018-01-01

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

A Simple low-cost device enables four epi-illumination techniques on standard light microscopes.

PubMed

Ishmukhametov, Robert R; Russell, Aidan N; Wheeler, Richard J; Nord, Ashley L; Berry, Richard M

2016-02-08

Back-scattering darkfield (BSDF), epi-fluorescence (EF), interference reflection contrast (IRC), and darkfield surface reflection (DFSR) are advanced but expensive light microscopy techniques with limited availability. Here we show a simple optical design that combines these four techniques in a simple low-cost miniature epi-illuminator, which inserts into the differential interference-contrast (DIC) slider bay of a commercial microscope, without further additions required. We demonstrate with this device: 1) BSDF-based detection of Malarial parasites inside unstained human erythrocytes; 2) EF imaging with and without dichroic components, including detection of DAPI-stained Leishmania parasite without using excitation or emission filters; 3) RIC of black lipid membranes and other thin films, and 4) DFSR of patterned opaque and transparent surfaces. We believe that our design can expand the functionality of commercial bright field microscopes, provide easy field detection of parasites and be of interest to many users of light microscopy.

A Simple low-cost device enables four epi-illumination techniques on standard light microscopes

NASA Astrophysics Data System (ADS)

Ishmukhametov, Robert R.; Russell, Aidan N.; Wheeler, Richard J.; Nord, Ashley L.; Berry, Richard M.

2016-02-01

Back-scattering darkfield (BSDF), epi-fluorescence (EF), interference reflection contrast (IRC), and darkfield surface reflection (DFSR) are advanced but expensive light microscopy techniques with limited availability. Here we show a simple optical design that combines these four techniques in a simple low-cost miniature epi-illuminator, which inserts into the differential interference-contrast (DIC) slider bay of a commercial microscope, without further additions required. We demonstrate with this device: 1) BSDF-based detection of Malarial parasites inside unstained human erythrocytes; 2) EF imaging with and without dichroic components, including detection of DAPI-stained Leishmania parasite without using excitation or emission filters; 3) RIC of black lipid membranes and other thin films, and 4) DFSR of patterned opaque and transparent surfaces. We believe that our design can expand the functionality of commercial bright field microscopes, provide easy field detection of parasites and be of interest to many users of light microscopy.

Phase Sensitive Demodulation in Multiphoton Microscopy

NASA Astrophysics Data System (ADS)

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

2002-06-01

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

Light-sheet microscopy for everyone? Experience of building an OpenSPIM to study flatworm development.

PubMed

Girstmair, Johannes; Zakrzewski, Anne; Lapraz, François; Handberg-Thorsager, Mette; Tomancak, Pavel; Pitrone, Peter Gabriel; Simpson, Fraser; Telford, Maximilian J

2016-06-30

Selective plane illumination microscopy (SPIM a type of light-sheet microscopy) involves focusing a thin sheet of laser light through a specimen at right angles to the objective lens. As only the thin section of the specimen at the focal plane of the lens is illuminated, out of focus light is naturally absent and toxicity due to light (phototoxicity) is greatly reduced enabling longer term live imaging. OpenSPIM is an open access platform (Pitrone et al. 2013 and OpenSPIM.org) created to give new users step-by-step instructions on building a basic configuration of a SPIM microscope, which can in principle be adapted and upgraded to each laboratory's own requirements and budget. Here we describe our own experience with the process of designing, building, configuring and using an OpenSPIM for our research into the early development of the polyclad flatworm Maritigrella crozieri - a non-model animal. Our OpenSPIM builds on the standard design with the addition of two colour laser illumination for simultaneous detection of two probes/molecules and dual sided illumination, which provides more even signal intensity across a specimen. Our OpenSPIM provides high resolution 3d images and time lapse recordings, and we demonstrate the use of two colour lasers and the benefits of two color dual-sided imaging. We used our microscope to study the development of the embryo of the polyclad flatworm M. crozieri. The capabilities of our microscope are demonstrated by our ability to record the stereotypical spiral cleavage pattern of M. crozieri with high-speed multi-view time lapse imaging. 3D and 4D (3D + time) reconstruction of early development from these data is possible using image registration and deconvolution tools provided as part of the open source Fiji platform. We discuss our findings on the pros and cons of a self built microscope. We conclude that home-built microscopes, such as an OpenSPIM, together with the available open source software, such as MicroManager and Fiji, make SPIM accessible to anyone interested in having continuous access to their own light-sheet microscope. However, building an OpenSPIM is not without challenges and an open access microscope is a worthwhile, if significant, investment of time and money. Multi-view 4D microscopy is more challenging than we had expected. We hope that our experience gained during this project will help future OpenSPIM users with similar ambitions.

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

PubMed Central

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

2011-01-01

We report replacement of one side of a static illumination, dual sided, thin-sheet laser imaging microscope (TSLIM) with an intensity modulated laser scanner in order to implement structured illumination (SI) and HiLo image demodulation techniques for background rejection. The new system is equipped with one static and one scanned light-sheet and is called a scanning thin-sheet laser imaging microscope (sTSLIM). It is an optimized version of a light-sheet fluorescent microscope that is designed to image large specimens (<15 mm in diameter). In this paper we describe the hardware and software modifications to TSLIM that allow for static and uniform light-sheet illumination with SI and HiLo image demodulation. The static light-sheet has a thickness of 3.2 µm; whereas, the scanned side has a light-sheet thickness of 4.2 µm. The scanned side images specimens with subcellular resolution (<1 µm lateral and <4 µm axial resolution) with a size up to 15 mm. SI and HiLo produce superior contrast compared to both the uniform static and scanned light-sheets. HiLo contrast was greater than SI and is faster and more robust than SI because as it produces images in two-thirds of the time and exhibits fewer intensity streaking artifacts. PMID:22254177

Virtual Microscopy: A Useful Tool for Meeting Evolving Challenges in the Veterinary Medical Curriculum

NASA Astrophysics Data System (ADS)

Kogan, Lori R.; Dowers, Kristy L.; Cerda, Jacey R.; Schoenfeld-Tacher, Regina M.; Stewart, Sherry M.

2014-12-01

Veterinary schools, similar to many professional health programs, face a myriad of evolving challenges in delivering their professional curricula including expansion of class size, costs to maintain expensive laboratories, and increased demands on veterinary educators to use curricular time efficiently and creatively. Additionally, exponential expansion of the knowledge base through ongoing biomedical research, educational goals to increase student engagement and clinical reasoning earlier in the curriculum, and students' desire to access course materials and enhance their educational experience through the use of technology all support the need to reassess traditional microscope laboratories within Professional Veterinary Medical (PVM) educational programs. While there is clear justification for teaching veterinary students how to use a microscope for clinical evaluation of cytological preparations (i.e., complete blood count, urinalysis, fecal analysis, fine needle aspirates, etc.), virtual microscopy may be a viable alternative to using light microscopy for teaching and learning fundamental histological concepts. This article discusses results of a survey given to assess Professional Veterinary Medical students' perceptions of using virtual microscope for learning basic histology/microscopic anatomy and implications of these results for using virtual microscopy as a pedagogical tool in teaching first-year Professional Veterinary Medical students' basic histology.

Site of potential operating microscope light-induced phototoxicity on the human retina during temporal approach eye surgery.

PubMed

Pavilack, M A; Brod, R D

2001-02-01

To determine the site of focal illumination on the retina of phakic human cadaver eyes from an operating microscope positioned for temporal approach eye surgery. Experimental study. A Zeiss OPMI-6SFR operating microscope (Zeiss Humphrey Systems, Dublin, CA) was positioned over two phakic human cadaver eyes to measure the site of the focal illumination on the retina by directly observing the illumination on the posterior scleral surface of the globe. External localization of the foveola was made by direct observation using scleral indentation and indirect ophthalmoscopy. Various combinations of microscope angulation and field of view were analyzed. Distance of focal illumination from the operating room microscope relative to the foveola was measured. The diameter of the "hot spot" of focal illumination on the retina was 4.0 mm. With the eye positioned straight ahead and the level operating room microscope positioned for temporal approach eye surgery, the center of retinal illumination was 0.9 and 1.4 mm nasal relative to the foveola when the microscope field of view was centered over the cornea and temporal limbus, respectively. With the microscope angled 5, 10, 15, and 20 degrees temporally (oculars tilted toward surgeon), the center of the illumination was displaced nasal to the foveola by 1.1, 1.5, 3.8, and 5.1 mm, respectively, when the field of view was centered over the cornea and 1.5, 2.6, 4.7, and 6.0 mm, respectively, nasal to the foveola when centered over the temporal limbus. Retinal illumination from an operating microscope positioned for temporal approach eye surgery has the potential for light-induced injury to the fovea. Angulation of the operating microscope by up to 10 degrees temporally when the microscope field of view was centered over the cornea and up to 5 degrees temporally when centered over the temporal limbus was not adequate to displace the focal illumination off the foveola when the eye was in the straight-ahead position. Tilting the operating microscope 15 degrees or more temporally when centered on the pupil and 10 degrees or more when centered over the temporal limbus should safely displace the retinal light exposure away from the fovea during temporal approach surgery. Suggestions for reducing the risk of iatrogenic phototoxicity are reviewed.

Microscopy imaging device with advanced imaging properties

DOEpatents

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

2015-11-24

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

Microscopy imaging device with advanced imaging properties

DOEpatents

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

2016-10-25

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

Microscopy imaging device with advanced imaging properties

DOEpatents

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

2016-11-22

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

Microscopy imaging device with advanced imaging properties

DOEpatents

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

2017-04-25

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

Green tea extract induces protective autophagy in A549 non-small lung cancer cell line.

PubMed

Izdebska, Magdalena; Klimaszewska-Wiśniewska, Anna; Hałas, Marta; Gagat, Maciej; Grzanka, Alina

2015-12-31

For many decades, polyphenols, including green tea extract catechins, have been reported to exert multiple anti-tumor activities. However, to date the mechanisms of their action have not been completely elucidated. Thus, the aim of this study was to assess the effect of green tea extract on non-small lung cancer A549 cells. A549 cells following treatment with GTE were analyzed using the inverted light and fluorescence microscope. In order to evaluate cell sensitivity and cell death, the MTT assay and Tali image-based cytometer were used, respectively. Ultrastructural alterations were assessed using a transmission electron microscope. The obtained data suggested that GTE, even at the highest dose employed (150 μM), was not toxic to A549 cells. Likewise, the treatment with GTE resulted in only a very small dose-dependent increase in the population of apoptotic cells. However, enhanced accumulation of vacuole-like structures in response to GTE was seen at the light and electron microscopic level. Furthermore, an increase in the acidic vesicular organelles and LC3-II puncta formation was observed under the fluorescence microscope, following GTE treatment. The analysis of the functional status of autophagy revealed that GTE-induced autophagy may provide self-protection against its own cytotoxicity, since we observed that the blockage of autophagy by bafilomycin A1 decreased the viability of A549 cells and potentiated necrotic cell death induction in response to GTE treatment. Collectively, our results revealed that A549 cells are insensitive to both low and high concentrations of the green tea extract, probably due to the induction of cytoprotective autophagy. These data suggest that a potential utility of GTE in lung cancer therapy may lie in its synergistic combinations with drugs or small molecules that target autophagy, rather than in monotherapy.

Upgrade of a Scanning Confocal Microscope to a Single-Beam Path STED Microscope

PubMed Central

Klauss, André; König, Marcelle; Hille, Carsten

2015-01-01

By overcoming the diffraction limit in light microscopy, super-resolution techniques, such as stimulated emission depletion (STED) microscopy, are experiencing an increasing impact on life sciences. High costs and technically demanding setups, however, may still hinder a wider distribution of this innovation in biomedical research laboratories. As far-field microscopy is the most widely employed microscopy modality in the life sciences, upgrading already existing systems seems to be an attractive option for achieving diffraction-unlimited fluorescence microscopy in a cost-effective manner. Here, we demonstrate the successful upgrade of a commercial time-resolved confocal fluorescence microscope to an easy-to-align STED microscope in the single-beam path layout, previously proposed as “easy-STED”, achieving lateral resolution < λ/10 corresponding to a five-fold improvement over a confocal modality. For this purpose, both the excitation and depletion laser beams pass through a commercially available segmented phase plate that creates the STED-doughnut light distribution in the focal plane, while leaving the excitation beam unaltered when implemented into the joint beam path. Diffraction-unlimited imaging of 20 nm-sized fluorescent beads as reference were achieved with the wavelength combination of 635 nm excitation and 766 nm depletion. To evaluate the STED performance in biological systems, we compared the popular phalloidin-coupled fluorescent dyes Atto647N and Abberior STAR635 by labeling F-actin filaments in vitro as well as through immunofluorescence recordings of microtubules in a complex epithelial tissue. Here, we applied a recently proposed deconvolution approach and showed that images obtained from time-gated pulsed STED microscopy may benefit concerning the signal-to-background ratio, from the joint deconvolution of sub-images with different spatial information which were extracted from offline time gating. PMID:26091552

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

PubMed

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

2014-05-01

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

Automated Diatom Analysis Applied to Traditional Light Microscopy: A Proof-of-Concept Study

NASA Astrophysics Data System (ADS)

Little, Z. H. L.; Bishop, I.; Spaulding, S. A.; Nelson, H.; Mahoney, C.

2017-12-01

Diatom identification and enumeration by high resolution light microscopy is required for many areas of research and water quality assessment. Such analyses, however, are both expertise and labor-intensive. These challenges motivate the need for an automated process to efficiently and accurately identify and enumerate diatoms. Improvements in particle analysis software have increased the likelihood that diatom enumeration can be automated. VisualSpreadsheet software provides a possible solution for automated particle analysis of high-resolution light microscope diatom images. We applied the software, independent of its complementary FlowCam hardware, to automated analysis of light microscope images containing diatoms. Through numerous trials, we arrived at threshold settings to correctly segment 67% of the total possible diatom valves and fragments from broad fields of view. (183 light microscope images were examined containing 255 diatom particles. Of the 255 diatom particles present, 216 diatoms valves and fragments of valves were processed, with 170 properly analyzed and focused upon by the software). Manual analysis of the images yielded 255 particles in 400 seconds, whereas the software yielded a total of 216 particles in 68 seconds, thus highlighting that the software has an approximate five-fold efficiency advantage in particle analysis time. As in past efforts, incomplete or incorrect recognition was found for images with multiple valves in contact or valves with little contrast. The software has potential to be an effective tool in assisting taxonomists with diatom enumeration by completing a large portion of analyses. Benefits and limitations of the approach are presented to allow for development of future work in image analysis and automated enumeration of traditional light microscope images containing diatoms.

Distribution of melanosomes across the retinal pigment epithelium of a hooded rat: implications for light damage

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

Howell, W.L.; Rapp, L.M.; Williams, T.P.

1982-02-01

Distribution of melanosomes across the retinal pigment epithelium of hooded rats (Long-Evans) is studied at the light microscopic and electron microscopic levels. This distribution is shown to be nonuniform: more melanosomes exist in the periphery than elsewhere and, importantly, there are very few melanosomes in a restricted area of the central portion of the superior hemisphere compared with the corresponding part of the inferior hemisphere. The region with fewest melanosomes is precisely the one that is highly susceptible to light damage. Because this region is the same in both pigmented and albino eyes, the paucity of melanin in this regionmore » is not the cause of its great sensitivity to light damage. Nor does light cause the nonuniform distribution of melanin. A possible explanation, involving a proposed vestigial tapetum, is given in order to explain the correlation of melanosome counts and sensitivity to light damage.« less

Calibrating excitation light fluxes for quantitative light microscopy in cell biology

PubMed Central

Grünwald, David; Shenoy, Shailesh M; Burke, Sean; Singer, Robert H

2011-01-01

Power output of light bulbs changes over time and the total energy delivered will depend on the optical beam path of the microscope, filter sets and objectives used, thus making comparison between experiments performed on different microscopes complicated. Using a thermocoupled power meter, it is possible to measure the exact amount of light applied to a specimen in fluorescence microscopy, regardless of the light source, as the light power measured can be translated into a power density at the sample. This widely used and simple tool forms the basis of a new degree of calibration precision and comparability of results among experiments and setups. Here we describe an easy-to-follow protocol that allows researchers to precisely estimate excitation intensities in the object plane, using commercially available opto-mechanical components. The total duration of this protocol for one objective and six filter cubes is 75 min including start-up time for the lamp. PMID:18974739

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

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

Nazaretski, Evgeny

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

Microscope-Based Fluid Physics Experiments in the Fluids and Combustion Facility on ISS

NASA Technical Reports Server (NTRS)

Doherty, Michael P.; Motil, Susan M.; Snead, John H.; Malarik, Diane C.

2000-01-01

At the NASA Glenn Research Center, the Microgravity Science Program is planning to conduct a large number of experiments on the International Space Station in both the Fluid Physics and Combustion Science disciplines, and is developing flight experiment hardware for use within the International Space Station's Fluids and Combustion Facility. Four fluids physics experiments that require an optical microscope will be sequentially conducted within a subrack payload to the Fluids Integrated Rack of the Fluids and Combustion Facility called the Light Microscopy Module, which will provide the containment, changeout, and diagnostic capabilities to perform the experiments. The Light Microscopy Module is planned as a fully remotely controllable on-orbit microscope facility, allowing flexible scheduling and control of experiments within International Space Station resources. This paper will focus on the four microscope-based experiments, specifically, their objectives and the sample cell and instrument hardware to accommodate their requirements.

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

NASA Astrophysics Data System (ADS)

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

2016-10-01

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

Effect of the angle of the operating microscope light beam on visual recovery after phacoemulsification: Randomized trial.

PubMed

Harman, Francesca E; Corbett, Melanie C; Stevens, Julian D

2010-08-01

To evaluate differences in visual recovery after phacoemulsification with direct or tilted surgical microscope illumination using a macular photostress test. Western Eye Hospital, Imperial College Health Care National Health Service Trust, London, United Kingdom. This randomized double-masked controlled trial enrolled patients presenting to a daycare unit for single-eye cataract surgery. Inclusion criteria were no ocular pathology other than cataract, corneal keratometric astigmatism less than 1.50 diopters, intended target of emmetropia in the operated eye, and cataract grade 1 to 3 (Lens Opacification Classification System II). Exclusion criteria were an abnormal preoperative photostress test. Patients were randomized to have phacoemulsification with the operating microscope angled 15 degrees nasal to the fovea (study group) or with the operating microscope directly overhead around the optic disc region (control group). The same surgeon performed all phacoemulsification procedures using a standardized technique and topical anesthesia. Outcome measures were uncorrected (UDVA) and corrected (CDVA) distance visual acuity 10 minutes and 60 minutes postoperatively. In the 30 patients evaluated, the mean UDVA 10 minutes postoperatively was 0.40 logMAR +/- 0.26 (SD) in the study group and 0.72 +/- 0.36 logMAR in the control group (P<.01). The mean CDVA was 0.18 +/- 0.26 logMAR and 0.44 +/- 0.30 logMAR, respectively (P = .016). There was no significant between-group difference in acuity at 60 minutes. Tilting the microscope beam away from the fovea resulted in faster visual recovery and less macular photic stress. No author has a financial or proprietary interest in any material or method mentioned. Copyright 2010 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

From Animaculum to single molecules: 300 years of the light microscope

PubMed Central

Wollman, Adam J. M.; Nudd, Richard; Hedlund, Erik G.; Leake, Mark C.

2015-01-01

Although not laying claim to being the inventor of the light microscope, Antonj van Leeuwenhoek (1632–1723) was arguably the first person to bring this new technological wonder of the age properly to the attention of natural scientists interested in the study of living things (people we might now term ‘biologists’). He was a Dutch draper with no formal scientific training. From using magnifying glasses to observe threads in cloth, he went on to develop over 500 simple single lens microscopes (Baker & Leeuwenhoek 1739 Phil. Trans. 41, 503–519. (doi:10.1098/rstl.1739.0085)) which he used to observe many different biological samples. He communicated his finding to the Royal Society in a series of letters (Leeuwenhoek 1800 The select works of Antony Van Leeuwenhoek, containing his microscopical discoveries in many of the works of nature, vol. 1) including the one republished in this edition of Open Biology. Our review here begins with the work of van Leeuwenhoek before summarizing the key developments over the last ca 300 years, which has seen the light microscope evolve from a simple single lens device of van Leeuwenhoek's day into an instrument capable of observing the dynamics of single biological molecules inside living cells, and to tracking every cell nucleus in the development of whole embryos and plants. PMID:25924631

Production of radially and azimuthally polarized polychromatic beams

NASA Astrophysics Data System (ADS)

Shoham, A.; Vander, R.; Lipson, S. G.

2006-12-01

We describe a system that efficiently provides radially or azimuthally polarized radiation from a randomly polarized source. It is constructed from two conical reflectors and a cylindrical sheet of polarizing film. Envisaged applications include a microscope illuminator for high-resolution surface plasmon resonance microscopy, illumination for high-resolution microlithography, and efficient coupling of a laser source to hollow optical fibers. The angular coherence function of light polarized by the device was measured to evaluate its usefulness for these applications.

Effects of photodynamic therapy on Gram-positive and Gram-negative bacterial biofilms by bioluminescence imaging and scanning electron microscopic analysis.

PubMed

Garcez, Aguinaldo S; Núñez, Silvia C; Azambuja, Nilton; Fregnani, Eduardo R; Rodriguez, Helena M H; Hamblin, Michael R; Suzuki, Hideo; Ribeiro, Martha S

2013-11-01

The aim of this study was to test photodynamic therapy (PDT) as an alternative approach to biofilm disruption on dental hard tissue, We evaluated the effect of methylene blue and a 660 nm diode laser on the viability and architecture of Gram-positive and Gram-negative bacterial biofilms. Ten human teeth were inoculated with bioluminescent Pseudomonas aeruginosa or Enterococcus faecalis to form 3 day biofilms in prepared root canals. Bioluminescence imaging was used to serially quantify and evaluate the bacterial viability, and scanning electron microscopic (SEM) imaging was used to assess architecture and morphology of bacterial biofilm before and after PDT employing methylene blue and 40 mW, 660 nm diode laser light delivered into the root canal via a 300 μm fiber for 240 sec, resulting in a total energy of 9.6 J. The data were statistically analyzed with analysis of variance (ANOVA) followed by Tukey test. The bacterial reduction showed a dose dependence; as the light energy increased, the bioluminescence decreased in both planktonic suspension and in biofilms. The SEM analysis showed a significant reduction of biofilm on the surface. PDT promoted disruption of the biofilm and the number of adherent bacteria was reduced. The photodynamic effect seems to disrupt the biofilm by acting both on bacterial cells and on the extracellular matrix.

Use of a night vision intensifier for direct visualization by eye of far-red and near-infrared fluorescence through an optical microscope.

PubMed

Siddiqi, M A; Kilduff, G M; Gearhart, J D

2003-11-01

We describe the design, construction and testing of a prototype device that allows the direct visualization by eye of far-red and near-infrared (NIR) fluorescence through an optical microscope. The device incorporates a gallium arsenide (GaAs) image intensifier, typically utilized in low-light or 'night vision' applications. The intensifier converts far-red and NIR light into electrons and then into green light, which is visible to the human eye. The prototype makes possible the direct, real-time viewing by eye of normally invisible far-red and NIR fluorescence from a wide variety of fluorophores, using the full field of view of the microscope to which it is applied. The high sensitivity of the image intensifier facilitates the viewing of a wide variety of photosensitive specimens, including live cells and embryos, at vastly reduced illumination levels in both fluorescence and bright-field microscopy. Modifications to the microscope are not required in order to use the prototype, which is fully compatible with all current fluorescence techniques. Refined versions of the prototype device will have broad research and clinical applications.

Cytotoxicity of lidocaine to human corneal endothelial cells in vitro.

PubMed

Yu, Hao-Ze; Li, Yi-Han; Wang, Rui-Xin; Zhou, Xin; Yu, Miao-Miao; Ge, Yuan; Zhao, Jun; Fan, Ting-Jun

2014-04-01

Lidocaine has been reported to induce apoptosis on rabbit corneal endothelial cells. However, the apoptotic effect and exact mechanism involved in cytotoxicity of lidocaine are not well-established in human corneal endothelial (HCE) cells. In this study, we investigated the apoptosis-inducing effect of lidocaine on HCE cells in vitro. After HCE cells were treated with lidocaine at concentrations of 0.15625-10.0 g/l, the morphology and ultrastructure of the cells were observed by inverted light microscope and transmission electron microscope (TEM). Cell viability was measured by MTT assay, and the apoptotic ratio was evaluated with flow cytometry and fluorescent microscopic counting after FITC-Annexin V/PI and AO/EB staining. DNA fragmentation was detected by electrophoresis, and the activation of caspases was evaluated by ELISA. In addition, changes in mitochondrial membrane potential were determined by JC-1 staining. Results suggest that lidocaine above 1.25 g/l reduced cellular viability and triggered apoptosis in HCE cells in a time- and dose-dependent manner. Diminishment of ΔΨm and the activation of caspases indicate that lidocaine-induced apoptosis was caspase dependent and may be related to mitochondrial pathway. © 2013 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

Microscopic video observation of capillary vessel systems using diffuse back lighting

NASA Astrophysics Data System (ADS)

Sakai, Minako; Arai, Hiroki; Iwai, Toshiaki

2017-04-01

We have been developing a simple and practical video microscopy system based on absorption spectra of biological substance to perform spectroscopic observation of living tissues. The diffuse backlighting effect is actively used in the developed system, which is generated by multiple light scattering in the tissue. It is demonstrated that the light specularly reflected from the skin surface can be completely suppressed in the microscopic observation and the biological activity of the capillary vessel systems distributed under the skin can be successfully observed. As a result, we can confirm the effectiveness of the video microscopy system using diffuse backlighting and the applicability of our developed system.

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

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

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

NASA Astrophysics Data System (ADS)

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

2010-03-01

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

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

ScienceCinema

Nazaretski, Evgeny

2018-06-13

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

Fiber optic light-scattering measurement system for evaluation of embryo viability: model experiment

NASA Astrophysics Data System (ADS)

Itoh, Harumi; Arai, Tsunenori; Kikuchi, Makoto

1996-05-01

We evaluated the particle density detectability and particle size detectivity of our fiber-optic light-scattering measurement system. In order to prevent the multiple pregnancy on current in vitro fertilization-embryo transfer, we have aimed to develop a new quantitative and non- invasive method to select a single viable human embryo. We employed the measurement of mitochondria localization in an embryo, which may have the correlation with development ability. We applied the angular distribution measurement of the light-scattering intensity from the embryo to obtain the information originated from the mitochondria. The latex spheres with a diameter of 1.0 micrometers were used to simulate the scattering intensity of the mitochondria. The measurement probes of our system consisted of two fibers for illumination and sensing. They were arranged at a right angle to a microscope optical axis to measure the angular distribution of the light-scattering intensity. We observed that the light-scattering intensity increased monotonically in the range from 106 to 1010 particles per ml. Since the mitochondria density in a human embryo corresponded to 2.5 X 107 per ml in the measurement chamber, we may measure the mitochondria density in the human embryo. The angular dependence of light-scattering intensity changed with the sphere diameters. This result showed the possibility of the selective measurement of the mitochondria density in the embryo in spite of the presence of the other cell organelle. We think that our light-scattering measurement system might be applicable to the evaluation method for the embryo viability.

Quantitative locomotion study of freely swimming micro-organisms using laser diffraction.

PubMed

Magnes, Jenny; Susman, Kathleen; Eells, Rebecca

2012-10-25

Soil and aquatic microscopic organisms live and behave in a complex three-dimensional environment. Most studies of microscopic organism behavior, in contrast, have been conducted using microscope-based approaches, which limit the movement and behavior to a narrow, nearly two-dimensional focal field.(1) We present a novel analytical approach that provides real-time analysis of freely swimming C. elegans in a cuvette without dependence on microscope-based equipment. This approach consists of tracking the temporal periodicity of diffraction patterns generated by directing laser light through the cuvette. We measure oscillation frequencies for freely swimming nematodes. Analysis of the far-field diffraction patterns reveals clues about the waveforms of the nematodes. Diffraction is the process of light bending around an object. In this case light is diffracted by the organisms. The light waves interfere and can form a diffraction pattern. A far-field, or Fraunhofer, diffraction pattern is formed if the screen-to-object distance is much larger than the diffracting object. In this case, the diffraction pattern can be calculated (modeled) using a Fourier transform.(2) C. elegans are free-living soil-dwelling nematodes that navigate in three dimensions. They move both on a solid matrix like soil or agar in a sinusoidal locomotory pattern called crawling and in liquid in a different pattern called swimming.(3) The roles played by sensory information provided by mechanosensory, chemosensory, and thermosensory cells that govern plastic changes in locomotory patterns and switches in patterns are only beginning to be elucidated.(4) We describe an optical approach to measuring nematode locomotion in three dimensions that does not require a microscope and will enable us to begin to explore the complexities of nematode locomotion under different conditions.

Phase-shifting interference microscope with extendable field of measurement

NASA Astrophysics Data System (ADS)

Lin, Shyh-Tsong; Hsu, Wei-Feng; Wang, Ming-Shiang

2018-04-01

An innovative phase-shifting interference microscope aimed at extending the field of measurement is proposed in this paper. The microscope comprises a light source module, a phase modulation module, and an interferometric module, which reconstructs the micro-structure contours of samples using the five-step phase-shifting algorithm. This paper discusses the measurement theory and outlines the configuration, experimental setup, and experimental results obtained using the proposed interference microscope. The results confirm the efficacy of the microscope, achieving a standard deviation of 2.4 nm from a step height of 86.2 nm in multiple examinations.

The Light Microscopy Module: An On-Orbit Multi-User Microscope Facility

NASA Technical Reports Server (NTRS)

Motil, Susan M.; Snead, John H.

2002-01-01

The Light Microscopy Module (LMM) is planned as a remotely controllable on-orbit microscope subrack facility, allowing flexible scheduling and operation of fluids and biology experiments within the Fluids and Combustion Facility (FCF) Fluids Integrated Rack (FIR) on the International Space Station (ISS). The LMM will be the first integrated payload with the FIR to conduct four fluid physics experiments. A description of the LMM diagnostic capabilities, including video microscopy, interferometry, laser tweezers, confocal, and spectrophotometry, will be provided.

Dual-mode optical microscope based on single-pixel imaging

NASA Astrophysics Data System (ADS)

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

2016-07-01

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

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

PubMed

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

2017-12-01

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

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

PubMed Central

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

2017-01-01

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

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

PubMed

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

2011-04-07

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

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

PubMed

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

2012-01-01

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

Low efficiency upconversion nanoparticles for high-resolution coalignment of near-infrared and visible light paths on a light microscope

PubMed Central

Sundaramoorthy, Sriramkumar; Badaracco, Adrian Garcia; Hirsch, Sophia M.; Park, Jun Hong; Davies, Tim; Dumont, Julien; Shirasu-Hiza, Mimi; Kummel, Andrew C.; Canman, Julie C.

2017-01-01

The combination of near infrared (NIR) and visible wavelengths in light microscopy for biological studies is increasingly common. For example, many fields of biology are developing the use of NIR for optogenetics, in which an NIR laser induces a change in gene expression and/or protein function. One major technical barrier in working with both NIR and visible light on an optical microscope is obtaining their precise coalignment at the imaging plane position. Photon upconverting particles (UCPs) can bridge this gap as they are excited by NIR light but emit in the visible range via an anti-Stokes luminescence mechanism. Here, two different UCPs have been identified, high-efficiency micro540-UCPs and lower efficiency nano545-UCPs, that respond to NIR light and emit visible light with high photostability even at very high NIR power densities (>25,000 Suns). Both of these UCPs can be rapidly and reversibly excited by visible and NIR light and emit light at visible wavelengths detectable with standard emission settings used for Green Fluorescent Protein (GFP), a commonly used genetically-encoded fluorophore. However, the high efficiency micro540-UCPs were suboptimal for NIR and visible light coalignment, due to their larger size and spatial broadening from particle-to-particle energy transfer consistent with a long lived excited state and saturated power dependence. In contrast, the lower efficiency nano-UCPs were superior for precise coalignment of the NIR beam with the visible light path (~2 µm versus ~8 µm beam broadening respectively) consistent with limited particle-to-particle energy transfer, superlinear power dependence for emission, and much smaller particle size. Furthermore, the nano-UCPs were superior to a traditional two-camera method for NIR and visible light path alignment in an in vivo Infrared-Laser-Evoked Gene Operator (IR-LEGO) optogenetics assay in the budding yeast S. cerevisiae. In summary, nano-UCPs are powerful new tools for coaligning NIR and visible light paths on a light microscope. PMID:28221018

A compact light-sheet microscope for the study of the mammalian central nervous system

PubMed Central

Yang, Zhengyi; Haslehurst, Peter; Scott, Suzanne; Emptage, Nigel; Dholakia, Kishan

2016-01-01

Investigation of the transient processes integral to neuronal function demands rapid and high-resolution imaging techniques over a large field of view, which cannot be achieved with conventional scanning microscopes. Here we describe a compact light sheet fluorescence microscope, featuring a 45° inverted geometry and an integrated photolysis laser, that is optimized for applications in neuroscience, in particular fast imaging of sub-neuronal structures in mammalian brain slices. We demonstrate the utility of this design for three-dimensional morphological reconstruction, activation of a single synapse with localized photolysis, and fast imaging of neuronal Ca2+ signalling across a large field of view. The developed system opens up a host of novel applications for the neuroscience community. PMID:27215692

HIGH TEMPERATURE MICROSCOPE AND FURNACE

DOEpatents

Olson, D.M.

1961-01-31

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

Colonization of cashew plants by Lasiodiplodia theobromae: Microscopical features

USDA-ARS?s Scientific Manuscript database

Lasiodiplodia theobromae is a phytopathogenic fungus causing gummosis, a threatening disease for cashew plants in Brazil. In an attempt to investigate the ultrastructural features of the pathogen colonization and its response to immunofluorescence labeling, light, confocal and electron microscope st...

Fluorescence-guided resection of intracranial VX2 tumor in a preclinical model using 5-aminolevulinic acid (ALA): preliminary results

NASA Astrophysics Data System (ADS)

Bogaards, Arjen; Varma, Abhay; Moriyama, Eduardo H.; Lin, Annie; Giles, Anoja; Bisland, Stuart K.; Lilge, Lothar D.; Bilbao, G. M.; Muller, Paul J.; Wilson, Brian C.

2003-06-01

Fluorescence-guided brain tumor resection may help the neurosurgeon to identify tumor margins that merge imperceptibly into the normal brain tissue and are difficult to identify under white light illumination even using an operating microscope. We compared the amount of residual tumor after white light resection using an operating microscope versus that after fluorescnece-guided resection of an intracranial VX2 tumor in a preclinical model using our previously developed co-axial fluorscence imaging and spectroscopy system, exciting and detecting PpIX fluorescence at 405nm and 635nm respectively. Preliminary results: No fluorescence was present in 3 non-tumor-bearing animals. Fluorescence was present in all 15 tumor-bearing animals after white light resection was completed. To date in 4 rabbits, a decrease in residual tumor was found when using additional fluorescence guided resection compared to white light resection only. Conclusions: ALA induced PpIX fluorescence detects tumor margins not seen under an operation microscope using while light. Using fluorescence imaging to guide tumor resection resulted in a 3-fold decrease in the amount of residual timor. However, these preliminary results indicate that also an additional amount of normal brain is resected, which will be further investigated.

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

NASA Astrophysics Data System (ADS)

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

2006-10-01

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

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

PubMed Central

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

2012-01-01

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

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

PubMed

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

2012-01-01

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

The e-evolution of microscopy in dental education.

PubMed

Farah, Camile S; Maybury, Terrence S

2009-08-01

Recent technological innovation has now made it possible to turn the computer into a microscope. This has entailed a shift from light microscopy to virtual microscopy. This development then foregrounds the issue of the pedagogy involved in this move from the analogue technology of the light microscope to the digital, computerized instance of virtual microscopy. In order to address this issue, undergraduate students enrolled in the Bachelor of Dental Science program at the University of Queensland School of Dentistry were surveyed to ascertain their preference for light or virtual microscopy. The value of this study is that it was conducted on the same cohort of students in two separate courses in 2006 and 2008, giving it longitudinal validity. The responses were overwhelmingly in favor of virtual microscopy. When it came to completely replacing the light microscope with virtual microscopy, however, students were much more ambivalent about such a wholesale change although this was less of an issue in the senior year. This shift from light to virtual microscopy signals larger changes in the tertiary sector from print-literate to electronic forms of knowledge and from teacher-centered to student-focused frames of learning. In short, we are in the midst of the e-evolution of microscopy in dental education.

Evaluation of bone response to titanium-coated polymethyl methacrylate resin (PMMA) implants by X-ray tomography.

PubMed

Shalabi, Manal M; Wolke, Johannes G C; Cuijpers, Vincent M J I; Jansen, John A

2007-10-01

High-resolution three-dimensional data about the bone response to oral implants can be obtained by using microfocus computer tomography. However, a disadvantage is that metallic implants cause streaking artifacts due to scattering of X-rays, which prevents an accurate evaluation of the interfacial bone-to-implant contact. It has been suggested that the use of thin titanium coatings deposited on polymeric implants can offer an alternative option for analyzing bone contact using micro-CT imaging. Consequently, the aim of the current study was to investigate bone behavior to titanium-coated polymethylmethacrylate (PMMA) implants by micro-CT and histological evaluation. For the experiment titanium-coated PMMA implants were used. The implants had a machined threaded appearance and were provided with a 400-500 nm thick titanium coating. The implants were inserted in the right or left tibia of 10 goats. After an implantation period of 12 weeks the implants were retrieved and prepared for micro-computer tomography (microCT), light microscopy, and X-ray microanalysis. The micro-CT showed that the screw-threads and typical implant configuration were well maintained through the installation procedure. Overall, histological responses showed that the titanium-coated implants were well tolerated and caused no atypical tissue response. In addition, the bone was seen in direct contact with the titanium-coated layer. The X-ray microanalysis results confirmed the light microscopical data. In conclusion, the obtained results proof the final use of titanium-coated PMMA implants for evaluation of the bone-implant response using microCT. However, this study also confirms that for a proper analysis of the bone-implant interface the additional use of microscopical techniques is still required.

Role of Caspase-3 in acute light damage to retina of rats.

PubMed

Wang, Xiao; Hu, Shi-Xing; Li, Wei; Lin, Shao-Chun

2007-03-01

To investigate the role of Caspase-3 in retinal damage caused by light exposure in rats. Light injury to retina was induced by persistent exposure to illumination (intensity: 30 000 +/- 50 lux) of operating microscope for 30 minutes in the right eyes of Sprague-Dawley rats. The pathological changes of retina were observed under optical and electron microscopies at different time points, which were 6 hours, 1, 3, 7, and 15 days after the light exposure. Apoptosis of retinal cells was analyzed by flow cytometry. The activity of Caspase-3 was evaluated by using the Caspase-3 assay kit. At the same time, the expression of Caspase-3 protease was determined with Western blot analysis. The examination results of optical and transmission electron microscopes showed that edema of inner and outer segments of the retina, especially the chondriosome inside the inner segment, became obvious 6 hours after the light exposure. The change was deteriorated along with the increasing time. The structures of the discoidal valve dissociated in the outer segment simultaneously. Disorderly arranged nuclei, karyopycnosis, and thinning in the outer nuclear layer were observed. The retinal pigment epithelium almost disappeared during the later stage. The staining results of Annexin-V combined with PI demonstrated that the proportion of apoptotic cells increased with time. The proportion between 7th day (82.7%) and 15th day (80.4%), however, showed no significant difference. Caspase-3 became remarkably active with the lapse of time, which increased from 0.02 at 6th hour to the peak of 9.8 at 7th day before it started to descend. The Western blot detected a expression of the active form of Caspase-3 at 7th day and 15th day. Apoptosis of photoreceptor cells is markedly involved in the light damage and Caspase-3 protease may play an important role in the apoptotic process of the retina after light exposure in rats.

Direction-division multiplexed holographic free-electron-driven light sources

NASA Astrophysics Data System (ADS)

Clarke, Brendan P.; MacDonald, Kevin F.; Zheludev, Nikolay I.

2018-01-01

We report on a free-electron-driven light source with a controllable direction of emission. The source comprises a microscopic array of plasmonic surface-relief holographic domains, each tailored to direct electron-induced light emission at a selected wavelength into a collimated beam in a prescribed direction. The direction-division multiplexed source is tested by driving it with the 30 kV electron beam of a scanning electron microscope: light emission, at a wavelength of 800 nm in the present case, is switched among different output angles by micron-scale repositioning of the electron injection point among domains. Such sources, with directional switching/tuning possible at picosecond timescales, may be applied to field-emission and surface-conduction electron-emission display technologies, optical multiplexing, and charged-particle-beam position metrology.

Enhanced optical coupling and Raman scattering via microscopic interface engineering

NASA Astrophysics Data System (ADS)

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

2017-11-01

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

Polarized light and scanning electron microscopic investigation of enamel hypoplasia in primary teeth.

PubMed

Sabel, Nina; Klingberg, Gunilla; Dietz, Wolfram; Nietzsche, Sandor; Norén, Jörgen G

2010-01-01

Enamel hypoplasia is a developmental disturbance during enamel formation, defined as a macroscopic defect in the enamel, with a reduction of the enamel thickness with rounded, smooth borders. Information on the microstructural level is still limited, therefore further studies are of importance to better understand the mechanisms behind enamel hypoplasia. To study enamel hypoplasia in primary teeth by means of polarized light microscopy and scanning electron microscopy. Nineteen primary teeth with enamel hypoplasia were examined in a polarized light microscope and in a scanning electron microscope. The cervical and incisal borders of the enamel hypoplasia had a rounded appearance, as the prisms in the rounded cervical area of the hypoplasia were bent. The rounded borders had a normal surface structure whereas the base of the defects appeared rough and porous. Morphological findings in this study indicate that the aetiological factor has a short duration and affects only certain ameloblasts. The bottom of the enamel hypoplasia is porous and constitutes possible pathways for bacteria into the dentin.

Retinal endoilluminator toxicity of xenon and light-emitting diode (LED) light source: rabbit model.

PubMed

Aydin, Bahri; Dinç, Erdem; Yilmaz, S Necat; Altiparmak, U Emrah; Yülek, Fatma; Ertekin, Sevda; Yilmaz, Mustafa; Yakın, Mehmet

2014-09-01

This study evaluates retinal toxicity due to endoillumination with the light-emitting diode (LED) light source in comparison to endoillumination with xenon light source. Twenty-five eyes of 14 New Zealand pigmented rabbits were used in the study. The LED light (Omesis Medical Systems, Turkey) group was composed of 7 right eyes, while the other 7 right eyes constituted the xenon group (420 nm filter, 357mW/cm(2)) (Bright Star; DORC, Zuidland, Netherlands). Eleven untreated left eyes composed the control group. Twenty gauge pars plana incision 1.5 mm behind the limbus was performed in the right eyes. Twenty gauge bullet type fiberoptic endoilluminator was inserted into the eye from the incision without any pars plana vitrectomy. Fiberoptic endoilluminator was placed in such a way that it was directed toward visual streak of the rabbit retina with a 5 mm distance to retinal surface. Endoillumination was then applied for 20 min with a maximum light intensity for LED and xenon light. In left control eyes, no surgical procedure and no endoillumination were performed. One week after the endoillumination procedure, both eyes of the rabbits were enucleated following electroretinography. Sections stained with hematoxylin and eosin to evaluate morphologic changes. Retina tissues were assessed by active caspase-3 staining. There was no difference in the shape of the waveforms recorded in the eyes endoilluminated with LED light and xenon light sources compared to control eyes both before and after endoillumination application (p > 0.05). Microscopic evaluation of the retinas with hematoxylin and eosin staining demonstrated that all study groups have normal histologic properties similar to control group. No apoptosis positive cells were found within all sections in all groups. When the LED light source is used with maximum power and limited duration for endoillumination in rabbit eyes it does not produce phototoxic effects that may be detectable by electrophysiology and histology similar to xenon light.

Determining the phonon energy of highly oriented pyrolytic graphite by scanning tunneling microscope light emission spectroscopy

NASA Astrophysics Data System (ADS)

Uehara, Yoichi; Michimata, Junichi; Watanabe, Shota; Katano, Satoshi; Inaoka, Takeshi

2018-03-01

We have investigated the scanning tunneling microscope (STM) light emission spectra of isolated single Ag nanoparticles lying on highly oriented pyrolytic graphite (HOPG). The STM light emission spectra exhibited two types of spectral structures (step-like and periodic). Comparisons of the observed structures and theoretical predictions indicate that the phonon energy of the ZO mode of HOPG [M. Mohr et al., Phys. Rev. B 76, 035439 (2007)] can be determined from the energy difference between the cutoff of STM light emission and the step in the former structure, and from the period of the latter structure. Since the role of the Ag nanoparticles does not depend on the substrate materials, this method will enable the phonon energies of various materials to be measured by STM light emission spectroscopy. The spatial resolution is comparable to the lateral size of the individual Ag nanoparticles (that is, a few nm).

Plasmon-resonance-enhanced visible-light photocatalytic activity of Ag quantum dots/TiO2 microspheres for methyl orange degradation

NASA Astrophysics Data System (ADS)

Yu, Xin; Shang, Liwei; Wang, Dongjun; An, Li; Li, Zhonghua; Liu, Jiawen; Shen, Jun

2018-06-01

We successfully prepared Ag quantum dots modified TiO2 microspheres by facile solvothermal and calcination method. The as-prepared Ag quantum dots/TiO2 microspheres were characterized by scanning electron microscope, transmission electron microscope, X-ray diffraction, X-ray photoelectron spectroscopy and UV-vis diffuse reflectance spectroscopy. The Ag quantum dots/TiO2 photocatalyst showed excellent visible light absorption and efficient photocatalytic activity for methyl orange degradation. And the sample with the molar ratio of 0.05 (Ag to Ti) showed the best visible light photocatalytic activity for methyl orange degradation, mainly because of the surface plasmon resonance (SPR) effects of Ag quantum dots to generate electron and hole pairs for enhanced visible light photocatalysis. Finally, possible visible light photocatalytic mechanism of Ag quantum dots/TiO2 microspheres for methyl orange degradation was proposed in detail.

3D widefield light microscope image reconstruction without dyes

NASA Astrophysics Data System (ADS)

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.

Study of factors affecting the appearance of colors under microscopes

NASA Astrophysics Data System (ADS)

Zakizadeh, Roshanak; Martinez-Garcia, Juan; Raja, Kiran B.; Siakidis, Christos

2013-11-01

The variation of colors in microscopy systems can be quite critical for some users. To address this problem, a study is conducted to analyze how different factors such as size of the sample, intensity of the microscope's light source and the characteristics of the material like chroma and saturation can affect the color appearance through the eyepiece of the microscope. To study the changes in colors considering these factors, the spectral reflectance of 24 colors of GretagMacbeth Classic ColorChecker® and Mini ColorChecker® which are placed under a Nikon ECLIPSE MA200 microscope®2 using dark filed and bright field illuminations which result in different intensity levels, is measured using a spectroradiometer®3 which was placed in front of the eyepiece of the microscope. The results are compared with the original data from N. Ohta1. The evaluation is done by observing the shift in colors in the CIE 1931 Chromaticity Diagram and the CIELAB space, also by applying a wide set of color-difference formulas, namely: CIELAB, CMC, BFD, CIE94, CIEDE2000, DIN99d and DIN99b. Furthermore, to emphasize on the color regions in which the highest difference is observed, the authors have obtained the results from another microscope; Olympus SZX10®4, which in this case the measurement is done by mounting the spectroradiometer to the camera port of the microscope. The experiment leads to some interesting results, among which is the consistency in the highest difference observed considering different factors or how the change in saturation of the samples of the same hue can affect the results.

Digital learning programs - competition for the classical microscope?

PubMed

Schmidt, Peter

2013-01-01

The development of digital media has been impressive in recent years which is also among the reason for their increasing use in academic teaching. This is especially true for teaching Anatomy and Histology in the first two years in medical and dental curricula. Modern digital technologies allow for efficient, affordable and easily accessible distribution of histological images in high quality. Microscopy depends almost exclusively on such images. Since 20 years numerous digital teaching systems have been developed for this purpose. Respective developments have changed the ways students acquire knowledge and prepare for exams. Teaching staff should adapt lectures, seminars and labs accordingly. As a first step, a collection of high resolution digital microscopic slides was made available for students at the Friedrich-Schiller-University in Jena. The aim of the present study was to evaluate the importance of conventional light microscopy and related technologies in current and future medical and dental education aswell. A survey was done among 172 medical and dental students at the Friedrich-Schiller-University Jena. 51% of students use now frequently new digital media for learning histology in contrast to 5% in the year 2000 [1]. Digital media including Internet, CD- based learning combined with social networks successfully compete with classical light microscopy.

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

PubMed

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

2018-04-19

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

Visualizing 3-D microscopic specimens

NASA Astrophysics Data System (ADS)

Forsgren, Per-Ola; Majlof, Lars L.

1992-06-01

The confocal microscope can be used in a vast number of fields and applications to gather more information than is possible with a regular light microscope, in particular about depth. Compared to other three-dimensional imaging devices such as CAT, NMR, and PET, the variations of the objects studied are larger and not known from macroscopic dissections. It is therefore important to have several complementary ways of displaying the gathered information. We present a system where the user can choose display techniques such as extended focus, depth coding, solid surface modeling, maximum intensity and other techniques, some of which may be combined. A graphical user interface provides easy and direct control of all input parameters. Motion and stereo are available options. Many three- dimensional imaging devices give recordings where one dimension has different resolution and sampling than the other two which requires interpolation to obtain correct geometry. We have evaluated algorithms with interpolation in object space and in projection space. There are many ways to simplify the geometrical transformations to gain performance. We present results of some ways to simplify the calculations.

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

PubMed Central

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

2012-01-01

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

Localization of nitric oxide synthase and NADPH-diaphorase in guinea pig and human cochleae.

PubMed

Ruan, R S; Leong, S K; Yeoh, K H

1997-01-01

The distributions of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) and nitric oxide synthase (NOS) in mammalian cochlea were studied at light and electron microscope levels by NADPH-d histochemistry and brain NOS (bNOS) immunohistochemistry. The cochleae from 15 albino guinea pigs were perilymphatically fixed with 2% periodate-lysine-paraformaldehyde, decalcified in 10% EDTA and processed for light and electron microscopy after NADPH-d or NOS staining in frozen and vibratome sections respectively. One human cochlea was available for light microscope examination of NADPH-d or bNOS stained sections. Light microscope results revealed that type I neurons and nerve fibers of the spiral ganglion cells were labeled by bNOS immunohistochemistry as well as NADPH-d histochemistry in both guinea pig and human cochleae. At subcellular level, NADPH-d reaction product was localized in the mitochondria of the neuronal cytoplasm and axoplasm and in the cytoplasm of the vascular endothelium. The immunoreaction products of bNOS were evenly distributed in the neuronal cytoplasm and axoplasm. Myelinated and unmyelinated fibers in the intraganglionic spiral bundle and the inner spiral and inner radial fibers below the inner hair cells were labeled for bNOS. The nerve endings below the outer hair cells were not stained. NOS immunoreaction product was also found in the outer hair cells, Schwann cells of myelinated nerve fibers, Deiter's cells, pillar cells and the tympanic lamina cells. No difference was found in the staining pattern of both NADPH-d and NOS reaction products between human and guinea pig cochleae at the light microscope level. The results suggest that NO plays an important role in the maintenance of auditory function in the mammal.

Evaluation of the human hair root for DNA typing subsequent to microscopic comparison.

PubMed

Linch, C A; Smith, S L; Prahlow, J A

1998-03-01

Telogen human hairs are one of the most common useful evidence findings at crime scenes and/or on homicide victims. Occasionally, the microscopic characterization of the found telogen hair is the only physical evidence association to a victim or suspect. Recently efforts to characterize these hairs by mitochondrial DNA (mtDNA) methods have progressed. The nature of the telogen hair root morphology and ultrastructure has, however, been largely ignored. Examiners have recognized these hairs are unlikely to be typable by nuclear DNA (nuDNA) methods. Most forensic biologists have little knowledge of the complex cellular composition of anagen, catagen, and telogen hair roots or their morphogenesis. This paper reviews ex situ human hair root morphology as it relates to the likelihood of successful nuclear DNA typing. Dermatology texts of hair root morphology always demonstrate their microscopic appearance in the skin. This study investigates the use of fluorescence in situ hybridization (FISH) methods to sex type telogen head hairs, and it further investigates hair root morphology as it relates to the potential nuclear DNA content of evidence hairs. There is a need for the use of appropriate, consensus terminology for describing hair root morphology. There is also a need for standardized laboratory light microscopic methods in evaluating a hair root for DNA typing. FISH was found to be an unsuitable technique for sex determination of telogen hair clubs. It was determined that anagen/catagen hair roots without translucent sheath material are excellent candidates for nuDNA PCR-based typing and that hairs with telogen club root material only should not be submitted for nuDNA typing attempts.

Hyperspectral microscopy to identify foodborne bacteria with optimum lighting source

USDA-ARS?s Scientific Manuscript database

Hyperspectral microscopy is an emerging technology for rapid detection of foodborne pathogenic bacteria. Since scattering spectral signatures from hyperspectral microscopic images (HMI) vary with lighting sources, it is important to select optimal lights. The objective of this study is to compare t...

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

PubMed

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

2009-09-01

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

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

PubMed

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.

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

NASA Astrophysics Data System (ADS)

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

1996-01-01

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

RGB digital lensless holographic microscopy

NASA Astrophysics Data System (ADS)

Garcia-Sucerquia, Jorge

2013-11-01

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

Optical forces, torques, and force densities calculated at a microscopic level using a self-consistent hydrodynamics method

NASA Astrophysics Data System (ADS)

Ding, Kun; Chan, C. T.

2018-04-01

The calculation of optical force density distribution inside a material is challenging at the nanoscale, where quantum and nonlocal effects emerge and macroscopic parameters such as permittivity become ill-defined. We demonstrate that the microscopic optical force density of nanoplasmonic systems can be defined and calculated using the microscopic fields generated using a self-consistent hydrodynamics model that includes quantum, nonlocal, and retardation effects. We demonstrate this technique by calculating the microscopic optical force density distributions and the optical binding force induced by external light on nanoplasmonic dimers. This approach works even in the limit when the nanoparticles are close enough to each other so that electron tunneling occurs, a regime in which classical electromagnetic approach fails completely. We discover that an uneven distribution of optical force density can lead to a light-induced spinning torque acting on individual particles. The hydrodynamics method offers us an accurate and efficient approach to study optomechanical behavior for plasmonic systems at the nanoscale.

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

PubMed Central

Weston, Anne E.; Armer, Hannah E. J.

2009-01-01

Modern cell biology is reliant on light and fluorescence microscopy for analysis of cells, tissues and protein localisation. However, these powerful techniques are ultimately limited in resolution by the wavelength of light. Electron microscopes offer much greater resolution due to the shorter effective wavelength of electrons, allowing direct imaging of sub-cellular architecture. The harsh environment of the electron microscope chamber and the properties of the electron beam have led to complex chemical and mechanical preparation techniques, which distance biological samples from their native state and complicate data interpretation. Here we describe recent advances in sample preparation and instrumentation, which push the boundaries of high-resolution imaging. Cryopreparation, cryoelectron microscopy and environmental scanning electron microscopy strive to image samples in near native state. Advances in correlative microscopy and markers enable high-resolution localisation of proteins. Innovation in microscope design has pushed the boundaries of resolution to atomic scale, whilst automatic acquisition of high-resolution electron microscopy data through large volumes is finally able to place ultrastructure in biological context. PMID:19916039

Light Microscopy's New Jobs

NASA Astrophysics Data System (ADS)

Ritsch-Marte, Monika

2009-04-01

300 years since the first glimpse through the earliest microscopes, light microscopy is still an active field of research, breaking new frontiers in optical imaging and even becoming a means of mechanical manipulation of microparticles.

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

PubMed

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

2013-01-01

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

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

PubMed

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

2007-09-03

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

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

NASA Astrophysics Data System (ADS)

Anderson, Benjamin; Bernhardt, Elizabeth; Kuzyk, Mark

2012-10-01

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

Electroluminescence of a polythiophene molecular wire suspended between a metallic surface and the tip of a scanning tunneling microscope.

PubMed

Reecht, Gaël; Scheurer, Fabrice; Speisser, Virginie; Dappe, Yannick J; Mathevet, Fabrice; Schull, Guillaume

2014-01-31

The electroluminescence of a polythiophene wire suspended between a metallic surface and the tip of a scanning tunneling microscope is reported. Under positive sample voltage, the spectral and voltage dependencies of the emitted light are consistent with the fluorescence of the wire junction mediated by localized plasmons. This emission is strongly attenuated for the opposite polarity. Both emission mechanism and polarity dependence are similar to what occurs in organic light emitting diodes (OLED) but at the level of a single molecular wire.

Camera array based light field microscopy

PubMed Central

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

2015-01-01

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

A line scanned light-sheet microscope with phase shaped self-reconstructing beams.

PubMed

Fahrbach, Florian O; Rohrbach, Alexander

2010-11-08

We recently demonstrated that Microscopy with Self-Reconstructing Beams (MISERB) increases both image quality and penetration depth of illumination beams in strongly scattering media. Based on the concept of line scanned light-sheet microscopy, we present an add-on module to a standard inverted microscope using a scanned beam that is shaped in phase and amplitude by a spatial light modulator. We explain technical details of the setup as well as of the holograms for the creation, positioning and scaling of static light-sheets, Gaussian beams and Bessel beams. The comparison of images from identical sample areas illuminated by different beams allows a precise assessment of the interconnection between beam shape and image quality. The superior propagation ability of Bessel beams through inhomogeneous media is demonstrated by measurements on various scattering media.

A method for fast automated microscope image stitching.

PubMed

Yang, Fan; Deng, Zhen-Sheng; Fan, Qiu-Hong

2013-05-01

Image stitching is an important technology to produce a panorama or larger image by combining several images with overlapped areas. In many biomedical researches, image stitching is highly desirable to acquire a panoramic image which represents large areas of certain structures or whole sections, while retaining microscopic resolution. In this study, we develop a fast normal light microscope image stitching algorithm based on feature extraction. At first, an algorithm of scale-space reconstruction of speeded-up robust features (SURF) was proposed to extract features from the images to be stitched with a short time and higher repeatability. Then, the histogram equalization (HE) method was employed to preprocess the images to enhance their contrast for extracting more features. Thirdly, the rough overlapping zones of the images preprocessed were calculated by phase correlation, and the improved SURF was used to extract the image features in the rough overlapping areas. Fourthly, the features were corresponded by matching algorithm and the transformation parameters were estimated, then the images were blended seamlessly. Finally, this procedure was applied to stitch normal light microscope images to verify its validity. Our experimental results demonstrate that the improved SURF algorithm is very robust to viewpoint, illumination, blur, rotation and zoom of the images and our method is able to stitch microscope images automatically with high precision and high speed. Also, the method proposed in this paper is applicable to registration and stitching of common images as well as stitching the microscope images in the field of virtual microscope for the purpose of observing, exchanging, saving, and establishing a database of microscope images. Copyright © 2013 Elsevier Ltd. All rights reserved.

Diffraction-Unlimited Fluorescence Imaging with an EasySTED Retrofitted Confocal Microscope.

PubMed

Klauss, André; Hille, Carsten

2017-01-01

The easySTED technology provides the means to retrofit a confocal microscope to a diffraction-unlimited stimulated emission depletion (STED) microscope.Although commercial STED systems are available today, for many users of confocal laser scanning microscopes the option of retrofitting their confocal system to a STED system ready for diffraction-unlimited imaging may present an attractive option. The easySTED principle allowing for a joint beam path of excitation and depletion light promises some advantages concerning technical complexity and alignment effort for such an STED upgrade. In the one beam path design of easySTED the use of a common laser source, either a supercontinuum source or two separate lasers coupled into the same single-mode fiber, becomes feasible. The alignment of the focal light distribution of the STED beam relative to that of the excitation beam in all three spatial dimensions is therefore omitted respectively reduced to coupling the STED laser into the common single-mode fiber. Thus, only minor modifications need to be applied to the beam path in the confocal microscope to be upgraded. Those comprise adding polarization control elements and the easySTED waveplate, and adapting the beamsplitter to the excitation/STED wavelength combination.

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

PubMed

Boruah, B R; Neil, M A A

2009-01-01

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

Electron-Beam-Lithographed Nanostructures as Reference Materials for Label-Free Scattered-Light Biosensing of Single Filoviruses.

PubMed

Agrawal, Anant; Majdi, Joseph; Clouse, Kathleen A; Stantchev, Tzanko

2018-05-23

Optical biosensors based on scattered-light measurements are being developed for rapid and label-free detection of single virions captured from body fluids. Highly controlled, stable, and non-biohazardous reference materials producing virus-like signals are valuable tools to calibrate, evaluate, and refine the performance of these new optical biosensing methods. To date, spherical polymer nanoparticles have been the only non-biological reference materials employed with scattered-light biosensing techniques. However, pathogens like filoviruses, including the Ebola virus, are far from spherical and their shape strongly affects scattered-light signals. Using electron beam lithography, we fabricated nanostructures resembling individual filamentous virions attached to a biosensing substrate (silicon wafer overlaid with silicon oxide film) and characterized their dimensions with scanning electron and atomic force microscopes. To assess the relevance of these nanostructures, we compared their signals across the visible spectrum to signals recorded from Ebola virus-like particles which exhibit characteristic filamentous morphology. We demonstrate the highly stable nature of our nanostructures and use them to obtain new insights into the relationship between virion dimensions and scattered-light signal.

Automated system for characterization and classification of malaria-infected stages using light microscopic images of thin blood smears.

PubMed

Das, D K; Maiti, A K; Chakraborty, C

2015-03-01

In this paper, we propose a comprehensive image characterization cum classification framework for malaria-infected stage detection using microscopic images of thin blood smears. The methodology mainly includes microscopic imaging of Leishman stained blood slides, noise reduction and illumination correction, erythrocyte segmentation, feature selection followed by machine classification. Amongst three-image segmentation algorithms (namely, rule-based, Chan-Vese-based and marker-controlled watershed methods), marker-controlled watershed technique provides better boundary detection of erythrocytes specially in overlapping situations. Microscopic features at intensity, texture and morphology levels are extracted to discriminate infected and noninfected erythrocytes. In order to achieve subgroup of potential features, feature selection techniques, namely, F-statistic and information gain criteria are considered here for ranking. Finally, five different classifiers, namely, Naive Bayes, multilayer perceptron neural network, logistic regression, classification and regression tree (CART), RBF neural network have been trained and tested by 888 erythrocytes (infected and noninfected) for each features' subset. Performance evaluation of the proposed methodology shows that multilayer perceptron network provides higher accuracy for malaria-infected erythrocytes recognition and infected stage classification. Results show that top 90 features ranked by F-statistic (specificity: 98.64%, sensitivity: 100%, PPV: 99.73% and overall accuracy: 96.84%) and top 60 features ranked by information gain provides better results (specificity: 97.29%, sensitivity: 100%, PPV: 99.46% and overall accuracy: 96.73%) for malaria-infected stage classification. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

Comparison of skin responses from macroscopic and microscopic UV challenges

NASA Astrophysics Data System (ADS)

Seo, InSeok; Bargo, Paulo R.; Chu, Melissa; Ruvolo, Eduardo; Kollias, Nikiforos

2011-03-01

The minimal erythema dose induced by solar-simulated radiation is a useful measure of UV sensitivity of skin. Most skin phototests have been conducted by projecting a flat field of UV radiation onto the skin in an area greater than 15 cm × 15 cm with an increment of radiation doses. In this study, we investigated the responses of human skin to solar-simulated radiation of different field sizes. Twelve human subjects of skin phototype I-IV were exposed to solar-simulated radiation (SSR) on their upper inner arm or on their lower back with a series of doses in increments of 20% in order to determine the threshold dose to induce a minimal perceptible erythema response (MED). Each dose was delivered with a liquid light guide (8 mm diameter on the back or 6 mm on the upper inner arm) and with quartz optical fibers of 200 μm diameter. The resulting skin responses were evaluated visually and investigated with a reflectance confocal microscope and imaging. The erythema response to the microscopic challenge was always diffuse with no clear boundaries extending to several times the exposed site diameter at doses greater than 2 MED. The skin returned to normal appearance from the microscopic challenge after two weeks of exposure while change in appearance for the larger areas persisted for several weeks to months. This new modality of testing provides the possibility to study skin at the microscopic level with a rapid recovery following challenge.

Excitation-scanning hyperspectral imaging system for microscopic and endoscopic applications

NASA Astrophysics Data System (ADS)

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

2016-04-01

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

Polarized Light Corridor Demonstrations.

ERIC Educational Resources Information Center

Davies, G. R.

1990-01-01

Eleven demonstrations of light polarization are presented. Each includes a brief description of the apparatus and the effect demonstrated. Illustrated are strain patterns, reflection, scattering, the Faraday Effect, interference, double refraction, the polarizing microscope, and optical activity. (CW)

Scanning Microscopes Using X Rays and Microchannels

NASA Technical Reports Server (NTRS)

Wang, Yu

2003-01-01

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

Three-dimensional scanning confocal laser microscope

DOEpatents

Anderson, R. Rox; Webb, Robert H.; Rajadhyaksha, Milind

1999-01-01

A confocal microscope for generating an image of a sample includes a first scanning element for scanning a light beam along a first axis, and a second scanning element for scanning the light beam at a predetermined amplitude along a second axis perpendicular to the first axis. A third scanning element scans the light beam at a predetermined amplitude along a third axis perpendicular to an imaging plane defined by the first and second axes. The second and third scanning element are synchronized to scan at the same frequency. The second and third predetermined amplitudes are percentages of their maximum amplitudes. A selector determines the second and third predetermined amplitudes such that the sum of the percentages is equal to one-hundred percent.

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

USDA-ARS?s Scientific Manuscript database

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

Integration of Histology Lectures and Practical Teaching in China

ERIC Educational Resources Information Center

Lu, Xiaoye; Cheng, Xin; Li, Ke; Lee, Kenneth Ka Ho; Yang, Xuesong

2016-01-01

Objectives: Human histology is a discipline concerning the study of microscopic structures of human tissues and organs--with the aid of light or electron microscopes. Traditional teaching of histology is composed of two separated components, theory and practice. The main disadvantage with traditional histology teaching is the detachment of theory…

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

PubMed

Maybury, Terrence; Farah, Camile S

2009-09-01

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

Effect of intraperitoneal selenium administration on liver glycogen levels in rats subjected to acute forced swimming.

PubMed

Akil, Mustafa; Bicer, Mursel; Kilic, Mehmet; Avunduk, Mustafa Cihat; Mogulkoc, Rasim; Baltaci, Abdulkerim Kasim

2011-03-01

There are a few of studies examining how selenium, which is known to reduce oxidative damage in exercise, influences glucose metabolism and exhaustion in physical activity. The present study aims to examine how selenium administration affects liver glycogen levels in rats subjected to acute swimming exercise. The study included 32 Sprague-Dawley type male rats, which were equally allocated to four groups: Group 1, general control; Group 2; selenium-supplemented control (6 mg/kg/day sodium selenite); Group 3, swimming control; Group 4, selenium-supplemented swimming (6 mg/kg/day sodium selenite). Liver tissue samples collected from the animals at the end of the study were fixed in 95% ethyl alcohol. From the tissue samples buried into paraffin, 5-µm cross-sections were obtained using a microtome, put on a microscope slide, and stained with PAS. Stained preparations were assessed using a Nikon Eclipse E400 light microscope. All images obtained with the light microscope were transferred to a PC and evaluated using Clemex PE 3.5 image analysis software. The highest liver glycogen levels were found in groups 1 and 2 (p < 0.05). The levels in group 4 were lower than those in groups 1 and 2 but higher than the levels in group 3 (p < 0.05). The lowest liver glycogen levels were obtained in group 3 (p < 0.05). Results of the study indicate that liver glycogen levels that decrease in acute swimming exercise can be restored by selenium administration. It can be argued that physiological doses of selenium administration can contribute to performance.

Longitudinal train dynamics model for a rail transit simulation system

DOE PAGES

Wang, Jinghui; Rakha, Hesham A.

2018-01-01

The paper develops a longitudinal train dynamics model in support of microscopic railway transportation simulation. The model can be calibrated without any mechanical data making it ideal for implementation in transportation simulators. The calibration and validation work is based on data collected from the Portland light rail train fleet. The calibration procedure is mathematically formulated as a constrained non-linear optimization problem. The validity of the model is assessed by comparing instantaneous model predictions against field observations, and also evaluated in the domains of acceleration/deceleration versus speed and acceleration/deceleration versus distance. A test is conducted to investigate the adequacy of themore » model in simulation implementation. The results demonstrate that the proposed model can adequately capture instantaneous train dynamics, and provides good performance in the simulation test. Thus, the model provides a simple theoretical foundation for microscopic simulators and will significantly support the planning, management and control of railway transportation systems.« less

Biosynthesis of ZnO nanoparticles using rambutan (Nephelium lappaceumL.) peel extract and their photocatalytic activity on methyl orange dye

NASA Astrophysics Data System (ADS)

Karnan, Thenmozhi; Selvakumar, Stanly Arul Samuel

2016-12-01

In the present study, describes the synthesis of ZnO nanoparticles from rambutan (Nephelium lappaceumL.) peel extract via bio synthesis method and developed a new low cost technology to prepare ZnO nanoparticles. During the synthesis, fruit peel extract act as a natural ligation agent. The successfully prepared product was analyzed with some standard characterization studies like X-Ray Diffraction (XRD), UV-VIS Diffuse reflectance spectra (UV-Vis DRS), Field Emission Scanning Electron Microscope (FESEM), High resolution transmittance electron microscope (HR-TEM), N2 adsorption-desorption isotherm and UV-Vis absorption Spectroscopy. The photocatalytic activity of ZnO nanoparticles was evaluated by photodegradation of methyl orange (MO) dye under UV light and the result depicts around 83.99% decolorisation efficiency at 120 min of illumination. In addition with photodecolorisation, mineralization was also achieved. The mineralization has been confirmed by measuring Chemical Oxygen Demand (COD) values.

Longitudinal train dynamics model for a rail transit simulation system

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

Wang, Jinghui; Rakha, Hesham A.

The paper develops a longitudinal train dynamics model in support of microscopic railway transportation simulation. The model can be calibrated without any mechanical data making it ideal for implementation in transportation simulators. The calibration and validation work is based on data collected from the Portland light rail train fleet. The calibration procedure is mathematically formulated as a constrained non-linear optimization problem. The validity of the model is assessed by comparing instantaneous model predictions against field observations, and also evaluated in the domains of acceleration/deceleration versus speed and acceleration/deceleration versus distance. A test is conducted to investigate the adequacy of themore » model in simulation implementation. The results demonstrate that the proposed model can adequately capture instantaneous train dynamics, and provides good performance in the simulation test. Thus, the model provides a simple theoretical foundation for microscopic simulators and will significantly support the planning, management and control of railway transportation systems.« less

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

PubMed

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

2015-07-13

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

The microscopes of Antoni van Leeuwenhoek.

PubMed

van Zuylen, J

1981-03-01

The seventeenth-century Dutch microscopist, Antoni van Leeuwenhoek, was the first man to make a protracted study of microscopical objects, and, unlike his contemporary Robert Hooke, he viewed by transmitted light. Leeuwenhoek made over 500 of his own, curious, simple microscopes, but now only nine are known to exist. The exact nature of the lenses Leeuwenhoek made, has for long been a puzzle. The existing microscopes have now been examined in detail, and their optical characteristics measured and tabulated. It is proposed that the lens of highest magnification, x 266, was made using a special blown bubble technique.

Rheological and structural properties of sea cucumber Stichopus japonicus during heat treatment

NASA Astrophysics Data System (ADS)

Gao, Xin; Xue, Dongmei; Zhang, Zhaohui; Xu, Jiachao; Xue, Changhu

2005-07-01

Changes in tissue structure, rheological properties and water content of raw and heated sea cucumber meat were studied. Sea cucumber Stichopus japonicus was heated at 25°C , 70°C and 100°C water for 5 min. The structural changes were observed using a light microscope and the rheological parameters (rupture strength, adhesive strength and deformation) determined using a texture meter. Microscopic photograph revealed that the structural change of heated meat was greater than that of raw meat. The rupture strength, adhesive strength and deformation of raw meat were smaller than those of the heated meat. Meanwhile, rheological parameters showed positive correlation with heating temperature. These changes are mainly caused by thermal denaturation and gelatinization of collagen during heating. These changes were also evidenced in observations using a light microscope and differential scanning calorimetry.

Sheet-scanned dual-axis confocal microscopy using Richardson-Lucy deconvolution.

PubMed

Wang, D; Meza, D; Wang, Y; Gao, L; Liu, J T C

2014-09-15

We have previously developed a line-scanned dual-axis confocal (LS-DAC) microscope with subcellular resolution suitable for high-frame-rate diagnostic imaging at shallow depths. Due to the loss of confocality along one dimension, the contrast (signal-to-background ratio) of a LS-DAC microscope is deteriorated compared to a point-scanned DAC microscope. However, by using a sCMOS camera for detection, a short oblique light-sheet is imaged at each scanned position. Therefore, by scanning the light sheet in only one dimension, a thin 3D volume is imaged. Both sequential two-dimensional deconvolution and three-dimensional deconvolution are performed on the thin image volume to improve the resolution and contrast of one en face confocal image section at the center of the volume, a technique we call sheet-scanned dual-axis confocal (SS-DAC) microscopy.

Fabrication and characterization of novel microsphere-embedded optical devices for enhancing microscopy resolution

NASA Astrophysics Data System (ADS)

Darafsheh, Arash

2018-02-01

Microsphere-assisted imaging can be incorporated onto conventional light microscopes allowing wide-field and flourescence imaging with enhanced resolution. We demonstrated that imaging of specimens containing subdiffraction-limited features is achievable through high-index microspheres embedded in a transparent thin film placed over the specimen. We fabricated novel microsphere-embedded microscope slides composed of barium titanate glass microspheres (with diameter 10-100 μm and refractive index 1.9-2.2) embedded in a transparent polydimethylsiloxane (PDMS) elastomer layer with controllable thickness. We characterized the imaging performance of such microsphere-embedded devices in white-light microscopies, by measuring the imaging resolution, field-of-view, and magnification as a function of microsphere size. Our results inform on the design of novel optical devices, such as microsphere-embedded microscope slides for imaging applications.

Digital image processing of bone - Problems and potentials

NASA Technical Reports Server (NTRS)

Morey, E. R.; Wronski, T. J.

1980-01-01

The development of a digital image processing system for bone histomorphometry and fluorescent marker monitoring is discussed. The system in question is capable of making measurements of UV or light microscope features on a video screen with either video or computer-generated images, and comprises a microscope, low-light-level video camera, video digitizer and display terminal, color monitor, and PDP 11/34 computer. Capabilities demonstrated in the analysis of an undecalcified rat tibia include the measurement of perimeter and total bone area, and the generation of microscope images, false color images, digitized images and contoured images for further analysis. Software development will be based on an existing software library, specifically the mini-VICAR system developed at JPL. It is noted that the potentials of the system in terms of speed and reliability far exceed any problems associated with hardware and software development.

Cardiac morphology after conditions of microgravity during Cosmos 2044

NASA Technical Reports Server (NTRS)

Goldstein, Margaret A.; Edwards, Robert J.; Schroeter, John P.

1992-01-01

Light- and electron-microscopic studies were performed on cardiac muscle from rats flown on Cosmos 2044 and from four control groups. Average cross-sectional area of myofibers was measured by video analysis of the light-microscopic images of papillary and ventricular muscle samples from all animals. This cross-sectional area was significantly decreased in flight rats (P = 0.03) compared with synchronous controls. Additional findings at the electron microscopic level consistent with this atrophy were obtained by stereological analysis and optical diffraction analysis of papillary muscle samples. Slightly higher mitochondrial volume density values and mitochondria-to-myofibril ratios as well as normal A-band spacings (d1,0) and Z-band spacings of myofibrils were observed in the tail-suspension and flight groups. General morphological features similar to those in ventricular samples from the previous Cosmos 1887 flight were observed.

Imaging Schwarzschild multilayer X-ray microscope

NASA Technical Reports Server (NTRS)

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

1993-01-01

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

Ultrastructural effects of silicone oil on the clear crystalline lens of the human eye.

PubMed

Soliman, Wael; Sharaf, Mohamed; Abdelazeem, Khaled; El-Gamal, Dalia; Nafady, Allam

2018-03-01

To evaluate light and electron microscopic changes of the anterior capsule and its epithelium after clear lens extraction of vitrectomized myopic eyes with silicone oil tamponade. This prospective, controlled, non-randomized, interventional study included 20 anterior lens capsular specimens that were excised during combined clear lens extraction and silicone oil removal from previously vitrectomized highly myopic patients with silicone oil tamponade for previous retinal detachment surgeries. The specimens were examined via light microscopy and electron microscopy and compared with 20 anterior capsule specimens removed during clear lens extraction of non-vitrectomized highly myopic eyes. Light microscopic examination of clear lens anterior capsule specimens of vitrectomized myopic eyes filled with silicone oil showed relatively more flat cells with irregular outline of lens' epithelial cells with wide intercellular spaces, deeply stained nuclei, and multiple intracytoplasmic vacuoles. Scanning electron microscopy revealed collagenous surfaces filled with multiple pits, depressions, and abnormal deposits. Transmission electron microscopy revealed lens epithelial cells with apoptotic changes, many cytoplasmic vacuoles, and filopodia-like protrusions between lens epithelial cells and the capsule. Epithelial proliferation and multilayering were also observed. silicone oil may play a role in the development of apoptotic and histopathological changes in clear lens epithelial cells. Clarity of the lens at the time of silicone oil removal does not indicate an absence of cataractous changes. We found justification of combined clear lens extraction and silicone oil removal or combined phacovitrectomy when silicone oil injection is planned, but further long-term studies with larger patient groups are required.

Total variation based image deconvolution for extended depth-of-field microscopy images

NASA Astrophysics Data System (ADS)

Hausser, F.; Beckers, I.; Gierlak, M.; Kahraman, O.

2015-03-01

One approach for a detailed understanding of dynamical cellular processes during drug delivery is the use of functionalized biocompatible nanoparticles and fluorescent markers. An appropriate imaging system has to detect these moving particles so as whole cell volumes in real time with high lateral resolution in a range of a few 100 nm. In a previous study Extended depth-of-field microscopy (EDF-microscopy) has been applied to fluorescent beads and tradiscantia stamen hair cells and the concept of real-time imaging has been proved in different microscopic modes. In principle a phase retardation system like a programmable space light modulator or a static waveplate is incorporated in the light path and modulates the wavefront of light. Hence the focal ellipsoid is smeared out and images seem to be blurred in a first step. An image restoration by deconvolution using the known point-spread-function (PSF) of the optical system is necessary to achieve sharp microscopic images of an extended depth-of-field. This work is focused on the investigation and optimization of deconvolution algorithms to solve this restoration problem satisfactorily. This inverse problem is challenging due to presence of Poisson distributed noise and Gaussian noise, and since the PSF used for deconvolution exactly fits in just one plane within the object. We use non-linear Total Variation based image restoration techniques, where different types of noise can be treated properly. Various algorithms are evaluated for artificially generated 3D images as well as for fluorescence measurements of BPAE cells.

Effect of 3C-SiC intermediate layer in GaN—based light emitting diodes grown on Si(111) substrate

NASA Astrophysics Data System (ADS)

Zhu, Youhua; Wang, Meiyu; Li, Yi; Tan, Shuxin; Deng, Honghai; Guo, Xinglong; Yin, Haihong; Egawa, Takashi

2017-03-01

GaN-based light emitting diodes (LEDs) have been grown by metalorganic chemical vapor deposition on Si(111) substrate with and without 3C-SiC intermediate layer (IL). Structural property has been characterized by means of atomic force microscope, X-ray diffraction, and transmission electron microscope measurements. It has been revealed that a significant improvement in crystalline quality of GaN and superlattice epitaxial layers can be achieved by using 3C-SiC as IL. Regarding of electrical and optical characteristics, it is clearly observed that the LEDs with its IL have a smaller leakage current and higher light output power comparing with the LEDs without IL. The better performance of LEDs using 3C-SiC IL can be contributed to both of the improvements in epitaxial layers quality and light extraction efficiency. As a consequence, in terms of optical property, a double enhancement of the light output power and external quantum efficiency has been realized.

Arrays of microscopic organic LEDs for high-resolution optogenetics

PubMed Central

Steude, Anja; Witts, Emily C.; Miles, Gareth B.; Gather, Malte C.

2016-01-01

Optogenetics is a paradigm-changing new method to study and manipulate the behavior of cells with light. Following major advances of the used genetic constructs over the last decade, the light sources required for optogenetic control are now receiving increased attention. We report a novel optogenetic illumination platform based on high-density arrays of microscopic organic light-emitting diodes (OLEDs). Because of the small dimensions of each array element (6 × 9 μm2) and the use of ultrathin device encapsulation, these arrays enable illumination of cells with unprecedented spatiotemporal resolution. We show that adherent eukaryotic cells readily proliferate on these arrays, and we demonstrate specific light-induced control of the ionic current across the membrane of individual live cells expressing different optogenetic constructs. Our work paves the way for the use of OLEDs for cell-specific optogenetic control in cultured neuronal networks and for acute brain slices, or as implants in vivo. PMID:27386540

Malaria Diagnosis Using a Mobile Phone Polarized Microscope

NASA Astrophysics Data System (ADS)

Pirnstill, Casey W.; Coté, Gerard L.

2015-08-01

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

Malaria Diagnosis Using a Mobile Phone Polarized Microscope

PubMed Central

Pirnstill, Casey W.; Coté, Gerard L.

2015-01-01

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

Estimation of safe exposure time from an ophthalmic operating microscope with regard to ultraviolet radiation and blue-light hazards to the eye

NASA Astrophysics Data System (ADS)

Michael, Ralph; Wegener, Alfred

2004-08-01

Hazards from the optical radiation of an operating microscope that cause damage at the corneal, lenticular, and retinal levels were investigated; we considered, in particular, ultraviolet radiation (UVR) and blue light. The spectral irradiance from a Zeiss operation microscope OPMI VISU 200 was measured in the corneal plane between 300 and 1100 nm. Effective irradiance and radiance were calculated with relative spectral effectiveness data from the American Conference for Governmental and Industrial Hygienists. Safe exposure time to avoid UVR injury to the lens and cornea was found to be 2 h without a filter, 4 h with a UVR filter, 200 h with a yellow filter, and 400 h with a filter combination. Safe exposure time to avoid retinal photochemical injury was found to be 3 min without a filter and with a UVR filter, 10 min with a yellow filter, and 49 min with a filter combination. The effective radiance limit for retinal thermal injury was not exceeded. The hazard due to the UVR component from the operating microscope is not critical, and operation time can be safely prolonged with the use of appropriate filters. The retinal photochemical hazard appears critical without appropriate filters, permitting only some minutes of safe exposure time. The calculated safe exposure times are for worst-case conditions and maximal light output and include a safety factor.

Operating microscope light-induced phototoxic maculopathy after transscleral sutured posterior chamber intraocular lens implantation.

PubMed

Kweon, Eui Yong; Ahn, Min; Lee, Dong Wook; You, In Cheon; Kim, Min Jung; Cho, Nam Chun

2009-01-01

The purpose of this study is to report the features of operating microscope light-induced retinal phototoxic maculopathy after transscleral sutured posterior chamber intraocular lens (TSS PC-IOL) implantation. The charts of 118 patients who underwent TSS PC-IOL implantation surgery at Chonbuk National University Hospital (Jeonju, Korea) between March 1999 and February 2008 were retrospectively reviewed. Fourteen patients underwent combined 3-port pars plana vitrectomy and TSS PC-IOL implantation (vitrectomy group), and 104 patients underwent TSS PC-IOL implantation only (nonvitrectomy group). All surgeries were performed under the same coaxial illuminated microscope. All diagnoses were confirmed through careful fundus examination and fluorescein angiography (FA). Diagnoses of retinal phototoxic maculopathy were established in 10 (8.47%) of 118 TSS PC-IOL implantation cases. Phototoxic maculopathy occurred more frequently in the vitrectomy group than in the nonvitrectomy group (6/14 versus 4/104, respectively; P < 0.001, chi-square = 24.21). Affected patients reported decreased vision and were found to have coarse alterations of the retinal pigment epithelium (RPE). In 5 of the phototoxic maculopathy cases (50%), the visual acuity was 20/200 or worse. Operating microscope light-induced retinal phototoxic maculopathy can occur more frequently after TSS PC-IOL implantation than after casual cataract surgery, especially when TSS PC-IOL is combined with vitrectomy surgery. Surgeons should take precautions to prevent retinal phototoxicity after TSS PC-IOL implantation and vitrectomy.

Estimation of safe exposure time from an ophthalmic operating microscope with regard to ultraviolet radiation and blue-light hazards to the eye.

PubMed

Michael, Ralph; Wegener, Alfred

2004-08-01

Hazards from the optical radiation of an operating microscope that cause damage at the corneal, lenticular, and retinal levels were investigated; we considered, in particular, ultraviolet radiation (UVR) and blue light. The spectral irradiance from a Zeiss operation microscope OPMI VISU 200 was measured in the corneal plane between 300 and 1100 nm. Effective irradiance and radiance were calculated with relative spectral effectiveness data from the American Conference for Governmental and Industrial Hygienists. Safe exposure time to avoid UVR injury to the lens and cornea was found to be 2 h without a filter, 4 h with a UVR filter, 200 a yellow filter, and 400 h with a filter combination. Safe exposure time to avoid retinal photochemical injury was found to be 3 min without a filter and with a UVR filter, 10 min with a yellow filter, and 49 min with a filter combination. The effective radiance limit for retinal thermal injury was not exceeded. The hazard due to the UVR component from the operating microscope is not critical, and operation time can be safely prolonged with the use of appropriate filters. The retinal photochemical hazard appears critical without appropriate filters, permitting only some minutes of safe exposure time. The calculated safe exposure times are for worst-case conditions and maximal light output and include a safety factor.

Innovative Strategies for Clinical Microscopy Instruction: Virtual Versus Light Microscopy.

PubMed

McDaniel, M Jane; Russell, Gregory B; Crandall, Sonia J

2018-06-01

The purpose of the study was to compare virtual microscopy with light microscopy to determine differences in learning outcomes and learner attitudes in teaching clinical microscopy to physician assistant (PA) students. A prospective, randomized, crossover design study was conducted with a convenience sample of 67 first-year PA students randomized to 2 groups. One group used light microscopes to find microscopic structures, whereas the other group used instructor-directed video streaming of microscopic elements. At the midpoint of the study, the groups switched instructional strategies. Learning outcomes were assessed via posttest after each section of the study, with comparison of final practical examination results to previous cohorts. Attitudes about the 2 educational strategies were assessed through a postcourse questionnaire with a Likert scale. Analysis of the first posttest demonstrated that students in the video-streamed group had significantly better learning outcomes than those in the light microscopy group (P = .004; Cohen's d = 0.74). Analysis of the posttest after crossover showed no differences between the 2 groups (P = .48). Between the 2 posttests, students first assigned to the light microscopy group scored a 6.6 mean point increase (±10.4 SD; p = .0011), whereas students first assigned to the virtual microscopy group scored a 1.3 mean point increase (±7.1 SD; p = .29). The light microscopy group improved more than the virtual microscopy group (P = .019). Analysis of practical examination data revealed higher scores for the study group compared with 5 previous cohorts of first-year students (P < .0001; Cohen's d = 0.66). Students preferred virtual microscopy to traditional light microscopy. Virtual microscopy is an effective educational strategy, and students prefer this method when learning to interpret images of clinical specimens.

eduSPIM: Light Sheet Microscopy in the Museum

PubMed Central

Schmid, Benjamin; Weber, Michael; Huisken, Jan

2016-01-01

Light Sheet Microscopy in the Museum Light sheet microscopy (or selective plane illumination microscopy) is an important imaging technique in the life sciences. At the same time, this technique is also ideally suited for community outreach projects, because it produces visually appealing, highly dynamic images of living organisms and its working principle can be understood with basic optics knowledge. Still, the underlying concepts are widely unknown to the non-scientific public. On the occasion of the UNESCO International Year of Light, a technical museum in Dresden, Germany, launched a special, interactive exhibition. We built a fully functional, educational selective plane illumination microscope (eduSPIM) to demonstrate how developments in microscopy promote discoveries in biology. Design Principles of an Educational Light Sheet Microscope To maximize educational impact, we radically reduced a standard light sheet microscope to its essential components without compromising functionality and incorporated stringent safety concepts beyond those needed in the lab. Our eduSPIM system features one illumination and one detection path and a sealed sample chamber. We image fixed zebrafish embryos with fluorescent vasculature, because the structure is meaningful to laymen and visualises the optical principles of light sheet microscopy. Via a simplified interface, visitors acquire fluorescence and transmission data simultaneously. The eduSPIM Design Is Tailored Easily to Fit Numerous Applications The universal concepts presented here may also apply to other scientific approaches that are communicated to laymen in interactive settings. The specific eduSPIM design is adapted easily for various outreach and teaching activities. eduSPIM may even prove useful for labs needing a simple SPIM. A detailed parts list and schematics to rebuild eduSPIM are provided. PMID:27560188

Sub-diffraction limit resolution in microscopy

NASA Technical Reports Server (NTRS)

Cheng, Ming (Inventor); Chen, Weinong (Inventor)

2007-01-01

A method and apparatus for visualizing sub-micron size particles employs a polarizing microscope wherein a focused beam of polarized light is projected onto a target, and a portion of the illuminating light is blocked from reaching the specimen, whereby to produce a shadow region, and projecting diffracted light from the target onto the shadow region.

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

PubMed

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

2012-10-01

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

Diffuse light-sheet microscopy for stripe-free calcium imaging of neural populations.

PubMed

Taylor, Michael A; Vanwalleghem, Gilles C; Favre-Bulle, Itia A; Scott, Ethan K

2018-06-19

Light-sheet microscopy is used extensively in developmental biology and neuroscience. One limitation of this approach is that absorption and scattering produces shadows in the illuminating light sheet, resulting in stripe artifacts. Here, we introduce diffuse light-sheet microscopes that use a line diffuser to randomize the light propagation within the image plane, allowing the light sheets to reform after obstacles. We incorporate diffuse light sheets in two existing configurations: selective plane illumination microscopy (SPIM) in which the sample is illuminated with a static sheet of light, and digitally scanned light sheet (DSLS) in which a thin Gaussian beam is scanned across the image plane during each acquisition. We compare diffuse light-sheet microscopes to their conventional counterparts for calcium imaging of neural activity in larval zebrafish. We show that stripe artifacts can cast deep shadows that conceal some neurons, and that the stripes can flicker, producing spurious signals that could be interpreted as biological activity. Diffuse light sheets mitigate these problems, illuminating the blind spots produced by stripes and removing artifacts produced by the stripes' movements. The upgrade to diffuse light sheets is simple and inexpensive, especially in the case of DSLS, where it requires the addition of one optical element. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Photochemical eradication of methicillin-resistant Staphylococcus aureus by blue light activation of riboflavin.

PubMed

Makdoumi, Karim; Goodrich, Ray; Bäckman, Anders

2017-08-01

To compare elimination of methicillin-resistant Staphylococcus aureus (MRSA) by exposure of blue light alone and with riboflavin. A reference strain of MRSA was cultured and diluted in PBS with and without riboflavin (0.01%). Fifteen microlitre was added on a microscope slide, creating a fluid layer with a thickness of around 400 microns. Both of the bacterial suspensions were exposed to blue light, and the effect between exposure with and without riboflavin was compared. Evaluation involved two different wavelengths (412 and 450 nm) of blue light with a lower (5.4 J/cm 2 ) and higher dose (approximately 28.5 J/cm 2 ). The effect of 412 nm light was also evaluated for a thicker fluid layer (1.17 mm). After exposure, colony-forming units (CFUs) were determined for each solution. All measurements were repeated eight times. The reductions in bacteria were similar for both wavelengths. With riboflavin, a statistically significant elimination was observed for both 412 and 450 nm (p < 0.001). At both dosages, the mean reduction was more pronounced with the presence of riboflavin than without it. Using the higher dose, CFU reduction was 99% and 98%, respectively, for 412 and 450 nm light. The bactericidal efficacy was high also in the deeper fluid layer (93%, higher dose). Riboflavin enhanced the antibacterial effect on the exposed MRSA strain of blue light for both 412 and 450 nm blue light. This indicates that blue light could be considered for possible implementation in deep corneal infections. © 2017 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Note: Evaluation of slurry particle size analyzers for chemical mechanical planarization process

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

Jang, Sunjae; Kulkarni, Atul; Qin, Hongyi

In the chemical mechanical planarization (CMP) process, slurry particle size is important because large particles can cause defects. Hence, selection of an appropriate particle measuring system is necessary in the CMP process. In this study, a scanning mobility particle sizer (SMPS) and dynamic light scattering (DLS) were compared for particle size distribution (PSD) measurements. In addition, the actual particle size and shape were confirmed by transmission electron microscope (TEM) results. SMPS classifies the particle size according to the electrical mobility, and measures the particle concentration (single particle measurement). On the other hand, the DLS measures the particle size distribution bymore » analyzing scattered light from multiple particles (multiple particle measurement). For the slurry particles selected for evaluation, it is observed that SMPS shows bi-modal particle sizes 30 nm and 80 nm, which closely matches with the TEM measurements, whereas DLS shows only single mode distribution in the range of 90 nm to 100 nm and showing incapability of measuring small particles. Hence, SMPS can be a better choice for the evaluation of CMP slurry particle size and concentration measurements.« less

A Photomicrography Primer.

ERIC Educational Resources Information Center

Davidson, Michael W.

1991-01-01

Describes techniques and equipment which allows school microscopes to perform crossed-polarized light microscopy, reflected light microscopy, and photomicrography. Provides information on using chemicals from a high school stockroom to view crystals, viewing integrated circuits, and capturing images on film. Lists possible independent student…

Imaging System for Vaginal Surgery.

PubMed

Taylor, G Bernard; Myers, Erinn M

2015-12-01

The vaginal surgeon is challenged with performing complex procedures within a surgical field of limited light and exposure. The video telescopic operating microscope is an illumination and imaging system that provides visualization during open surgical procedures with a limited field of view. The imaging system is positioned within the surgical field and then secured to the operating room table with a maneuverable holding arm. A high-definition camera and Xenon light source allow transmission of the magnified image to a high-definition monitor in the operating room. The monitor screen is positioned above the patient for the surgeon and assistants to view real time throughout the operation. The video telescopic operating microscope system was used to provide surgical illumination and magnification during total vaginal hysterectomy and salpingectomy, midurethral sling, and release of vaginal scar procedures. All procedures were completed without complications. The video telescopic operating microscope provided illumination of the vaginal operative field and display of the magnified image onto high-definition monitors in the operating room for the surgeon and staff to simultaneously view the procedures. The video telescopic operating microscope provides high-definition display, magnification, and illumination during vaginal surgery.

Cell-phone-based platform for biomedical device development and education applications.

PubMed

Smith, Zachary J; Chu, Kaiqin; Espenson, Alyssa R; Rahimzadeh, Mehdi; Gryshuk, Amy; Molinaro, Marco; Dwyre, Denis M; Lane, Stephen; Matthews, Dennis; Wachsmann-Hogiu, Sebastian

2011-03-02

In this paper we report the development of two attachments to a commercial cell phone that transform the phone's integrated lens and image sensor into a 350x microscope and visible-light spectrometer. The microscope is capable of transmission and polarized microscopy modes and is shown to have 1.5 micron resolution and a usable field-of-view of 150 x 50 with no image processing, and approximately 350 x 350 when post-processing is applied. The spectrometer has a 300 nm bandwidth with a limiting spectral resolution of close to 5 nm. We show applications of the devices to medically relevant problems. In the case of the microscope, we image both stained and unstained blood-smears showing the ability to acquire images of similar quality to commercial microscope platforms, thus allowing diagnosis of clinical pathologies. With the spectrometer we demonstrate acquisition of a white-light transmission spectrum through diffuse tissue as well as the acquisition of a fluorescence spectrum. We also envision the devices to have immediate relevance in the educational field.

Cell-Phone-Based Platform for Biomedical Device Development and Education Applications

PubMed Central

Smith, Zachary J.; Chu, Kaiqin; Espenson, Alyssa R.; Rahimzadeh, Mehdi; Gryshuk, Amy; Molinaro, Marco; Dwyre, Denis M.; Lane, Stephen; Matthews, Dennis; Wachsmann-Hogiu, Sebastian

2011-01-01

In this paper we report the development of two attachments to a commercial cell phone that transform the phone's integrated lens and image sensor into a 350× microscope and visible-light spectrometer. The microscope is capable of transmission and polarized microscopy modes and is shown to have 1.5 micron resolution and a usable field-of-view of 150×150 with no image processing, and approximately 350×350 when post-processing is applied. The spectrometer has a 300 nm bandwidth with a limiting spectral resolution of close to 5 nm. We show applications of the devices to medically relevant problems. In the case of the microscope, we image both stained and unstained blood-smears showing the ability to acquire images of similar quality to commercial microscope platforms, thus allowing diagnosis of clinical pathologies. With the spectrometer we demonstrate acquisition of a white-light transmission spectrum through diffuse tissue as well as the acquisition of a fluorescence spectrum. We also envision the devices to have immediate relevance in the educational field. PMID:21399693

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

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

A relic of the Wellcome Tropical Research Laboratories in Khartoum (1903-34).

PubMed

Adeel, Ahmed Awad

2016-01-01

This article explores the origins of an old brass monocular microscope in the Central Laboratory in Khartoum, which used to be the Wellcome Tropical Research Laboratory in Khartoum (1903-1934). Examination of the microscope and review of published literature gave clues to the historical background of this microscope. Identical microscopes were first manufactured by R and J Beck in 1898, and continued to be advertised in 1899. The microscope was probably among the instruments provided by Wellcome for the initial establishment of the laboratories in 1902-1903. The article includes a brief review of the development of light microscopy. The need for preservation and proper restoration of old relics of the Wellcome laboratories in Khartoum is emphasized.

A relic of the Wellcome Tropical Research Laboratories in Khartoum (1903–34)

PubMed Central

2016-01-01

This article explores the origins of an old brass monocular microscope in the Central Laboratory in Khartoum, which used to be the Wellcome Tropical Research Laboratory in Khartoum (1903–1934). Examination of the microscope and review of published literature gave clues to the historical background of this microscope. Identical microscopes were first manufactured by R and J Beck in 1898, and continued to be advertised in 1899. The microscope was probably among the instruments provided by Wellcome for the initial establishment of the laboratories in 1902–1903. The article includes a brief review of the development of light microscopy. The need for preservation and proper restoration of old relics of the Wellcome laboratories in Khartoum is emphasized. PMID:27651557

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

PubMed

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

2007-05-15

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

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

NASA Astrophysics Data System (ADS)

Steidtner, Jens; Pettinger, Bruno

2007-10-01

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

Digital learning programs - competition for the classical microscope?

PubMed Central

Schmidt, Peter

2013-01-01

The development of digital media has been impressive in recent years which is also among the reason for their increasing use in academic teaching. This is especially true for teaching Anatomy and Histology in the first two years in medical and dental curricula. Modern digital technologies allow for efficient, affordable and easily accessible distribution of histological images in high quality. Microscopy depends almost exclusively on such images. Since 20 years numerous digital teaching systems have been developed for this purpose. Respective developments have changed the ways students acquire knowledge and prepare for exams. Teaching staff should adapt lectures, seminars and labs accordingly. As a first step, a collection of high resolution digital microscopic slides was made available for students at the Friedrich-Schiller-University in Jena. The aim of the present study was to evaluate the importance of conventional light microscopy and related technologies in current and future medical and dental education aswell. A survey was done among 172 medical and dental students at the Friedrich-Schiller-University Jena. 51% of students use now frequently new digital media for learning histology in contrast to 5% in the year 2000 [1]. Digital media including Internet, CD- based learning combined with social networks successfully compete with classical light microscopy. PMID:23467698

Effects of hydrogen peroxide on the light reflectance and morphology of bovine enamel.

PubMed

Kwon, Y H; Huo, M S; Kim, K H; Kim, S K; Kim, Y J

2002-05-01

The purpose of this study was to examine the effects of a bleaching agent (30% hydrogen peroxide) on the surface of bovine enamel using a scanning electron microscope and a UV-VIS-NIR spectrophotometer. Five non-carious bovine incisors were bleached for 0, 1, 2 and 3 days using 30% hydrogen peroxide. The light reflectance spectrum was measured using a spectrophotometer with diffuse reflectance mode. Colour values and colour differences in the teeth were evaluated from the reflectance measurements with the CIE L*a*b* colour coordinate system. Surface alterations in the bleached and unbleached teeth were studied using a scanning electron microscope. The change of reflectance in the teeth was related to the change of colour. Most reflectance change occurred within a 1-day bleaching, and this result was confirmed by a CIE L*a*b* colour coordinate system. The colour differences in the bleached teeth were significant enough to be perceived by the observer's eye. The comparison of bleached to unbleached bovine enamel revealed that the bleached surface showed non-uniform slight morphological alterations, and it developed varying degrees of surface porosity. This study indicates that the bleached bovine teeth showed apparent colour differences as well as slight morphological alterations after bleaching.

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

PubMed

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

2017-07-01

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

Assessment of incomplete clipping of aneurysms intraoperatively by a near-infrared indocyanine green-video angiography (Niicg-Va) integrated microscope.

PubMed

Imizu, S; Kato, Y; Sangli, A; Oguri, D; Sano, H

2008-08-01

The objective of this article was to assess the clinical use and the completeness of clipping with total occlusion of the aneurysmal lumen, real-time assessment of vascular patency in the parent, branching and perforating vessels, intraoperative assessment of blood flow, image quality, spatial resolution and clinical value in difficult aneurysms using near infrared indocyanine green video angiography integrated on to an operative Pentero neurosurgical microscope (Carl Zeiss, Oberkochen Germany). Thirteen patients with aneurysms were operated upon. An infrared camera with near infrared technology was adapted on to the OPMI Pentero microscope with a special filter and infrared excitation light to illuminate the operating field which was designed to allow passage of the near infrared light required for excitation of indocyanine green (ICG) which was used as the intravascular marker. The intravascular fluorescence was imaged with a video camera attached to the microscope. ICG fluorescence (700-850 nm) from a modified microscope light source on to the surgical field and passage of ICG fluorescence (780-950 nm) from the surgical field, back into the optical path of the microscope was used to detect the completeness of aneurysmal clipping Incomplete clipping in three patients (1 female and 2 males) with unruptured complicated aneurysms was detected using indocyanine green video angiography. There were no adverse effects after injection of indocyanine green. The completeness of clipping was inadequately detected by Doppler ultrasound miniprobe and rigid endoscopy and was thus complemented by indocyanine green video angiography. The operative microscope-integrated ICG video angiography as a new intraoperative method for detecting vascular flow, was found to be quick, reliable, cost-effective and possibly a substitute or adjunct for Doppler ultrasonography or intraoperative DSA, which is presently the gold standard. The simplicity of the method, the speed with which the investigation can be performed, the quality of the images, and the outcome of surgical procedures have all reduced the need for angiography. This technique may be useful during routine aneurysm surgery as an independent form of angiography and/or as an adjunct to intraoperative or postoperative DSA.

Polarized Light Microscopy in Reproductive and Developmental Biology

PubMed Central

KOIKE-TANI, MAKI; TANI, TOMOMI; MEHTA, SHALIN B.; VERMA, AMITABH; OLDENBOURG, RUDOLF

2016-01-01

SUMMARY The polarized light microscope reveals orientational order in native molecular structures inside living cells, tissues, and whole organisms. It is a powerful tool used to monitor and analyze the early developmental stages of organisms that lend themselves to microscopic observations. In this article, we briefly discuss the components specific to a traditional polarizing microscope and some historically important observations on: chromosome packing in the sperm head, the first zygote division of the sea urchin, and differentiation initiated by the first asymmetric cell division in the sand dollar. We then introduce the LC-PolScope and describe its use for measuring birefringence and polarized fluorescence in living cells and tissues. Applications range from the enucleation of mouse oocytes to analyzing the polarized fluorescence of the water strider acrosome. We end with new results on the birefringence of the developing chick brain, which we analyzed between developmental stages of days 12–20. PMID:23901032

Holographic photolysis of caged neurotransmitters

PubMed Central

Lutz, Christoph; Otis, Thomas S.; DeSars, Vincent; Charpak, Serge; DiGregorio, David A.; Emiliani, Valentina

2009-01-01

Stimulation of light-sensitive chemical probes has become a powerful tool for the study of dynamic signaling processes in living tissue. Classically, this approach has been constrained by limitations of lens–based and point-scanning illumination systems. Here we describe a novel microscope configuration that incorporates a nematic liquid crystal spatial light modulator (LC-SLM) to generate holographic patterns of illumination. This microscope can produce illumination spots of variable size and number and patterns shaped to precisely match user-defined elements in a specimen. Using holographic illumination to photolyse caged glutamate in brain slices, we demonstrate that shaped excitation on segments of neuronal dendrites and simultaneous, multi-spot excitation of different dendrites enables precise spatial and rapid temporal control of glutamate receptor activation. By allowing the excitation volume shape to be tailored precisely, the holographic microscope provides an extremely flexible method for activation of various photosensitive proteins and small molecules. PMID:19160517

Standoff detection of explosives: a challenging approach for optical technologies

NASA Astrophysics Data System (ADS)

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

2011-06-01

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

The evolution of structured illumination microscopy in studies of HIV.

PubMed

Marno, Kelly; Al'Zoubi, Lara; Pearson, Matthew; Posch, Markus; McKnight, Áine; Wheeler, Ann P

2015-10-15

The resolution limit of conventional light microscopy has proven to be limiting for many biological structures such as viruses including Human immunodeficiency virus (HIV). Individual HIV virions are impossible to study using confocal microscopy as they are well below the 200 nm resolution limit of conventional light microscopes. Structured illumination microscopy (SIM) allows a twofold enhancement in image resolution compared to standard widefield illumination and so provides an excellent tool for study of HIV. Viral capsids (CAs) vary between 110 and 146 nm so this study challenges the performance of SIM microscopes. SIM microscopy was first developed in 2000, commercialised in 2007 and rapidly developed. Here we present the changes in capabilities of the SIM microscopes for study of HIV localisation as the instrumentation for structured illumination microscopy has evolved over the past 8 years. Copyright © 2015. Published by Elsevier Inc.

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

NASA Astrophysics Data System (ADS)

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.

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

NASA Astrophysics Data System (ADS)

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

2007-02-01

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

Neuronal nitric oxide synthase immunopositive neurons in cat claustrum--a light and electron microscopic study.

PubMed

Hinova-Palova, Dimka; Edelstein, Lawrence; Paloff, Adrian; Hristov, Stanislav; Papantchev, Vassil; Ovtscharoff, Wladimir

2008-08-01

Nitric oxide is a unique neurotransmitter, which participates in many physiological and pathological processes in the organism. Nevertheless there are little data about the neuronal Nitric Oxide Synthase immunoreactive (nNOS-ir) neurons and fibers in the dorsal claustrum (DC) of a cat. In this respect the aims of this study were: (1) to demonstrate nNOS-ir in the neurons and fibers of the DC; (2) to describe their light microscopic morphology and distribution; (3) to investigate and analyze the ultrastructure of the nNOS-ir neurons, fibers and synaptic terminals; (4) to verify whether the nNOS-ir neurons consist a specific subpopulation of claustral neurons; (5) to verify whether the nNOS-ir neurons have a specific pattern of organization throughout the DC. For demonstration of the nNOS-ir the Avidin-Biotin-Peroxidase Complex method was applied. Immunopositive for nNOS neurons and fibers were present in all parts of DC. On the light microscope level nNOS-ir neurons were different in shape and size. According to the latter they were divided into three groups-small (with diameter under 15 microm), medium-sized (with diameter from 16 to 20 microm) and large (with diameter over 21 microm). Some of nNOS-ir neurons were lightly-stained while others were darkly-stained. On the electron microscope level the immunoproduct was observed in neurons, dendrites and terminal boutons. Different types of nNOS-ir neurons differ according to their ultrastructural features. Three types of nNOS-ir synaptic boutons were found. As a conclusion we hope that the present study will contribute to a better understanding of the functioning of the DC in cat and that some of the data presented could be extrapolated to other mammals, including human.

Strength and Deformability of Light-toned Layered Deposits Observed by MER Opportunity: Eagle to Erebus Craters

NASA Astrophysics Data System (ADS)

Okubo, C. H.; Schultz, R. A.; Nahm, A. L.

2007-07-01

The strength and deformability of light-toned layered deposits are estimated based on measurements of porosity from Microscopic Imager data acquired by MER Opportunity during its traverse from Eagle Crater to Erebus Crater.

3D real-time visualization of blood flow in cerebral aneurysms by light field particle image velocimetry

NASA Astrophysics Data System (ADS)

Carlsohn, Matthias F.; Kemmling, André; Petersen, Arne; Wietzke, Lennart

2016-04-01

Cerebral aneurysms require endovascular treatment to eliminate potentially lethal hemorrhagic rupture by hemostasis of blood flow within the aneurysm. Devices (e.g. coils and flow diverters) promote homeostasis, however, measurement of blood flow within an aneurysm or cerebral vessel before and after device placement on a microscopic level has not been possible so far. This would allow better individualized treatment planning and improve manufacture design of devices. For experimental analysis, direct measurement of real-time microscopic cerebrovascular flow in micro-structures may be an alternative to computed flow simulations. An application of microscopic aneurysm flow measurement on a regular basis to empirically assess a high number of different anatomic shapes and the corresponding effect of different devices would require a fast and reliable method at low cost with high throughout assessment. Transparent three dimensional 3D models of brain vessels and aneurysms may be used for microscopic flow measurements by particle image velocimetry (PIV), however, up to now the size of structures has set the limits for conventional 3D-imaging camera set-ups. On line flow assessment requires additional computational power to cope with the processing large amounts of data generated by sequences of multi-view stereo images, e.g. generated by a light field camera capturing the 3D information by plenoptic imaging of complex flow processes. Recently, a fast and low cost workflow for producing patient specific three dimensional models of cerebral arteries has been established by stereo-lithographic (SLA) 3D printing. These 3D arterial models are transparent an exhibit a replication precision within a submillimeter range required for accurate flow measurements under physiological conditions. We therefore test the feasibility of microscopic flow measurements by PIV analysis using a plenoptic camera system capturing light field image sequences. Averaging across a sequence of single double or triple shots of flashed images enables reconstruction of the real-time corpuscular flow through the vessel system before and after device placement. This approach could enable 3D-insight of microscopic flow within blood vessels and aneurysms at submillimeter resolution. We present an approach that allows real-time assessment of 3D particle flow by high-speed light field image analysis including a solution that addresses high computational load by image processing. The imaging set-up accomplishes fast and reliable PIV analysis in transparent 3D models of brain aneurysms at low cost. High throughput microscopic flow assessment of different shapes of brain aneurysms may therefore be possibly required for patient specific device designs.

The Light-Velocity Postulate: The Essential Difference between the Theories of Lorentz-Poincare and Einstein

ERIC Educational Resources Information Center

Abiko, Seiya

2005-01-01

Einstein, who had already developed the light-quantum theory, knew the inadequacy of Maxwell's theory in the microscopic sphere. Therefore, in writing his paper on special relativity, he had to set up the light-velocity postulate independently of the relativity postulate in order to make the electromagnetic foundation of physics compatible with…

Microscopic fluorescence spectral analysis of basal cell carcinomas

NASA Astrophysics Data System (ADS)

He, Qingli; Lui, Harvey; Zloty, David; Cowan, Bryce; Warshawski, Larry; McLean, David I.; Zeng, Haishan

2007-05-01

Background and Objectives. Laser-induced autofluorescence (LIAF) is a promising tool for cancer diagnosis. This method is based on the differences in autofluorescence spectra between normal and cancerous tissues, but the underlined mechanisms are not well understood. The objective of this research is to study the microscopic origins and intrinsic fluorescence properties of basal cell carcinoma (BCC) for better understanding of the mechanism of in vivo fluorescence detection and margin delineation of BCCs on skin patients. A home-made micro- spectrophotometer (MSP) system was used to image the fluorophore distribution and to measure the fluorescence spectra of various microscopic structures and regions on frozen tissue sections. Materials and Methods. BCC tissue samples were obtained from 14 patients undergoing surgical resections. After surgical removal, each tissue sample was immediately embedded in OCT medium and snap-frozen in liquid nitrogen. The frozen tissue block was then cut into 16-μm thickness sections using a cryostat microtome and placed on microscopic glass slides. The sections for fluorescence study were kept unstained and unfixed, and then analyzed by the MSP system. The adjacent tissue sections were H&E stained for histopathological examination and also served to help identify various microstructures on the adjacent unstained sections. The MSP system has all the functions of a conventional microscope, plus the ability of performing spectral analysis on selected micro-areas of a microscopic sample. For tissue fluorescence analysis, 442nm He-Cd laser light is used to illuminate and excite the unstained tissue sections. A 473-nm long pass filter was inserted behind the microscope objective to block the transmitted laser light while passing longer wavelength fluorescence signal. The fluorescence image of the sample can be viewed through the eyepieces and also recorded by a CCD camera. An optical fiber is mounted onto the image plane of the photograph port of the microscope to collect light from a specific micro area of the sample. The collected light is transmitted via the fiber to a disperserve type CCD spectrometer for spectral analysis. Results. The measurement results showed significant spectral differences between normal and cancerous tissues. For normal tissue regions, the spectral results agreed with our previous findings on autofluorescence of normal skin sections. For the cancerous regions, the epidermis showed very weak fluorescence signal, while the stratum corneum exhibited fluorescence emissions peaking at about 510 nm. In the dermis, the basal cell island and a band of surrounding areas showed very weak fluorescence signal, while distal dermis above and below the basal cell island showed greater fluorescence signal but with different spectral shapes. The very weak autofluorescence from the basal cell island and its surrounding area may be attributed to their degenerative properties that limited the production of collagens. Conclusions. The obtained microscopic results very well explain the in vivo fluorescence properties of BCC lesions in that they have decreased fluorescence intensity compared to the surrounding normal skin. The intrinsic spectra of various microstructures and the microscopic fluorescence images (corresponding fluorophore distribution in tissue) obtained in this study will be used for further theoretical modeling of in vivo fluorescence spectroscopy and imaging of skin cancers.

Identification of powdered Chinese herbal medicines by fluorescence microscopy, Part 1: Fluorescent characteristics of mechanical tissues, conducting tissues, and ergastic substances.

PubMed

Wang, Ya-Qiong; Liang, Zhi-Tao; Li, Qin; Yang, Hua; Chen, Hu-Biao; Zhao, Zhong-Zhen; Li, Ping

2011-03-01

The light microscope has been successfully used in identification of Chinese herbal medicines (CHMs) for more than a century. However, positive identification is not always possible. Given the popularity of fluorescence microscopy in bioanalysis, researchers dedicated to finding new ways to identify CHMs more effectively are now turning to fluorescence microscopy for authentication purposes. Some studies on distinguishing confused species from the same genus and on exploring distributions of chemicals in tissues of CHMs by fluorescence microscopy have been reported; however, no systematic investigations on fluorescent characteristics of powdered CHMs have been reported. Here, 46 samples of 16 CHMs were investigated. Specifically, the mechanical tissues including stone cells and fibers, the conducting tissues including three types of vessels, and ergastic substances including crystals of calcium oxalate and secretions, in various powdered CHMs were investigated by both light microscope and fluorescence microscope. The results showed many microscopic features emit fluorescence that makes them easily observed, even against complex backgrounds. Under the fluorescence microscope, different microscopic features from the same powdered CHM or some same features from different powdered CHMs emitted the different fluorescence, making this information very helpful for the authentication of CHMs in powder form. Moreover, secretions with unique chemical profiles from different powdered CHMs showed different fluorescent characteristics. Hence, fluorescence microscopy could be a useful additional method for the authentication of powdered CHMs if the fluorescent characteristics of specific CHMs are known. Copyright © 2010 Wiley-Liss, Inc.

Remote Sensing of Ocean Color

NASA Astrophysics Data System (ADS)

Dierssen, Heidi M.; Randolph, Kaylan

The oceans cover over 70% of the earth's surface and the life inhabiting the oceans play an important role in shaping the earth's climate. Phytoplankton, the microscopic organisms in the surface ocean, are responsible for half of the photosynthesis on the planet. These organisms at the base of the food web take up light and carbon dioxide and fix carbon into biological structures releasing oxygen. Estimating the amount of microscopic phytoplankton and their associated primary productivity over the vast expanses of the ocean is extremely challenging from ships. However, as phytoplankton take up light for photosynthesis, they change the color of the surface ocean from blue to green. Such shifts in ocean color can be measured from sensors placed high above the sea on satellites or aircraft and is called "ocean color remote sensing." In open ocean waters, the ocean color is predominantly driven by the phytoplankton concentration and ocean color remote sensing has been used to estimate the amount of chlorophyll a, the primary light-absorbing pigment in all phytoplankton. For the last few decades, satellite data have been used to estimate large-scale patterns of chlorophyll and to model primary productivity across the global ocean from daily to interannual timescales. Such global estimates of chlorophyll and primary productivity have been integrated into climate models and illustrate the important feedbacks between ocean life and global climate processes. In coastal and estuarine systems, ocean color is significantly influenced by other light-absorbing and light-scattering components besides phytoplankton. New approaches have been developed to evaluate the ocean color in relationship to colored dissolved organic matter, suspended sediments, and even to characterize the bathymetry and composition of the seafloor in optically shallow waters. Ocean color measurements are increasingly being used for environmental monitoring of harmful algal blooms, critical coastal habitats (e.g., seagrasses, kelps), eutrophication processes, oil spills, and a variety of hazards in the coastal zone.

Resolving the depth of fluorescent light by structured illumination and shearing interferometry

NASA Astrophysics Data System (ADS)

Schindler, Johannes; Elmaklizi, Ahmed; Voit, Florian; Hohmann, Ansgar; Schau, Philipp; Brodhag, Nicole; Krauter, Philipp; Frenner, Karsten; Kienle, Alwin; Osten, Wolfgang

2016-03-01

A method for the depth-sensitive detection of fluorescent light is presented. It relies on a structured illumination restricting the excitation volume and on an interferometric detection of the wave front curvature. The illumination with two intersecting beams of a white-light laser separated in a Sagnac interferometer coupled to the microscope provides a coarse confinement in lateral and axial direction. The depth reconstruction is carried out by evaluating shearing interferograms produced with a Michelson interferometer. This setup can also be used with spatially and temporally incoherent light as emitted by fluorophores. A simulation workflow of the method was developed using a combination of a solution of Maxwell's equations with the Monte Carlo method. These simulations showed the principal feasibility of the method. The method is validated by measurements at reference samples with characterized material properties, locations and sizes of fluorescent regions. It is demonstrated that sufficient signal quality can be obtained for materials with scattering properties comparable to dental enamel while maintaining moderate illumination powers in the milliwatt range. The depth reconstruction is demonstrated for a range of distances and penetration depths of several hundred micrometers.

Miniature objective lens for array digital pathology: design improvement based on clinical evaluation

NASA Astrophysics Data System (ADS)

McCall, Brian; Pierce, Mark; Graviss, Edward A.; Richards-Kortum, Rebecca R.; Tkaczyk, Tomasz S.

2016-03-01

A miniature objective designed for digital detection of Mycobacterium tuberculosis (MTB) was evaluated for diagnostic accuracy. The objective was designed for array microscopy, but fabricated and evaluated at this stage of development as a single objective. The counts and diagnoses of patient samples were directly compared for digital detection and standard microscopy. The results were found to be correlated and highly concordant. The evaluation of this lens by direct comparison to standard fluorescence sputum smear microscopy presented unique challenges and led to some new insights in the role played by the system parameters of the microscope. The design parameters and how they were developed are reviewed in light of these results. New system parameters are proposed with the goal of easing the challenges of evaluating the miniature objective and maintaining the optical performance that produced the agreeable results presented without over-optimizing. A new design is presented that meets and exceeds these criteria.

Spectral analysis of scattered light from flowers' petals

NASA Astrophysics Data System (ADS)

Ozawa, Atsumi; Uehara, Tomomi; Sekiguchi, Fumihiko; Imai, Hajime

2009-07-01

A new method was developed for studying absorption characteristics of opaque samples based on the light scattering spectroscopy. Measurements were made in white, red and violet petals of Petunia hybrida, and gave the absorption spectra in a non-destructive manner without damaging the cell structures of the petal. The red petal has absorption peak at 550 nm and the violet has three absorption peaks: at 450, 670, and 550 nm. The results were discussed in correlation with the microscopic cell structures of the petal observed with optical microscope and transmission electron microscopy (TEM). Only the cells placed in the surface have the pigments giving the color of the petal.

Characterisation of a resolution enhancing image inversion interferometer.

PubMed

Wicker, Kai; Sindbert, Simon; Heintzmann, Rainer

2009-08-31

Image inversion interferometers have the potential to significantly enhance the lateral resolution and light efficiency of scanning fluorescence microscopes. Self-interference of a point source's coherent point spread function with its inverted copy leads to a reduction in the integrated signal for off-axis sources compared to sources on the inversion axis. This can be used to enhance the resolution in a confocal laser scanning microscope. We present a simple image inversion interferometer relying solely on reflections off planar surfaces. Measurements of the detection point spread function for several types of light sources confirm the predicted performance and suggest its usability for scanning confocal fluorescence microscopy.

Detection of fungal hyphae using smartphone and pocket magnifier: going cellular.

PubMed

Agarwal, Tushar; Bandivadekar, Pooja; Satpathy, Gita; Sharma, Namrata; Titiyal, Jeewan S

2015-03-01

The aim of this study was to detect fungal hyphae in a corneal scraping sample using a cost-effective assembly of smartphone and pocket magnifier. In this case report, a tissue sample was obtained by conventional corneal scraping from a clinically suspicious case of mycotic keratitis. The smear was stained with Gram stain, and a 10% potassium hydroxide mount was prepared. It was imaged using a smartphone coupled with a compact pocket magnifier and integrated light-emitting diode assembly at point-of-care. Photographs of multiple sections of slides were viewed using smartphone screen and pinch-to-zoom function. The same slides were subsequently screened under a light microscope by an experienced microbiologist. The scraping from the ulcer was also inoculated on blood agar and Sabouraud dextrose agar. Smartphone-based digital imaging revealed the presence of gram-positive organism with hyphae. Examination under a light microscope also yielded similar findings. Fusarium was cultured from the corneal scraping, confirming the diagnosis of mycotic keratitis. The patient responded to topical 5% natamycin therapy, with resolution of the ulcer after 4 weeks. Smartphones can be successfully used as novel point-of-care, cost-effective, reliable microscopic screening tools.

Photonic Microhand with Autonomous Action.

PubMed

Martella, Daniele; Nocentini, Sara; Nuzhdin, Dmitry; Parmeggiani, Camilla; Wiersma, Diederik S

2017-11-01

Grabbing and holding objects at the microscale is a complex function, even for microscopic living animals. Inspired by the hominid-type hand, a microscopic equivalent able to catch microelements is engineered. This microhand is light sensitive and can be either remotely controlled by optical illumination or can act autonomously and grab small particles on the basis of their optical properties. Since the energy is delivered optically, without the need for wires or batteries, the artificial hand can be shrunk down to the micrometer scale. Soft material is used, in particular, a custom-made liquid-crystal network that is patterned by a photolithographic technique. The elastic reshaping properties of this material allow finger movement, using environmental light as the only energy source. The hand can be either controlled externally (via the light field), or else the conditions in which it autonomously grabs a particle in its vicinity can be created. This microrobot has the unique feature that it can distinguish between particles of different colors and gray levels. The realization of this autonomous hand constitutes a crucial element in the development of microscopic creatures that can perform tasks without human intervention and self-organized automation at the micrometer scale. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Aligning Arrays of Lenses and Single-Mode Optical Fibers

NASA Technical Reports Server (NTRS)

Liu, Duncan

2004-01-01

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

Hyperspectral microscopic analysis of normal, benign and carcinoma microarray tissue sections

NASA Astrophysics Data System (ADS)

Maggioni, Mauro; Davis, Gustave L.; Warner, Frederick J.; Geshwind, Frank B.; Coppi, Andreas C.; DeVerse, Richard A.; Coifman, Ronald R.

2006-02-01

We apply a unique micro-optoelectromechanical tuned light source and new algorithms to the hyper-spectral microscopic analysis of human colon biopsies. The tuned light prototype (Plain Sight Systems Inc.) transmits any combination of light frequencies, range 440nm 700nm, trans-illuminating H and E stained tissue sections of normal (N), benign adenoma (B) and malignant carcinoma (M) colon biopsies, through a Nikon Biophot microscope. Hyper-spectral photomicrographs, randomly collected 400X magnication, are obtained with a CCD camera (Sensovation) from 59 different patient biopsies (20 N, 19 B, 20 M) mounted as a microarray on a single glass slide. The spectra of each pixel are normalized and analyzed to discriminate among tissue features: gland nuclei, gland cytoplasm and lamina propria/lumens. Spectral features permit the automatic extraction of 3298 nuclei with classification as N, B or M. When nuclei are extracted from each of the 59 biopsies the average classification among N, B and M nuclei is 97.1%; classification of the biopsies, based on the average nuclei classification, is 100%. However, when the nuclei are extracted from a subset of biopsies, and the prediction is made on nuclei in the remaining biopsies, there is a marked decrement in performance to 60% across the 3 classes. Similarly the biopsy classification drops to 54%. In spite of these classification differences, which we believe are due to instrument and biopsy normalization issues, hyper-spectral analysis has the potential to achieve diagnostic efficiency needed for objective microscopic diagnosis.

[Development of non-invasive clinical examination methods for the anterior segment of the eye and their clinical significance].

PubMed

Sawa, Mitsuru

2011-03-01

1. Slit-lamp microscopy is a principal ophthalmic clinical method, because it provides microscopic findings of the anterior segment of the eye noninvasively. Its findings, however, are qualitative and there are large inter-observer variations in their evaluation. Furthermore, slit-lamp microscopy provides morphological findings, but a functional evaluation is difficult. We developed two novel methods that establish a qualitative methodology of the slit-lamp microscope and the pathophysiology of the anterior segment of the eye. One is the flare-cell photometer to evaluate flare and cells in the aqueous humor of the eye and the other is an immunohistochemical examination method using tear fluid to evaluate ocular surface disorders. The comprehensive evaluation of these studies is herein overviewed. 2. INNOVATION OF THE FLARE-CELL PHOTOMETER AND ITS CLINICAL SIGNIFICANCE: The breakdown of the blood-aqueous barrier (BAB) causes an increase in protein (flare) and leakage of blood cells (cell) into the aqueous humor of the eye and the severity of BAB breakdown has a positive correlation with the intensity of flare and cells. The flare and cells in the aqueous can be observed qualitatively by slit-lamp microscopy. These findings are primarily distinguished in optics by light scattering. Therefore, detection of the intensity of light scattering due to flare and cells can evaluate the BAB function. The flare-cell photometer comprises 3 novel components: a laser beam system as an incident light, a photomultiplier to detect scattered light intensity and a computer-assisted system to operate the whole system and analyze detected scattered light signals due to flare and cells. The instrument enables us to quantitatively analyze the flare and cells non-invasively and accurately with a wide dynamic measurement range, resulting in a repeated examination of each individual case. It also enables the evaluation of inflammation in the aqueous not only postoperatively but also in endogenous uveitis, evaluation of the effects of anti-inflammatory drugs on BAB and evaluation of aqueous humor dynamics. Furthermore, repeating the examination can minimize inter-individual variations and reduce the number of animals in animal experiments. 3. Sampling of tears can be performed noninvasively, but the obtainable volume is limited. Therefore, a determination of targeting biomarkers and a development of their micro-volume analysis methods play a crucial role in pathophysiological studies of the ocular surface. Targeting biomarkers should be determined according to the various specified bioactive substances such as eosinophil cationic protein (ECP), cytokines and others. A number of microvolume analysis methods, such as chemiluminescent enzyme immunoassay, immunochromatography, micro-array system and polymerase chain reaction method are used. Objective disorders in the studies include allergic conjunctivitis and infectious diseases such as herpetic keratitis. Quantitative evaluation methods for ECP concentration, antigen-specific secretory IgA in allergic diseases and herpetic keratitis, herpes simplex virus-DNA and cytokine and chemokine profile in tear fluid sampled by filter paper method were investigated. We developed a clinically applicable quantitative immunochemical method for ECP concentration in tear fluid. The results revealed that tear fluid analysis using the above mentioned methods is a clinically useful to investigate the pathophysiology of the ocular surface. 4. Laser flare-cell photometer and tear fluid analysis are potent clinical quantitative methods to investigate the pathophysiology of the anterior segment of the eye.

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

PubMed Central

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

2007-01-01

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

Detection of microstructural defects in chalcopyrite Cu(In,Ga)Se2 solar cells by spectrally-filtered electroluminescence

NASA Astrophysics Data System (ADS)

Skvarenina, L.; Gajdos, A.; Macku, R.; Skarvada, P.

2017-12-01

The aim of this research is to detect and localize microstructural defects by using an electrically excited light emission from a forward/reverse-bias stressed pn-junction in thin-film Cu(In; Ga)Se2 solar cells with metal wrap through architecture. A different origin of the local light emission from intrinsic/extrinsic imperfections in these chalcopyrite-based solar cells can be distinguished by a spectrally-filtered electroluminescence mapping. After a light emission mapping and localization of the defects in a macro scale is performed a micro scale exploration of the solar cell surface by a scanning electron microscope which follows the particular defects obtained by an electroluminescence. In particular, these macroscopic/microscopic examinations are performed independently, then the searching of the corresponding defects in the micro scale is rather difficult due to a diffused light emission obtained from the macro scale localization. Some of the defects accompanied by a highly intense light emission very often lead to a strong local overheating. Therefore, the lock-in infrared thermography is also performed along with an electroluminescence mapping.

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

PubMed Central

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

2014-01-01

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

Control and acquisition systems for new scanning transmission x-ray microscopes at Advanced Light Source (abstract)

NASA Astrophysics Data System (ADS)

Tyliszczak, T.; Hitchcock, P.; Kilcoyne, A. L. D.; Ade, H.; Hitchcock, A. P.; Fakra, S.; Steele, W. F.; Warwick, T.

2002-03-01

Two new scanning x-ray transmission microscopes are being built at beamline 5.3.2 and beamline 7.0 of the Advanced Light Source that have novel aspects in their control and acquisition systems. Both microscopes use multiaxis laser interferometry to improve the precision of pixel location during imaging and energy scans as well as to remove image distortions. Beam line 5.3.2 is a new beam line where the new microscope will be dedicated to studies of polymers in the 250-600 eV energy range. Since this is a bending magnet beam line with lower x-ray brightness than undulator beam lines, special attention is given to the design not only to minimize distortions and vibrations but also to optimize the controls and acquisition to improve data collection efficiency. 5.3.2 microscope control and acquisition is based on a PC computer running WINDOWS 2000. All mechanical stages are moved by stepper motors with rack mounted controllers. A dedicated counter board is used for counting and timing and a multi-input/output board is used for analog acquisition and control of the focusing mirror. A three axis differential laser interferometer is being used to improve stability and precision by careful tracking of the relative positions of the sample and zone plate. Each axis measures the relative distance between a mirror placed on the sample stage and a mirror attached to the zone plate holder. Agilent Technologies HP 10889A servo-axis interferometer boards are used. While they were designed to control servo motors, our tests show that they can be used to directly control the piezo stage. The use of the interferometer servo-axis boards provides excellent point stability for spectral measurements. The interferometric feedback also provides active vibration isolation which reduces deleterious impact of mechanical vibrations up to 20-30 Hz. It also can improve the speed and precision of image scans. Custom C++ software has been written to provide user friendly control of the microscope and integration with visual light microscopy indexing of the samples. The beam line 7.0 microscope upgrade is a new design which will replace the existing microscope. The design is similar to that of beam line 5.3.2, including interferometric position encoding. However the acquisition and control is based on VXI systems, a Sun computer, and LABVIEW™ software. The main objective of the BL 7.0 microscope upgrade is to achieve precise image scans at very high speed (pixel dwells as short as 10 μs) to take full advantage of the high brightness of the 7.0 undulator beamline. Results of tests and a discussion of the benefits of our scanning microscope designs will be presented.

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

NASA Astrophysics Data System (ADS)

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

2011-09-01

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

78 FR 64916 - Application(s) for Duty-Free Entry of Scientific Instruments

Federal Register 2010, 2011, 2012, 2013, 2014

2013-10-30

...., light to heat), crystallization, melting, phase transformations, fracture, and other dynamic events. The... Sciences University, 1120 15th Street, Augusta, GA 30912. Instrument: Imaging System/Digital Microscope... the instrument include fast wavelength change, a dichromotome system, and two different light sources...

Integrated light and scanning electron microscopy of GFP-expressing cells.

PubMed

Peddie, Christopher J; Liv, Nalan; Hoogenboom, Jacob P; Collinson, Lucy M

2014-01-01

Integration of light and electron microscopes provides imaging tools in which fluorescent proteins can be localized to cellular structures with a high level of precision. However, until recently, there were few methods that could deliver specimens with sufficient fluorescent signal and electron contrast for dual imaging without intermediate staining steps. Here, we report protocols that preserve green fluorescent protein (GFP) in whole cells and in ultrathin sections of resin-embedded cells, with membrane contrast for integrated imaging. Critically, GFP is maintained in a stable and active state within the vacuum of an integrated light and scanning electron microscope. For light microscopists, additional structural information gives context to fluorescent protein expression in whole cells, illustrated here by analysis of filopodia and focal adhesions in Madin Darby canine kidney cells expressing GFP-Paxillin. For electron microscopists, GFP highlights the proteins of interest within the architectural space of the cell, illustrated here by localization of the conical lipid diacylglycerol to cellular membranes. © 2014 Elsevier Inc. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

2018-05-01

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

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

PubMed

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

2011-01-01

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

Imaging, microscopic analysis, and modeling of a CdTe module degraded by heat and light

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

Johnston, Steve; Albin, David; Hacke, Peter

Photoluminescence (PL), electroluminescence (EL), and dark lock-in thermography are collected during stressing of a CdTe module under one-Sun light at an elevated temperature of 100 degrees C. The PL imaging system is simple and economical. The PL images show differing degrees of degradation across the module and are less sensitive to effects of shunting and resistance that appear on the EL images. Regions of varying degradation are chosen based on avoiding pre-existing shunt defects. These regions are evaluated using time-of-flight secondary ion-mass spectrometry and Kelvin probe force microscopy. Reduced PL intensity correlates to increased Cu concentration at the front interface.more » Numerical modeling and measurements agree that the increased Cu concentration at the junction also correlates to a reduced space charge region.« less

Imaging, microscopic analysis, and modeling of a CdTe module degraded by heat and light

DOE PAGES

Johnston, Steve; Albin, David; Hacke, Peter; ...

2018-01-12

Photoluminescence (PL), electroluminescence (EL), and dark lock-in thermography are collected during stressing of a CdTe module under one-Sun light at an elevated temperature of 100 degrees C. The PL imaging system is simple and economical. The PL images show differing degrees of degradation across the module and are less sensitive to effects of shunting and resistance that appear on the EL images. Regions of varying degradation are chosen based on avoiding pre-existing shunt defects. These regions are evaluated using time-of-flight secondary ion-mass spectrometry and Kelvin probe force microscopy. Reduced PL intensity correlates to increased Cu concentration at the front interface.more » Numerical modeling and measurements agree that the increased Cu concentration at the junction also correlates to a reduced space charge region.« less

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

PubMed

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. Cluster-analyses of data from Raman microscopic imaging reconstructed histo-anatomical features in comparison to the light microscopic image and finally, by application of principal component analyses (PCA), it was possible to see a clear distinction between forensic and archaeological bone samples. Hence, the spectral characterization of inorganic and organic compounds by the afore mentioned techniques, followed by analyses such as multivariate imaging analysis (MIAs) and principal component analyses (PCA), appear to be suitable for the post mortem interval (PMI) estimation of human skeletal remains.

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

PubMed Central

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. Cluster-analyses of data from Raman microscopic imaging reconstructed histo-anatomical features in comparison to the light microscopic image and finally, by application of principal component analyses (PCA), it was possible to see a clear distinction between forensic and archaeological bone samples. Hence, the spectral characterization of inorganic and organic compounds by the afore mentioned techniques, followed by analyses such as multivariate imaging analysis (MIAs) and principal component analyses (PCA), appear to be suitable for the post mortem interval (PMI) estimation of human skeletal remains. PMID:28334006

Evaluating Red Reflex and Surgeon Preference Between Nearly-Collimated and Focused Beam Microscope Illumination Systems.

PubMed

Cionni, Robert J; Pei, Ron; Dimalanta, Ramon; Lubeck, David

2015-08-01

To evaluate the intensity and stability of the red reflex produced by ophthalmic surgical microscopes with nearly-collimated versus focused illumination systems and to assess surgeon preference in a simulated surgical setting. This two-part evaluation consisted of postproduction surgical video analysis of red reflex intensity and a microscope use and preference survey completed by 13 experienced cataract surgeons. Survey responses were based on bench testing and experience in a simulated surgical setting. A microscope with nearly-collimated beam illumination and two focused beam microscopes were assessed. Red reflex intensity and stability were greater with the nearly-collimated microscope illumination system. In the bench testing survey, surgeons reported that the red reflex was maintained over significantly greater distances away from pupillary center, and depth of focus was numerically greater with nearly-collimated illumination relative to focused illumination. Most participating surgeons (≥64%) reported a preference for the microscope with nearly-collimated illumination with regard to red reflex stability, depth of focus, visualization, surgical working distance, and perceived patient comfort. The microscope with nearly-collimated illumination produced a more intense and significantly more stable red reflex and was preferred overall by more surgeons. This is the first report of an attempt to quantify red reflex intensity and stability and to evaluate surgically-relevant parameters between microscope systems. The data and methods presented here may provide a basis for future studies attempting to quantify differences between surgical microscopes that may affect surgeon preference and microscope use in ophthalmic surgery.

Optical improvements in the diagnosis of bladder cancer: implications for clinical practice.

PubMed

Schubert, Tina; Rausch, Steffen; Fahmy, Omar; Gakis, Georgios; Stenzl, Arnulf

2017-11-01

For over 100 years white-light cystoscopy has remained the gold-standard technique for the detection of bladder cancer (BCa). Some limitations in the detection of flat lesions (CIS), the differentiation between inflammation and malignancy, the inaccurate determination of the tumor margin status as well as the tumor depth, have led to a variety of technological improvements. The aim of this review is to evaluate the impact of these improvements in the diagnosis of BCa and their effectiveness in clinical practice. A systematic literature search was conducted according to the PRISMA statement to identify studies reporting on imaging modalities in the diagnosis of NMIBC between 2000 and 2017. A two-stage selection process was utilized to determine eligible studies. A total of 74 studies were considered for final analysis. Optical imaging technologies have emerged as an adjunct to white-light cystoscopy and can be classified according to their scope as macroscopic, microscopic and molecular. Macroscopic techniques including photodynamic diagnosis (PDD), narrow-band imaging (NBI) and the Storz Professional Image Enhancement System (IMAGE1 S, formerly known as SPIES) are similar to white-light cystoscopy, but are superior in the detection of bladder tumors by means of contrast enhancement. Especially the detection rate of very mute lesions in the bladder mucosa (CIS) could be significantly increased by the use of these methods. Microscopic imaging techniques like confocal laser endomicroscopy and optical coherence tomography permit a real-time high-resolution assessment of the bladder mucosa at a cellular and sub-cellular level with spatial resolutions similar to histology, enabling the surgeon to perform an 'optical biopsy'. Molecular techniques are based on the combination of optical imaging technologies with fluorescence labeling of cancer-specific molecular agents like antibodies. This labeling is intended to favor an optical distinction between benign and malignant tissue. Optical improvements of the standard white-light cystoscopy have proven their benefit in the detection of BCa and have found their way into clinical practice. Especially the combination of macroscopic and microscopic techniques may improve diagnostic accuracy. Nevertheless, HAL-PDD guided cystoscopy is the only approach approved for routine use in the diagnosis of BCa by most urological associations in the EU and USA to date.

Macro-microscopic anatomy: obtaining a composite view of barrier zone formation in Acer saccharum

Treesearch

Kenneth Dudzik

1988-01-01

The technique for constructing a montage of large wood sections cut on a sliding microtome is discussed. Briefly, the technique involves photographing many serial micrographs in a pattern under a light microscope similar to the way flight lines are run in aerial photography. Assembly of the resulting overlapping photographs requires careful trimming. A composite of...

Early quantitative method for measuring germination in non-green spores of Dryopteris paleacea using an epifluorescence-microscope technique

NASA Technical Reports Server (NTRS)

Scheuerlein, R.; Wayne, R.; Roux, S. J.

1988-01-01

A method is described to determine germination by blue-light excited red fluorescence in the positively photoblastic spores of Dryopteris paleacea Sw. This fluorescence is due to chlorophyll as evidenced from 1) a fluorescence-emission spectrum in vivo, where a bright fluorescence around 675 nm is obtained only in red light (R)-irradiated spores and 2) in vitro measurements with acetone extracts prepared from homogenized spores. Significant amounts of chlorophyll can be found only in R-treated spores; this chlorophyll exhibits an emission band around 668 nm, when irradiated with 430 nm light at 21 degrees C. Compared to other criteria for germination, such as swelling of the cell, coat splitting, greening, and rhizoid formation, which require longer periods after induction for their expression, chlorophyll fluorescence can be used to quantify germination after two days. This result is confirmed by fluence-response curves for R-induced spore germination; the same relationship between applied R and germination is obtained by the evaluation with the epifluorescence method 2 days after the light treatment as compared with the evaluation with bright-field microscopy 5 days after the inducing R. Using this technique we show for the first time that Ca2+ contributes to the signal-transduction chain in phytochrome-mediated chlorophyll synthesis in spores of Dryopteris paleacea.

Sensing of Streptococcus mutans by microscopic imaging ellipsometry

NASA Astrophysics Data System (ADS)

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

2017-05-01

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

Telepathology. Long-distance diagnosis.

PubMed

Weinstein, R S; Bloom, K J; Rozek, L S

1989-04-01

Telepathology is defined as the practice of pathology at a distance, by visualizing an image on a video monitor rather than viewing a specimen directly through a microscope. Components of a telepathology system include the following: (1) a workstation equipped with a high-resolution video camera attached to a remote-controlled light microscope; (2) a pathologist workstation incorporating controls for manipulating the robotic microscope as well as a high-resolution video monitor; and (3) a telecommunications link. Progress has been made in designing and constructing telepathology workstations and fully motorized, computer-controlled light microscopes suitable for telepathology. In addition, components such as video signal digital encoders and decoders that produce remarkably stable, high-color fidelity, and high-resolution images have been incorporated into the workstations. Resolution requirements for the video microscopy component of telepathology have been formally examined in receiver operator characteristic (ROC) curve analyses. Test-of-concept demonstrations have been completed with the use of geostationary satellites as the broadband communication linkages for 750-line resolution video. Potential benefits of telepathology include providing a means of conveniently delivering pathology services in real-time to remote sites or underserviced areas, time-sharing of pathologists' services by multiple institutions, and increasing accessibility to specialty pathologists.

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

PubMed Central

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 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. PMID:26819828

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

PubMed

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.

Sub-micrometer resolution proximity X-ray microscope with digital image registration

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

Chkhalo, N. I.; Salashchenko, N. N.; Sherbakov, A. V., E-mail: SherbakovAV@ipm.sci-nnov.ru

A compact laboratory proximity soft X-ray microscope providing submicrometer spatial resolution and digital image registration is described. The microscope consists of a laser-plasma soft X-ray radiation source, a Schwarzschild objective to illuminate the test sample, and a two-coordinate detector for image registration. Radiation, which passes through the sample under study, generates an absorption image on the front surface of the detector. Optical ceramic YAG:Ce was used to convert the X-rays into visible light. An image was transferred from the scintillator to a charge-coupled device camera with a Mitutoyo Plan Apo series lens. The detector’s design allows the use of lensesmore » with numerical apertures of NA = 0.14, 0.28, and 0.55 without changing the dimensions and arrangement of the elements of the device. This design allows one to change the magnification, spatial resolution, and field of view of the X-ray microscope. A spatial resolution better than 0.7 μm and an energy conversion efficiency of the X-ray radiation with a wavelength of 13.5 nm into visible light collected by the detector of 7.2% were achieved with the largest aperture lens.« less

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

PubMed

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

1994-01-01

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

Combined reflection and transmission microscope for telemedicine applications in field settings.

PubMed

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 (<4.8 ounces) and utilizes a simple light emitting diode (LED) to illuminate the sample of interest using a beam-splitter cube that is positioned above the object plane. This LED illumination is then partially reflected from the sample to be collected by two lenses, creating a reflection 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

Do Uric Acid Deposits in Zooxanthellae Function as Eye-Spots?

PubMed Central

Yamashita, Hiroshi; Kobiyama, Atsushi; Koike, Kazuhiko

2009-01-01

The symbiosis between zooxanthellae (dinoflagellate genus Symbiodinium) and corals is a fundamental basis of tropical marine ecosystems. However the physiological interactions of the hosts and symbionts are poorly understood. Recently, intracellular crystalline deposits in Symbiodinium were revealed to be uric acid functioning for nutrient storage. This is the first exploration of these enigmatic crystalline materials that had previously been misidentified as oxalic acid, providing new insights into the nutritional strategies of Symbiodinium in oligotrophic tropical waters. However, we believe these deposits also function as eye-spots on the basis of light and electron microscopic observations of motile cells of cultured Symbiodinium. The cells possessed crystalline deposit clusters in rows with each row 100–150 nm thick corresponding to 1/4 the wavelength of light and making them suitable for maximum wave interference and reflection of light. Crystalline clusters in cells observed with a light microscope strongly refracted and polarized light, and reflected or absorbed short wavelength light. The facts that purines, including uric acid, have been identified as the main constituents of light reflectors in many organisms, and that the photoreceptor protein, opsin, was detected in our Symbiodinium strain, support the idea that uric acid deposits in Symbiodinium motile cells may function as a component of an eye-spot. PMID:19609449

Do uric acid deposits in zooxanthellae function as eye-spots?

PubMed

Yamashita, Hiroshi; Kobiyama, Atsushi; Koike, Kazuhiko

2009-07-17

The symbiosis between zooxanthellae (dinoflagellate genus Symbiodinium) and corals is a fundamental basis of tropical marine ecosystems. However the physiological interactions of the hosts and symbionts are poorly understood. Recently, intracellular crystalline deposits in Symbiodinium were revealed to be uric acid functioning for nutrient storage. This is the first exploration of these enigmatic crystalline materials that had previously been misidentified as oxalic acid, providing new insights into the nutritional strategies of Symbiodinium in oligotrophic tropical waters. However, we believe these deposits also function as eye-spots on the basis of light and electron microscopic observations of motile cells of cultured Symbiodinium. The cells possessed crystalline deposit clusters in rows with each row 100-150 nm thick corresponding to 1/4 the wavelength of light and making them suitable for maximum wave interference and reflection of light. Crystalline clusters in cells observed with a light microscope strongly refracted and polarized light, and reflected or absorbed short wavelength light. The facts that purines, including uric acid, have been identified as the main constituents of light reflectors in many organisms, and that the photoreceptor protein, opsin, was detected in our Symbiodinium strain, support the idea that uric acid deposits in Symbiodinium motile cells may function as a component of an eye-spot.

Liver and chorion cytochemistry.

PubMed

Roels, F; De Prest, B; De Pestel, G

1995-01-01

Microscopic visualization of peroxisomes in chorionic villus cytotrophoblast and in biopsy and autopsy samples of liver and kidney, the presence of enlarged liver macrophages containing lipid droplets insoluble in acetone and n-hexane as well as polarizing inclusions formed by stacks of trilamellar sheets are of diagnostic value in peroxisomal disorders. Methods are presented for evaluating these structures by light microscopy; trilamellar inclusions are only detected by electron microscopy. Macrophage features are preserved in archival paraffin blocks. In adrenal cortex, insoluble lipid, polarizing inclusions and trilamellar structures should be looked for. The stains are easily reproducible, and all reagents are commercially available.

Endoscopic Optical Coherence Tomography for Clinical Gastroenterology

PubMed Central

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

2014-01-01

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

Evaluation of a miniature microscope objective designed for fluorescence array microscopy detection of Mycobacterium tuberculosis.

PubMed

McCall, Brian; Olsen, Randall J; Nelles, Nicole J; Williams, Dawn L; Jackson, Kevin; Richards-Kortum, Rebecca; Graviss, Edward A; Tkaczyk, Tomasz S

2014-03-01

A prototype miniature objective that was designed for a point-of-care diagnostic array microscope for detection of Mycobacterium tuberculosis and previously fabricated and presented in a proof of concept is evaluated for its effectiveness in detecting acid-fast bacteria. To evaluate the ability of the microscope to resolve submicron features and details in the image of acid-fast microorganisms stained with a fluorescent dye, and to evaluate the accuracy of clinical diagnoses made with digital images acquired with the objective. The lens prescription data for the microscope design are presented. A test platform is built by combining parts of a standard microscope, a prototype objective, and a digital single-lens reflex camera. Counts of acid-fast bacteria made with the prototype objective are compared to counts obtained with a standard microscope over matched fields of view. Two sets of 20 smears, positive and negative, are diagnosed by 2 pathologists as sputum smear positive or sputum smear negative, using both a standard clinical microscope and the prototype objective under evaluation. The results are compared to a reference diagnosis of the same sample. More bacteria are counted in matched fields of view in digital images taken with the prototype objective than with the standard clinical microscope. All diagnostic results are found to be highly concordant. An array microscope built with this miniature lens design will be able to detect M tuberculosis with high sensitivity and specificity.

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

PubMed

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.

A setup for combined multiphoton laser scanning microscopic and multi-electrode patch clamp experiments on brain slices

NASA Astrophysics Data System (ADS)

Helm, P. Johannes; Reppen, Trond; Heggelund, Paul

2009-02-01

Multi Photon Laser Scanning Microscopy (MPLSM) appears today as one of the most powerful experimental tools in cellular neurophysiology, notably in studies of the functional dynamics of signal processing in single neurons. Simultaneous recording of fluorescence signals at high spatial and temporal resolution and electric signals by means of multi electrode patch clamp techniques have provided new paths for the systematic investigation of neuronal mechanisms. In particular, this approach has opened for direct studies of dendritic signal processing in neurons. We report about a setup optimized for simultaneous electrophysiological multi electrode patch clamp and multi photon laser scanning fluorescence microscopic experiments on brain slices. The microscopic system is based on a modified commercially available confocal scanning laser microscope (CLSM). From a technical and operational point of view, two developments are important: Firstly, in order to reduce the workload for the experimentalist, who in general is forced to concentrate on controlling the electrophysiological parameters during the recordings, a system of shutters has been installed together with dedicated electronic modules protecting the photo detectors against destructive light levels caused by erroneous opening or closing of microscopic light paths by the experimentalist. Secondly, the standard detection unit has been improved by installing the photomultiplier tubes (PMT) in a Peltier cooled thermal box shielding the detector from both room temperature and distortions caused by external electromagnetic fields. The electrophysiological system is based on an industrial standard multi patch clamp unit ergonomically arranged around the microscope stage. The electrophysiological and scanning processes can be time coordinated by standard trigger electronics.

A high-resolution multimode digital microscope system.

PubMed

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

2013-01-01

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

A surface science compatible epifluorescence microscope for inspection of samples under ultra high vacuum and cryogenic conditions.

PubMed

Marquardt, Christian; Paulheim, Alexander; Rohbohm, Nils; Merkel, Rudolf; Sokolowski, Moritz

2017-08-01

We modified an epi-illumination light microscope and mounted it on an ultra high vacuum chamber for investigating samples used in a surface science experiment. For easy access and bake out, all optical components are placed outside the vacuum and the sample is imaged through a glass window. The microscope can be operated in reflection brightfield or epifluorescence mode to image the sample surface or fluorescent dye molecules adsorbed on it. The homemade sample mounting was made compatible for the use under the microscope; sample temperatures as low as 6 K can be achieved. The performance of the microscope is demonstrated on two model samples: Brightfield-images of a well-prepared Ag(100) surface show a macroscopic corrugation of the surface, although low energy electron diffraction data indicate a highly ordered crystalline surface. The surface shows macroscopic protrusions with flat regions, about 20-200 μm in diameter, in between. Fluorescence images of diluted 3,4,9,10-perylene tetracarboxylicacid dianhydride (PTCDA) molecules adsorbed on an ultrathin epitaxial KCl film on the Ag(100) surface show a shading effect at surface protrusions due to an inclined angle of incidence of the PTCDA beam during deposition. For some preparations, the distribution of the fluorescence intensity is inhomogeneous and shows a dense network of bright patches about 5 μm in diameter related to the macroscopic corrugation of the surface. We propose that such a light microscope can aid many surface science experiments, especially those dealing with epitaxial growth or fluorescent materials.

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

PubMed

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

2003-05-01

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

Synthesis, characterization, and application of novel biodegradable self-assembled 2-(N-phthalimido) ethyl-palmitate nanoparticles for cancer therapy

NASA Astrophysics Data System (ADS)

Kasoju, Naresh; Bora, Debajeet K.; Bhonde, Ramesh R.; Bora, Utpal

2010-03-01

We report the synthesis of novel biodegradable nanoparticles (NPs) which can kill the cancer cells without any additional drug loading. The NP was a self-assembled form of a phthalimide based conjugate, in which the phthalimide moiety was responsible for the anticancer activity. We describe the synthesis of a novel 2-(N-phthalimido) ethyl palmitate (PHEP-Pal) conjugate and subsequent preparation of NPs by a simple self assembly process. The successful synthesis of conjugate was confirmed by various characterization studies including nuclear magnetic resonance spectroscope, Fourier transform infrared spectroscope, TOF-liquid chromatography mass spectroscope, differential scanning calorimetry, and X-ray diffraction unit. The synthesis, shape, size, and size distribution of PHEP-Pal NPs were determined by transmission electron microscope, atomic force microscope, and dynamic light scattering technique. Finally, cell culture studies using A549 and HeLa cells were done to evaluate the anticancer effect of PHEP-Pal NPs, which demonstrated the potency of these NPs for use in cancer chemotherapy.

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

NASA Astrophysics Data System (ADS)

Kettel, Johannes; Reinecke, Holger; Müller, Claas

2016-04-01

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

Fiber optic biofluorometer for physiological research on muscle slices

NASA Astrophysics Data System (ADS)

Belz, Mathias; Dendorfer, Andreas; Werner, Jan; Lambertz, Daniel; Klein, Karl-Friedrich

2016-03-01

A focus of research in cell physiology is the detection of Ca2+, NADH, FAD, ATPase activity or membrane potential, only to name a few, in muscle tissues. In this work, we report on a biofluorometer using ultraviolet light emitting diodes (UV-LEDs), optical fibers and two photomultipliers (PMTs) using synchronized fluorescence detection with integrated background correction to detect free calcium, Ca2+, in cardiac muscle tissue placed in a horizontal tissue bath and a microscope setup. Fiber optic probes with imaging optics have been designed to transport excitation light from the biofluorometer's light output to a horizontal tissue bath and to collect emission light from a tissue sample of interest to two PMTs allowing either single excitation / single emission or ratiometric, dual excitation / single emission or single excitation / dual emission fluorescence detection of indicator dyes or natural fluorophores. The efficient transport of light from the excitation LEDs to the tissue sample, bleaching effects of the excitation light in both, polymer and fused silica-based fibers will be discussed. Furthermore, a new approach to maximize light collection of the emission light using high NA fibers and high NA coupling optics will be shown. Finally, first results on Ca2+ measurements in cardiac muscle slices in a traditional microscope setup and a horizontal tissue bath using fiber optic probes will be introduced and discussed.

Generation of multiple Bessel beams for a biophotonics workstation.

PubMed

Cizmár, T; Kollárová, V; Tsampoula, X; Gunn-Moore, F; Sibbett, W; Bouchal, Z; Dholakia, K

2008-09-01

We present a simple method using an axicon and spatial light modulator to create multiple parallel Bessel beams and precisely control their individual positions in three dimensions. This technique is tested as an alternative to classical holographic beam shaping commonly used now in optical tweezers. Various applications of precise control of multiple Bessel beams are demonstrated within a single microscope giving rise to new methods for three-dimensional positional control of trapped particles or active sorting of micro-objects as well as "focus-free" photoporation of living cells. Overall this concept is termed a 'biophotonics workstation' where users may readily trap, sort and porate material using Bessel light modes in a microscope.

Light and Life in Baltimore—and Beyond

PubMed Central

Edidin, Michael

2015-01-01

Baltimore has been the home of numerous biophysical studies using light to probe cells. One such study, quantitative measurement of lateral diffusion of rhodopsin, set the standard for experiments in which recovery after photobleaching is used to measure lateral diffusion. Development of this method from specialized microscopes to commercial scanning confocal microscopes has led to widespread use of the technique to measure lateral diffusion of membrane proteins and lipids, and as well diffusion and binding interactions in cell organelles and cytoplasm. Perturbation of equilibrium distributions by photobleaching has also been developed into a robust method to image molecular proximity in terms of fluorescence resonance energy transfer between donor and acceptor fluorophores. PMID:25650914

A stress-controlled shear cell for small-angle light scattering and microscopy.

PubMed

Aime, S; Ramos, L; Fromental, J M; Prévot, G; Jelinek, R; Cipelletti, L

2016-12-01

We develop and test a stress-controlled, parallel plates shear cell that can be coupled to an optical microscope or a small angle light scattering setup, for simultaneous investigation of the rheological response and the microscopic structure of soft materials under an imposed shear stress. In order to minimize friction, the cell is based on an air bearing linear stage, the stress is applied through a contactless magnetic actuator, and the strain is measured through optical sensors. We discuss the contributions of inertia and of the small residual friction to the measured signal and demonstrate the performance of our device in both oscillating and step stress experiments on a variety of viscoelastic materials.

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

PubMed

Antosiewicz, Tomasz J; Szoplik, Tomasz

2007-08-20

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

Membranous nephropathy (bubbling appearance and spike formation) without immunoglobulin deposition in a patient with systemic lupus erythematosus.

PubMed

Miura, Naoto; Mori, Yuki; Yoshino, Masabumi; Suga, Norihiro; Kitagawa, Wataru; Yamada, Harutaka; Nishikawa, Kazuhiro; Imai, Hirokazu

2008-12-01

A 53-year-old Japanese man with systemic lupus erythematosus developed proteinuria and hematuria after a urinary stone episode. A light microscopic study of a kidney biopsy specimen demonstrated a bubbling appearance and spike formation of the basement membrane. Immunofluorescent studies revealed that there were no significant depositions of immunoglobulins, such as IgG (-), IgA (-), IgM (+/-), kappa light chain (+/-), lambda light chain (+/-), or C3 (-) in the glomerular capillary wall, though C1q was present as one-plus positive staining in mesangial areas. Electron microscopic studies showed that the thickness of the basement membrane varied from thin to thick without electron dense deposits, and that the cellular components of the podocyte were irregularly present in the basement membrane. Urinary protein decreased after the usage of prednisolone and mizoribine; however, proteinuria aggravated after an episode of urinary stone during the same treatment.

Development of UV-curable liquid for in-liquid fluorescence alignment in ultraviolet nanoimprint lithography

NASA Astrophysics Data System (ADS)

Ochiai, Kento; Kikuchi, Eri; Ishito, Yota; Kumagai, Mari; Nakamura, Takahiro; Nakagawa, Masaru

2018-06-01

We studied a fluorescent UV-curable resin suitable for fluorescence alignment in UV nanoimprinting. The addition of a cationic fluorescent dye caused radical photopolymerization of a UV-curable resin by exposure to visible excitation light for fluorescence microscope observation. The microscope observation of a resin film prepared by pressing resin droplets on a silica substrate with a fluorinated silica superstrate revealed that the cationic dye molecules were preferably adsorbed onto the silica surface. It was indicated that the dye molecules concentrated on the silica surface may cause the photocuring. A nonionic fluorescent dye was selected owing to its low polar symmetrical structure and its solubility parameter close to monomers. The fluorescent UV-curable resin with the nonionic dye showed uncured stability to exposure to visible excitation light for 30 min with a light intensity of 8.5 mW cm‑2 detected at 530 nm.

Maintenance on the Advanced Colloids Experiment Module

NASA Image and Video Library

2018-04-16

iss055e035366 (April 16, 2018) --- NASA astronaut Ricky Arnold performs maintenance on the Advanced Colloids Experiment Module located inside the Light Microscopy Module which is a modified commercial, highly flexible, state-of-the-art light imaging microscope facility that provides researchers with powerful diagnostic hardware and software in microgravity.

Ophthalmic applications of confocal microscopy: diagnostics, refractive surgery, and eye banking

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1990-11-01

Confocal microscopy of ocular tissue provides two advantages over traditional imaging techniques: increased range and transverse resolution and increased contrast. The semitransparent cornea and ocular lens in the living eye can be optically sectioned and observed by reflected light confocal microscopy. Within the cornea we observed various cell components nerve fibers nerve cell bodies and fibrous networks. The confocal microscopic images from the in-situ ocular lens show the lens capsule the lens epithelium and the individual lens fibrils. All of the reflected light confocal microscopic images have high contrast and high resolution. Some of the applications of confocal imaging in ophthalmology include: diagnostics of the cornea and the ocular lens examination prior to and after refractive surgery examination of intraocular lenses (IOL) and examination of eye bank material. Other ophthalmic uses of confocal imaging include: studies of wound healing therapeutics and the effects of contact lenses on the cornea. The proposed features of a clinical confocal microscope are reviewed. 2.

Self-referenced axial chromatic dispersion measurement in multiphoton microscopy through 2-color THG imaging.

PubMed

Du, Yu; Zhuang, Ziwei; He, Jiexing; Liu, Hongji; Qiu, Ping; Wang, Ke

2018-05-16

With tunable excitation light, multiphoton microscopy (MPM) is widely used for imaging biological structures at subcellular resolution. Axial chromatic dispersion, present in virtually every transmissive optical system including the multiphoton microscope, leads to focal (and the resultant image) plane separation. Here we demonstrate experimentally a technique to measure the axial chromatic dispersion in a multiphoton microscope, using simultaneous 2-color third-harmonic generation (THG) imaging excited by a 2-color soliton source with tunable wavelength separation. Our technique is self-referenced, eliminating potential measurement error when 1-color tunable excitation light is used which necessitates reciprocating motion of the mechanical translation stage. Using this technique, we demonstrate measured axial chromatic dispersion with 2 different objective lenses in a multiphoton microscope. Further measurement in a biological sample also indicates that this axial chromatic dispersion, in combination with 2-color imaging, may open up opportunity for simultaneous imaging of two different axial planes. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Learning a cost function for microscope image segmentation.

PubMed

Nilufar, Sharmin; Perkins, Theodore J

2014-01-01

Quantitative analysis of microscopy images is increasingly important in clinical researchers' efforts to unravel the cellular and molecular determinants of disease, and for pathological analysis of tissue samples. Yet, manual segmentation and measurement of cells or other features in images remains the norm in many fields. We report on a new system that aims for robust and accurate semi-automated analysis of microscope images. A user interactively outlines one or more examples of a target object in a training image. We then learn a cost function for detecting more objects of the same type, either in the same or different images. The cost function is incorporated into an active contour model, which can efficiently determine optimal boundaries by dynamic programming. We validate our approach and compare it to some standard alternatives on three different types of microscopic images: light microscopy of blood cells, light microscopy of muscle tissue sections, and electron microscopy cross-sections of axons and their myelin sheaths.

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

PubMed Central

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

2009-01-01

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

Adaptation of commercial microscopes for advanced imaging applications

NASA Astrophysics Data System (ADS)

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.

Space-resolved diffusing wave spectroscopy measurements of the macroscopic deformation and the microscopic dynamics in tensile strain tests

NASA Astrophysics Data System (ADS)

Nagazi, Med-Yassine; Brambilla, Giovanni; Meunier, Gérard; Marguerès, Philippe; Périé, Jean-Noël; Cipelletti, Luca

2017-01-01

We couple a laser-based, space-resolved dynamic light scattering apparatus to a universal traction machine for mechanical extensional tests. We perform simultaneous optical and mechanical measurements on polyether ether ketone, a semi-crystalline polymer widely used in the industry. Due to the high turbidity of the sample, light is multiply scattered by the sample and the diffusing wave spectroscopy (DWS) formalism is used to interpret the data. Space-resolved DWS yields spatial maps of the sample strain and of the microscopic dynamics. An excellent agreement is found between the strain maps thus obtained and those measured by a conventional stereo-digital image correlation technique. The microscopic dynamics reveals both affine motion and plastic rearrangements. Thanks to the extreme sensitivity of DWS to displacements as small as 1 nm, plastic activity and its spatial localization can be detected at an early stage of the sample strain, making the technique presented here a valuable complement to existing material characterization methods.

Evaluating Red Reflex and Surgeon Preference Between Nearly-Collimated and Focused Beam Microscope Illumination Systems

PubMed Central

Cionni, Robert J.; Pei, Ron; Dimalanta, Ramon; Lubeck, David

2015-01-01

Purpose To evaluate the intensity and stability of the red reflex produced by ophthalmic surgical microscopes with nearly-collimated versus focused illumination systems and to assess surgeon preference in a simulated surgical setting. Methods This two-part evaluation consisted of postproduction surgical video analysis of red reflex intensity and a microscope use and preference survey completed by 13 experienced cataract surgeons. Survey responses were based on bench testing and experience in a simulated surgical setting. A microscope with nearly-collimated beam illumination and two focused beam microscopes were assessed. Results Red reflex intensity and stability were greater with the nearly-collimated microscope illumination system. In the bench testing survey, surgeons reported that the red reflex was maintained over significantly greater distances away from pupillary center, and depth of focus was numerically greater with nearly-collimated illumination relative to focused illumination. Most participating surgeons (≥64%) reported a preference for the microscope with nearly-collimated illumination with regard to red reflex stability, depth of focus, visualization, surgical working distance, and perceived patient comfort. Conclusions The microscope with nearly-collimated illumination produced a more intense and significantly more stable red reflex and was preferred overall by more surgeons. Translational Relevance This is the first report of an attempt to quantify red reflex intensity and stability and to evaluate surgically-relevant parameters between microscope systems. The data and methods presented here may provide a basis for future studies attempting to quantify differences between surgical microscopes that may affect surgeon preference and microscope use in ophthalmic surgery. PMID:26290778

Effects of electrohydraulic extracorporeal shock wave lithotripsy on submandibular gland in the rat: electron microscopic evaluation.

PubMed

Bayar, Nuray; Kaymaz, F Figen; Apan, Alpaslan; Yilmaz, Erdal; Cakar, A Nur

2002-05-15

Extracorporeal shockwave lithotripsy (ESWL) has been applied in sialolithiasis as a new treatment modality. The aim of this experimental study is to investigate the local effects of electrohydraulic ESWL applied to the right submandibular gland of the rats. This prospective study was conveyed in four groups; groups I, II, III and IV; each group consisting of 20, 20, 18 and 9 rats, respectively, with a randomized distribution. Groups I, II, III and IV received 250, 500, 1000 and 2000 shock waves at 14-16 kV (average 15.1 kV), respectively, to the right submandibular glands on the 0th day. In groups I, II, III, right submandibular glands of the rats were removed on the 0th, 1st, 7th and 15th days; in group IV, this procedure could be managed only on the 0th and 7th days. Light and electron microscopic evaluation were assessed. Using the light microscopic changes, severity of damage score of the glands (SDS) was found. Statistical analysis was done using SDSs. Light and electron microscopic observations have shown that the damage produced by the shock waves were confined to focal areas in the acinar cells (AC), granulated convoluted tubule (GCT) cells and blood vessels at all doses applied. Vacuolization in the cytoplasms of the AC and GCT cells, disintegration of membranes, alteration in the cytoplasmic organization, swelling of the mitochondria and loss of the features were observed on electron microscopy. Increase in the secretion rate; stasis and dilatation in the blood vessels; blebbing and loss of features in the cytoplasm of the endothelial cells were observed. According to the result of the statistical analysis using SDSs; at 250 shock wave dose, a statistically significant difference between the SDSs of the days (0th, 1st, 7th and 15th) was found (P<0.05). The SDS on the 0th day was found to have the lowest value among the other days. And also a statistically significant difference was found on the 0th day between the SDSs at doses of 250, 500, 1000 and 2000 shock waves (P<0.05). The SDS at 250 and 500 shock waves was found to have the lower value than the SDS at the 2000 shock wave. It was observed that produced damage was less prominent by small doses (250, 500 doses) initially (0th day). Electrohydraulic ESWL caused a "patchy type" generalized pathology on submandibular glands of the rats and damaged focal areas were widespread all through the gland from the 1st day on. Formation of the damage was concluded to be related to the direct effect of the shock waves rather than the dose used. Electrohydraulic lithotripters are not suitable for sialolithiasis because of the focus problems, local tissue damage and the risk of the damage to the adjacent structures.

Evaluation of transurethral and transperineal tin ethyl etiopurpurin-photodynamic therapy on the canine prostate one week after drug injection

NASA Astrophysics Data System (ADS)

Selman, Steven H.; Keck, Rick W.; Kondo, Sandy; Albrecht, Detlef

1999-06-01

We have been investigating the potential applicability of photodynamic therapy for the treatment of benign and malignant disease of the prostate. Both transurethral and transperineal approaches to the delivery of light to the tin ethyl etiopurpurin sensitized canine prostate have been studied. Pharmacologic studies were performed and suggested that delaying light treatment for 7 days after drug administration would maximize the desired effect on the targeted prostatic tissue while minimizing the damage to surrounding bladder and rectum. A total of 12 dogs were treated with transurethral light alone (n=6) or the combination of transurethral light and transperineal light one week after tin ethyl etiopurpurin administration. (Previous studies have shown that light alone has no effect on prostate size or histology.) Animals were euthanized 48 hours and 3 weeks after completion of treatment (drug, 1mg/kg day 0, light [400mw/750sec]day 7). Tissue response was determined by gross and microscopic examination. Additionally, pre- and post- treatment transrectal ultrasounds were compared to assess changes in prostate volume and tissue echogenicity. The combination of transurethral and transperineal light results in extensive destruction of glandular epithelium with minimal damage to surrounding structures. Prostate volumes decreased by an average of 52%. Untreated areas were found to lie greater than 0.5 cm from the light diffuser. These studies have encouraged us to continue to investigate this modality as a technique for total ablation of prostatic glandular epithelium.

Adding an Extra Dimension to What Students See through the Light Microscope: A Lab Exercise Demonstrating Critical Analysis for Microscopy Students

ERIC Educational Resources Information Center

Garrill, Ashley

2011-01-01

This article describes an undergraduate lab exercise that demonstrates the importance of students thinking critically about what they see through a microscope. The students are given growth data from tip-growing organisms that suggest the cells grow in a pulsatile manner. The students then critique this data in several exercises that incorporate…

COMPACT NON-CONTACT TOTAL EMISSION DETECTION FOR IN-VIVO MULTI-PHOTON EXCITATION MICROSCOPY

PubMed Central

Glancy, Brian; Karamzadeh, Nader S.; Gandjbakhche, Amir H.; Redford, Glen; Kilborn, Karl; Knutson, Jay R.; Balaban, Robert S.

2014-01-01

Summary We describe a compact, non-contact design for a Total Emission Detection (c-TED) system for intra-vital multi-photon imaging. To conform to a standard upright two-photon microscope design, this system uses a parabolic mirror surrounding a standard microscope objective in concert with an optical path that does not interfere with normal microscope operation. The non-contact design of this device allows for maximal light collection without disrupting the physiology of the specimen being examined. Tests were conducted on exposed tissues in live animals to examine the emission collection enhancement of the c-TED device compared to heavily optimized objective-based emission collection. The best light collection enhancement was seen from murine fat (5×-2× gains as a function of depth), while murine skeletal muscle and rat kidney showed gains of over two and just under two-fold near the surface, respectively. Gains decreased with imaging depth (particularly in the kidney). Zebrafish imaging on a reflective substrate showed close to a two-fold gain throughout the entire volume of an intact embryo (approximately 150 μm deep). Direct measurement of bleaching rates confirmed that the lower laser powers (enabled by greater light collection efficiency) yielded reduced photobleaching in vivo. The potential benefits of increased light collection in terms of speed of imaging and reduced photo-damage, as well as the applicability of this device to other multi-photon imaging methods is discussed. PMID:24251437

Modular low-light microscope for imaging cellular bioluminescence and radioluminescence

PubMed Central

Kim, Tae Jin; Türkcan, Silvan; Pratx, Guillem

2017-01-01

Low-light microscopy methods are receiving increased attention as new applications have emerged. One such application is to allow longitudinal imaging of light-sensitive cells with no phototoxicity and no photobleaching of fluorescent biomarkers. Another application is for imaging signals that are inherently dim and undetectable using standard microscopy, such as bioluminescence, chemiluminescence, or radioluminescence. In this protocol, we provide instructions on how to build a modular low-light microscope (1-4 d) by coupling two microscope objective lenses, back-to-back from each other, using standard optomechanical components. We also provide directions on how to image dim signals such as radioluminescence (1-1.5 h), bioluminescence (∼30 min) and low-excitation fluorescence (∼15 min). In particular, radioluminescence microscopy is explained in detail as it is a newly developed technique, which enables the study of small molecule transport (eg. radiolabeled drugs, metabolic precursors, and nuclear medicine contrast agents) by single cells without perturbing endogenous biochemical processes. In this imaging technique, a scintillator crystal (eg. CdWO4) is placed in close proximity to the radiolabeled cells, where it converts the radioactive decays into optical flashes detectable using a sensitive camera. Using the image reconstruction toolkit provided in this protocol, the flashes can be reconstructed to yield high-resolution image of the radiotracer distribution. With appropriate timing, the three aforementioned imaging modalities may be performed altogether on a population of live cells, allowing the user to perform parallel functional studies of cell heterogeneity at the single-cell level. PMID:28426025

Healing properties of implants inserted concomitantly with anorganic bovine bone. A histomorphometric human study.

PubMed

Menicucci, G; Mussano, F; Schierano, G; Rizzati, A; Aimetti, M; Gassino, G; Traini, T; Carossa, S

2013-03-01

The present prospective, randomized, double-blind study evaluated the bone-forming process around implants inserted simultaneously with anorganic bovine bone (ABB) in sinus grafting. A total of 18 threaded mini-implants with Osseotite (O) and Nanotite (N) surfaces were placed in seven patients (nine sites). After 12 months, the implants were retrieved and processed for histological analysis. A total of 18 cutting and grinding sections were investigated with bright-field light microscopy, circularly polarized light microscopy (CPLM), confocal scanning laser microscope (CSLM), and scanning electron microscope (SEM) with energy dispersive spectrometer (EDS). The bone-to-implant contact rate in native crestal bone was 62.6 ± 0.4% for N implants and 54.3 ± 0.5% for the O implants (p = 0.001). The collagen fibre density, as assessed by CPLM, was 79.8 ± 6.0 nm for the N group and 74.6 ± 4.6 nm for the O group (p < 0.05). Line scan EDS starting from ABB to newly formed bone showed a decrease in calcium content and an increase of carbon while phosphorus content was constant. While the N surface improved the peri-implant endosseous healing properties in the native bone, when compared to the O surface, it did not improve the healing properties in the bone-graft area. © 2013 Australian Dental Association.

Thulium-170-labeled microparticles for local radiotherapy: preliminary studies.

PubMed

Polyak, Andras; Das, Tapas; Chakraborty, Sudipta; Kiraly, Reka; Dabasi, Gabriella; Joba, Robert Peter; Jakab, Csaba; Thuroczy, Julianna; Postenyi, Zita; Haasz, Veronika; Janoki, Gergely; Janoki, Gyozo A; Pillai, Maroor R A; Balogh, Lajos

2014-10-01

The present article describes the preparation, characterization, and biological evaluation of Thulium-170 ((170)Tm) [T1/2 = 128.4 days; Eβmax = 968 keV; Eγ = 84 keV (3.26%)] labeled tin oxide microparticles for its possible use in radiation synovectomy (RSV) of medium-sized joints. (170)Tm was produced by irradiation of natural thulium oxide target. 170Tm-labeled microparticles were synthesized with high yield and radionuclidic purity (> 99%) along with excellent in vitro stability by following a simple process. Particle sizes and morphology of the radiolabeled particles were examined by light microscope, dynamic light scattering, and transmission electron microscope and found to be of stable spherical morphology within the range of 1.4-3.2 μm. The preparation was injected into the knee joints of healthy Beagle dogs intraarticularly for biological studies. Serial whole-body and regional images were taken by single-photon-emission computed tomography (SPECT) and SPECT-CT cameras up to 9 months postadministration, which showed very low leakage (< 8% of I.D.) of the instilled particles. The majority of leaked radiocolloid particles were found in inguinal lymph nodes during the 9 months of follow-up. All the animals tolerated the treatment well; the compound did not show any possible radiotoxicological effect. These preliminary studies showed that 170Tm-labeled microparticles could be a promising nontoxic and effective radiopharmaceutical for RSV applications or later local antitumor therapy.

Automated measurement of diatom size

USGS Publications Warehouse

Spaulding, Sarah A.; Jewson, David H.; Bixby, Rebecca J.; Nelson, Harry; McKnight, Diane M.

2012-01-01

Size analysis of diatom populations has not been widely considered, but it is a potentially powerful tool for understanding diatom life histories, population dynamics, and phylogenetic relationships. However, measuring cell dimensions on a light microscope is a time-consuming process. An alternative technique has been developed using digital flow cytometry on a FlowCAM® (Fluid Imaging Technologies) to capture hundreds, or even thousands, of images of a chosen taxon from a single sample in a matter of minutes. Up to 30 morphological measures may be quantified through post-processing of the high resolution images. We evaluated FlowCAM size measurements, comparing them against measurements from a light microscope. We found good agreement between measurement of apical cell length in species with elongated, straight valves, including small Achnanthidium minutissimum (11-21 µm) and largeDidymosphenia geminata (87–137 µm) forms. However, a taxon with curved cells, Hannaea baicalensis (37–96 µm), showed differences of ~ 4 µm between the two methods. Discrepancies appear to be influenced by the choice of feret or geodesic measurement for asymmetric cells. We describe the operating conditions necessary for analysis of size distributions and present suggestions for optimal instrument conditions for size analysis of diatom samples using the FlowCAM. The increased speed of data acquisition through use of imaging flow cytometers like the FlowCAM is an essential step for advancing studies of diatom populations.

Automated Microscopy: Macro Language Controlling a Confocal Microscope and its External Illumination: Adaptation for Photosynthetic Organisms.

PubMed

Steinbach, Gábor; Kaňa, Radek

2016-04-01

Photosynthesis research employs several biophysical methods, including the detection of fluorescence. Even though fluorescence is a key method to detect photosynthetic efficiency, it has not been applied/adapted to single-cell confocal microscopy measurements to examine photosynthetic microorganisms. Experiments with photosynthetic cells may require automation to perform a large number of measurements with different parameters, especially concerning light conditions. However, commercial microscopes support custom protocols (through Time Controller offered by Olympus or Experiment Designer offered by Zeiss) that are often unable to provide special set-ups and connection to external devices (e.g., for irradiation). Our new system combining an Arduino microcontroller with the Cell⊕Finder software was developed for controlling Olympus FV1000 and FV1200 confocal microscopes and the attached hardware modules. Our software/hardware solution offers (1) a text file-based macro language to control the imaging functions of the microscope; (2) programmable control of several external hardware devices (light sources, thermal controllers, actuators) during imaging via the Arduino microcontroller; (3) the Cell⊕Finder software with ergonomic user environment, a fast selection method for the biologically important cells and precise positioning feature that reduces unwanted bleaching of the cells by the scanning laser. Cell⊕Finder can be downloaded from http://www.alga.cz/cellfinder. The system was applied to study changes in fluorescence intensity in Synechocystis sp. PCC6803 cells under long-term illumination. Thus, we were able to describe the kinetics of phycobilisome decoupling. Microscopy data showed that phycobilisome decoupling appears slowly after long-term (>1 h) exposure to high light.

Growth stimulation of Bacillus cereus and Pseudomonas putida using nanostructured ZnO thin film as transducer element

NASA Astrophysics Data System (ADS)

Loukanov, Alexandre; Filipov, Chavdar; Valcheva, Violeta; Lecheva, Marta; Emin, Saim

2015-04-01

The semiconductor zinc oxide nanomaterial (ZnO or ZnO:H) is widely used in advanced biosensor technology for the design of highly-sensitive detector elements for various applications. In the attempt to evaluate its effect on common microorganisms, two types of nanostructured transducer films have been used (average diameter 600-1000 nm). They have been prepared by using both wet sol-gel method and magnetron sputtering. Their polycrystalline structure and specific surface features have been analyzed by X-ray diffraction (XRD), scanning electron microscope, and atomic force microscope. The assessment of growth stimulation of bacteria was determined using epifluorescent microscope by cell staining with Live/Dead BacLight kit. In our experiments, the growth stimulation of Gram-positive and Gram-negative bacteria on nanostructured ZnO film is demonstrated by Bacillus cereus and Pseudomonas putida. These two bacterial species have been selected, because they are well known and studied in biosensor technologies, with structural difference of their cell walls. These pathogens are easy for with common source in the liquid food or some commercial products. Our data has revealed that the method of transducer film preparation influences strongly bacterial inhibition and division. These results present the transforming signal precisely, when ZnO is used in biosensor applications.

Measurement of glomerulus diameter and Bowman's space width of renal albino rats.

PubMed

Kotyk, Taras; Dey, Nilanjan; Ashour, Amira S; Balas-Timar, Dana; Chakraborty, Sayan; Ashour, Ahmed S; Tavares, João Manuel R S

2016-04-01

Glomerulus diameter and Bowman's space width in renal microscopic images indicate various diseases. Therefore, the detection of the renal corpuscle and related objects is a key step in histopathological evaluation of renal microscopic images. However, the task of automatic glomeruli detection is challenging due to their wide intensity variation, besides the inconsistency in terms of shape and size of the glomeruli in the renal corpuscle. Here, a novel solution is proposed which includes the Particles Analyzer technique based on median filter for morphological image processing to detect the renal corpuscle objects. Afterwards, the glomerulus diameter and Bowman's space width are measured. The solution was tested with a dataset of 21 rats' renal corpuscle images acquired using light microscope. The experimental results proved that the proposed solution can detect the renal corpuscle and its objects efficiently. As well as, the proposed solution has the ability to manage any input images assuring its robustness to the deformations of the glomeruli even with the glomerular hypertrophy cases. Also, the results reported significant difference between the control and affected (due to ingested additional daily dose (14.6mg) of fructose) groups in terms of glomerulus diameter (97.40±19.02μm and 177.03±54.48μm, respectively). Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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.

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

PubMed

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

2015-09-01

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

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

PubMed

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

2017-06-01

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

Film thickness measurement based on nonlinear phase analysis using a Linnik microscopic white-light spectral interferometer.

PubMed

Guo, Tong; Chen, Zhuo; Li, Minghui; Wu, Juhong; Fu, Xing; Hu, Xiaotang

2018-04-20

Based on white-light spectral interferometry and the Linnik microscopic interference configuration, the nonlinear phase components of the spectral interferometric signal were analyzed for film thickness measurement. The spectral interferometric signal was obtained using a Linnik microscopic white-light spectral interferometer, which includes the nonlinear phase components associated with the effective thickness, the nonlinear phase error caused by the double-objective lens, and the nonlinear phase of the thin film itself. To determine the influence of the effective thickness, a wavelength-correction method was proposed that converts the effective thickness into a constant value; the nonlinear phase caused by the effective thickness can then be determined and subtracted from the total nonlinear phase. A method for the extraction of the nonlinear phase error caused by the double-objective lens was also proposed. Accurate thickness measurement of a thin film can be achieved by fitting the nonlinear phase of the thin film after removal of the nonlinear phase caused by the effective thickness and by the nonlinear phase error caused by the double-objective lens. The experimental results demonstrated that both the wavelength-correction method and the extraction method for the nonlinear phase error caused by the double-objective lens improve the accuracy of film thickness measurements.

Dry Eye Following Phacoemulsification Surgery and its Relation to Associated Intraoperative Risk Factors.

PubMed

Sahu, P K; Das, G K; Malik, Aman; Biakthangi, Laura

2015-01-01

The purpose was to study dry eye following phacoemulsification surgery and analyze its relation to associated intra-operative risk factors. A prospective observational study was carried out on 100 eyes of 100 patients without preoperative dry eye. Schirmer's Test I, tear meniscus height, tear break-up time, and lissamine green staining of cornea and conjunctiva were performed preoperatively and at 5 days, 10 days, 1-month, and 2 months after phacoemulsification surgery, along with the assessment of subjective symptoms, using the dry eye questionnaire. The correlations between these values and the operating microscope light exposure time along with the cumulative dissipated energy (CDE) were investigated. There was a significant deterioration of all dry eye test values following phacoemulsification surgery along with an increase in subjective symptoms. These values started improving after 1-month postoperatively, but preoperative levels were not achieved till 2 months after surgery. Correlations of dry eye test values were noted with the operating microscope light exposure time and CDE, but they were not significant. Phacoemulsification surgery is capable of inducing dry eye, and patients should be informed accordingly prior to surgery. The clinician should also be cognizant that increased CDE can induce dry eyes even in eyes that were healthy preoperatively. In addition, intraoperative exposure to the microscopic light should be minimized.

Photodynamic treatment of epithelial tissue derived from patients with endometrial cancer: a contribution to the role of laminin and epidermal growth factor receptor in photodynamic therapy

NASA Astrophysics Data System (ADS)

Ziolkowski, Piotr P.; Symonowicz, Krzysztof; Osiecka, Beata J.; Rabczynski, Jerzy; Gerber, Jerzy

1999-07-01

Photodynamic therapy (PDT) was used to treat endometrial G1 cancer tissue derived from patients who had undergone a total hysterectomy and bilateral salpingo-oophorectomy. After surgical treatment the cancerous tissue was kept in a medium containing Dulbecco solution, fetal calf serum, and antibiotics. The tissue was then exposed to hematoporphyrin derivative (0.1 mg/l) and 24 h later exposed to light (total light dose--18 J/sq cm). Necrosis depth was evaluated 24 h later using a light microscope. In order to assess the possible role of the basal membrane component laminin, as well as epidermal growth factor receptor susceptibility to PDT, immunohistochemical studies were carried out. Additionally, nucleolar organizer regions evaluation was performed. Our experiment confirmed that PDT results in the necrosis in the treated endometrial cancer, while not affecting the laminin in the cancerous tissue. In contrast, PDT strongly affects the epidermal growth factor receptor and nucleolar organizer regions in cancer cells. We suggest that laminin may contribute to the prevention of cancer dissemination in the cases where PDT has to be repeated, and that after PDT the cells become less susceptible to a mitogen, like, e.g., epidermal growth factor.

Significant reduction of inflammation and sebaceous glands size in acne vulgaris lesions after intense pulsed light treatment.

PubMed

Barakat, Manal T; Moftah, Noha H; El Khayyat, Mohammad A M; Abdelhakim, Zainab A

2017-01-01

Intense pulsed light (IPL) has been used for years in treatment of acne vulgaris. However, quantitative evaluation of histopathological changes after its use as a sole therapy was poorly investigated. Accordingly, this study aims to objectively evaluate inflammatory infiltrate and sebaceous glands in acne vulgaris after IPL. Twenty-four patients of acne were treated with six IPL sessions. Clinical evaluation was done at 2 weeks after last session by counting acne lesions. Patient satisfaction using Cardiff Acne Disability Index (CADI) was recorded at baseline, 2 weeks and 3 months after IPL. Using histopathological and computerized morphometric analysis, quantitative evaluation of inflammatory infiltrate and measurement of surface area of sebaceous glands were performed for skin biopsies at baseline and 2 weeks after last session. After IPL, there was significant reduction of all acne lesions especially inflammatory variety with significant decrease of CADI score at 2 weeks and 3 months after IPL (p < .05). Microscopically, there was significant decrease in density of inflammatory infiltrate and surface area of sebaceous glands (p < .05). So, IPL is fairly effective therapy in acne vulgaris especially inflammatory variety. The results suggest that IPL could improve acne lesions through targeting both inflammation and sebaceous glands. © 2016 Wiley Periodicals, Inc.

In situ microscopy for on-line determination of biomass.

PubMed

Bittner, C; Wehnert, G; Scheper, T

1998-10-05

A sensor is presented, which allows on-line microscopic observation of microorganisms during fermentations in bioreactors. This sensor, an In Situ Microscope (ISM) consists of a direct-light microscope with a measuring chamber, integrated in a 25 mm stainless steel tube, two CCD-cameras, and two frame-grabbers. The data obtained are processed by an automatic image analysis system. The ISM is connected with the bioreactor via a standard port, and it is immersed directly in the culture liquid-in our case Saccharomyces cerevisiae in a synthetic medium. The microscopic examination of the liquid is performed in the measuring chamber, which is situated near the front end of the sensor head. The measuring chamber is opened and closed periodically. In the open state, the liquid in the bioreactor flows unrestricted through the chamber. In closing, a defined volume of 2,2. 10(-8) mL of the liquid becomes enclosed. After a few seconds, when the movement of the cells in the enclosed culture has stopped, they are examined with the microscope. The microscopic images of the cells are registered with the CCD-cameras and are visualized on a monitor, allowing a direct view of the cell population. After detection, the measuring chamber reopens, and the enclosed liquid is released. The images obtained are evaluated as to cell concentration, cell size, cell volume, biomass, and other relevant parameters simultaneously by automatic image analysis. With a PC (486/33 MHz), image processing takes about 15 s per image. The detection range tested when measuring cells of S. cerevisiae is about 10(6) to 10(9) cells/mL (equivalent to a biomass of 0.01 g/L to 12 g/L). The calculated biomass values correlate very well with those obtained using dry weight analysis. Furthermore, histograms can be calculated, which are comparable to those obtained by flow cytometry. Copyright 1998 John Wiley & Sons, Inc.

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

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

Anti-proliferation effect of blue light-emitting diodes against antibiotic-resistant Helicobacter pylori.

PubMed

Ma, Jianwei; Hiratsuka, Takahiro; Etoh, Tsuyoshi; Akada, Junko; Fujishima, Hajime; Shiraishi, Norio; Yamaoka, Yoshio; Inomata, Masafumi

2017-12-07

Infection by Helicobacter pylori is implicated in a wide range of upper gastrointestinal diseases. Owing to the rapid emergence of antibiotic-resistant strains of H. pylori, the development of novel treatment modalities for antibiotic-resistant H. pylori infection is a key priority. Blue light-emitting diodes (LED) may represent a unique option owing to their antimicrobial effect. In this study, we aimed to evaluate the anti-proliferative effect of blue LED against antibiotic-resistant H. pylori. Ten antibiotic-resistant strains and one sensitive H. pylori strain were used in this study. After irradiation by blue LED along time course, the viability of H. pylori was evaluated by enumerating colony forming units. Morphological changes in H. pylori were observed using a scanning electron microscope. Reductase activity was measured as an indicator of bacterial cellular activity. Total reactive oxygen species was monitored using fluorescence intensity and fluorescence microscope imaging. After irradiation by blue LED, the numbers of H. pylori in all the strains were significantly reduced compared with control group. The H. pylori exhibited a short rod-shaped morphology after irradiation; no such change was observed in H. pylori not exposed to blue LED. Re-irradiation of surviving strain after the initial irradiation also exhibited the same anti-proliferation effect. After blue LED irradiation, bacterial cellular activity was lower, and total reactive oxygen species production was significantly higher in blue LED group, compared with that in control. Blue LED could be a new treatment to eradicate infection with antibiotic-resistant H. pylori. © 2017 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

[Simple evaluation of numbers of asbestos bodies in bronchoalveolar lavage fluid under light microscopy: analysis of 35 pulmonary nodular lesions].

PubMed

Kawahara, Kunimitsu; Kawasumi, Hiromi; Nagano, Teruaki; Sasada, Shinji; Okamoto, Norio

2008-04-01

More than 1 asbestos body (AB) per ml of bronchoalveolar lavage fluid (BALF) under light microscopy was defined as AB positive (ABP) and suggests an occupational asbestos exposure. We microscopically evaluated the AB number per one ml of BALF, which we defined as the AB concentration (ABC), using bronchoalveolar lavage (BAL) cytocentrifuge slides obtained from 35 patients having pulmonary nodular lesions (20 carcinoma and 15 nonneoplastic disease) and examined the correlation between ABC and clinicopathological data including findings on Helical computed tomography scan (HCTS) and occupational history of asbestos exposure (OHAE). BAL was performed by the standard technique without removing mucous with a gauze filter. AB was microscopically defined as a structure consisting of a core of transparent asbestos surrounded by an iron-protein coat. Twenty of 35 patients were ABP (ABP rate; 57%) and ABC ranged from 0 to 207.98/ml (mean ABC; 11.33/ml). Mean ABC was significantly higher in patients with OHAE (15.04/ml) compared to that in patients without OHAE (3.23/ml). Twenty-two of 35 patients (63%) lacked abnormality on HCTS and among these, 12 patients (55%) were ABP. In 20 pulmonary carcinoma patients, the ABP rate was 85% and ABC ranged from 0 to 31.1/ml (Mean ABC; 2.99/ml). The ABP rate of pulmonary carcinoma patients was 40% (8 patients) and among these, 5 patients (63%) did not show any abnormality on HCTS. In conclusion, our method was simple and useful and should be applied to patients with pulmonary nodular lesions and OHAE, even if there are no abnormalities on HCTS.

Critical dimensional linewidth calibration using UV microscope and laser interferometry

NASA Astrophysics Data System (ADS)

Li, Qi; Gao, Si-tian; Li, Wei; Lu, Ming-zhen; Zhang, Ming-kai

2013-10-01

In order to calibrate the critical dimensional (CD) uncertainty of lithography masks in semiconductor manufacturing, NIM is building a two dimensional metrological UV microscope which has traceable measurement ability for nanometer linewidths and pitches. The microscope mainly consists of UV light receiving components, piezoelectric ceramics (PZT) driven stage and interferometer calibration framework. In UV light receiving components they include all optical elements on optical path. The UV light originates from Köhler high aperture transmit/reflect illumination sources; then goes through objective lens to UV splitting optical elements; after that, one part of light attains UV camera for large range calibration, the other part of light passes through a three dimensional adjusted pinhole and is collected by PMT for nanoscale scanning. In PZT driven stage, PZT stick actuators with closed loop control are equipped to push/pull a flexural hinge based platform. The platform has a novel designed compound flexural hinges which nest separate X, Y direction moving mechanisms within one layer but avoiding from mutual cross talk, besides this, the hinges also contain leverage structures to amplify moving distance. With these designs, the platform can attain 100 μm displacement ranges as well as 1 nm resolution. In interferometer framework a heterodyne multi-pass interferometer is mounted on the platform, which measures X-Y plane movement and Z axis rotation, through reference mirror mounted on objective lens tube and Zerodur mirror mounted on PZT platform, the displacement is traced back to laser wavelength. When development is finished, the apparatus can offer the capability to calibrate one dimensional linewidths and two dimensional pitches ranging from 200nm to 50μm with expanded uncertainty below 20nm.

Microtextured metals for stray-light suppression in the Clementine startracker

NASA Technical Reports Server (NTRS)

Johnson, E. A.

1993-01-01

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

Morphology of the leather defect light flecks and spots.

PubMed

Nafstad, O; Wisløff, H; Grønstøl, H

2001-01-01

The skin histology and the scanning electron microscope morphology of the hide defect light flecks and spots after tanning were studied in 11 steers infested with biting lice (Damalinia bovis). Nine steers from herds free of lice were used as controls. Skin biopsies from 6 of the animals in the lice infested group showed mild to moderate hyperkeratosis and moderate perivascular to diffuse dermatitis with infiltration of mainly mononuclear cells and some eosinophilic granulocytes. The steers were slaughtered at an age of 18 to 23 months. Light flecks and spots occurred on all examined hides from the infested group after tanning. No examined hides from the control group demonstrated similar damage. Both light microscopic examination of sections of tanned hide with light flecks and spots and scanning electron microscopy of the same defects showed superficial grain loss and craters with a irregular fibre base encircled by smooth and intact grain. The association between louse infestation at an early age and damage of hides following slaughter 6 to 15 months later, suggested that louse infestations lead to a prolonged or lifelong weakening in the dermis. This weakening may cause superficial grain loss during the tanning process.

Morphology of the Leather Defect Light Flecks and Spots

PubMed Central

Nafstad, O; Wisløff, H; Grønstøl, H

2001-01-01

The skin histology and the scanning electron microscope morphology of the hide defect light flecks and spots after tanning were studied in 11 steers infested with biting lice (Damalinia bovis). Nine steers from herds free of lice were used as controls. Skin biopsies from 6 of the animals in the lice infested group showed mild to moderate hyperkeratosis and moderate perivascular to diffuse dermatitis with infiltration of mainly mononuclear cells and some eosinophilic granulocytes. The steers were slaughtered at an age of 18 to 23 months. Light flecks and spots occurred on all examined hides from the infested group after tanning. No examined hides from the control group demonstrated similar damage. Both light microscopic examination of sections of tanned hide with light flecks and spots and scanning electron microscopy of the same defects showed superficial grain loss and craters with a irregular fibre base encircled by smooth and intact grain. The association between louse infestation at an early age and damage of hides following slaughter 6 to 15 months later, suggested that louse infestations lead to a prolonged or lifelong weakening in the dermis. This weakening may cause superficial grain loss during the tanning process. PMID:11455890

Shearing interference microscope for step-height measurements.

PubMed

Trịnh, Hưng-Xuân; Lin, Shyh-Tsong; Chen, Liang-Chia; Yeh, Sheng-Lih; Chen, Chin-Sheng; Hoang, Hong-Hai

2017-05-01

A shearing interference microscope using a Savart prism as the shear plate is proposed for inspecting step-heights. Where the light beam propagates through the Savart prism and microscopic system to illuminate the sample, it then turns back to re-pass through the Savart prism and microscopic system to generate a shearing interference pattern on the camera. Two measurement modes, phase-shifting and phase-scanning, can be utilized to determine the depths of the step-heights on the sample. The first mode, which employs a narrowband source, is based on the five-step phase-shifting algorithm and has a measurement range of a quarter-wavelength. The second mode, which adopts a broadband source, is based on peak-intensity identification technology and has a measurement range up to a few micrometres. This paper is to introduce the configuration and measurement theory of this microscope, perform a setup used to implement it, and present the experimental results from the uses of the setup. The results not only verify the validity but also confirm the high measurement repeatability of the proposed microscope. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

A new computerized moving stage for optical microscopes

NASA Astrophysics Data System (ADS)

Hatiboglu, Can Ulas; Akin, Serhat

2004-06-01

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

Biobeam—Multiplexed wave-optical simulations of light-sheet microscopy

PubMed Central

Weigert, Martin; Bundschuh, Sebastian T.

2018-01-01

Sample-induced image-degradation remains an intricate wave-optical problem in light-sheet microscopy. Here we present biobeam, an open-source software package that enables simulation of operational light-sheet microscopes by combining data from 105–106 multiplexed and GPU-accelerated point-spread-function calculations. The wave-optical nature of these simulations leads to the faithful reproduction of spatially varying aberrations, diffraction artifacts, geometric image distortions, adaptive optics, and emergent wave-optical phenomena, and renders image-formation in light-sheet microscopy computationally tractable. PMID:29652879

Oxygen Nanobubble Tracking by Light Scattering in Single Cells and Tissues.

PubMed

Bhandari, Pushpak; Wang, Xiaolei; Irudayaraj, Joseph

2017-03-28

Oxygen nanobubbles (ONBs) have significant potential in targeted imaging and treatment in cancer diagnosis and therapy. Precise localization and tracking of single ONBs is demonstrated based on hyperspectral dark-field microscope (HSDFM) to image and track single oxygen nanobubbles in single cells. ONBs were proposed as promising contrast-generating imaging agents due to their strong light scattering generated from nonuniformity of refractive index at the interface. With this powerful platform, we have revealed the trajectories and quantities of ONBs in cells, and demonstrated the relation between the size and diffusion coefficient. We have also evaluated the presence of ONBs in the nucleus with respect to an increase in incubation time and have quantified the uptake in single cells in ex vivo tumor tissues. Our results demonstrate that HSDFM can be a versatile platform to detect and measure cellulosic nanoparticles at the single-cell level and to assess the dynamics and trajectories of this delivery system.

Influence of irradiance on Knoop hardness, degree of conversion, and polymerization shrinkage of nanofilled and microhybrid composite resins.

PubMed

Fugolin, Ana Paula Piovezan; Correr-Sobrinho, Lourenço; Correr, Américo Bortolazzo; Sinhoreti, Mário Alexandre Coelho; Guiraldo, Ricardo Danil; Consani, Simonides

2016-01-01

The purpose of this study was to investigate the influence of the irradiance emitted by a light-curing unit on microhardness, degree of conversion (DC), and gaps resulting from shrinkage of 2 dental composite resins. Cylinders of nanofilled and microhybrid composites were fabricated and light cured. After 24 hours, the tops and bottoms of the specimens were evaluated via indentation testing and Fourier transform infrared spectroscopy to determine Knoop hardness number (KHN) and DC, respectively. Gap width (representing polymerization shrinkage) was measured under a scanning electron microscope. The nanofilled composite specimens presented significantly greater KHNs than did the microhybrid specimens (P < 0.05). The microhybrid composite resin exhibited significantly greater DC and gap width than the nanofilled material (P < 0.05). Irradiance had a mostly material-dependent influence on the hardness and DC, but not the polymerization shrinkage, of composite resins.

Mueller matrix microscope: a quantitative tool to facilitate detections and fibrosis scorings of liver cirrhosis and cancer tissues.

PubMed

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

2016-07-01

Today the increasing cancer incidence rate is becoming one of the biggest threats to human health.Among all types of cancers, liver cancer ranks in the top five in both frequency and mortality rate all over the world. During the development of liver cancer, fibrosis often evolves as part of a healing process in response to liver damage, resulting in cirrhosis of liver tissues. In a previous study, we applied the Mueller matrix microscope to pathological liver tissue samples and found that both the Mueller matrix polar decomposition (MMPD) and Mueller matrix transformation (MMT) parameters are closely related to the fibrous microstructures. In this paper,we take this one step further to quantitatively facilitate the fibrosis detections and scorings of pathological liver tissue samples in different stages from cirrhosis to cancer using the Mueller matrix microscope. The experimental results of MMPD and MMT parameters for the fibrotic liver tissue samples in different stages are measured and analyzed. We also conduct Monte Carlo simulations based on the sphere birefringence model to examine in detail the influence of structural changes in different fibrosis stages on the imaging parameters. Both the experimental and simulated results indicate that the polarized light microscope and transformed Mueller matrix parameter scan provide additional quantitative information helpful for fibrosis detections and scorings of liver cirrhosis and cancers. Therefore, the polarized light microscope and transformed Mueller matrix parameters have a good application prospect in liver cancer diagnosis.

Mueller matrix microscope: a quantitative tool to facilitate detections and fibrosis scorings of liver cirrhosis and cancer tissues

NASA Astrophysics Data System (ADS)

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

2016-07-01

Today the increasing cancer incidence rate is becoming one of the biggest threats to human health. Among all types of cancers, liver cancer ranks in the top five in both frequency and mortality rate all over the world. During the development of liver cancer, fibrosis often evolves as part of a healing process in response to liver damage, resulting in cirrhosis of liver tissues. In a previous study, we applied the Mueller matrix microscope to pathological liver tissue samples and found that both the Mueller matrix polar decomposition (MMPD) and Mueller matrix transformation (MMT) parameters are closely related to the fibrous microstructures. In this paper, we take this one step further to quantitatively facilitate the fibrosis detections and scorings of pathological liver tissue samples in different stages from cirrhosis to cancer using the Mueller matrix microscope. The experimental results of MMPD and MMT parameters for the fibrotic liver tissue samples in different stages are measured and analyzed. We also conduct Monte Carlo simulations based on the sphere birefringence model to examine in detail the influence of structural changes in different fibrosis stages on the imaging parameters. Both the experimental and simulated results indicate that the polarized light microscope and transformed Mueller matrix parameters can provide additional quantitative information helpful for fibrosis detections and scorings of liver cirrhosis and cancers. Therefore, the polarized light microscope and transformed Mueller matrix parameters have a good application prospect in liver cancer diagnosis.

Application of microscopy in authentication of traditional Tibetan medicinal plants of five Rhodiola (Crassulaceae) alpine species by comparative anatomy and micromorphology.

PubMed

Li, Tao; Zhang, Hao

2008-06-01

A comparative analysis was undertaken to conduct an anatomical and micromorphological study of five species of Rhodiola-R. kirilowii, R. yunnanensis, R. crenulata, R. fastigata, and R. quadrifida-collected from the western Sichuan province plateau of China. Rhodiola plants are a popularly used ethnodrug from the Qinghai-Tibetan plateau of China. Modern studies have shown that the plants of Rhodiola possess different pharmacological activities, chemical constituents, and efficiencies in clinical application. To distinguish five main species of Rhodiola and ensure their safety and efficacy, microscopic characteristics of roots, rhizomes, and stems, including transverse sections, stem and foliar epidermis, as well as the crude drug powder, were observed. The fixed, sectioned, and stained plant materials, as well as the crude powder, were studied using a light microscope according to the usual microscopic techniques. The results of the microscopic features were systematically and comparatively described and illustrated. The five species have distinct microscopic characteristic differences, thus allowing us to distinguish between the species. Also, semi-quantitative and quantitative micrographic parameter tables were simultaneously presented. Further, a key to the five species and a comparative chart of the key authentication parameters based on these anatomic characteristics analyzed was drawn up and is presented for the Rhodiola species studied. The study indicated that light microscopy and related techniques provide a method that is convenient, feasible, and can be unambiguously applied to the authentication of species of Rhodiola. (c) 2008 Wiley-Liss, Inc.

Identification of mycobacterium tuberculosis in sputum smear slide using automatic scanning microscope

NASA Astrophysics Data System (ADS)

Rulaningtyas, Riries; Suksmono, Andriyan B.; Mengko, Tati L. R.; Saptawati, Putri

2015-04-01

Sputum smear observation has an important role in tuberculosis (TB) disease diagnosis, because it needs accurate identification to avoid high errors diagnosis. In development countries, sputum smear slide observation is commonly done with conventional light microscope from Ziehl-Neelsen stained tissue and it doesn't need high cost to maintain the microscope. The clinicians do manual screening process for sputum smear slide which is time consuming and needs highly training to detect the presence of TB bacilli (mycobacterium tuberculosis) accurately, especially for negative slide and slide with less number of TB bacilli. For helping the clinicians, we propose automatic scanning microscope with automatic identification of TB bacilli. The designed system modified the field movement of light microscope with stepper motor which was controlled by microcontroller. Every sputum smear field was captured by camera. After that some image processing techniques were done for the sputum smear images. The color threshold was used for background subtraction with hue canal in HSV color space. Sobel edge detection algorithm was used for TB bacilli image segmentation. We used feature extraction based on shape for bacilli analyzing and then neural network classified TB bacilli or not. The results indicated identification of TB bacilli that we have done worked well and detected TB bacilli accurately in sputum smear slide with normal staining, but not worked well in over staining and less staining tissue slide. However, overall the designed system can help the clinicians in sputum smear observation becomes more easily.

Evaluation of Light-Activated Provisional Resin Materials for Periodontal Soft Tissue Management

PubMed Central

Jun, Soo-Kyung; Lee, Hae-Hyoung

2016-01-01

The purpose of this study was to determine mechanical properties using a compressive test with cylinder specimen (h = 6 mm and ϕ = 4 mm) as well as cytotoxicity using elutes from disk specimen (ϕ = 10 mm and h = 2 mm) against human gingival fibroblasts and oral keratinocytes with light-activated provisional resin materials (Revotek LC and Luxatemp Solar) compared to chemically activated counterpart (Snap, Trim II, and Jet). Significantly increased compressive strength (210~280 MPa) was detected in light-activated products compared to chemically activated ones (20~65 MPa, P < 0.05) and similar compressive modulus was detected in both types (0.8~1.5 and 0.5~1.3 GPa). Simultaneously, the light-activated products showed less adverse effects on the periodontal soft tissue cells in any polymerization stage compared to the chemically activated products. Particularly, chemically activated products had significantly greater adverse effects during the “polymerizing” phase compared to those that were “already set” (P < 0.05), as shown in confocal microscopic images of live and dead cells. In conclusion, light-activated provisional resin materials have better mechanical properties as well as biocompatibility against two tested types of oral cells compared to the chemically activated counterpart, which are considered as more beneficial choice for periodontal soft tissue management. PMID:27672651

Evaluation of Light-Activated Provisional Resin Materials for Periodontal Soft Tissue Management.

PubMed

Jun, Soo-Kyung; Lee, Hae-Hyoung; Lee, Jung-Hwan

The purpose of this study was to determine mechanical properties using a compressive test with cylinder specimen ( h = 6 mm and ϕ = 4 mm) as well as cytotoxicity using elutes from disk specimen ( ϕ = 10 mm and h = 2 mm) against human gingival fibroblasts and oral keratinocytes with light-activated provisional resin materials (Revotek LC and Luxatemp Solar) compared to chemically activated counterpart (Snap, Trim II, and Jet). Significantly increased compressive strength (210~280 MPa) was detected in light-activated products compared to chemically activated ones (20~65 MPa, P < 0.05) and similar compressive modulus was detected in both types (0.8~1.5 and 0.5~1.3 GPa). Simultaneously, the light-activated products showed less adverse effects on the periodontal soft tissue cells in any polymerization stage compared to the chemically activated products. Particularly, chemically activated products had significantly greater adverse effects during the "polymerizing" phase compared to those that were "already set" ( P < 0.05), as shown in confocal microscopic images of live and dead cells. In conclusion, light-activated provisional resin materials have better mechanical properties as well as biocompatibility against two tested types of oral cells compared to the chemically activated counterpart, which are considered as more beneficial choice for periodontal soft tissue management.

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

PubMed

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

2013-10-21

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

Fractal propagation method enables realistic optical microscopy simulations in biological tissues

PubMed Central

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

2017-01-01

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

Plum pudding random medium model of biological tissue toward remote microscopy from spectroscopic light scattering

PubMed Central

Xu, Min

2017-01-01

Biological tissue has a complex structure and exhibits rich spectroscopic behavior. There has been no tissue model until now that has been able to account for the observed spectroscopy of tissue light scattering and its anisotropy. Here we present, for the first time, a plum pudding random medium (PPRM) model for biological tissue which succinctly describes tissue as a superposition of distinctive scattering structures (plum) embedded inside a fractal continuous medium of background refractive index fluctuation (pudding). PPRM faithfully reproduces the wavelength dependence of tissue light scattering and attributes the “anomalous” trend in the anisotropy to the plum and the powerlaw dependence of the reduced scattering coefficient to the fractal scattering pudding. Most importantly, PPRM opens up a novel venue of quantifying the tissue architecture and microscopic structures on average from macroscopic probing of the bulk with scattered light alone without tissue excision. We demonstrate this potential by visualizing the fine microscopic structural alterations in breast tissue (adipose, glandular, fibrocystic, fibroadenoma, and ductal carcinoma) deduced from noncontact spectroscopic measurement. PMID:28663913

Dynamic views of living cell fine structure revealed by birefringence imaging

NASA Astrophysics Data System (ADS)

Oldenbourg, Rudolf

2001-11-01

We have been developing and applying a new type of polarized light microscope, the new Pol-Scope, which dramatically enhances the unique capabilities of the traditional polarizing microscope. In living cells, without applying exogenous dyes or florescent labels, we have studied the dynamic organization of filamentous actin in neuronal growth cones and improved the efficiency of spindle imaging for in-vitro fertilization and enucleation procedures.

In vivo imaging of cerebral hemodynamics and tissue scattering in rat brain using a surgical microscope camera system

NASA Astrophysics Data System (ADS)

Nishidate, Izumi; Kanie, Takuya; Mustari, Afrina; Kawauchi, Satoko; Sato, Shunichi; Sato, Manabu; Kokubo, Yasuaki

2018-02-01

We investigated a rapid imaging method to monitor the spatial distribution of total hemoglobin concentration (CHbT), the tissue oxygen saturation (StO2), and the scattering power b in the expression of musp=a(lambda)^-b as the scattering parameters in cerebral cortex using a digital red-green-blue camera. In the method, Monte Carlo simulation (MCS) for light transport in brain tissue is used to specify a relation among the RGB-values and the concentration of oxygenated hemoglobin (CHbO), that of deoxygenated hemoglobin (CHbR), and the scattering power b. In the present study, we performed sequential recordings of RGB images of in vivo exposed brain of rats while changing the fraction of inspired oxygen (FiO2), using a surgical microscope camera system. The time courses of CHbO, CHbR, CHbT, and StO2 indicated the well-known physiological responses in cerebral cortex. On the other hand, a fast decrease in the scattering power b was observed immediately after the respiratory arrest, which is similar to the negative deflection of the extracellular DC potential so-called anoxic depolarization. It is said that the DC shift coincident with a rise in extracellular potassium and can evoke cell deformation generated by water movement between intracellular and extracellular compartments, and hence the light scattering by tissue. Therefore, the decrease in the scattering power b after the respiratory arrest is indicative of changes in light scattering by tissue. The results in this study indicate potential of the method to evaluate the pathophysiological conditions and loss of tissue viability in brain tissue.

Improvement of two-photon microscopic imaging in deep regions of living mouse brains by utilizing a light source based on an electrically controllable gain-switched laser diode

NASA Astrophysics Data System (ADS)

Sawada, Kazuaki; Kawakami, Ryosuke; Fang, Yi-Cheng; Hung, Jui-Hung; Kozawa, Yuichi; Otomo, Kohei; Sato, Shunichi; Yokoyama, Hiroyuki; Nemoto, Tomomi

2018-02-01

In vivo two-photon microscopy is an advantageous technique for observing living mouse brains at high spatial resolutions. We previously used a 1064 nm high-power light source based on an electrically controllable gain-switched laser diode (maximum power: 4 W, repetition rate: 10 MHz, pulse width: 7.5 picoseconds) and successfully visualized EYFP expressing neurons at deeper regions in H-line mouse brains under living conditions. However, severe damages were frequently observed when the laser power after the objective lens was over 600 mW, suggesting that a higher average power might not be suitable for visualizing neural structures and functions at deep regions. To increase fluorescent signals as a strategy to avoid such invasions, here, we evaluated the effects of the excitation laser parameters such as the repetition rate (5 - 10 MHz), or the peak power, at the moderate average powers (10 - 500 mW), by taking the advantage that this electrically controllable light source could be used to change the repetition rate independently from the average power or the pulse width. The fluorescent signals of EYFP at layer V of the cerebral cortex were increased by approximately twofold when the repetition rate was decreased from 10 MHz to 5 MHz at the same average power. We also confirmed similar effects in the EYFP solution (335 μM) and fixed brain slices. These results suggest that in vivo two-photon microscopic imaging might be improved by increasing the peak power at the same average power while avoiding the severe damages in living brains.

Modulus design multiwavelength polarization microscope for transmission Mueller matrix imaging

NASA Astrophysics Data System (ADS)

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

2018-01-01

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

Light field creating and imaging with different order intensity derivatives

NASA Astrophysics Data System (ADS)

Wang, Yu; Jiang, Huan

2014-10-01

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

Bessel light sheet structured illumination microscopy

NASA Astrophysics Data System (ADS)

Noshirvani Allahabadi, Golchehr

Biomedical study researchers using animals to model disease and treatment need fast, deep, noninvasive, and inexpensive multi-channel imaging methods. Traditional fluorescence microscopy meets those criteria to an extent. Specifically, two-photon and confocal microscopy, the two most commonly used methods, are limited in penetration depth, cost, resolution, and field of view. In addition, two-photon microscopy has limited ability in multi-channel imaging. Light sheet microscopy, a fast developing 3D fluorescence imaging method, offers attractive advantages over traditional two-photon and confocal microscopy. Light sheet microscopy is much more applicable for in vivo 3D time-lapsed imaging, owing to its selective illumination of tissue layer, superior speed, low light exposure, high penetration depth, and low levels of photobleaching. However, standard light sheet microscopy using Gaussian beam excitation has two main disadvantages: 1) the field of view (FOV) of light sheet microscopy is limited by the depth of focus of the Gaussian beam. 2) Light-sheet images can be degraded by scattering, which limits the penetration of the excitation beam and blurs emission images in deep tissue layers. While two-sided sheet illumination, which doubles the field of view by illuminating the sample from opposite sides, offers a potential solution, the technique adds complexity and cost to the imaging system. We investigate a new technique to address these limitations: Bessel light sheet microscopy in combination with incoherent nonlinear Structured Illumination Microscopy (SIM). Results demonstrate that, at visible wavelengths, Bessel excitation penetrates up to 250 microns deep in the scattering media with single-side illumination. Bessel light sheet microscope achieves confocal level resolution at a lateral resolution of 0.3 micron and an axial resolution of 1 micron. Incoherent nonlinear SIM further reduces the diffused background in Bessel light sheet images, resulting in confocal quality images in thick tissue. The technique was applied to live transgenic zebra fish tg(kdrl:GFP), and the sub-cellular structure of fish vasculature genetically labeled with GFP was captured in 3D. The superior speed of the microscope enables us to acquire signal from 200 layers of a thick sample in 4 minutes. The compact microscope uses exclusively off-the-shelf components and offers a low-cost imaging solution for studying small animal models or tissue samples.

Optimisation approaches for concurrent transmitted light imaging during confocal microscopy.

PubMed

Collings, David A

2015-01-01

The transmitted light detectors present on most modern confocal microscopes are an under-utilised tool for the live imaging of plant cells. As the light forming the image in this detector is not passed through a pinhole, out-of-focus light is not removed. It is this extended focus that allows the transmitted light image to provide cellular and organismal context for fluorescence optical sections generated confocally. More importantly, the transmitted light detector provides images that have spatial and temporal registration with the fluorescence images, unlike images taken with a separately-mounted camera. Because plants often provide difficulties for taking transmitted light images, with the presence of pigments and air pockets in leaves, this study documents several approaches to improving transmitted light images beginning with ensuring that the light paths through the microscope are correctly aligned (Köhler illumination). Pigmented samples can be imaged in real colour using sequential scanning with red, green and blue lasers. The resulting transmitted light images can be optimised and merged in ImageJ to generate colour images that maintain registration with concurrent fluorescence images. For faster imaging of pigmented samples, transmitted light images can be formed with non-absorbed wavelengths. Transmitted light images of Arabidopsis leaves expressing GFP can be improved by concurrent illumination with green and blue light. If the blue light used for YFP excitation is blocked from the transmitted light detector with a cheap, coloured glass filters, the non-absorbed green light will form an improved transmitted light image. Changes in sample colour can be quantified by transmitted light imaging. This has been documented in red onion epidermal cells where changes in vacuolar pH triggered by the weak base methylamine result in measurable colour changes in the vacuolar anthocyanin. Many plant cells contain visible levels of pigment. The transmitted light detector provides a useful tool for documenting and measuring changes in these pigments while maintaining registration with confocal imaging.

Light and life in Baltimore--and beyond.

PubMed

Edidin, Michael

2015-02-03

Baltimore has been the home of numerous biophysical studies using light to probe cells. One such study, quantitative measurement of lateral diffusion of rhodopsin, set the standard for experiments in which recovery after photobleaching is used to measure lateral diffusion. Development of this method from specialized microscopes to commercial scanning confocal microscopes has led to widespread use of the technique to measure lateral diffusion of membrane proteins and lipids, and as well diffusion and binding interactions in cell organelles and cytoplasm. Perturbation of equilibrium distributions by photobleaching has also been developed into a robust method to image molecular proximity in terms of fluorescence resonance energy transfer between donor and acceptor fluorophores. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

[Electron microscopic study of the An-750 strain of Powassan virus isolated in the Soviet Union].

PubMed

Sobolev, S G; Shestopalova, N M; Linev, M B; Rubin, S G

1978-01-01

Electron microscopic examinations of brains of white mice inoculated with the An 750 strain isolated for the first time from adult mosquitoes and with the prototype LB strain of Powassan virus were carried out. The method of combination of light and electron microscopy used in the study permitted to compare ultrastructural changes in one cell with the results of light microscopy. Sizes of virions and their localizations in the brain cells were determined. Virus particles were found in large and small neurons as well as in glial elements. Subcellular changes in neurons associated with virus multiplication are described. The causes of differences in sizes of virions measured in ultrathin sections are discussed.

Three-dimensional microscopic tomographic imagings of the cataract in a human lens in vivo

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1998-10-01

The problem of three-dimensional visualization of a human lens in vivo has been solved by a technique of volume rendering a transformed series of 60 rotated Scheimpflug (a dual slit reflected light microscope) digital images. The data set was obtained by rotating the Scheimpflug camera about the optic axis of the lens in 3 degree increments. The transformed set of optical sections were first aligned to correct for small eye movements, and then rendered into a volume reconstruction with volume rendering computer graphics techniques. To help visualize the distribution of lens opacities (cataracts) in the living, human lens the intensity of light scattering was pseudocolor coded and the cataract opacities were displayed as a movie.

Effect of storage temperature on quality of light and full-fat ice cream.

PubMed

Buyck, J R; Baer, R J; Choi, J

2011-05-01

Ice cream quality is dependent on many factors including storage temperature. Currently, the industry standard for ice cream storage is -28.9 °C. Ice cream production costs may be decreased by increasing the temperature of the storage freezer, thus lowering energy costs. The first objective of this research was to evaluate the effect of 4 storage temperatures on the quality of commercial vanilla-flavored light and full-fat ice cream. Storage temperatures used were -45.6, -26.1, and -23.3 °C for the 3 treatments and -28.9 °C as the control or industry standard. Ice crystal sizes were analyzed by a cold-stage microscope and image analysis at 1, 19.5, and 39 wk of storage. Ice crystal size did not differ among the storage temperatures of light and full-fat ice creams at 19.5 or 39 wk. An increase in ice crystal size was observed between 19.5 and 39 wk for all storage temperatures except -45.6 °C. Coldness intensity, iciness, creaminess, and storage/stale off-flavor of the light and full-fat ice creams were evaluated at 39 wk of storage. Sensory evaluation indicated no difference among the different storage temperatures for light and full-fat ice creams. In a second study, light and full-fat ice creams were heat shocked by storing at -28.9 °C for 35 wk and then alternating between -23.3 and -12.2 °C every 24h for 4 wk. Heat-shocked ice creams were analyzed at 2 and 4 wk of storage for ice crystal size and were evaluated by the sensory panel. A difference in ice crystal size was observed for light and full-fat ice creams during heat-shock storage; however, sensory results indicated no differences. In summary, storage of light or full-fat vanilla-flavored ice creams at the temperatures used within this research did not affect quality of the ice creams. Therefore, ice cream manufacturers could conserve energy by increasing the temperature of freezers from -28.9 to -26.1 °C. Because freezers will typically fluctuate from the set temperature, usage of -26.1 °C allows for a safety factor, even though storage at -23.3 °C did not affect ice cream quality. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Non-label bioimaging utilizing scattering lights

NASA Astrophysics Data System (ADS)

Watanabe, Tomonobu M.; Ichimura, Taro; Fujita, Hideaki

2017-04-01

Optical microscopy is an indispensable tool for medical and life sciences. Especially, the microscopes utilized with scattering light offer a detailed internal observation of living specimens in real time because of their non-labeling and non-invasive capability. We here focus on two kinds of scattering lights, Raman scattering light and second harmonic generation light. Raman scattering light includes the information of all the molecular vibration modes of the molecules, and can be used to distinguish types and/or state of cell. Second harmonic generation light is derived from electric polarity of proteins in the specimen, and enables to detect their structural change. In this conference, we would like to introduce our challenges to extract biological information from those scattering lights.

Subsite Awareness in Neuropathology Evaluation of National Toxicology Program (NTP) Studies: A Review of Select Neuroanatomical Structures with their Functional Significance in Rodents

PubMed Central

Rao, Deepa B.; Little, Peter B.; Sills, Robert

2013-01-01

This review manuscript is designed to serve as an introductory guide in neuroanatomy for toxicologic pathologists evaluating general toxicity studies. The manuscript provides an overview of approximately 50 neuroanatomical subsites and their functional significance across seven coronal sections of the brain. Also reviewed are three sections of the spinal cord, cranial and peripheral nerves (trigeminal and sciatic respectively), and intestinal autonomic ganglia. The review is limited to the evaluation of hematoxylin and eosin (H&E) stained tissue sections, as light microscopic evaluation of these sections is an integral part of the first-tier toxicity screening of environmental chemicals, drugs, and other agents. Prominent neuroanatomical sites associated with major neurological disorders are noted. This guide, when used in conjunction with detailed neuroanatomic atlases may aid in an understanding of the significance of functional neuroanatomy, thereby improving the characterization of neurotoxicity in general toxicity and safety evaluation studies. PMID:24135464

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

PubMed

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

2017-02-01

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

Middle School Science Notes

ERIC Educational Resources Information Center

School Science Review, 1976

1976-01-01

Describes a lighted demonstration apparatus for representing the distribution of electrons, protons and neutrons in an atom. Also includes experiments with ice, forces, microscopes, spectra, and geological modeling. (CS)

Generation of drugs coated iron nanoparticles through high energy ball milling

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

Radhika Devi, A.; Murty, B. S.; Chelvane, J. A.

The iron nanoparticles coated with oleic acid and drugs such as folic acid/Amoxicillin were synthesized by high energy ball milling and characterized by X-ray diffraction, Transmission electron microscope, zeta potential, dynamic light scattering, Fourier Transform Infra red (FT-IR) measurements, and thermo gravimetric analysis (TGA). FT-IR and TGA measurements show good adsorption of drugs on oleic acid coated nanoparticles. Magnetic measurements indicate that saturation magnetization is larger for amoxicillin coated particles compared to folic acid coated particles. The biocompatibility of the magnetic nanoparticles prepared was evaluated by in vitro cytotoxicity assay using L929 cells as model cells.

Aerophagia induced by the nasal obstruction on experimental animals.

PubMed

Nakajima, K; Ohi, G

1977-04-01

The excessive accumulation of gas in the gastrointestinal tracts was invariably induced on experimental animals (mice, rats, guinea pigs, hamsters and rabbits) by simply obstructing nasal passages. The analysis of the gas showed the almost identical composition to the ambient air or flutus which was largely due to swallowed air. Also the numerous small foams were found on and underneath the epithelial lining of small intestine. The pathological evaluation was done both macroscopically and microscopically. Dying animals after nasal obstruction showed hemorrhagic and necrotic changes in the jejunum and ileum. This observation may cast some light to the pathogenesis of necrotizing enterocolitis in human neonatal.

Fluorescein sodium-guided surgery of a brain abscess: A case report

PubMed Central

Höhne, Julius; Brawanski, Alexander; Schebesch, Karl-Michael

2016-01-01

Background: Up to now, the feasibility and benefit of using fluorescein sodium under a dedicated surgical microscope filter (YE560, YELLOW 560 nm filter, Carl Zeiss Meditec, Germany) has never been clinically evaluated in infectious disorders of the brain. Case Description: Here, we report the case of a male patient with a brain abscess in the right parietal lobe that was removed under fluorescence-guidance (intravenous administration of fluorescein sodium 10%, 5 mg/kg bodyweight). The abscess capsule showed intensive yellow fluorescent staining, while − under white light − the cortex appeared normal. Conclusion: This technique may improve the identification and surgical removal of brain abscesses. PMID:28031990

Comparison of the killing effects between nitrogen-doped and pure TiO2 on HeLa cells with visible light irradiation

PubMed Central

2013-01-01

The killing effect of nitrogen-doped titanium dioxide (N-TiO2) nanoparticles on human cervical carcinoma (HeLa) cells by visible light photodynamic therapy (PDT) was higher than that of TiO2 nanoparticles. To study the mechanism of the killing effect, the reactive oxygen species produced by the visible-light-activated N-TiO2 and pure-TiO2 were evaluated and compared. The changes of the cellular parameters, such as the mitochondrial membrane potential (MMP), intracellular Ca2+, and nitrogen monoxide (NO) concentrations after PDT were measured and compared for N-TiO2- and TiO2-treated HeLa cells. The N-TiO2 resulted in more loss of MMP and higher increase of Ca2+ and NO in HeLa cells than pure TiO2. The cell morphology changes with time were also examined by a confocal microscope. The cells incubated with N-TiO2 exhibited serious distortion and membrane breakage at 60 min after the PDT. PMID:23433090

Light Microscopy Module Imaging Tested and Demonstrated

NASA Technical Reports Server (NTRS)

Gati, Frank

2004-01-01

The Fluids Integrated Rack (FIR), a facility-class payload, and the Light Microscopy Module (LMM), a subrack payload, are integrated research facilities that will fly in the U.S. Laboratory module, Destiny, aboard the International Space Station. Both facilities are being engineered, designed, and developed at the NASA Glenn Research Center by Northrop Grumman Information Technology. The FIR is a modular, multiuser scientific research facility that is one of two racks that make up the Fluids and Combustion Facility (the other being the Combustion Integrated Rack). The FIR has a large volume dedicated for experimental hardware; easily reconfigurable diagnostics, power, and data systems that allow for unique experiment configurations; and customizable software. The FIR will also provide imagers, light sources, power management and control, command and data handling for facility and experiment hardware, and data processing and storage. The first payload in the FIR will be the LMM. The LMM integrated with the FIR is a remotely controllable, automated, on-orbit microscope subrack facility, with key diagnostic capabilities for meeting science requirements--including video microscopy to observe microscopic phenonema and dynamic interactions, interferometry to make thin-film measurements with nanometer resolution, laser tweezers to manipulate micrometer-sized particles, confocal microscopy to provide enhanced three-dimensional visualization of structures, and spectrophotometry to measure the photonic properties of materials. Vibration disturbances were identified early in the LMM development phase as a high risk for contaminating the science microgravity environment. An integrated FIR-LMM test was conducted in Glenn's Acoustics Test Laboratory to assess mechanical sources of vibration and their impact to microscopic imaging. The primary purpose of the test was to characterize the LMM response at the sample location, the x-y stage within the microscope, to vibration emissions from the FIR and LMM support structures.

Wide-field computational imaging of pathology slides using lens-free on-chip microscopy.

PubMed

Greenbaum, Alon; Zhang, Yibo; Feizi, Alborz; Chung, Ping-Luen; Luo, Wei; Kandukuri, Shivani R; Ozcan, Aydogan

2014-12-17

Optical examination of microscale features in pathology slides is one of the gold standards to diagnose disease. However, the use of conventional light microscopes is partially limited owing to their relatively high cost, bulkiness of lens-based optics, small field of view (FOV), and requirements for lateral scanning and three-dimensional (3D) focus adjustment. We illustrate the performance of a computational lens-free, holographic on-chip microscope that uses the transport-of-intensity equation, multi-height iterative phase retrieval, and rotational field transformations to perform wide-FOV imaging of pathology samples with comparable image quality to a traditional transmission lens-based microscope. The holographically reconstructed image can be digitally focused at any depth within the object FOV (after image capture) without the need for mechanical focus adjustment and is also digitally corrected for artifacts arising from uncontrolled tilting and height variations between the sample and sensor planes. Using this lens-free on-chip microscope, we successfully imaged invasive carcinoma cells within human breast sections, Papanicolaou smears revealing a high-grade squamous intraepithelial lesion, and sickle cell anemia blood smears over a FOV of 20.5 mm(2). The resulting wide-field lens-free images had sufficient image resolution and contrast for clinical evaluation, as demonstrated by a pathologist's blinded diagnosis of breast cancer tissue samples, achieving an overall accuracy of ~99%. By providing high-resolution images of large-area pathology samples with 3D digital focus adjustment, lens-free on-chip microscopy can be useful in resource-limited and point-of-care settings. Copyright © 2014, American Association for the Advancement of Science.

Direct observation of the actin filament by tip-scan atomic force microscopy

PubMed Central

Narita, Akihiro; Usukura, Eiji; Yagi, Akira; Tateyama, Kiyohiko; Akizuki, Shogo; Kikumoto, Mahito; Matsumoto, Tomoharu; Maéda, Yuichiro; Ito, Shuichi; Usukura, Jiro

2016-01-01

Actin filaments, the actin–myosin complex and the actin–tropomyosin complex were observed by a tip-scan atomic force microscope (AFM), which was recently developed by Olympus as the AFM part of a correlative microscope. This newly developed AFM uses cantilevers of similar size as stage-scan AFMs to improve substantially the spatial and temporal resolution. Such an approach has previously never been possible by a tip-scan system, in which a cantilever moves in the x, y and z directions. We evaluated the performance of this developed tip-scan AFM by observing the molecular structure of actin filaments and the actin–tropomyosin complex. In the image of the actin filament, the molecular interval of the actin subunits (∼5.5 nm) was clearly observed as stripes. From the shape of the stripes, the polarity of the actin filament was directly determined and the results were consistent with the polarity determined by myosin binding. In the image of the actin–tropomyosin complex, each tropomyosin molecule (∼2 nm in diameter) on the actin filament was directly observed without averaging images of different molecules. Each tropomyosin molecule on the actin filament has never been directly observed by AFM or electron microscopy. Thus, our developed tip-scan AFM offers significant potential in observing purified proteins and cellular structures at nanometer resolution. Current results represent an important step in the development of a new correlative microscope to observe nm-order structures at an acceptable frame rate (∼10 s/frame) by AFM at the position indicated by the fluorescent dye observed under a light microscope. PMID:27242058

Optimising sulfuric acid hard coat anodising for an Al-Mg-Si wrought aluminium alloy

NASA Astrophysics Data System (ADS)

Bartolo, N.; Sinagra, E.; Mallia, B.

2014-06-01

This research evaluates the effects of sulfuric acid hard coat anodising parameters, such as acid concentration, electrolyte temperature, current density and time, on the hardness and thickness of the resultant anodised layers. A small scale anodising facility was designed and set up to enable experimental investigation of the anodising parameters. An experimental design using the Taguchi method to optimise the parameters within an established operating window was performed. Qualitative and quantitative methods of characterisation of the resultant anodised layers were carried out. The anodised layer's thickness, and morphology were determined using a light optical microscope (LOM) and field emission gun scanning electron microscope (FEG-SEM). Hardness measurements were carried out using a nano hardness tester. Correlations between the various anodising parameters and their effect on the hardness and thickness of the anodised layers were established. Careful evaluation of these effects enabled optimum parameters to be determined using the Taguchi method, which were verified experimentally. Anodised layers having hardness varying between 2.4-5.2 GPa and a thickness of between 20-80 μm were produced. The Taguchi method was shown to be applicable to anodising. This finding could facilitate on-going and future research and development of anodising, which is attracting remarkable academic and industrial interest.

Pathological Evaluation of Radiation-Induced Vascular Lesions of the Brain: Distinct from De Novo Cavernous Hemangioma

PubMed Central

Cha, Yoon Jin; Nahm, Ji Hae; Ko, Ji Eun; Shin, Hyun Joo; Chang, Jong-Hee; Cho, Nam Hoon

2015-01-01

Purpose We aimed to evaluate the histologic and radiologic findings of vascular lesions after stereotactic radiosurgery (SRS) categorized as radiation-induced cavernous hemangioma (RICH). Materials and Methods Among 89 patients who underwent neurosurgery for cavernous hemangioma, eight RICHs from 7 patients and 10 de novo CHs from 10 patients were selected for histopathological and radiological comparison. Results Histologically, RICHs showed hematoma-like gross appearance. Microscopically, RICH exhibited a hematoma-like area accompanied by proliferation of thin-walled vasculature with fibrin deposits and infiltrating foamy macrophages. In contrast, CHs demonstrated localized malformed vasculature containing fresh and old clotted blood on gross examination. Typically, CHs consisted of thick, ectatic hyalinized vessels lined by endothelium under a light microscope. Magnetic resonance imaging of RICHs revealed some overlapping but distinct features with CHs, including enhancing cystic and solid components with absence or incomplete popcorn-like appearance and partial hemosiderin rims. Conclusion Together with histologic and radiologic findings, RICH may result from blood-filled space after tissue destruction by SRS, accompanied with radiation-induced reactive changes rather than vascular malformation. Thus, the term "RICH" would be inappropriate, because it is more likely to be an inactive organizing hematoma rather than proliferation of malformed vasculature. PMID:26446658

Large-scale tissue clearing (PACT): Technical evaluation and new perspectives in immunofluorescence, histology, and ultrastructure.

PubMed

Neckel, Peter H; Mattheus, Ulrich; Hirt, Bernhard; Just, Lothar; Mack, Andreas F

2016-09-29

Novel techniques, like CLARITY and PACT, render large tissue specimens transparent and thereby suitable for microscopic analysis. We used these techniques to evaluate their potential in the intestine as an exemplary organ with a complex tissue composition. Immunohistochemistry, light sheet-, and confocal scanning-microscopy enabled us to follow complex three-dimensional structures, like nerve fibers, vessels, and epithelial barriers throughout the entire organ. Moreover, in a systematic electron microscopic study, we analyzed the morphology and preservation of tissue on ultrastructural level during the clearing process. We also connect tissue clearing with classical histology and demonstrate that cleared tissues can be stained with Hematoxylin-Eosin and Heidenhain's Azan stain, suggesting potential use in histopathology. These experiments showed that a neutral pH during the clearing process results in much better preservation of tissue ultrastructure and standard stainability. Volume changes of specimens were monitored and quantified during the course of the protocol. Additionally, we employed the technique to visualize the enteric nervous system and the epithelial barrier in post mortem human gut preparations. Our data show the high potential of tissue clearing throughout different tissue types supporting its usefulness in research and diagnosis, and contribute to the technical discussion of ultrastructural tissue-retention.

Large-scale tissue clearing (PACT): Technical evaluation and new perspectives in immunofluorescence, histology, and ultrastructure

PubMed Central

Neckel, Peter H.; Mattheus, Ulrich; Hirt, Bernhard; Just, Lothar; Mack, Andreas F.

2016-01-01

Novel techniques, like CLARITY and PACT, render large tissue specimens transparent and thereby suitable for microscopic analysis. We used these techniques to evaluate their potential in the intestine as an exemplary organ with a complex tissue composition. Immunohistochemistry, light sheet-, and confocal scanning-microscopy enabled us to follow complex three-dimensional structures, like nerve fibers, vessels, and epithelial barriers throughout the entire organ. Moreover, in a systematic electron microscopic study, we analyzed the morphology and preservation of tissue on ultrastructural level during the clearing process. We also connect tissue clearing with classical histology and demonstrate that cleared tissues can be stained with Hematoxylin-Eosin and Heidenhain’s Azan stain, suggesting potential use in histopathology. These experiments showed that a neutral pH during the clearing process results in much better preservation of tissue ultrastructure and standard stainability. Volume changes of specimens were monitored and quantified during the course of the protocol. Additionally, we employed the technique to visualize the enteric nervous system and the epithelial barrier in post mortem human gut preparations. Our data show the high potential of tissue clearing throughout different tissue types supporting its usefulness in research and diagnosis, and contribute to the technical discussion of ultrastructural tissue-retention. PMID:27680942

Synthesis of carbon-doped nanosheets m-BiVO4 with three-dimensional (3D) hierarchical structure by one-step hydrothermal method and evaluation of their high visible-light photocatalytic property

NASA Astrophysics Data System (ADS)

Zhao, Deqiang; Zong, Wenjuan; Fan, Zihong; Fang, Yue-Wen; Xiong, Shimin; Du, Mao; Wu, Tianhui; Ji, Fangying; Xu, Xuan

2017-04-01

To achieve an efficient visible-light absorption and degradation of bismuth vanadate (BiVO4), in this paper, a carbon-doped (C-doped) nanosheets monoclinic BiVO4 (m-BiVO4), with thicknesses within 19.86 ± 8.48 nm, was synthesized using polyvinylpyrrolidone K-30 (PVP) as a template and l-carbonic as the carbon source by one-step hydrothermal synthesis method. This C-doped BiVO4 in three-dimensional (3D) hierarchical structure enjoys high visible-light photocatalytic property. The samples were characterized using x-ray diffraction, scanning electron microscope, Raman spectra, energy dispersive spectrometer, transmission electron microscope, x-ray photoelectron spectroscopy, UV-Vis diffused reflectance spectroscopy, specific surface area, electron spin resonance, and transient photocurrent response, photoluminescence spectra, and incident-photon-to-current conversion efficiency, respectively. What is more, we studied the C-doping effect on the band-gap energy of BiVO4 based on First-principles. X-ray diffraction analysis showed that all photocatalysts were in the same single monoclinic scheelite structure. According to the other characterization results, the element C was successfully doped in BiVO4, resulting in the 3D hierarchical structure of C-doped BiVO4 (P-L-BiVO4). We speculated that it could be the directional coalescence mechanism by which the l-cysteine promoted the two-dimensional growth and C-doping process of BiVO4, thus leading to the formation of nanosheets which were then promoted into 3D self-assembly by PVP and the shortening of the band gap. Among all samples, P-L-BiVO4 can make the highest removal ratio of rhodamine B under visible-light irradiation. The stability of P-L-BiVO4 was verified by recycle experiments. It showed that P-L-BiVO4 had strong visible-light absorption behavior and high electron-hole separation efficiency and stability, making a significant advantage in actual situation.

Confocal Fluorescence Microscopy of Mung Beanleaves

NASA Astrophysics Data System (ADS)

Chen, Zhiwei; Liu, Dongwu

Recently, confocal microscope has become a routine technique and indispensable tool for cell biological studies and molecular investigations. The light emitted from the point out-of-focus is blocked by the pinhole and can not reach the detector, which is one of the critical features of the confocal microscope. In present studies, the probes acridine orange (AO) and rhodamine-123 were used to research stoma and mitochondria of mung bean leaves, respectively. The results indicated that the stomatal guard cells and mitochondria were clearly seen in epidermic tissue of mung bean leaves. Taken together, it is a good method to research plant cells with confocal microscope and fluorescence probes.

Hyperlens-array-implemented optical microscopy

NASA Astrophysics Data System (ADS)

Iwanaga, Masanobu

2014-08-01

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

Plasma sprayed Fe(76)Nd(16)B(8) permanent magnets

NASA Technical Reports Server (NTRS)

Overfelt, R. A.; Anderson, C. D.; Flanagan, W. F.

1986-01-01

Thin coatings (0.16 mm) and thick coatings (0.50 mm) of Fe(76)Nd(16)B(8) were deposited on stainless-steel substrates by low pressure plasma spraying. Microscopic examination of the coatings in a light microscope revealed excessive porosity, but good bonding to the substrate. Fracture cross sections examined in a scanning electron microscope showed the grains to be equiaxed and approximately 1 micron or less in diameter in the as-sprayed condition. The intrinsic coercivities of the as-sprayed coatings varied from 5.8 to 10.9 kOe. The effects of postspray heat treatments on the intrinsic coercivity are also given.

Parallel-multiplexed excitation light-sheet microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Xu, Dongli; Zhou, Weibin; Peng, Leilei

2017-02-01

Laser scanning light-sheet imaging allows fast 3D image of live samples with minimal bleach and photo-toxicity. Existing light-sheet techniques have very limited capability in multi-label imaging. Hyper-spectral imaging is needed to unmix commonly used fluorescent proteins with large spectral overlaps. However, the challenge is how to perform hyper-spectral imaging without sacrificing the image speed, so that dynamic and complex events can be captured live. We report wavelength-encoded structured illumination light sheet imaging (λ-SIM light-sheet), a novel light-sheet technique that is capable of parallel multiplexing in multiple excitation-emission spectral channels. λ-SIM light-sheet captures images of all possible excitation-emission channels in true parallel. It does not require compromising the imaging speed and is capable of distinguish labels by both excitation and emission spectral properties, which facilitates unmixing fluorescent labels with overlapping spectral peaks and will allow more labels being used together. We build a hyper-spectral light-sheet microscope that combined λ-SIM with an extended field of view through Bessel beam illumination. The system has a 250-micron-wide field of view and confocal level resolution. The microscope, equipped with multiple laser lines and an unlimited number of spectral channels, can potentially image up to 6 commonly used fluorescent proteins from blue to red. Results from in vivo imaging of live zebrafish embryos expressing various genetic markers and sensors will be shown. Hyper-spectral images from λ-SIM light-sheet will allow multiplexed and dynamic functional imaging in live tissue and animals.

Light Microscopy Module (LMM)-Emulator

NASA Technical Reports Server (NTRS)

Levine, Howard G.; Smith, Trent M.; Richards, Stephanie E.

2016-01-01

The Light Microscopy Module (LMM) is a microscope facility developed at Glenn Research Center (GRC) that provides researchers with powerful imaging capability onboard the International Space Station (ISS). LMM has the ability to have its hardware recongured on-orbit to accommodate a wide variety of investigations, with the capability of remotely acquiring and downloading digital images across multiple levels of magnication.

Multimodal Spectral Imaging of Cells Using a Transmission Diffraction Grating on a Light Microscope

PubMed Central

Isailovic, Dragan; Xu, Yang; Copus, Tyler; Saraswat, Suraj; Nauli, Surya M.

2011-01-01

A multimodal methodology for spectral imaging of cells is presented. The spectral imaging setup uses a transmission diffraction grating on a light microscope to concurrently record spectral images of cells and cellular organelles by fluorescence, darkfield, brightfield, and differential interference contrast (DIC) spectral microscopy. Initially, the setup was applied for fluorescence spectral imaging of yeast and mammalian cells labeled with multiple fluorophores. Fluorescence signals originating from fluorescently labeled biomolecules in cells were collected through triple or single filter cubes, separated by the grating, and imaged using a charge-coupled device (CCD) camera. Cellular components such as nuclei, cytoskeleton, and mitochondria were spatially separated by the fluorescence spectra of the fluorophores present in them, providing detailed multi-colored spectral images of cells. Additionally, the grating-based spectral microscope enabled measurement of scattering and absorption spectra of unlabeled cells and stained tissue sections using darkfield and brightfield or DIC spectral microscopy, respectively. The presented spectral imaging methodology provides a readily affordable approach for multimodal spectral characterization of biological cells and other specimens. PMID:21639978

Light and scanning electron microscope investigations comparing calculus removal using an Er:YAG laser and a frequency-doubled alexandrite laser

NASA Astrophysics Data System (ADS)

Rechmann, Peter; Hennig, Thomas; Sadegh, Hamid M. M.; Goldin, Dan S.

1997-05-01

With respect to lasers emitting within the mid-IR spectral domain fiber applicators are being developed. Intended is the use of these lasers in periodontal therapy and their application inside the gingival pocket. Aim of the study presented here is to compare the effect of an Er:YAG laser on dental calculus with the results following irradiation with a frequency doubled Alexandrite laser. The surface of freshly extracted wisdom teeth and of extracted teeth suffering from severe periodontitis were irradiated with both laser wavelengths using a standardized application protocol. Calculus on the enamel surface, at the enamel cementum junction and on the root surface was irradiated. For light microscope investigations undecalcified histological sections were prepared after treatment. For the scanning electron microscope teeth were dried in alcohol and sputtered with gold. Investigations revealed that with both laser systems calculus can be removed. Using the frequency doubled Alexandrite laser selective removal of calculus is possible while engaging the Er:YAG laser even at lowest energies necessary for calculus removal healthy cementum is ablated without control.

Microgravity Foam Structure and Rheology

NASA Technical Reports Server (NTRS)

Durian, Douglas J.

1997-01-01

To exploit rheological and multiple-light scattering techniques, and ultimately microgravity conditions, in order to quantify and elucidate the unusual elastic character of foams in terms of their underlying microscopic structure and dynamics. Special interest is in determining how this elastic character vanishes, i.e. how the foam melts into a simple viscous liquid, as a function of both increasing liquid content and shear strain rate. The unusual elastic character of foams will be quantified macroscopically by measurement of the shear stress as a function of static shear strain, shear strain rate, and time following a step strain; such data will be analyzed in terms of a yield stress, a static shear modulus, and dynamical time scales. Microscopic information about bubble packing and rearrangement dynamics, from which these macroscopic non-Newtonian properties presumably arise, will be obtained non-invasively by novel multiple-light scattering diagnostics such as Diffusing-Wave Spectroscopy (DWS). Quantitative trends with materials parameters, such as average bubble size, and liquid content, will be sought in order to elucidate the fundamental connection between the microscopic structure and dynamics and the macroscopic rheology.

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

NASA Astrophysics Data System (ADS)

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

2017-05-01

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

Light microscopic image analysis system to quantify immunoreactive terminal area apposed to nerve cells.

PubMed

Wu, L C; D'Amelio, F; Fox, R A; Polyakov, I; Daunton, N G

1997-06-06

The present report describes a desktop computer-based method for the quantitative assessment of the area occupied by immunoreactive terminals in close apposition to nerve cells in relation to the perimeter of the cell soma. This method is based on Fast Fourier Transform (FFT) routines incorporated in NIH-Image public domain software. Pyramidal cells of layer V of the somatosensory cortex outlined by GABA immunolabeled terminals were chosen for our analysis. A Leitz Diaplan light microscope was employed for the visualization of the sections. A Sierra Scientific Model 4030 CCD camera was used to capture the images into a Macintosh Centris 650 computer. After preprocessing, filtering was performed on the power spectrum in the frequency domain produced by the FFT operation. An inverse FFT with filter procedure was employed to restore the images to the spatial domain. Pasting of the original image to the transformed one using a Boolean logic operation called 'AND'ing produced an image with the terminals enhanced. This procedure allowed the creation of a binary image using a well-defined threshold of 128. Thus, the terminal area appears in black against a white background. This methodology provides an objective means of measurement of area by counting the total number of pixels occupied by immunoreactive terminals in light microscopic sections in which the difficulties of labeling intensity, size, shape and numerical density of terminals are avoided.

Light microscopic image analysis system to quantify immunoreactive terminal area apposed to nerve cells

NASA Technical Reports Server (NTRS)

Wu, L. C.; D'Amelio, F.; Fox, R. A.; Polyakov, I.; Daunton, N. G.

1997-01-01

The present report describes a desktop computer-based method for the quantitative assessment of the area occupied by immunoreactive terminals in close apposition to nerve cells in relation to the perimeter of the cell soma. This method is based on Fast Fourier Transform (FFT) routines incorporated in NIH-Image public domain software. Pyramidal cells of layer V of the somatosensory cortex outlined by GABA immunolabeled terminals were chosen for our analysis. A Leitz Diaplan light microscope was employed for the visualization of the sections. A Sierra Scientific Model 4030 CCD camera was used to capture the images into a Macintosh Centris 650 computer. After preprocessing, filtering was performed on the power spectrum in the frequency domain produced by the FFT operation. An inverse FFT with filter procedure was employed to restore the images to the spatial domain. Pasting of the original image to the transformed one using a Boolean logic operation called 'AND'ing produced an image with the terminals enhanced. This procedure allowed the creation of a binary image using a well-defined threshold of 128. Thus, the terminal area appears in black against a white background. This methodology provides an objective means of measurement of area by counting the total number of pixels occupied by immunoreactive terminals in light microscopic sections in which the difficulties of labeling intensity, size, shape and numerical density of terminals are avoided.

Generic Features of Tertiary Chromatin Structure as Detected in Natural Chromosomes

PubMed Central

Müller, Waltraud G.; Rieder, Dietmar; Kreth, Gregor; Cremer, Christoph; Trajanoski, Zlatko; McNally, James G.

2004-01-01

Knowledge of tertiary chromatin structure in mammalian interphase chromosomes is largely derived from artificial tandem arrays. In these model systems, light microscope images reveal fibers or beaded fibers after high-density targeting of transactivators to insertional domains spanning several megabases. These images of fibers have lent support to chromonema fiber models of tertiary structure. To assess the relevance of these studies to natural mammalian chromatin, we identified two different ∼400-kb regions on human chromosomes 6 and 22 and then examined light microscope images of interphase tertiary chromatin structure when the regions were transcriptionally active and inactive. When transcriptionally active, these natural chromosomal regions elongated, yielding images characterized by a series of adjacent puncta or “beads”, referred to hereafter as beaded images. These elongated structures required transcription for their maintenance. Thus, despite marked differences in the density and the mode of transactivation, the natural and artificial systems showed similarities, suggesting that beaded images are generic features of transcriptionally active tertiary chromatin. We show here, however, that these images do not necessarily favor chromonema fiber models but can also be explained by a radial-loop model or even a simple nucleosome affinity, random-chain model. Thus, light microscope images of tertiary structure cannot distinguish among competing models, although they do impose key constraints: chromatin must be clustered to yield beaded images and then packaged within each cluster to enable decondensation into adjacent clusters. PMID:15485905

Quantitative luminescence imaging system

DOEpatents

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

1990-08-14

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

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

NASA Astrophysics Data System (ADS)

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

2005-08-01

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

Wavefront image sensor chip

PubMed Central

Cui, Xiquan; Ren, Jian; Tearney, Guillermo J.; Yang, Changhuei

2010-01-01

We report the implementation of an image sensor chip, termed wavefront image sensor chip (WIS), that can measure both intensity/amplitude and phase front variations of a light wave separately and quantitatively. By monitoring the tightly confined transmitted light spots through a circular aperture grid in a high Fresnel number regime, we can measure both intensity and phase front variations with a high sampling density (11 µm) and high sensitivity (the sensitivity of normalized phase gradient measurement is 0.1 mrad under the typical working condition). By using WIS in a standard microscope, we can collect both bright-field (transmitted light intensity) and normalized phase gradient images. Our experiments further demonstrate that the normalized phase gradient images of polystyrene microspheres, unstained and stained starfish embryos, and strongly birefringent potato starch granules are improved versions of their corresponding differential interference contrast (DIC) microscope images in that they are artifact-free and quantitative. Besides phase microscopy, WIS can benefit machine recognition, object ranging, and texture assessment for a variety of applications. PMID:20721059

Light-activated photocurrent degradation and self-healing in perovskite solar cells

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

Nie, Wanyi; Blancon, Jean-Christophe; Neukirch, Amanda J.

Solution-processed organometallic perovskite solar cells have emerged as one of the most promising thin-film photovoltaic technology. But, a key challenge is their lack of stability over prolonged solar irradiation. Few studies have investigated the effect of light soaking on hybrid perovskites and have attributed the degradation in the optoelectronic properties to photochemical or field-assisted ion migration. We show that the slow photocurrent degradation in thin-film photovoltaic devices is due to the formation of light-activated meta-stable deep-level trap states. However, the devices can self-heal completely by resting them in the dark for <1 min or the degradation can be completely preventedmore » by operating the devices at 0 °C. Here, we investigate several physical mechanisms to explain the microscopic origin for the formation of these trap states, among which the creation of small polaronic states involving localized cooperative lattice strain and molecular orientations emerges as a credible microscopic mechanism requiring further detailed studies.« less

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

PubMed Central

Vohnsen, Brian

2014-01-01

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

Gastroesophageal junction of Anatolian shepherd dog; a study by topographic anatomy, scanning electron and light microscopy.

PubMed

Alsafy, M A M; El-Gendy, S A A

2012-03-01

The aim of this study was to cast a spotlight on the topography and to point out the clinical importance of the gastroesophageal junction (GEJ) in Anatolian Shepherd dogs. Nine Anatolian Shepherd dogs were used to study the morphology of the GEJ. The esophagus was appeared has a portion within the thoracic cavity while no portion of the esophagus presented within the abdominal cavity that documented the absence of the intra-abdominal portion in all studied dogs. The topographic anatomy, scanning electron and light microscopic examinations revealed that the gastroesophageal junction was located at the level of the phrenico-esophageal ligament (PEL) inside the esophageal hiatus. Our results were distinguished the morphology of the esophageal and gastric cardiac mucosa at the level of the gastroesophageal junction by the scanning electron micrographs. The light microscopical examination was explained the PEL attached to the esophageal side in one dog and to the gastric cardiac side in three dogs.

Light-activated photocurrent degradation and self-healing in perovskite solar cells

DOE PAGES

Nie, Wanyi; Blancon, Jean-Christophe; Neukirch, Amanda J.; ...

2016-05-16

Solution-processed organometallic perovskite solar cells have emerged as one of the most promising thin-film photovoltaic technology. But, a key challenge is their lack of stability over prolonged solar irradiation. Few studies have investigated the effect of light soaking on hybrid perovskites and have attributed the degradation in the optoelectronic properties to photochemical or field-assisted ion migration. We show that the slow photocurrent degradation in thin-film photovoltaic devices is due to the formation of light-activated meta-stable deep-level trap states. However, the devices can self-heal completely by resting them in the dark for <1 min or the degradation can be completely preventedmore » by operating the devices at 0 °C. Here, we investigate several physical mechanisms to explain the microscopic origin for the formation of these trap states, among which the creation of small polaronic states involving localized cooperative lattice strain and molecular orientations emerges as a credible microscopic mechanism requiring further detailed studies.« less

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

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

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

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

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

DOE PAGES

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

2016-10-25

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

Simultaneous multiview capture and fusion improves spatial resolution in wide-field and light-sheet microscopy

PubMed Central

Wu, Yicong; Chandris, Panagiotis; Winter, Peter W.; Kim, Edward Y.; Jaumouillé, Valentin; Kumar, Abhishek; Guo, Min; Leung, Jacqueline M.; Smith, Corey; Rey-Suarez, Ivan; Liu, Huafeng; Waterman, Clare M.; Ramamurthi, Kumaran S.; La Riviere, Patrick J.; Shroff, Hari

2016-01-01

Most fluorescence microscopes are inefficient, collecting only a small fraction of the emitted light at any instant. Besides wasting valuable signal, this inefficiency also reduces spatial resolution and causes imaging volumes to exhibit significant resolution anisotropy. We describe microscopic and computational techniques that address these problems by simultaneously capturing and subsequently fusing and deconvolving multiple specimen views. Unlike previous methods that serially capture multiple views, our approach improves spatial resolution without introducing any additional illumination dose or compromising temporal resolution relative to conventional imaging. When applying our methods to single-view wide-field or dual-view light-sheet microscopy, we achieve a twofold improvement in volumetric resolution (~235 nm × 235 nm × 340 nm) as demonstrated on a variety of samples including microtubules in Toxoplasma gondii, SpoVM in sporulating Bacillus subtilis, and multiple protein distributions and organelles in eukaryotic cells. In every case, spatial resolution is improved with no drawback by harnessing previously unused fluorescence. PMID:27761486

Mechanism of Prism-Coupled Scanning Tunneling Microscope Light Emission

NASA Astrophysics Data System (ADS)

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

2011-09-01

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

Soft x-ray imaging with incoherent sources

NASA Astrophysics Data System (ADS)

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

2017-05-01

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

Methyl green and nitrotetrazolium blue chloride co-expression in colon tissue: A hyperspectral microscopic imaging analysis

NASA Astrophysics Data System (ADS)

Li, Qingli; Liu, Hongying; Wang, Yiting; Sun, Zhen; Guo, Fangmin; Zhu, Jianzhong

2014-12-01

Histological observation of dual-stained colon sections is usually performed by visual observation under a light microscope, or by viewing on a computer screen with the assistance of image processing software in both research and clinical settings. These traditional methods are usually not sufficient to reliably differentiate spatially overlapping chromogens generated by different dyes. Hyperspectral microscopic imaging technology offers a solution for these constraints as the hyperspectral microscopic images contain information that allows differentiation between spatially co-located chromogens with similar but different spectra. In this paper, a hyperspectral microscopic imaging (HMI) system is used to identify methyl green and nitrotetrazolium blue chloride in dual-stained colon sections. Hyperspectral microscopic images are captured and the normalized score algorithm is proposed to identify the stains and generate the co-expression results. Experimental results show that the proposed normalized score algorithm can generate more accurate co-localization results than the spectral angle mapper algorithm. The hyperspectral microscopic imaging technology can enhance the visualization of dual-stained colon sections, improve the contrast and legibility of each stain using their spectral signatures, which is helpful for pathologist performing histological analyses.

SPIM-fluid: open source light-sheet based platform for high-throughput imaging

PubMed Central

Gualda, Emilio J.; Pereira, Hugo; Vale, Tiago; Estrada, Marta Falcão; Brito, Catarina; Moreno, Nuno

2015-01-01

Light sheet fluorescence microscopy has recently emerged as the technique of choice for obtaining high quality 3D images of whole organisms/embryos with low photodamage and fast acquisition rates. Here we present an open source unified implementation based on Arduino and Micromanager, which is capable of operating Light Sheet Microscopes for automatized 3D high-throughput imaging on three-dimensional cell cultures and model organisms like zebrafish, oriented to massive drug screening. PMID:26601007

Coherence and diffraction limited resolution in microscopic OCT by a unified approach for the correction of dispersion and aberrations

NASA Astrophysics Data System (ADS)

Schulz-Hildebrandt, H.; Münter, Michael; Ahrens, M.; Spahr, H.; Hillmann, D.; König, P.; Hüttmann, G.

2018-03-01

Optical coherence tomography (OCT) images scattering tissues with 5 to 15 μm resolution. This is usually not sufficient for a distinction of cellular and subcellular structures. Increasing axial and lateral resolution and compensation of artifacts caused by dispersion and aberrations is required to achieve cellular and subcellular resolution. This includes defocus which limit the usable depth of field at high lateral resolution. OCT gives access the phase of the scattered light and hence correction of dispersion and aberrations is possible by numerical algorithms. Here we present a unified dispersion/aberration correction which is based on a polynomial parameterization of the phase error and an optimization of the image quality using Shannon's entropy. For validation, a supercontinuum light sources and a costume-made spectrometer with 400 nm bandwidth were combined with a high NA microscope objective in a setup for tissue and small animal imaging. Using this setup and computation corrections, volumetric imaging at 1.5 μm resolution is possible. Cellular and near cellular resolution is demonstrated in porcine cornea and the drosophila larva, when computational correction of dispersion and aberrations is used. Due to the excellent correction of the used microscope objective, defocus was the main contribution to the aberrations. In addition, higher aberrations caused by the sample itself were successfully corrected. Dispersion and aberrations are closely related artifacts in microscopic OCT imaging. Hence they can be corrected in the same way by optimization of the image quality. This way microscopic resolution is easily achieved in OCT imaging of static biological tissues.

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

NASA Astrophysics Data System (ADS)

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.

Structure and function of embryonic rat retinal sheet transplants.

PubMed

Peng, Qing; Thomas, Biju B; Aramant, Robert B; Chen, Zhenhai; Sadda, Srinivas R; Seiler, Magdalene J

2007-09-01

To evaluate retinal sheet transplants in S334ter-line-3 retinal degenerate rats by comparing visual responses recorded electrophysiologically with morphology based on light and electron microscopy. S334ter-line-3 retinal degenerate rats (n = 7) received retinal sheet transplants between postnatal days 28 and 31. The donor tissue was derived from transgenic embryonic day 19 (E19) rat retinae expressing human placental alkaline phosphatase (hPAP). Fresh retinal sheets were gently transplanted into the subretinal space of the left eye with the help of a custom-made implantation tool. Selected rats (n = 5) were subjected to electrophysiologic evaluation of visual responses from the superior colliculus about 84-121 days after surgery. Transplanted eyes were processed for light microscopy (LM) and electron microscopy (EM) evaluations. All the transplanted rats that were evaluated for visual responses in the brain showed responses to very low light stimulation (-3.42 to -2.8 log cd/m(2)) of the eye in a small area of the superior colliculus corresponding with the placement of the transplant in the host retina. Histologic evaluation showed that most of the transplants contained well-laminated areas with correct polarity in the subretinal space. Inside the transplant areas, rosettes of photoreceptors with inner and outer segments were found. In the laminated areas, the outer segments of photoreceptors were facing the host retinal pigment epithelium (RPE). Immunohistochemical evaluation of hPAP donor cells revealed areas with specific staining of the transplants in the subretinal space. Electron microscopic evaluation showed a glial demarcation membrane between the host and the transplant, however, processes originating from the transplant were observed inside the host retina. Sheets of E19 rat retina transplanted into the subretinal space of S334ter-line-3 rats survived without immune rejection and continued to show visual function when tested after 3 months. Well-developed photoreceptors and many synapse types were seen within the transplants. hPAP staining showed a certain degree of integration between the host retina and the transplant suggesting that transplanted photoreceptors contributed to the restored light sensitivity.

Evaluation of the hepatoprotective effect of combination between hinokiflavone and Glycyrrhizin against CCl4 induced toxicity in rats.

PubMed

Abdel-Kader, Maged S; Abulhamd, Ashraf T; Hamad, Abubaker M; Alanazi, Abdullah H; Ali, Rizwan; Alqasoumi, Saleh I

2018-05-01

Liver diseases are one of the fatal syndromes due to the vital role of the liver. Most of the effective treatment of liver conditions are of natural origin. Silymarin (SI) is the standard drug used for treatment of impaired liver functions. Two natural compounds possessing promising liver protection and with different chemical structures namely; the bioflavonoid hinokiflavone (HF) isolated from Junipers phoenicea family Cupressaceae and the sweet saponin Glycyrrhizin (GL) present in  Glycyrrhiza glabra  (liquorice) were selected for the current study. Since the two compounds are of different nature, they may act by different mechanisms and express synergistic effect. Combination of the two compounds using to dose levels were challenged with single doses of HF, GL and SI as well. The comparison was monitored via measuring serum biochemical parameters including, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyltranspeptidase (GGT), alkaline phosphatase (ALP) and total bilirubin, tissue parameters such as MDA, NP-SH and TP, histopathological study using light and electron microscope. Protective effect on kidney was also monitored histopathologically and biochemically through observing the levels of LDH, creatinine, creatinine-kinase, urea and uric acid. The combinations of HF and GL showed protective effect more than the used single doses of HF and GL alone. However, SI was superior to the used combination in the two used doses in all the measured parameters. The liver and kidney cells appearance under normal and electron microscope showed that SI treated groups showed almost normal cells with slight toxic signs. Cells from group treated with the higher doses of the combination of HF and GL showed slight signs of intoxication under light and electron microscope indicating good level of protection. Although the combination of HF and GL expressed good protection in the higher dose, however, the combination did not exceed the protective effect of SI.

Blueberries Inside Popcorn

NASA Image and Video Library

2004-08-18

This view from the microscopic imager on NASA Mars Exploration Rover Opportunity shows a type of light-colored, rough-textured spherules scientists call popcorn in contrast to the darker, smoother spherules called blueberries.

No-anesthesia clear corneal phacoemulsification versus topical and topical plus intracameral anesthesia. Randomized clinical trial.

PubMed

Pandey, S K; Werner, L; Apple, D J; Agarwal, A; Agarwal, A; Agarwal, S

2001-10-01

To compare the intraoperative pain scores during clear corneal phacoemulsification under no anesthesia, topical anesthesia, and topical plus intracameral anesthesia. Dr. Agarwal's Eye Hospital and Eye Research Center, Chennai, India. Seventy-five patients were randomized to have phacoemulsification under no anesthesia, topical anesthesia, or topical plus intracameral anesthesia. Uncooperative or illiterate patients and those with hard cataract, a shallow anterior chamber, or small pupils were excluded. A protocol was established for supplemental anesthesia in case of breakthrough pain during the surgery. Each patient was asked to grade the overall severity of intraoperative pain immediately after surgery on a 10-point visual analog scale. Also evaluated were the general discomfort during surgery, discomfort from the microscope lights, surgeon stress during surgery, and total surgical time. Comparison among the 3 groups was performed using an analysis of variance. No supplemental anesthesia was required in any group. No significant difference was noted in the mean scores of the subjective sensation of pain with or without topical anesthesia (P =.610). The mean scores of patient discomfort from the microscope lights and surgical time were also statistically insignificant. Patient discomfort and surgeon stress during surgery were significantly greater in the no-anesthesia group than in the topical and topical plus intracameral groups (P =.0235 and P = 0.0206, respectively). No-anesthesia clear corneal phacoemulsification was performed by a highly experienced, skilled surgeon without causing an unacceptable level of pain. However, this technique is not suitable for every cataract surgeon or patient.

Early development of fern gametophytes in microgravity

NASA Technical Reports Server (NTRS)

Roux, Stanley J.; Chatterjee, Ani; Hillier, Sheila; Cannon, Tom

2003-01-01

Dormant spores of the fern Ceratopteris richardii were flown on Shuttle mission STS-93 to evaluate the effects of micro-g on their development and on their pattern of gene expression. Prior to flight the spores were sterilized and sown into one of two environments: (1) Microscope slides in a video-microscopy module; and (2) Petri dishes. All spores were then stored in darkness until use. Spore germination was initiated on orbit after exposure to light. For the spores on microscope slides, cell level changes were recorded through the clear spore coat of the spores by video microscopy. After their exposure to light, spores in petri dishes were frozen in orbit at four different time points during which on earth gravity fixes the polarity of their development. Spores were then stored frozen in Biological Research in Canister units until recovery on earth. The RNAs from these cells and from 1-g control cells were extracted and analyzed on earth after flight to assay changes in gene expression. Video microscopy results revealed that the germinated spores developed normally in microgravity, although the polarity of their development, which is guided by gravity on earth, was random in space. Differential Display-PCR analyses of RNA extracted from space-flown cells showed that there was about a 5% change in the pattern of gene expression between cells developing in micro-g compared to those developing on earth. c2002 Published by Elsevier Science Ltd on behalf of COSPAR.

Structural refinement, band-gap analysis and optical properties of GdAlO3 nanophosphors influenced by Dy3+ ion concentrations for white light emitting device applications

NASA Astrophysics Data System (ADS)

Jisha, P. K.; Naik, Ramachandra; Prashantha, S. C.; Nagaswarupa, H. P.; Nagabhushana, H.; Basavaraj, R. B.; Sharma, S. C.; Prasad, Daruka

2016-04-01

Nanosized GdAlO3 phosphors activated with Dy3+ were prepared by a combustion method. Synthesized phosphors were calcined at 1000 °C for 3 h in order to achieve crystallinity. Powder x-ray diffraction (XRD), scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis was used to characterize the prepared product. The orthorhombic phase was observed in the XRD pattern. The particle size of the samples was calculated as around 25 nm. The SEM images show an irregular shape of the prepared nanophosphor. Functional groups of the phosphors were examined by Fourier transform infrared (FTIR) spectroscopy. Photoluminescence (PL) properties of Dy3+ doped GdAlO3 for near-ultraviolet excitation (352 nm) were studied in order to investigate the possibility of its use in white light emitting device applications. Judd-Ofelt intensity parameters, radiative transition rate (A T) and radiative lifetimes (τ rad) were evaluated from the emission spectrum by adopting a standard procedure. The Commission International de l’Eclairage (CIE) color coordinates and correlated color temperature (CCT) are studied for the optimized phosphor. It is found that the color coordinates of Dy3+ doped GdAlO3 powders fall in the white region of the CIE diagram, and the average CCT value was found to be about 6276 K. Therefore, the present phosphor is highly useful for display applications.

Motion compensation for in vivo subcellular optical microscopy.

PubMed

Lucotte, B; Balaban, R S

2014-04-01

In this review, we focus on the impact of tissue motion on attempting to conduct subcellular resolution optical microscopy, in vivo. Our position is that tissue motion is one of the major barriers in conducting these studies along with light induced damage, optical probe loading as well as absorbing and scattering effects on the excitation point spread function and collection of emitted light. Recent developments in the speed of image acquisition have reached the limit, in most cases, where the signal from a subcellular voxel limits the speed and not the scanning rate of the microscope. Different schemes for compensating for tissue displacements due to rigid body and deformation are presented from tissue restriction, gating, adaptive gating and active tissue tracking. We argue that methods that minimally impact the natural physiological motion of the tissue are desirable because the major reason to perform in vivo studies is to evaluate normal physiological functions. Towards this goal, active tracking using the optical imaging data itself to monitor tissue displacement and either prospectively or retrospectively correct for the motion without affecting physiological processes is desirable. Critical for this development was the implementation of near real time image processing in conjunction with the control of the microscope imaging parameters. Clearly, the continuing development of methods of motion compensation as well as significant technological solutions to the other barriers to tissue subcellular optical imaging in vivo, including optical aberrations and overall signal-to-noise ratio, will make major contributions to the understanding of cell biology within the body.

Visualizing individual microtubules by bright field microscopy

NASA Astrophysics Data System (ADS)

Gutiérrez-Medina, Braulio; Block, Steven M.

2010-11-01

Microtubules are slender (˜25 nm diameter), filamentous polymers involved in cellular structure and organization. Individual microtubules have been visualized via fluorescence imaging of dye-labeled tubulin subunits and by video-enhanced, differential interference-contrast microscopy of unlabeled polymers using sensitive CCD cameras. We demonstrate the imaging of unstained microtubules using a microscope with conventional bright field optics in conjunction with a webcam-type camera and a light-emitting diode illuminator. The light scattered by microtubules is image-processed to remove the background, reduce noise, and enhance contrast. The setup is based on a commercial microscope with a minimal set of inexpensive components, suitable for implementation in a student laboratory. We show how this approach can be used in a demonstration motility assay, tracking the gliding motions of microtubules driven by the motor protein kinesin.

Electron microscopic identification of the intestinal protozoan flagellates of the xylophagous cockroach Parasphaeria boleiriana from Brazil.

PubMed

Brugerolle, G; Silva-Neto, I D; Pellens, R; Grandcolas, P

2003-06-01

Flagellate protozoa of the hindgut of the xylophagous blattid Parasphaeria boleiriana were examined by light and electron microscopy. This species harbours two oxymonad species of the genera Monocercomonoides and Polymastix, the latter bearing Fusiformis bacteria on its surface. A diplomonad was present and has features of the genus Hexamita rather than Spironucleus. In addition, two trichomonads of the genera Monocercomonas and Tetratrichomastix were identified. A precise comparison with species of blattids and other insects was difficult because most of these flagellates have been described only by light microscopy after cell staining and there are few electron microscope studies and no molecular studies. None of the flagellates contained wood fragments in their food vacuoles and so evidently do not participate in the digestion of wood or cellulose.

Waveguide bends from nanometric silica wires

NASA Astrophysics Data System (ADS)

Tong, Limin; Lou, Jingyi; Mazur, Eric

2005-02-01

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

High-resolution confocal Raman microscopy using pixel reassignment.

PubMed

Roider, Clemens; Ritsch-Marte, Monika; Jesacher, Alexander

2016-08-15

We present a practical modification of fiber-coupled confocal Raman scanning microscopes that is able to provide high confocal resolution in conjunction with high light collection efficiency. For this purpose, the single detection fiber is replaced by a hexagonal lenslet array in combination with a hexagonally packed round-to-linear multimode fiber bundle. A multiline detector is used to collect individual Raman spectra for each fiber. Data post-processing based on pixel reassignment allows one to improve the lateral resolution by up to 41% compared to a single fiber of equal light collection efficiency. We present results from an experimental implementation featuring seven collection fibers, yielding a resolution improvement of about 30%. We believe that our implementation represents an attractive upgrade for existing confocal Raman microscopes that employ multi-line detectors.

Laser ablated hard coating for microtools

DOEpatents

McLean, II, William; Balooch, Mehdi; Siekhaus, Wigbert J.

1998-05-05

Wear-resistant coatings composed of laser ablated hard carbon films, are deposited by pulsed laser ablation using visible light, on instruments such as microscope tips and micro-surgical tools. Hard carbon, known as diamond-like carbon (DLC), films produced by pulsed laser ablation using visible light enhances the abrasion resistance, wear characteristics, and lifetimes of small tools or instruments, such as small, sharp silicon tips used in atomic probe microscopy without significantly affecting the sharpness or size of these devices. For example, a 10-20 nm layer of diamond-like carbon on a standard silicon atomic force microscope (AFM) tip, enables the useful operating life of the tip to be increased by at least twofold. Moreover, the low inherent friction coefficient of the DLC coating leads to higher resolution for AFM tips operating in the contact mode.

Office-Based Diagnosis of Demodex Using Smartphone.

PubMed

Kaya, Abdullah; Gürdal, Canan

2018-06-25

Demodex is an important pathogen in ophthalmology. It is believed to cause a variety of eyelid and eyelash diseases. Currently, light microscopes are being used for imaging demodex. However, microscopes are not available everywhere. Also, it is not cost-effective to perform light microscopy in every case. In this case, we demonstrate a new method: imaging demodex using cell phone. A 90-diopter noncontact double aspheric lens was attached to the posterior camera of the smartphone with clear tape. An eyelash of a patient with blepharitis was removed. A video was taken using smartphone. There was a moving demodex parasite in the root of the eyelash. A clear video image could be taken using the smartphone. A smartphone and a 90-diopter lens are adequate for the imaging and diagnosis of demodex.

Differential dynamic microscopy to characterize Brownian motion and bacteria motility

NASA Astrophysics Data System (ADS)

Germain, David; Leocmach, Mathieu; Gibaud, Thomas

2016-03-01

We have developed a lab module for undergraduate students, which involves the process of quantifying the dynamics of a suspension of microscopic particles using Differential Dynamic Microscopy (DDM). DDM is a relatively new technique that constitutes an alternative method to more classical techniques such as dynamic light scattering (DLS) or video particle tracking (VPT). The technique consists of imaging a particle dispersion with a standard light microscope and a camera and analyzing the images using a digital Fourier transform to obtain the intermediate scattering function, an autocorrelation function that characterizes the dynamics of the dispersion. We first illustrate DDM in the textbook case of colloids under Brownian motion, where we measure the diffusion coefficient. Then we show that DDM is a pertinent tool to characterize biological systems such as motile bacteria.

Analysis of thick brain sections by obverse-reverse computer microscopy: application of a new, high clarity Golgi-Nissl stain.

PubMed

Glaser, E M; Van der Loos, H

1981-08-01

Exceptionally clear Golgi-Nissl sections of 300 micron thickness have been morphometrically studied by light microscopy using oil immersion objectives. The clarity results from a new variation of a staining procedure that combines Golgi and Nissl images in one section. A viewing technique has been developed that permits a histologic preparation to be examined from its obverse (or normally viewed) side and its reverse (or under) side. The technique was designed for use with a computer microscope but can be employed with any light microscope whose stage position can be measured within 100 micron. Sections thicker than 300 micron can be studied dependent on the working distance of the objective lens, provided that the clarity of the material permits it.

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

NASA Astrophysics Data System (ADS)

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.

3D geometric phase analysis and its application in 3D microscopic morphology measurement

NASA Astrophysics Data System (ADS)

Zhu, Ronghua; Shi, Wenxiong; Cao, Quankun; Liu, Zhanwei; Guo, Baoqiao; Xie, Huimin

2018-04-01

Although three-dimensional (3D) morphology measurement has been widely applied on the macro-scale, there is still a lack of 3D measurement technology on the microscopic scale. In this paper, a microscopic 3D measurement technique based on the 3D-geometric phase analysis (GPA) method is proposed. In this method, with machine vision and phase matching, the traditional GPA method is extended to three dimensions. Using this method, 3D deformation measurement on the micro-scale can be realized using a light microscope. Simulation experiments were conducted in this study, and the results demonstrate that the proposed method has a good anti-noise ability. In addition, the 3D morphology of the necking zone in a tensile specimen was measured, and the results demonstrate that this method is feasible.

AOTF microscope for imaging with increased speed and spectral versatility.

PubMed Central

Wachman, E S; Niu, W; Farkas, D L

1997-01-01

We have developed a new fluorescence microscope that addresses the spectral and speed limitations of current light microscopy instrumentation. In the present device, interference and neutral density filters normally used for fluorescence excitation and detection are replaced by acousto-optic tunable filters (AOTFs). Improvements are described, including the use of a dispersing prism in conjunction with the imaging AOTF and an oblique-illumination excitation scheme, which together enable the AOTF microscope to produce images comparable to those obtained with conventional fluorescence instruments. The superior speed and spectral versatility of the AOTF microscope are demonstrated by a ratio image pair acquired in 3.5 ms and a micro-spectral absorbance measurement of hemoglobin through a cranial window in a living mouse. Images FIGURE 1 FIGURE 2 FIGURE 4 FIGURE 5 FIGURE 6 FIGURE 7 PMID:9284289

The erosive potential of soft drinks on enamel surface substrate: an in vitro scanning electron microscopy investigation.

PubMed

Owens, Barry M; Kitchens, Michael

2007-11-01

Using scanning electron and light microscopy, this study qualitatively evaluated the erosive potential of carbonated cola beverages as well as sports and high-energy drinks on enamel surface substrate. Beverages used in this study included: Coca Cola Classic, Diet Coke, Gatorade sports drink, Red Bull high-energy drink, and tap water (control). Extracted human permanent molars free of hypocalcification and/or caries were used in this study. The coronal portion of each tooth was removed and sectioned longitudinally from the buccal to the lingual surface. The crown sections were embedded in acrylic resin, leaving the enamel surfaces exposed. Following finishing and polishing of all surfaces, one side was covered with red nail varnish while the remaining side was exposed to individual beverage immersion for 14 days, 24 hours per day, at 37 degrees C. The specimens were evaluated for enamel surface changes using scanning electron and light microscopy. Enamel specimens exhibited visual surface changes following immersion in the test beverages with Red Bull and Gatorade revealing the most striking surface morphological changes. Specimens subjected to Coca Cola Classic and Diet Coke immersion also displayed irregular post-treatment surface morphology. As verified by microscopic evaluation, all test beverages displayed enamel dissolution in the following order: Red Bull>Gatorade>Coca-Cola Classic>Diet Coke.

Characterization and identification of microorganisms by FT-IR microspectrometry

NASA Astrophysics Data System (ADS)

Ngo-Thi, N. A.; Kirschner, C.; Naumann, D.

2003-12-01

We report on a novel FT-IR approach for microbial characterization/identification based on a light microscope coupled to an infrared spectrometer which offers the possibility to acquire IR-spectra of microcolonies containing only few hundred cells. Microcolony samples suitable for FT-IR microspectroscopic measurements were obtained by a replica technique with a stamping device that transfers spatially accurate cells of microcolonies growing on solid culture plates to a special, IR-transparent or reflecting stamping plate. High quality spectra could be recorded either by applying the transmission/absorbance or the reflectance/absorbance mode of the infrared microscope. Signal to noise ratios higher than 1000 were obtained for microcolonies as small as 40 μm in diameter. Reproducibility levels were established that allowed species and strain identification. The differentiation and classification capacity of the FT-IR microscopic technique was tested for different selected microorganisms. Cluster and factor analysis methods were used to evaluate the complex spectral data. Excellent discrimination between bacteria and yeasts, and at the same time Gram-negative and Gram-positive bacterial strains was obtained. Twenty-two selected strains of different species within the genus Staphylococcus were repetitively measured and could be grouped into correct species cluster. Moreover, the results indicated that the method allows also identifications at the subspecies level. Additionally, the new approach allowed spectral mapping analysis of single colonies which provided spatially resolved characterization of growth heterogeneity within complex microbial populations such as colonies.

Implementing digital technology to enhance student learning of pathology.

PubMed

Farah, C S; Maybury, T

2009-08-01

The introduction of digital technologies into the dental curriculum is an ongoing feature of broader changes going on in tertiary education. This report examines the introduction of digital virtual microscopy technology into the curriculum of the School of Dentistry at the University of Queensland (UQ) in Brisbane, Australia. Sixty students studying a course in pathology in 2005 were introduced to virtual microscopy technology alongside the more traditional light microscope and then asked to evaluate their own learning outcomes from this technology via a structured 5-point LIKART survey. A wide variety of questions dealing the pedagogic implications of the introduction of virtual microscopy into pathology were asked of students with the overall result being that it positively enhanced their learning of pathology via digital microscopic means. The success of virtual microscopy in dentistry at UQ is then discussed in the larger context of changes going on in tertiary education. In particular, the change from the print-literate tradition to the electronic one, that is from 'literacy to electracy'. Virtual microscopy is designated as a component of this transformation to electracy. Whilst traditional microscopic skills may still be valued in dental curricula, the move to virtual microscopy and computer-assisted, student-centred learning of pathology appears to enhance the learning experience in relation to its effectiveness in helping students engage and interact with the course material.

Evaluation of the antioxidant effects of melatonin on the larynx mucosa of rats exposed to environmental tobacco smoke.

PubMed

Donmez, Z; Yigit, Ö; Bilici, S; Dursun, N; Gul, M; Dastan, S D; Uzun, H

2016-06-01

This study's aim was to investigate the effect of melatonin in terms of mitigating the effects of smoking on the laryngeal mucosa of rats exposed to environmental tobacco smoke. Rats were divided into four groups: Melatonin + Smoking group exposed to smoke with melatonin; Smoking group exposed to smoke without melatonin; Saline group not exposed to smoke without melatonin; Melatonin group not exposed to smoke with melatonin. CuZn-superoxide dismutase (CuZn-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities were evaluated in plasma and tissues. Tissues were also examined the changes of squamous hyperplasia, keratosis, parakeratosis and epithelial hyperplasia by light microscope and the ultrastructural changes by electron microscope. Tissue SOD, CAT and GSH-Px activities were significantly higher in Saline and Melatonin groups than Melatonin + Smoking and Smoking groups. Plasma CuZn-SOD and CAT activities were significantly higher in Saline and Melatonin groups than Smoking group. Plasma GSH-Px showed no significant difference. The rate of epithelial hyperplasia was significantly higher in Smoking group than the other groups. The rate of parakeratosis was significantly higher in Smoking group than the other groups. The epithelial cells in Melatonin + Smoking group displayed, normal cell structure similar to those in Saline group under electron microscope. The study shows that smoking induces substantial pathological changes in the laryngeal mucosa and melatonin may have some beneficial effects in partially reversing smoking-induced laryngeal injury by inducing the expression of antioxidants; biochemical and histological outcomes also support these findings due to preventing tissue damage in laryngeal mucosa exposed to smoke. © 2015 John Wiley & Sons Ltd.

Bond strength and interactions of machined titanium-based alloy with dental cements.

PubMed

Wadhwani, Chandur; Chung, Kwok-Hung

2015-11-01

The most appropriate luting agent for restoring cement-retained implant restorations has yet to be determined. Leachable chemicals from some types of cement designed for teeth may affect metal surfaces. The purpose of this in vitro study was to evaluate the shear bond strength and interactions of machined titanium-based alloy with dental luting agents. Eight dental luting agents representative of 4 different compositional classes (resin, polycarboxylate, glass ionomer, and zinc oxide-based cements) were used to evaluate their effect on machined titanium-6 aluminum-4 vanadium (Ti-6Al-4V) alloy surfaces. Ninety-six paired disks were cemented together (n=12). After incubation in a 37°C water bath for 7 days, the shear bond strength was measured with a universal testing machine (Instron) and a custom fixture with a crosshead speed of 5 mm/min. Differences were analyzed statistically with 1-way ANOVA and Tukey HSD tests (α=.05). The debonded surfaces of the Ti alloy disks were examined under a light microscope at ×10 magnification to record the failure pattern, and the representative specimens were observed under a scanning electron microscope. The mean ±SD of shear failure loads ranged from 3.4 ±0.5 to 15.2 ±2.6 MPa. The retention provided by both polycarboxylate cements was significantly greater than that of all other groups (P<.05). The scanning electron microscope examination revealed surface pits only on the bonded surface cemented with the polycarboxylate cements. Cementation with polycarboxylate cement obtained higher shear bond strength. Some chemical interactions occurred between the machined Ti-6Al-4V alloy surface and polycarboxylate cements during cementation. Copyright © 2015 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Biodegradable polymer film as a source for formation of human fetal retinal pigment epithelium spheroids.

PubMed

Rezai, K A; Farrokh-Siar, L; Botz, M L; Godowski, K C; Swanbom, D D; Patel, S C; Ernest, J T

1999-05-01

To evaluate the attachment of human fetal rctinal pigment epithelial (HFRPE) cells to a biodegradable polymer film with subsequent formation of spheroids in vitro. Ten biodegradable polymer films with different compositions were examined for their physical properties and ease of manipulation under a dissecting microscope. The film with the most suitable handling characteristics was chosen, and a purely isolated sheet of HFRPE cells was attached to it. The purity of the cells was assessed by their pigmentation and expression of cytokeratin. Proliferation was assessed by incorporation of 5-bromo-2'-deoxyuridine (BrdtJ). Cellular structure was analyzed under light and electron microscopes, and the functional capability of the cells was evaluated by rod outer segment (ROS) phagocytosis. The polymer film with composition 50:50 poly (DL-lactide) (PLA)/poly (DL-lactide-co-glycolide) (PLG) with an inherent viscosity of 1.03 dl/g was found to be the most suitable for handling under the microscope. Sheets of HFRPE cells attached to the polymer films within 48 hours and began to form spheroids. All the isolated cells were pigmented and expressed cytokeratin. They possessed a cuboidal morphology, numerous apical microvilli, and no sign of dedifferentiation. HFRPE cells produced extracellular matrix (collagen filaments) on their basal side, filling the cavities of the polymer film. The cells subsequently proliferated, incorporated BrdU, migrated onto the culture plate to form monolayers, and phagocytized ROS. Biodegradable polymer films can be used as a scaffold for the adhesion of the HFRPE sheet and formation of spheroids. Spheroids represent a source of high density and well-differentiated HFRPE cells that are easy to transfer. Furthermore, the stricture of the membrane makes it suitable for additional applications.

Study of Te Inclusion and Related Point Defects in THM-Growth CdMnTe Crystal

NASA Astrophysics Data System (ADS)

Mao, Yifei; Zhang, Jijun; Min, Jiahua; Liang, Xiaoyan; Huang, Jian; Tang, Ke; Ling, Liwen; Li, Ming; Zhang, Ying; Wang, Linjun

2018-02-01

This study establishes a model for describing the interaction between Te inclusions, dislocations and point defects in CdMnTe crystals. The role of the complex environment surrounding the formation of Te inclusions was analyzed. Images of Te inclusions captured by scanning electron microscope and infrared microscope were used to observe the morphology of Te inclusions. The morphology of Te inclusions is discussed in light of crystallography, from the crystal growth temperature at 900°C to the melting temperature of Te inclusions using the traveling heater method. The dislocation nets around Te inclusions were calculated by counting lattice mismatches between the Te inclusions and the bulk CdMnTe at 470°C. The point defects of Te antisites were found to be gathered around Te inclusions, with dislocation climb during the cooling phase of crystal growth from 470°C to room temperature. The Te inclusions, dislocation nets and surrounding point defects are considered to be an entirety for evaluating the effect of Te inclusions on CdMnTe detector performance, and an effective mobility-lifetime product (μτ) was obtained.

Real-Time Confocal Imaging Of The Living Eye

NASA Astrophysics Data System (ADS)

Jester, James V.; Cavanagh, H. Dwight; Essepian, John; Shields, William J.; Lemp, Michael A.

1989-12-01

In 1986, we adapted the Tandem Scanning Reflected Light Microscope of Petran and Hadraysky to permit non-invasive, confocal imaging of the living eye in real-time. We were first to obtain stable, confocal optical sections in vivo, from human and animal eyes. Using confocal imaging systems we have now studied living, normal volunteers, rabbits, cats and primates sequentially, non-invasively, and in real-time. The continued development of real-time confocal imaging systems will unlock the door to a new field of cell biology involving for the first time the study of dynamic cellular processes in living organ systems. Towards this end we have concentrated our initial studies on three areas (1) evaluation of confocal microscope systems for real-time image acquisition, (2) studies of the living normal cornea (epithelium, stroma, endothelium) in human and other species; and (3) sequential wound-healing responses in the cornea in single animals to lamellar-keratectomy injury (cellular migration, inflammation, scarring). We believe that this instrument represents an important, new paradigm for research in cell biology and pathology and that it will fundamentally alter all experimental and clinical approaches in future years.

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

PubMed

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

2002-10-07

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

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

PubMed

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.

Experiments on terahertz 3D scanning microscopic imaging

NASA Astrophysics Data System (ADS)

Zhou, Yi; Li, Qi

2016-10-01

Compared with the visible light and infrared, terahertz (THz) radiation can penetrate nonpolar and nonmetallic materials. There are many studies on the THz coaxial transmission confocal microscopy currently. But few researches on the THz dual-axis reflective confocal microscopy were reported. In this paper, we utilized a dual-axis reflective confocal scanning microscope working at 2.52 THz. In contrast with the THz coaxial transmission confocal microscope, the microscope adopted in this paper can attain higher axial resolution at the expense of reduced lateral resolution, revealing more satisfying 3D imaging capability. Objects such as Chinese characters "Zhong-Hua" written in paper with a pencil and a combined sheet metal which has three layers were scanned. The experimental results indicate that the system can extract two Chinese characters "Zhong," "Hua" or three layers of the combined sheet metal. It can be predicted that the microscope can be applied to biology, medicine and other fields in the future due to its favorable 3D imaging capability.

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

NASA Astrophysics Data System (ADS)

Pal, Robert; Beeby, Andrew

2014-09-01

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.

Modulus design multiwavelength polarization microscope for transmission Mueller matrix imaging.

PubMed

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

2018-01-01

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

Thermal effects of white light illumination during microsurgery: clinical pilot study on the application safety of surgical microscopes.

PubMed

Hibst, Raimund; Saal, David; Russ, Detlef; Kunzi-Rapp, Karin; Kienle, Alwin; Stock, Karl

2010-01-01

Modern operating microscopes offer high power illumination to ensure optimal visualization, but can also cause thermal damage. The aim of our study is to quantify the thermal effects in vivo and discuss conditions for safe use. In a pilot study on volunteers, we measured the temperature at the skin surface during microscope illumination, including the influence of anaesthesia and the effects of staining, draping, or moistening of the skin. Irradiation within the limit given by safety regulations (200 mW/cm(2)) results in skin surface temperature of 43 degrees C. Higher intensities (forearm 335 mW/cm(2), back 250 mW/cm(2)) are tolerated, resulting in reversible hyperaemia. At a very high illumination intensity (750 mW/cm(2)), pain occurs within 30 s at temperatures of 46 degrees C+/-1 degrees C (hand and forearm), and 43 degrees C+/-2 degrees C (back), respectively. Anaesthesia has no distinct effect on the temperature, whereas staining and drapes result in much higher temperatures (>100 degrees C). Moistening at practicable flow rates can reduce temperature efficiently when combined with a light absorbing and water absorbent drape. In conclusion, surgeons must be aware that surgical microscope illumination without protective means can cause skin temperatures to rise much above pain threshold, which in our study serves as a (conservative) benchmark for potential damage.

Thermal effects of white light illumination during microsurgery: clinical pilot study on the application safety of surgical microscopes

NASA Astrophysics Data System (ADS)

Hibst, Raimund; Saal, David; Russ, Detlef; Kunzi-Rapp, Karin; Kienle, Alwin; Stock, Karl

2010-07-01

Modern operating microscopes offer high power illumination to ensure optimal visualization, but can also cause thermal damage. The aim of our study is to quantify the thermal effects in vivo and discuss conditions for safe use. In a pilot study on volunteers, we measured the temperature at the skin surface during microscope illumination, including the influence of anaesthesia and the effects of staining, draping, or moistening of the skin. Irradiation within the limit given by safety regulations (200 mW/cm2) results in skin surface temperature of 43 °C. Higher intensities (forearm 335 mW/cm2, back 250 mW/cm2) are tolerated, resulting in reversible hyperaemia. At a very high illumination intensity (750 mW/cm2), pain occurs within 30 s at temperatures of 46 °C+/-1 °C (hand and forearm), and 43 °C+/-2 °C (back), respectively. Anaesthesia has no distinct effect on the temperature, whereas staining and drapes result in much higher temperatures (>100 °C). Moistening at practicable flow rates can reduce temperature efficiently when combined with a light absorbing and water absorbent drape. In conclusion, surgeons must be aware that surgical microscope illumination without protective means can cause skin temperatures to rise much above pain threshold, which in our study serves as a (conservative) benchmark for potential damage.

Adding an extra dimension to what students see through the light microscope: a lab exercise demonstrating critical analysis for microscopy students.

PubMed

Garrill, Ashley

2011-01-01

This article describes an undergraduate lab exercise that demonstrates the importance of students thinking critically about what they see through a microscope. The students are given growth data from tip-growing organisms that suggest the cells grow in a pulsatile manner. The students then critique this data in several exercises that incorporate aspects of a problem-based learning approach, envisaging growth not just in two dimensions, but in three dimensions. For some cells, what appears to be pulsatile growth could also be explained by growth at a constant rate up and down in the z-axis. Depending on the diffraction pattern generated by the tip of the cell, this movement in the z-axis could go undetected. This raises the possibility that pulsatile growth seen in some species may be an artifact generated by the limitations of the light microscope. Students were subsequently asked to rate their awareness of the need to think critically about what they see through a microscope, using a scale of 1 (unaware) to 5 (very much aware). Prior to doing the lab exercise, the mean rating was 2.7; this increased to 4.4 after the lab. The students also indicated a likelihood of being more critical in their thinking in other aspects of their biology curriculum.