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.

A LEGO Mindstorms Brewster angle microscope

NASA Astrophysics Data System (ADS)

Fernsler, Jonathan; Nguyen, Vincent; Wallum, Alison; Benz, Nicholas; Hamlin, Matthew; Pilgram, Jessica; Vanderpoel, Hunter; Lau, Ryan

2017-09-01

A Brewster Angle Microscope (BAM) built from a LEGO Mindstorms kit, additional LEGO bricks, and several standard optics components, is described. The BAM was built as part of an undergraduate senior project and was designed, calibrated, and used to image phospholipid, cholesterol, soap, and oil films on the surface of water. A BAM uses p-polarized laser light reflected off a surface at the Brewster angle, which ideally yields zero reflectivity. When a film of different refractive index is added to the surface a small amount of light is reflected, which can be imaged in a microscope camera. Films of only one molecule (approximately 1 nm) thick, a monolayer, can be observed easily in the BAM. The BAM was used in a junior-level Physical Chemistry class to observe phase transitions of a monolayer and the collapse of a monolayer deposited on the water surface in a Langmuir trough. Using a photometric calculation, students observed a change in thickness of a monolayer during a phase transition of 7 Å, which was accurate to within 1 Å of the value determined by more advanced methods. As supplementary material, we provide a detailed manual on how to build the BAM, software to control the BAM and camera, and image processing software.

Mechanical characterization at material interfaces through dark field Brillouin microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Fiore, Antonio; Scarcelli, Giuliano

2017-02-01

Brillouin microscopy allows high-resolution mapping of the mechanical properties of a sample by measuring the spectra of acoustically induced light scattering therein, and thus has been widely investigated for biomedical application. Measuring the Brillouin spectral shift is challenging when the light is focused onto the interfaces between two materials of different refractive index, because a sizeable portion of the incident light is Fresnel-reflected into the Brillouin spectrometer. To address this need, here, we designed a Brillouin confocal microscope in which the specular reflection at the interface between two materials is physically rejected without significant loss to the Brillouin signal. To achieve this goal, we illuminate the sample with a small-diameter Gaussian beam focused by a high numerical aperture objective lens. In the collection path, the beam reflected from the sample has the same diameter as the incident beam, while the scattered light beam is as large as the clear aperture of the microscope objective. Therefore, using a small blocking filter allows to efficiently reject the reflected light. We calculated the tradeoff between extinction improvement and signal loss when the diameter of the blocking filter is changed. Experimentally, we demonstrated extinction improvement of over 60dB with only 30% signal loss while achieving submicron resolutions. This innovation can be useful for in vivo measurements of the cornea to avoid artifacts in the epithelium and anterior portions of the stroma, as well as to investigate cells cultured on glass coverslips without necessity of index-matching materials.

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

NASA Astrophysics Data System (ADS)

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

2013-03-01

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

Specimen illumination apparatus with optical cavity for dark field illumination

DOEpatents

Pinkel, Daniel; Sudar, Damir; Albertson, Donna

1999-01-01

An illumination apparatus with a specimen slide holder, an illumination source, an optical cavity producing multiple reflection of illumination light to a specimen comprising a first and a second reflective surface arranged to achieve multiple reflections of light to a specimen is provided. The apparatus can further include additional reflective surfaces to achieve the optical cavity, a slide for mounting the specimen, a coverslip which is a reflective component of the optical cavity, one or more prisms for directing light within the optical cavity, antifading solutions for improving the viewing properties of the specimen, an array of materials for analysis, fluorescent components, curved reflective surfaces as components of the optical cavity, specimen detection apparatus, optical detection equipment, computers for analysis of optical images, a plane polarizer, fiberoptics, light transmission apertures, microscopic components, lenses for viewing the specimen, and upper and lower mirrors above and below the specimen slide as components of the optical cavity. Methods of using the apparatus are also provided.

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.

Shack-Hartmann reflective micro profilometer

NASA Astrophysics Data System (ADS)

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

2018-01-01

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

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.

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.

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.

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.

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.

Reflectance Spectra of Peacock Feathers and the Turning Angles of Melanin Rods in Barbules.

PubMed

Okazaki, Toshio

2018-02-01

I analyzed the association between the reflectance spectra and melanin rod arrangement in barbules of the eyespot of peacock feathers. The reflectance spectra from the yellow-green feather of the eyespot indicated double peaks of 430 and 540 nm. The maximum reflectance spectrum of the blue feather was 480 nm, and that of the dark blue feather was 420 nm. The reflectance spectra from brown feathers indicated double peaks of 490 and 610 nm. Transmission electron microscopic analysis confirmed that melanin rods were arranged fanwise in the outer layer toward the barbule tips. In addition, using polarized light microscope, I attempted to determine whether the turning angles of melanin rods in the barbules reflected different colors. The turning angle of the polarizing axis of the barbules was supported by that of the melanin rods, observed using transmission electron microscopic images. To compare the turning angle of melanin rods in the respective barbules, I calculated the opening width of the fanwise melanin rods by dividing the width of the barbules by the turning angle of the polarizing axis of barbules and obtained a positive correlation between the reflectance spectra and opening width of the fanwise melanin rods. Moreover, the widely spreading reflection from the barbules may occur because of the fanwise melanin rod arrangement.

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

Optical anisotropy and domain structure of multiferroic Ni-Mn-Ga and Co-Ni-Ga Heusler-type alloys

NASA Astrophysics Data System (ADS)

Ivanova, A. I.; Gasanov, O. V.; Kaplunova, E. I.; Kalimullina, E. T.; Zalyotov, A. B.; Grechishkin, R. M.

2015-03-01

A study is made of the reflectance anisotropy of martensitic and magnetic domains in ferromagnetic shape memory alloys (FSMA) Ni-Mn-Ga and Co-Ni-Ga. The reflectance of metallographic sections of these alloys was measured in the visible with the aid of standard inverted polarized light microscope with a 360° rotatable specimen stage. Calculations are presented for the estimation of image contrast values between neighboring martensite twins. Qualitative and quantitative observations and angular measurements in reflected polarized light proved to be useful for the analysis of specific features of the martensite microstructure of multiferroic materials.

Wide-field high spatial frequency domain imaging of tissue microstructure

NASA Astrophysics Data System (ADS)

Lin, Weihao; Zeng, Bixin; Cao, Zili; Zhu, Danfeng; Xu, M.

2018-02-01

Wide-field tissue imaging is usually not capable of resolving tissue microstructure. We present High Spatial Frequency Domain Imaging (HSFDI) - a noncontact imaging modality that spatially maps the tissue microscopic scattering structures over a large field of view. Based on an analytical reflectance model of sub-diffusive light from forward-peaked highly scattering media, HSFDI quantifies the spatially-resolved parameters of the light scattering phase function from the reflectance of structured light modulated at high spatial frequencies. We have demonstrated with ex vivo cancerous tissue to validate the robustness of HSFDI in significant contrast and differentiation of the microstructutral parameters between different types and disease states of tissue.

Phase resolved and coherence gated en face reflection imaging of multilayered embryonal carcinoma cells

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Fukami, Tadashi; Iwai, Hidenao; Yamashita, Yutaka

2012-03-01

Embryonal carcinoma (EC) cells, which are cell lines derived from teratocarcinomas, have characteristics in common with stem cells and differentiate into many kinds of functional cells. Similar to embryonic stem (ES) cells, undifferentiated EC cells form multi-layered spheroids. In order to visualize the three-dimensional structure of multilayered EC cells without labeling, we employed full-field interference microscopy with the aid of a low-coherence quantitative phase microscope, which is a reflection-type interference microscope employing the digital holographic technique with a low-coherent light source. Owing to the low-coherency of the light-source (halogen lamp), only the light reflected from reflective surface at a specific sectioning height generates an interference image on the CCD camera. P19CL6 EC cells, derived from mouse teratocarcinomas, formed spheroids that are about 50 to 200 micrometers in diameter. Since the height of each cell is around 10 micrometers, it is assumed that each spheroid has 5 to 20 cell layers. The P19CL6 spheroids were imaged in an upright configuration and the horizontally sectioned reflection images of the sample were obtained by sequentially and vertically scanning the zero-path-length height. Our results show the threedimensional structure of the spheroids, in which plasma and nuclear membranes were distinguishably imaged. The results imply that our technique is further capable of imaging induced pluripotent stem (iPS) cells for the assessment of cell properties including their pluripotency.

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.

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.

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

Near-Field Magneto-Optical Microscope

DOEpatents

Vlasko-Vlasov, Vitalii; Welp, Ulrich; and Crabtree, George W.

2005-12-06

A device and method for mapping magnetic fields of a sample at a resolution less than the wavelength of light without altering the magnetic field of the sample is disclosed. A device having a tapered end portion with a magneto-optically active particle positioned at the distal end thereof in communication with a fiber optic for transferring incoming linearly polarized light from a source thereof to the particle and for transferring reflected light from the particle is provided. The fiber optic has a reflective material trapping light within the fiber optic and in communication with a light detector for determining the polarization of light reflected from the particle as a function of the strength and direction of the magnetic field of the sample. Linearly polarized light from the source thereof transferred to the particle positioned proximate the sample is affected by the magnetic field of the sample sensed by the particle such that the difference in polarization of light entering and leaving the particle is due to the magnetic field of the sample. Relative movement between the particle and sample enables mapping.

Near Field Magneto-Optical Microscope

DOEpatents

Vlasko-Vlasov, Vitalii K.; Welp, Ulrich; Crabtree, George W.

2005-12-06

A device and method for mapping magnetic fields of a sample at a resolution less than the wavelength of light without altering the magnetic field of the sample is disclosed. A device having a tapered end portion with a magneto-optically active particle positioned at the distal end thereof in communication with a fiber optic for transferring incoming linearly polarized light from a source thereof to the particle and for transferring reflected light from the particle is provided. The fiber optic has a reflective material trapping light within the fiber optic and in communication with a light detector for determining the polarization of light reflected from the particle as a function of the strength and direction of the magnetic field of the sample. Linearly polarized light from the source thereof transferred to the particle positioned proximate the sample is affected by the magnetic field of the sample sensed by the particle such that the difference in polarization of light entering and leaving the particle is due to the magnetic field of the sample. Relative movement between the particle and sample enables mapping.

Far-infrared Beamline at the Canadian Light Source

NASA Astrophysics Data System (ADS)

Zhao, Jianbao; Billinghurst, Brant

2017-06-01

Far-infrared is a particularly useful technique for studies on lattice modes as they generally appear in the Far-infrared region. Far-infrared is also an important tool for gathering information on the electrical transport properties of metallic materials and the band gap of semiconductors. This poster will describe the horizontal microscope that has recently been built in the Far-infrared beamline at the Canadian Light Source Inc. (CLS). This microscope is specially designed for high-pressure Far-infrared absorbance and reflectance spectroscopic studies. The numerical aperture (0.5) and the long working distance (82.1 mm) in the microscope are good fits for Diamond Anvil Cell (DAC). The spectra are recorded using liquid helium cooled Si bolometer or Ge:Cu detector. The pressure in the DAC can be determined by using the fluorescence spectrometer available onsite. The Far-infrared beamline at CLS is a state-of-the-art synchrotron facility, offering significantly more brightness than conventional sources. Because of the high brightness of the synchrotron radiation, we can obtain the Far-infrared reflectance/absorbance spectra on the small samples with more throughput than with a conventional source. The Far-infrared beamline is open to users through peer review.

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.

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

PHOTONICS AND NANOTECHNOLOGY Microscopic theory of optical properties of composite media with chaotically distributed nanoparticles

NASA Astrophysics Data System (ADS)

Shalin, A. S.

2010-12-01

The boundary problem of light reflection and transmission by a film with chaotically distributed nanoinclusions is considered. Based on the proposed microscopic approach, analytic expressions are derived for distributions inside and outside the nanocomposite medium. Good agreement of the results with exact calculations and (at low concentrations of nanoparticles) with the integral Maxwell-Garnett effective-medium theory is demonstrated. It is shown that at high nanoparticle concentrations, averaging the dielectric constant in volume as is done within the framework of the effective-medium theory yields overestimated values of the optical film density compared to the values yielded by the proposed microscopic approach. We also studied the dependence of the reflectivity of a system of gold nanoparticles on their size, the size dependence of the plasmon resonance position along the wavelength scale, and demonstrated a good agreement with experimental data.

Standoff detection of explosives: a challenging approach for optical technologies

NASA Astrophysics Data System (ADS)

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

2011-06-01

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

Silvical characteristics of white ash (Fraxinus americana)

Treesearch

Jonathan W. Wright

1959-01-01

White ash (Fraxinus americana L.) derives its common name from the white under-surface of the leaf; the white effect is created by microscopic papillae with a high light-reflecting capacity. The specific name americana was given to the species because of its range in America.

Stress Measurement by Geometrical Optics

NASA Technical Reports Server (NTRS)

Robinson, R. S.; Rossnagel, S. M.

1986-01-01

Fast, simple technique measures stresses in thin films. Sample disk bowed by stress into approximately spherical shape. Reflected image of disk magnified by amount related to curvature and, therefore, stress. Method requires sample substrate, such as cheap microscope cover slide, two mirrors, laser light beam, and screen.

Reflection of a polarized light cone

NASA Astrophysics Data System (ADS)

Brody, Jed; Weiss, Daniel; Berland, Keith

2013-01-01

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

Low-cost cryo-light microscopy stage fabrication for correlated light/electron microscopy.

PubMed

Carlson, David B; Evans, James E

2011-06-05

The coupling of cryo-light microscopy (cryo-LM) and cryo-electron microscopy (cryo-EM) poses a number of advantages for understanding cellular dynamics and ultrastructure. First, cells can be imaged in a near native environment for both techniques. Second, due to the vitrification process, samples are preserved by rapid physical immobilization rather than slow chemical fixation. Third, imaging the same sample with both cryo-LM and cryo-EM provides correlation of data from a single cell, rather than a comparison of "representative samples". While these benefits are well known from prior studies, the widespread use of correlative cryo-LM and cryo-EM remains limited due to the expense and complexity of buying or building a suitable cryogenic light microscopy stage. Here we demonstrate the assembly, and use of an inexpensive cryogenic stage that can be fabricated in any lab for less than $40 with parts found at local hardware and grocery stores. This cryo-LM stage is designed for use with reflected light microscopes that are fitted with long working distance air objectives. For correlative cryo-LM and cryo-EM studies, we adapt the use of carbon coated standard 3-mm cryo-EM grids as specimen supports. After adsorbing the sample to the grid, previously established protocols for vitrifying the sample and transferring/handling the grid are followed to permit multi-technique imaging. As a result, this setup allows any laboratory with a reflected light microscope to have access to direct correlative imaging of frozen hydrated samples.

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.

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

PubMed

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

1994-01-01

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

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.

Selective colors reflection from stratified aragonite calcium carbonate plates of mollusk shells.

PubMed

Lertvachirapaiboon, Chutiparn; Parnklang, Tewarak; Pienpinijtham, Prompong; Wongravee, Kanet; Thammacharoen, Chuchaat; Ekgasit, Sanong

2015-08-01

An interaction between the incident light and the structural architecture within the shell of Asian green mussel (Perna viridis) induces observable pearlescent colors. In this paper, we investigate the influence of the structural architecture on the expressed colors. After a removal of the organic binder, small flakes from crushed shells show vivid rainbow reflection under an optical microscope. An individual flake expresses vivid color under a bright-field illumination while become transparent under a dark-field illumination. The expressed colors of the aragonite flakes are directly associated with its structural architecture. The flakes with aragonite thickness of 256, 310, and 353 nm, respectively, appear blue, green, and red under an optical microscope. The spectral simulation corroborates the experimentally observed optical effects as the flakes with thicker aragonite layers selectively reflected color with longer wavelengths. Flakes with multiple aragonite thicknesses expressed multi-color as the upper aragonite layers allow reflected colors from the lower layers to be observed. Copyright © 2015 Elsevier Inc. All rights reserved.

Deciphering the Origin of Plume-Textured Geodes.

ERIC Educational Resources Information Center

Garlick, George Donald; Jones, Francis Tucker

1990-01-01

Presented is an interpretation of the inward and outward growth and formation of plume textured geodes available from southern Brazil. Field occurrence, morphology of vesicles, growth history, closure of the agate shell, microscopic features, coherent reflection of light from convoluted surfaces, and accessory minerals of the inner cavity are…

Manipulation of Micro Scale Particles in Optical Traps Using Programmable Spatial Light Modulation

NASA Technical Reports Server (NTRS)

Seibel, Robin E.; Decker, Arthur J. (Technical Monitor)

2003-01-01

1064 nm light, from an Nd:YAG laser, was polarized and incident upon a programmable parallel aligned liquid crystal spatial light modulator (PAL-SLM), where it was phase modulated according to the program controlling the PAL-SLM. Light reflected from the PAL-SLM was injected into a microscope and focused. At the focus, multiple optical traps were formed in which 9.975 m spheres were captured. The traps and the spheres were moved by changing the program of the PAL-SLM. The motion of ordered groups of micro particles was clearly demonstrated.

Improving confocal microscopy with solid-state semiconductor excitation sources

NASA Astrophysics Data System (ADS)

Sivers, Nelson L.

To efficiently excite the fluorescent dyes used in imaging biological samples with a confocal microscope, the wavelengths of the exciting laser must be near the fluorochrome absorption peak. However, this causes imaging problems when the fluorochrome absorption and emission spectra overlap significantly, i.e. have small Stokes shifts, which is the case for most fluorochromes that emit in the red to infrared. As a result, the reflected laser excitation cannot be distinguished from the information-containing fluorescence signal. However, cryogenically cooling the exciting laser diode enabled the laser emission wavelengths to be tuned to shorter wavelengths, decreasing the interference between the laser and the fluorochrome's fluorescence. This reduced the amount of reflected laser light in the confocal image. However, the cooled laser diode's shorter wavelength signal resulted in slightly less efficient fluorochrome excitation. Spectrophotometric analysis showed that as the laser diodes were cooled, their output power increased, which more than compensated for the lower fluorochrome excitation and resulted in significantly more intense fluorescence. Thus, by tuning the laser diode emission wavelengths away from the fluorescence signal, less reflected laser light and more fluorescence information reached the detector, creating images with better signal to noise ratios. Additionally, new, high, luminous flux, light-emitting diodes (LEDs) are now powerful enough to create confocal fluorescence signals comparable to those produced by the traditional laser excitation sources in fluorescence confocal microscopes. The broader LED spectral response effectively excited the fluorochrome, yet was spectrally limited enough for standard filter sets to separate the LED excitation from the fluorochrome fluorescence signal. Spectrophotometric analysis of the excitation and fluorescence spectra of several fluorochromes showed that high-powered, LED-induced fluorescence contained the same spectral information and could be more intense than that produced by lasers. An alternative, LED-based, confocal microscope is proposed in this thesis that would be capable of exciting multiple fluorochromes in a single specimen, producing images of several distinct cellular components simultaneously. The inexpensive, LED-based, confocal microscope would require lower peak excitation intensities to produce fluorescence signals equal to those produced by laser excitation, reducing cellular damage and slowing fluorochrome photobleaching.

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

DOEpatents

Mariella, Jr., Raymond P.

2000-01-01

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

Polarization-sensitive color in butterfly scales: polarization conversion from ridges with reflecting elements.

PubMed

Zhang, Ke; Tang, Yiwen; Meng, Jinsong; Wang, Ge; Zhou, Han; Fan, Tongxiang; Zhang, Di

2014-11-03

Polarization-sensitive color originates from polarization-dependent reflection or transmission, exhibiting abundant light information, including intensity, spectral distribution, and polarization. A wide range of butterflies are physiologically sensitive to polarized light, but the origins of polarized signal have not been fully understood. Here we systematically investigate the colorful scales of six species of butterfly to reveal the physical origins of polarization-sensitive color. Microscopic optical images under crossed polarizers exhibit their polarization-sensitive characteristic, and micro-structural characterizations clarify their structural commonality. In the case of the structural scales that have deep ridges, the polarization-sensitive color related with scale azimuth is remarkable. Periodic ridges lead to the anisotropic effective refractive indices in the parallel and perpendicular grating orientations, which achieves form-birefringence, resulting in the phase difference of two different component polarized lights. Simulated results show that ridge structures with reflecting elements reflect and rotate the incident p-polarized light into s-polarized light. The dimensional parameters and shapes of grating greatly affect the polarization conversion process, and the triangular deep grating extends the outstanding polarization conversion effect from the sub-wavelength period to the period comparable to visible light wavelength. The parameters of ridge structures in butterfly scales have been optimized to fulfill the polarization-dependent reflection for secret communication. The structural and physical origin of polarization conversion provides a more comprehensive perspective on the creation of polarization-sensitive color in butterfly wing scales. These findings show great potential in anti-counterfeiting technology and advanced optical material design.

Analysis of thin-film polymers using attenuated total internal reflection-Raman microspectroscopy.

PubMed

Tran, Willie; Tisinger, Louis G; Lavalle, Luis E; Sommer, André J

2015-01-01

Two methods commonly employed for molecular surface analysis and thin-film analysis of microscopic areas are attenuated total reflection infrared (ATR-IR) microspectroscopy and confocal Raman microspectroscopy. In the former method, the depth of the evanescent probe beam can be controlled by the wavelength of light, the angle of incidence, or the refractive index of the internal reflection element. Because the penetration depth is proportional to the wavelength of light, one could interrogate a smaller film thickness by moving from the mid-infrared region to the visible region employing Raman spectroscopy. The investigation of ATR Raman microspectroscopy, a largely unexplored technique available to Raman microspectroscopy, was carried out. A Renishaw inVia Raman microscope was externally modified and used in conjunction with a solid immersion lens (SIL) to perform ATR Raman experiments. Thin-film polymer samples were analyzed to explore the theoretical sampling depth for experiments conducted without the SIL, with the SIL, and with the SIL using evanescent excitation. The feasibility of micro-ATR Raman was examined by collecting ATR spectra from films whose thickness measured from 200 to 60 nm. Films of these thicknesses were present on a much thicker substrate, and features from the underlying substrate did not become visible until the thin film reached a thickness of 68 nm.

Basic study of charring detection at the laser catheter-tip using back scattering light measurement during therapeutic laser irradiation in blood.

PubMed

Takahashi, Mei; Ito, Arisa; Kajihara, Takuro; Matsuo, Hiroki; Arai, Tsunenori

2010-01-01

The purpose of this study is to investigate transient process of the charring at the laser catheter-tip in blood during therapeutic laser irradiation by the back scattering light measurement to detect precursor state of the charring. We took account of using photodynamic therapy for arrhythmia in blood through the laser catheter. We observed the influence of the red laser irradiation (λ=663 nm) upon the shape of red blood cells (RBCs). The RBCs aggregation, round formation, and hemolysis were took place sequentially before charring. With a model blood sandwiched between glass plates simulated as a catheter-tip boundary, we measured diffuse-reflected-light power and transmitted-light power simultaneously and continuously by a microscopic optics during the laser irradiation. We found that measured light power changes were originated with RBCs shape change induced by temperature rise due to the laser irradiation. A gentle peak following a slow descending was observed in the diffuse-reflected-light power history. This history might indicate the precursor state of the charring, in which the hemolysis might be considered to advance rapidly. We think that the measurement of diffuse-reflected-light power history might be able to detect precursor state of charring at the catheter-tip in blood.

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.

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.

[The research of UV-responsive sensitivity enhancement of fluorescent coating films by MgF2 layer].

PubMed

Lu, Zhong-Rong; Ni, Zheng-Ji; Tao, Chun-Xian; Hong, Rui-Jin; Zhang, Da-Wei; Huang, Yuan-Shen

2014-03-01

A low cost and less complicated expansion approach of wavelength responses with a Lumogen phosphor coating was adopted, as they increased the quantum efficiency of CCD and CMOS detectors in ultra-violet by absorbing UV light and then re emitting visible light. In this paper, the sensitivity enhancement of fluorescence coatings was studied by adding an anti-reflection film or barrier film to reduce the loss of the scattering and reflection on the incident interface. The Lumogen and MgF2/Lumogen film were deposited on quartz glasses by physical vacuum deposition. The surface morphology, transmittance spectrum, reflectance spectrum and fluorescence emission spectrum were obtained by atomic force microscope (AFM), spectrophotometer and fluorescence spectrometer, respectively. The results indicated that MgF2 film had obvious positive effect on reducing scattering and reflection loss in 500-700 nm, and enhancing the absorption of Lumogen coating in ultraviolet spectrum. Meanwhile, the fluorescent emission intensity had a substantial increase by smoothing the film surface and thus reducing the light scattering. At the same time, the MgF2 layer could protect Lumogen coating from damaging and contamination, which give a prolong lifetime of the UV-responsive CCD sensors with fluorescent coatings.

Multi-spectral digital holographic microscopy for enhanced quantitative phase imaging of living cells

NASA Astrophysics Data System (ADS)

Kemper, Björn; Kastl, Lena; Schnekenburger, Jürgen; Ketelhut, Steffi

2018-02-01

Main restrictions of using laser light in digital holographic microscopy (DHM) are coherence induced noise and parasitic reflections in the experimental setup which limit resolution and measurement accuracy. We explored, if coherence properties of partial coherent light sources can be generated synthetically utilizing spectrally tunable lasers. The concept of the method is demonstrated by label-free quantitative phase imaging of living pancreatic tumor cells and utilizing an experimental configuration including a commercial microscope and a laser source with a broad tunable spectral range of more than 200 nm.

Linear polarization-discriminatory state inverter fabricated by oblique angle deposition.

PubMed

Park, Yong Jun; Sobahan, K M A; Kim, Jin Joo; Hwangbo, Chang Kwon

2009-06-22

In this paper, we report a linear polarization-discriminatory state inverter made of three-layer sculpture thin film fabricated by oblique angle deposition technique. The first and third layers are quarter-wave plates of zigzag structure and the middle of them is a circular Bragg reflector of left-handed helical structure. It is found that the normal incidence of P-polarized light on this polarization-discriminatory state inverter becomes the S-polarized light at output, while the incident S-polarized light of wavelength lying in the Bragg regime is reflected. The microstructure of the linear polarization-discriminatory state inverter is also investigated by using a scanning electron microscope.

Fano Description of Single-Hydrocarbon Fluorescence Excited by a Scanning Tunneling Microscope.

PubMed

Kröger, Jörg; Doppagne, Benjamin; Scheurer, Fabrice; Schull, Guillaume

2018-06-13

The detection of fluorescence with submolecular resolution enables the exploration of spatially varying photon yields and vibronic properties at the single-molecule level. By placing individual polycyclic aromatic hydrocarbon molecules into the plasmon cavity formed by the tip of a scanning tunneling microscope and a NaCl-covered Ag(111) surface, molecular light emission spectra are obtained that unravel vibrational progression. In addition, light spectra unveil a signature of the molecule even when the tunneling current is injected well separated from the molecular emitter. This signature exhibits a distance-dependent Fano profile that reflects the subtle interplay between inelastic tunneling electrons, the molecular exciton and localized plasmons in at-distance as well as on-molecule fluorescence. The presented findings open the path to luminescence of a different class of molecules than investigated before and contribute to the understanding of single-molecule luminescence at surfaces in a unified picture.

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

NASA Astrophysics Data System (ADS)

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

2017-03-01

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

Eliminating Unwanted Far-Field Excitation in Objective-Type TIRF. Part I. Identifying Sources of Nonevanescent Excitation Light

PubMed Central

Brunstein, Maia; Teremetz, Maxime; Hérault, Karine; Tourain, Christophe; Oheim, Martin

2014-01-01

Total internal reflection fluorescence microscopy (TIRFM) achieves subdiffraction axial sectioning by confining fluorophore excitation to a thin layer close to the cell/substrate boundary. However, it is often unknown how thin this light sheet actually is. Particularly in objective-type TIRFM, large deviations from the exponential intensity decay expected for pure evanescence have been reported. Nonevanescent excitation light diminishes the optical sectioning effect, reduces contrast, and renders TIRFM-image quantification uncertain. To identify the sources of this unwanted fluorescence excitation in deeper sample layers, we here combine azimuthal and polar beam scanning (spinning TIRF), atomic force microscopy, and wavefront analysis of beams passing through the objective periphery. Using a variety of intracellular fluorescent labels as well as negative staining experiments to measure cell-induced scattering, we find that azimuthal beam spinning produces TIRFM images that more accurately portray the real fluorophore distribution, but these images are still hampered by far-field excitation. Furthermore, although clearly measureable, cell-induced scattering is not the dominant source of far-field excitation light in objective-type TIRF, at least for most types of weakly scattering cells. It is the microscope illumination optical path that produces a large cell- and beam-angle invariant stray excitation that is insensitive to beam scanning. This instrument-induced glare is produced far from the sample plane, inside the microscope illumination optical path. We identify stray reflections and high-numerical aperture aberrations of the TIRF objective as one important source. This work is accompanied by a companion paper (Pt.2/2). PMID:24606927

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

Guo, Wei, E-mail: wguo2@ncsu.edu; Kirste, Ronny; Bryan, Zachary

Enhanced light extraction efficiency was demonstrated on nanostructure patterned GaN and AlGaN/AlN Multiple-Quantum-Well (MQW) structures using mass production techniques including natural lithography and interference lithography with feature size as small as 100 nm. Periodic nanostructures showed higher light extraction efficiency and modified emission profile compared to non-periodic structures based on integral reflection and angular-resolved transmission measurement. Light extraction mechanism of macroscopic and microscopic nanopatterning is discussed, and the advantage of using periodic nanostructure patterning is provided. An enhanced photoluminescence emission intensity was observed on nanostructure patterned AlGaN/AlN MQW compared to as-grown structure, demonstrating a large-scale and mass-producible pathway to higher lightmore » extraction efficiency in deep-ultra-violet light-emitting diodes.« less

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

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

Total internal reflection and dynamic light scattering microscopy of gels

NASA Astrophysics Data System (ADS)

Gregor, Brian F.

Two different techniques which apply optical microscopy in novel ways to the study of biological systems and materials were built and applied to several samples. The first is a system for adapting the well-known technique of dynamic light scattering (DLS) to an optical microscope. This can detect and scatter light from very small volumes, as compared to standard DLS which studies light scattering from volumes 1000x larger. The small scattering volume also allows for the observation of nonergodic dynamics in appropriate samples. Porcine gastric mucin (PGM) forms a gel at low pH which lines the epithelial cell layer and acts as a protective barrier against the acidic stomach environment. The dynamics and microscopic viscosity of PGM at different pH levels is studied using polystyrene microspheres as tracer particles. The microscopic viscosity and microrheological properties of the commercial basement membrane Matrigel are also studied with this instrument. Matrigel is frequently used to culture cells and its properties remain poorly determined. Well-characterized and purely synthetic Matrigel substitutes will need to have the correct rheological and morphological characteristics. The second instrument designed and built is a microscope which uses an interferometry technique to achieve an improvement in resolution 2.5x better in one dimension than the Abbe diffraction limit. The technique is based upon the interference of the evanescent field generated on the surface of a prism by a laser in a total internal reflection geometry. The enhanced resolution is demonstrated with fluorescent samples. Additionally. Raman imaging microscopy is demonstrated using the evanescent field in resonant and non-resonant samples, although attempts at applying the enhanced resolution technique to the Raman images were ultimately unsuccessful. Applications of this instrument include high resolution imaging of cell membranes and macroscopic structures in gels and proteins. Finally, a third section incorporating previous research on simulations of complex fluids is included. Two dimensional simulations of oil, water, and surfactant mixtures were computed with a lattice gas method. The simulated systems were randomly mixed and then the temperature was quenched to a predetermined point. Spontaneous micellization is observed for a narrow range of temperature quenches, and the overall growth rate of macroscopic structure is found to follow a Vogel-Fulcher growth law.

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.

An Ingenious Super Light Trapping Surface Templated from Butterfly Wing Scales

NASA Astrophysics Data System (ADS)

Han, Zhiwu; Li, Bo; Mu, Zhengzhi; Yang, Meng; Niu, Shichao; Zhang, Junqiu; Ren, Luquan

2015-08-01

Based on the super light trapping property of butterfly Trogonoptera brookiana wings, the SiO2 replica of this bionic functional surface was successfully synthesized using a simple and highly effective synthesis method combining a sol-gel process and subsequent selective etching. Firstly, the reflectivity of butterfly wing scales was carefully examined. It was found that the whole reflectance spectroscopy of the butterfly wings showed a lower level (less than 10 %) in the visible spectrum. Thus, it was confirmed that the butterfly wings possessed a super light trapping effect. Afterwards, the morphologies and detailed architectures of the butterfly wing scales were carefully investigated using the ultra-depth three-dimensional (3D) microscope and field emission scanning electronic microscopy (FESEM). It was composed by the parallel ridges and quasi-honeycomb-like structure between them. Based on the biological properties and function above, an exact SiO2 negative replica was fabricated through a synthesis method combining a sol-gel process and subsequent selective etching. At last, the comparative analysis of morphology feature size and the reflectance spectroscopy between the SiO2 negative replica and the flat plate was conducted. It could be concluded that the SiO2 negative replica inherited not only the original super light trapping architectures, but also the super light trapping characteristics of bio-template. This work may open up an avenue for the design and fabrication of super light trapping materials and encourage people to look for more super light trapping architectures in nature.

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. %.

Atmospheric scanning electron microscope for correlative microscopy.

PubMed

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

2012-01-01

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

In vivo bioluminescence and reflectance imaging of multiple organs in bioluminescence reporter mice by bundled-fiber-coupled microscopy

PubMed Central

Ando, Yoriko; Sakurai, Takashi; Koida, Kowa; Tei, Hajime; Hida, Akiko; Nakao, Kazuki; Natsume, Mistuo; Numano, Rika

2016-01-01

Bioluminescence imaging (BLI) is used in biomedical research to monitor biological processes within living organisms. Recently, fiber bundles with high transmittance and density have been developed to detect low light with high resolution. Therefore, we have developed a bundled-fiber-coupled microscope with a highly sensitive cooled-CCD camera that enables the BLI of organs within the mouse body. This is the first report of in vivo BLI of the brain and multiple organs in luciferase-reporter mice using bundled-fiber optics. With reflectance imaging, the structures of blood vessels and organs can be seen clearly with light illumination, and it allowed identification of the structural details of bioluminescence images. This technique can also be applied to clinical diagnostics in a low invasive manner. PMID:27231601

Characterization of the structure of low-e substrates and consequences for IR transflection measurements

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

DeVetter, Brent M.; Kenkel, Seth; Mittal, Shachi

Spectral distortions caused by the electric field standing wave effect were investigated for two commonly used reflective substrates: low-emissivity glass and gold-coated glass. Our analytical calculations showed that spectral distortions may arise for both incoherent and coherent light sources when performing transflectance measurements. We experimentally confirmed our predictions using a commercial mid-infrared quantum cascade laser microscope and an interferometric infrared imaging system.

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.

Integration of a high-NA light microscope in a scanning electron microscope.

PubMed

Zonnevylle, A C; Van Tol, R F C; Liv, N; Narvaez, A C; Effting, A P J; Kruit, P; Hoogenboom, J P

2013-10-01

We present an integrated light-electron microscope in which an inverted high-NA objective lens is positioned inside a scanning electron microscope (SEM). The SEM objective lens and the light objective lens have a common axis and focal plane, allowing high-resolution optical microscopy and scanning electron microscopy on the same area of a sample simultaneously. Components for light illumination and detection can be mounted outside the vacuum, enabling flexibility in the construction of the light microscope. The light objective lens can be positioned underneath the SEM objective lens during operation for sub-10 μm alignment of the fields of view of the light and electron microscopes. We demonstrate in situ epifluorescence microscopy in the SEM with a numerical aperture of 1.4 using vacuum-compatible immersion oil. For a 40-nm-diameter fluorescent polymer nanoparticle, an intensity profile with a FWHM of 380 nm is measured whereas the SEM performance is uncompromised. The integrated instrument may offer new possibilities for correlative light and electron microscopy in the life sciences as well as in physics and chemistry. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

Effect of operating microscope light on brain temperature during craniotomy.

PubMed

Gayatri, Parthasarathi; Menon, Girish G; Suneel, Puthuvassery R

2013-07-01

Operating microscopes used during neurosurgery are fitted with xenon light. Burn injuries have been reported because of xenon microscope lighting as the intensity of xenon light is 300 W. We designed this study to find out if the light of operating microscope causes an increase in temperature of the brain tissue, which is exposed underneath. Twenty-one adult patients scheduled for elective craniotomies were enrolled. Distal esophageal temperature (T Eso), brain temperature under the microscope light (T Brain), and brain temperature under dura mater (T Dura) were measured continuously at 15-minute intervals during microscope use. The irrigation fluid temperature, room temperature, intensity of the microscope light, and the distance of the microscope from the brain surface were kept constant. The average age of the patients was 44±15 years (18 males and 3 females). The mean duration of microscope use was 140±39 minutes. There were no significant changes in T Brain and T Dura and T Eso over time. T Dura was significantly lower than T Brain both at time 0 and 60 minutes but not at 90 minutes. T Brain was significantly lower than T Eso both at time 0 and 60 minutes but not at 90 minutes. The T Dura remained significantly lower than T Eso at 0, 60, and 90 minutes. Our study shows that there is no significant rise in brain temperature under xenon microscope light up to 120 minutes duration, at intensity of 60% to 70%, from a distance of 20 to 25 cm from the brain surface.

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.

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.

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.

Epifluorescence light collection for multiphoton microscopic endoscopy

NASA Astrophysics Data System (ADS)

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

2011-03-01

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

Laser interferometry force-feedback sensor for an interfacial force microscope

DOEpatents

Houston, Jack E.; Smith, William L.

2004-04-13

A scanning force microscope is provided with a force-feedback sensor to increase sensitivity and stability in determining interfacial forces between a probe and a sample. The sensor utilizes an interferometry technique that uses a collimated light beam directed onto a deflecting member, comprising a common plate suspended above capacitor electrodes situated on a substrate forming an interference cavity with a probe on the side of the common plate opposite the side suspended above capacitor electrodes. The probe interacts with the surface of the sample and the intensity of the reflected beam is measured and used to determine the change in displacement of the probe to the sample and to control the probe distance relative to the surface of the sample.

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

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Iwai, Hidenao; Yamashita, Yutaka

2011-11-01

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

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

NASA Astrophysics Data System (ADS)

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

2017-06-01

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

In Situ Visualization of the Phase Behavior of Oil Samples Under Refinery Process Conditions.

PubMed

Laborde-Boutet, Cedric; McCaffrey, William C

2017-02-21

To help address production issues in refineries caused by the fouling of process units and lines, we have developed a setup as well as a method to visualize the behavior of petroleum samples under process conditions. The experimental setup relies on a custom-built micro-reactor fitted with a sapphire window at the bottom, which is placed over the objective of an inverted microscope equipped with a cross-polarizer module. Using reflection microscopy enables the visualization of opaque samples, such as petroleum vacuum residues, or asphaltenes. The combination of the sapphire window from the micro-reactor with the cross-polarizer module of the microscope on the light path allows high-contrast imaging of isotropic and anisotropic media. While observations are carried out, the micro-reactor can be heated to the temperature range of cracking reactions (up to 450 °C), can be subjected to H2 pressure relevant to hydroconversion reactions (up to 16 MPa), and can stir the sample by magnetic coupling. Observations are typically carried out by taking snapshots of the sample under cross-polarized light at regular time intervals. Image analyses may not only provide information on the temperature, pressure, and reactive conditions yielding phase separation, but may also give an estimate of the evolution of the chemical (absorption/reflection spectra) and physical (refractive index) properties of the sample before the onset of phase separation.

Characterization of silver halide fiber optics and hollow silica waveguides for use in the construction of a mid-infrared attenuated total reflection fourier transform infrared (ATR FT-IR) spectroscopy probe.

PubMed

Damin, Craig A; Sommer, André J

2013-11-01

Advances in fiber optic materials have allowed for the construction of fibers and waveguides capable of transmitting infrared radiation. An investigation of the transmission characteristics associated with two commonly used types of infrared-transmitting fibers/waveguides for prospective use in a fiber/waveguide-coupled attenuated total internal reflection (ATR) probe was performed. Characterization of silver halide polycrystalline fiber optics and hollow silica waveguides was done on the basis of the transmission of infrared light using a conventional fiber optic coupling accessory and an infrared microscope. Using the fiber optic coupling accessory, the average percent transmission for three silver halide fibers was 18.1 ± 6.1% relative to a benchtop reflection accessory. The average transmission for two hollow waveguides (HWGs) using the coupling accessory was 8.0 ± 0.3%. (Uncertainties in the relative percent transmission represent the standard deviations.) Reduced transmission observed for the HWGs was attributed to the high numerical aperture of the coupling accessory. Characterization of the fibers/waveguides using a zinc selenide lens objective on an infrared microscope indicated 24.1 ± 7.2% of the initial light input into the silver halide fibers was transmitted. Percent transmission obtained for the HWGs was 98.7 ± 0.1%. Increased transmission using the HWGs resulted from the absence or minimization of insertion and scattering losses due to the hollow air core and a better-matched numerical aperture. The effect of bending on the transmission characteristics of the fibers/waveguides was also investigated. Significant deviations in the transmission of infrared light by the solid-core silver halide fibers were observed for various bending angles. Percent transmission greater than 98% was consistently observed for the HWGs at the bending angles. The combined benefits of high percent transmission, reproducible instrument responses, and increased bending tolerance indicated HWGs should be preferred in the construction of a fiber/waveguide-coupled ATR probe.

Transportable and vibration-free full-field low-coherent quantitative phase microscope

NASA Astrophysics Data System (ADS)

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

2018-02-01

We developed a transportable Linnik-type full-field low-coherent quantitative phase microscope that is able to compensate for optical path length (OPL) disturbance due to environmental mechanical noises. Though two-beam interferometers such as Linnik ones suffer from unstable OPL difference, we overcame this problem with a mechanical feedback system based on digital signal-processing that controls the OPL difference in sub-nanometer resolution precisely with a feedback bandwidth of 4 kHz. The developed setup has a footprint of 200 mm by 200 mm, a height of 500 mm, and a weight of 4.5 kilograms. In the transmission imaging mode, cells were cultured on a reflection-enhanced glass-bottom dish, and we obtained interference images sequentially while performing stepwise quarter-wavelength phase-shifting. Real-time image processing, including retrieval of the unwrapped phase from interference images and its background correction, along with the acquisition of interference images, was performed on a laptop computer. Emulation of the phase contrast (PhC) images and the differential interference contrast (DIC) images was also performed in real time. Moreover, our setup was applied for full-field cell membrane imaging in the reflection mode, where the cells were cultured on an anti-reflection (AR)-coated glass-bottom dish. The phase and intensity of the light reflected by the membrane revealed the outer shape of the cells independent of the refractive index. In this paper, we show imaging results on cultured cells in both transmission and reflection modes.

Fine structural dependence of ultraviolet reflections in the King Penguin beak horn.

PubMed

Dresp, Birgitta; Langley, Keith

2006-03-01

The visual perception of many birds extends into the near-ultraviolet (UV) spectrum and ultraviolet is used by some to communicate. The beak horn of the King Penguin (Aptenodytes patagonicus) intensely reflects in the ultraviolet and this appears to be implicated in partner choice. In a preliminary study, we recently demonstrated that this ultraviolet reflectance has a structural basis, resulting from crystal-like photonic structures, capable of reflecting in the near-UV. The present study attempted to define the origin of the photonic elements that produce the UV reflectance and to better understand how the UV signal is optimized by their fine structure. Using light and electron microscopic analysis combined with new spectrophotometric data, we describe here in detail the fine structure of the entire King Penguin beak horn in addition to that of its photonic crystals. The data obtained reveal a one-dimensional structural periodicity within this tissue and demonstrate a direct relationship between its fine structure and its function. In addition, they suggest how the photonic structures are produced and how they are stabilized. The measured lattice dimensions of the photonic crystals, together with morphological data on its composition, permit predictions of the wavelength of reflected light. These correlate well with experimentally observed values. The way the UV signal is optimized by the fine structure of the beak tissue is discussed with regard to its putative biological role.

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

The calibration of photographic and spectroscopic films: 1: A microscopic analysis of IIaO films. 2: The effects of agitation and soaking on IIaO films. 3: The effects of electric field on IIaO films. 4: The effects of X-ray radiation on IIaO films

NASA Technical Reports Server (NTRS)

Hammond, E. C., Jr.; Peters, K.; Boone, K.

1978-01-01

The grain structure of the emulsion using both reflected and transmission light was examined along with the effects of soaking. The effect of a static charge by a Tesla-coil, and the effects of airport equipment, and dental X-rays on the film were also analyzed.

Differential high-speed digital micromirror device based fluorescence speckle confocal microscopy.

PubMed

Jiang, Shihong; Walker, John

2010-01-20

We report a differential fluorescence speckle confocal microscope that acquires an image in a fraction of a second by exploiting the very high frame rate of modern digital micromirror devices (DMDs). The DMD projects a sequence of predefined binary speckle patterns to the sample and modulates the intensity of the returning fluorescent light simultaneously. The fluorescent light reflecting from the DMD's "on" and "off" pixels is modulated by correlated speckle and anticorrelated speckle, respectively, to form two images on two CCD cameras in parallel. The sum of the two images recovers a widefield image, but their difference gives a near-confocal image in real time. Experimental results for both low and high numerical apertures are shown.

Design of a dynamic biofilm imaging cell for white-light interferometric microscopy

NASA Astrophysics Data System (ADS)

Larimer, Curtis; Brann, Michelle; Suter, Jonathan D.; Addleman, R. Shane

2017-11-01

In microbiology research, there is a strong need for next-generation imaging and sensing instrumentation that will enable minimally invasive and label-free investigation of soft, hydrated structures, such as in bacterial biofilms. White-light interferometry (WLI) can provide high-resolution images of surface topology without the use of fluorescent labels but is not typically used to image biofilms because there is insufficient refractive index contrast to induce reflection from the biofilm's interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ, especially in a nondestructive manner. We build on our prior description of static biofilm imaging and describe the design of a dynamic growth flow cell that enables monitoring of the thickness and topology of live biofilms over time using a WLI microscope. The microfluidic system is designed to grow biofilms in dynamic conditions and to create a reflective interface on the surface while minimizing disruption of fragile structures. The imaging cell was also designed to accommodate limitations imposed by the depth of focus of the microscope's objective lens. Example images of live biofilm samples are shown to illustrate the ability of the flow cell and WLI instrument to (1) support bacterial growth and biofilm development, (2) image biofilm structure that reflects growth in flow conditions, and (3) monitor biofilm development over time nondestructively. In future work, the apparatus described here will enable surface metrology measurements (roughness, surface area, etc.) of biofilms and may be used to observe changes in biofilm structure in response to changes in environmental conditions (e.g., flow velocity, availability of nutrients, and presence of biocides). This development will open opportunities for the use of WLI in bioimaging.

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.

Hybrid fluorescence and electron cryo-microscopy for simultaneous electron and photon imaging.

PubMed

Iijima, Hirofumi; Fukuda, Yoshiyuki; Arai, Yoshihiro; Terakawa, Susumu; Yamamoto, Naoki; Nagayama, Kuniaki

2014-01-01

Integration of fluorescence light and transmission electron microscopy into the same device would represent an important advance in correlative microscopy, which traditionally involves two separate microscopes for imaging. To achieve such integration, the primary technical challenge that must be solved regards how to arrange two objective lenses used for light and electron microscopy in such a manner that they can properly focus on a single specimen. To address this issue, both lateral displacement of the specimen between two lenses and specimen rotation have been proposed. Such movement of the specimen allows sequential collection of two kinds of microscopic images of a single target, but prevents simultaneous imaging. This shortcoming has been made up by using a simple optical device, a reflection mirror. Here, we present an approach toward the versatile integration of fluorescence and electron microscopy for simultaneous imaging. The potential of simultaneous hybrid microscopy was demonstrated by fluorescence and electron sequential imaging of a fluorescent protein expressed in cells and cathodoluminescence imaging of fluorescent beads. Copyright © 2013 Elsevier Inc. All rights reserved.

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.

PubMed Central

SERDAROGLU OFLAZER, P.; DEYMEER, F.; PARMAN, Y.

2011-01-01

SUMMARY In a muscle biopsy based study, only 9 out of 5450 biopsy samples, received from all parts of greater Istanbul area, had typical clinical and most suggestive light microscopic sporadic-inclusion body myositis (s-IBM) findings. Two other patients with and ten further patients without characteristic light microscopic findings had referring diagnosis of s-IBM. As the general and the ageadjusted populations of Istanbul in 2010 were 13.255.685 and 2.347.300 respectively, the calculated corresponding ‘estimated prevalences' of most suggestive s-IBM in the Istanbul area were 0.679 X 10-6 and 3.834 X 10-6. Since Istanbul receives heavy migration from all regions of Turkey and ours is the only muscle pathology laboratory in Istanbul, projection of these figures to the Turkish population was considered to be reasonable and an estimate of the prevalence of s-IBM in Turkey was obtained. The calculated ‘estimated prevalence' of s-IBM in Turkey is lower than the previously reported rates from other countries. The wide variation in the prevalence rates of s-IBM may reflect different genetic, immunogenetic or environmental factors in different populations. PMID:21842592

Back focal plane microscopic ellipsometer with internal reflection geometry

NASA Astrophysics Data System (ADS)

Otsuki, Soichi; Murase, Norio; Kano, Hiroshi

2013-05-01

A back focal plane (BFP) ellipsometer is presented to measure a thin film on a cover glass using an oil-immersion high-numerical-aperture objective lens. The internal reflection geometry lowers the pseudo Brewster angle (ϕB) to the range over which the light distribution is observed in BFP of the objective. A calculation based on Mueller matrix was developed to compute ellipsometric parameters from the intensity distribution on BFP. The center and radius of the partial reflection region below the critical angle were determined and used to define a polar coordinate on BFP. Harmonic components were computed from the intensities along the azimuth direction and transformed to ellipsometric parameters at multiple incident angles around ϕB. The refractive index and thickness of the film and the contributions of the objective effect were estimated at the same time by fitting.

Omnidirectional anti-reflection properties of vertically align SiO2 nanorod films prepared by electron beam evaporation with glancing angle deposition

NASA Astrophysics Data System (ADS)

Prachachet, R.; Samransuksamer, B.; Horprathum, M.; Eiamchai, P.; Limwichean, S.; Chananonnawathorn, C.; Lertvanithphol, T.; Muthitamongkol, P.; Boonruang, S.; Buranasiri, P.

2018-03-01

Omnidirectional anti-reflection coating nanostructure film have attracted enormous attention for the developments of the optical coating, lenses, light emitting diode, display and photovoltaic. However, fabricated of the omnidirectional antireflection nanostructure film on glass substrate in large area was a challenge topic. In the past two decades, the invention of glancing angle deposition technique as a growth of well-controlled two and three-dimensional morphologies has gained significant attention because of it is simple, fast, cost-effective and high mass production capability. In this present work, the omnidirectional anti-reflection nanostructure coating namely silicon dioxide (SiO2) nanorods has been investigated for optimized high transparent layer at all light incident angle. The SiO2 nanorod films of an optimally low refractive index have been fabricated by electron beam evaporation with the glancing angle deposition technique. The morphological of the prepared sampled were characterized by field-emission scanning electron microscope (FE-SEM) and high-resolution transmission electron microscope (HRTEM). The optical transmission and omnidirectional property of the SiO2 nanorod films were investigated by UV-Vis-NIR spectrophotometer. The measurement were performed at normal incident angle and a full spectral range of 200 - 2000 nm. The angle dependent transmission measure were investigated by rotating the specimen, with incidence angle defined relative to the surface normal of the prepared samples. The morphological characterization results showed that when the glancing angle deposition technique was applied, the vertically align SiO2 nanorods with partially isolated columnar structure can be constructed due to the enhanced shadowing and limited addtom diffusion effect. The average transmission of the vertically align SiO2 nanorods were higher than the glass substrate reference sample over the visible wavelength range at all incident angle due to the transition in the refractive index profile from air to the nanostructure layer that improved the anti-reflection characteristics.

Confocal reflectance quantitative phase microscope system for cellular membranes dynamics study (Conference Presentation)

NASA Astrophysics Data System (ADS)

Singh, Vijay Raj; Yaqoob, Zahid; So, Peter T. C.

2017-02-01

Quantitative phase microscopy (QPM) techniques developed so far primarily belongs to high speed transmitted light based systems that has enough sensitivity to resolve membrane fluctuations and dynamics, but has no depth resolution. Therefore, most biomechanics studies using QPM today is confined to simple cells, such as RBCs, without internal organelles. An important instrument that will greatly extend the biomedical applications of QPM is to develop next generation microscope with 3D capability and sufficient temporal resolution to study biomechanics of complex eukaryotic cells including the mechanics of their internal compartments. For eukaryotic cells, the depth sectioning capability is critical and should be sufficient to distinguish nucleic membrane fluctuations from plasma membrane fluctuations. Further, this microscope must provide high temporal resolution since typical eukaryotes membranes are substantially stiffer than RBCs. A confocal reflectance quantitative phase microscope is presented based on multi-pinhole scanning, with the capabilities of higher temporal resolution and sensitivity for nucleic and plasma membranes of eukaryotic cells. System hardware is developed based on an array of confocal pinhole generated by using the `ON' state of subset of micro-mirrors of digital micro-mirror device (DMD, from Texas Instruments) and high-speed raster scanning provides 14ms imaging speed in wide-field mode. A common path interferometer is integrated at the imaging arm for detection of specimens' quantitative phase information. Theoretical investigation of quantitative phase reconstructed from system is investigated and application of system is presented for dimensional fluctuations measurements of both cellular plasma and nucleic membranes of embryonic stem cells.

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.

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.

Segregation of colloidal swimmers by their activity

NASA Astrophysics Data System (ADS)

Ferrari, Melissa; Youssef, Mena; Driscoll, Michelle; Sacanna, Stefano; Pine, David; Chaikin, Paul

We study a system of micron sized self-propelled colloidal swimmers whose activity can be switched on or off with the flick of a light switch. We have designed a system where an external LED source reflects light off of an array with hundreds of thousands of independently controlled tiny mirrors, through an optical microscope, and onto the plane of the swimmers. By exposing a collection of particles to a spatial or dynamic light field, we have the ability to control the speed of a particle based on its position, and therefore the density of the collection of particles in space. Theoreticians in the field have been building a framework that describes systems which are out-of-equilibrium and we will show how our system can be useful tool in mapping these theories to experiment. Center for Bio-inspired Energy Science.

Photocatalytic activity of Fe-doped CaTiO₃ under UV-visible light.

PubMed

Yang, He; Han, Chong; Xue, Xiangxin

2014-07-01

The photocatalytic degradation of methylene blue (MB) over Fe-doped CaTiO₃ under UV-visible light was investigated. The as-prepared samples were characterized using X-ray diffraction (XRD), scanning electron microscope (SEM) equipped with an energy dispersive spectrometer (EDS) system, Fourier transform infrared spectra (FT-IR), and UV-visible diffuse reflectance spectroscopy (DRS). The results show that the doping with Fe significantly promoted the light absorption ability of CaTiO₃ in the visible light region. The Fe-doped CaTiO₃ exhibited higher photocatalytic activity than CaTiO₃ for the degradation of MB. However, the photocatalytic activity of the Fe-doped CaTiO₃ was greatly influenced by the calcination temperature during the preparation process. The Fe-doped CaTiO₃ prepared at 500°C exhibited the best photocatalytic activity, with degradation of almost 100% MB (10ppm) under UV-visible light for 180 min. Copyright © 2014. Published by Elsevier B.V.

Absorption and Reflection Experiments on High-Mobility 2DEGs in the Regime of Microwave-Induced Resistance Oscillations

NASA Astrophysics Data System (ADS)

Studenikin, S. A.; Potemski, M.; Sachrajda, A. S.; Hilke, M.; Pfeiffer, L. N.; West, K. W.

2005-04-01

We have performed microwave absorption and near-field reflection experiments on a high mobility GaAs/AlGaAs heterostructure for the same conditions for which Microwave-Induced Resistance Oscillations (MIROs) are observed. It is shown that the electrodynamic aspect of the problem is important in these experiments. In the absorption experiments a broad CR line was observed due to a large reflection from the highly conductive electron gas. There were no additional features observed related to absorption at harmonics of the cyclotron resonance. In near-field reflection experiments a very different oscillation pattern was revealed when compared to MIROs. The oscillation pattern observed in the reflection experiments is probably due to plasma effects occurring in a finite-size sample. The whole microscopic picture of MIROs is more complicated than simply a resonant absorption at harmonics of the cyclotron resonance. Nevertheless, the experimental observations are in good agreement with the model by Durst et al. involving the photo-assisted scattering in the presence of a crossed magnetic field and dc bias. The observed damping factor of MIROs may be attributed to a change in the electron mobility as a function of temperature. MIROs may be considered as a light-induced drift effect, a broad class of phenomena associated with a light-induced asymmetry in the velocity distribution function.

Absorption and Reflection Experiments on High-Mobility 2DEGs in the Regime of Microwave-Induced Resistance Oscillations

NASA Astrophysics Data System (ADS)

Studenikin, S. A.; Potemski, M.; Sachrajda, A. S.; Hilke, M.; Pfeiffer, L. N.; West, K. W.

We have performed microwave absorption and near-field reflection experiments on a high mobility GaAs/AlGaAs heterostructure for the same conditions for which Microwave-Induced Resistance Oscillations (MIROs) are observed. It is shown that the electrodynamic aspect of the problem is important in these experiments. In the absorption experiments a broad CR line was observed due to a large reflection from the highly conductive electron gas. There were no additional features observed related to absorption at harmonics of the cyclotron resonance. In near-field reflection experiments a very different oscillation pattern was revealed when compared to MIROs. The oscillation pattern observed in the reflection experiments is probably due to plasma effects occurring in a finite-size sample. The whole microscopic picture of MIROs is more complicated than simply a resonant absorption at harmonics of the cyclotron resonance. Nevertheless, the experimental observations are in good agreement with the model by Durst et al. involving the photo-assisted scattering in the presence of a crossed magnetic field and dc bias. The observed damping factor of MIROs may be attributed to a change in the electron mobility as a function of temperature. MIROs may be considered as a light-induced drift effect, a broad class of phenomena associated with a light-induced asymmetry in the velocity distribution function.

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

Capturing latent fingerprints from metallic painted surfaces using UV-VIS spectroscope

NASA Astrophysics Data System (ADS)

Makrushin, Andrey; Scheidat, Tobias; Vielhauer, Claus

2015-03-01

In digital crime scene forensics, contactless non-destructive detection and acquisition of latent fingerprints by means of optical devices such as a high-resolution digital camera, confocal microscope, or chromatic white-light sensor is the initial step prior to destructive chemical development. The applicability of an optical sensor to digitalize latent fingerprints primarily depends on reflection properties of a substrate. Metallic painted surfaces, for instance, pose a problem for conventional sensors which make use of visible light. Since metallic paint is a semi-transparent layer on top of the surface, visible light penetrates it and is reflected off of the metallic flakes randomly disposed in the paint. Fingerprint residues do not impede light beams making ridges invisible. Latent fingerprints can be revealed, however, using ultraviolet light which does not penetrate the paint. We apply a UV-VIS spectroscope that is capable of capturing images within the range from 163 to 844 nm using 2048 discrete levels. We empirically show that latent fingerprints left behind on metallic painted surfaces become clearly visible within the range from 205 to 385 nm. Our proposed streakiness score feature determining the proportion of a ridge-valley pattern in an image is applied for automatic assessment of a fingerprint's visibility and distinguishing between fingerprint and empty regions. The experiments are carried out with 100 fingerprint and 100 non-fingerprint samples.

Macular photostress and visual experience between microscope and intracameral illumination during cataract surgery.

PubMed

Seo, Hyejin; Nam, Dong Heun; Lee, Jong Yeon; Park, Su Jin; Kim, Yu Jeong; Kim, Seong-Woo; Chung, Tae-Young; Inoue, Makoto; Kim, Terry

2018-02-01

To evaluate macular photostress and visual experience between coaxial microscope illumination versus oblique intracameral illumination during cataract surgery. Gachon University Gil Hospital, Incheon, South Korea. Prospective case series. Consecutive patients who had cataract surgery using microscope illumination and intracameral illumination were included. The patients were asked to complete a questionnaire (seeing strong lights, feeling photophobia, feeling startled (fright) when seeing lights, seeing any colors, seeing any instruments or surgical procedures, and estimating intraoperative visual function) designed to describe their cataract surgery experience. The images projected on the retina of the model eye (rear view) with artificial opaque fragments in the anterior chamber during simulating cataract surgery were compared between the 2 illumination types. Sixty patients completed the questionnaire. Scores for strong lights, photophobia, fright, and color perception were significantly higher with microscope illumination than with intracameral illumination (all P < .001). More patients preferred the intracameral illumination (45 [75.0%]) to the microscope illumination (13 [21.7%]). In the rear-view images created in a model eye, only the bright microscope light in the center was seen without any lens image in the microscope illumination. However, in the intracameral illumination, the less bright light from the light pipe in the periphery and the lens fragments were seen more clearly. In a view of the patients' visual experience, oblique intracameral illumination caused less subjective photostress and was preferred over coaxial microscope illumination. Objective findings from the model-eye experiment correlated to the result of visual experience. Copyright © 2018 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

Are those bugs reflective? Non-destructive biofilm imaging with white light interferometry

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

Larimer, Curtis J.; Brann, Michelle R.; Suter, Jonathan D.

White light interferometry (WLI) is not typically used to image bacterial biofilms that are immersed in water because there is insufficient refractive index contrast to induce reflection from the biofilm’s interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ by any means, especially in a non-destructive manner. Here we describe a new method for measuring and monitoring the thickness and topology of live biofilms using a WLI microscope. A microfluidic system was used to create a reflective interface on the surface of biofilms. Live biofilm samples were monitored non-destructively over time.more » The method enables surface metrology measurements (roughness, surface area) and a novel approach to measuring thickness of the thin hydrated biofilms. Increase in surface roughness preceded observable increase in biofilm thickness, indicating that this measure may be used to predict future development of biofilms. We have also developed a flow cell that enables WLI biofilm imaging in a dynamic environment. We have used this flow cell to observe changes in biofilm structure in response to changes in environmental conditions - flow velocity, availability of nutrients, and presence of biocides.« less

Advanced spinning disk-TIRF microscopy for faster imaging of the cell interior and the plasma membrane.

PubMed

Zobiak, Bernd; Failla, Antonio Virgilio

2018-03-01

Understanding the cellular processes that occur between the cytosol and the plasma membrane is an important task for biological research. Till now, however, it was not possible to combine fast and high-resolution imaging of both the isolated plasma membrane and the surrounding intracellular volume. Here, we demonstrate the combination of fast high-resolution spinning disk (SD) and total internal reflection fluorescence (TIRF) microscopy for specific imaging of the plasma membrane. A customised SD-TIRF microscope was used with specific design of the light paths that allowed, for the first time, live SD-TIRF experiments at high acquisition rates. A series of experiments is shown to demonstrate the feasibility and performance of our setup. © 2017 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.

Free-standing GaN grating couplers and rib waveguide for planar photonics at telecommunication wavelength

NASA Astrophysics Data System (ADS)

Liu, Qifa; Wang, Wei

2018-01-01

Gallium Nitride (GaN) free-standing planar photonic device at telecommunication wavelength based on GaN-on-silicon platform was presented. The free-standing structure was realized by particular double-side fabrication process, which combining GaN front patterning, Si substrate back releasing and GaN slab etching. The actual device parameters were identified via the physical characterizations employing scanning electron microscope (SEM), atomic force microscope (AFM) and reflectance spectra testing. High coupling efficiency and good light confinement properties of the gratings and rib waveguide at telecommunication wavelength range were verified by finite element method (FEM) simulation. This work illustrates the potential of new GaN photonic structure which will enable new functions for planar photonics in communication and sensing applications, and is favorable for the realization of integrated optical circuit.

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

PubMed Central

Kelner, Roy; Katz, Barak; Rosen, Joseph

2014-01-01

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

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.

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.

Intraoperative Fluorescence Cerebral Angiography by Laser Surgical Microscopy: Comparison With Xenon Microscopy and Simultaneous Observation of Cerebral Blood Flow and Surrounding Structures.

PubMed

Ito, Yuhei; Suzuki, Kyouichi; Ichikawa, Tsuyoshi; Watanabe, Yoichi; Sato, Taku; Sakuma, Jun; Saito, Kiyoshi

2018-06-12

Laser surgical microscopes should enable uniform illumination of the operative field, and require less luminous energy compared with existing xenon surgical microscopes. To examine the utility of laser illumination in fluorescence cerebral angiography. Fluorescein sodium (fluorescein) was used as a fluorescent dye. We first compared the clarity of cerebral blood flow images collected by fluorescence angiography between the laser illumination and xenon illumination methods. We then assessed use of the laser illuminator for simultaneous observation of blood flow and surrounding structures during fluorescence angiography. Furthermore, the study was designed to evaluate usefulness of the thus determined excitation light in clinical cases. Fluorescence angiography using blue light laser for excitation provided higher clarity and contrast blood flow images compared with using blue light generated from a xenon lamp. Further, illumination with excitation light consisting of a combination of 3 types of laser (higher level of blue light, no green light, and lower level of red light) enabled both blood flow and surrounding structures to be observed through the microscope directly by the surgeon. Laser-illuminated fluorescence angiography provides high clarity and contrast images of cerebral blood flow. Further, a laser providing strong blue light and weak red light for excitation light enables simultaneous visual observation of fluorescent blood flow and surrounding structures by the surgeon using a surgical microscope. Overall, these data suggest that laser surgical microscopes are useful for both ordinary operative manipulations and fluorescence angiography.

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

In vitro near-infrared imaging of natural secondary caries.

PubMed

Simon, Jacob C; Lucas, Seth; Lee, Robert; Darling, Cynthia L; Staninec, Michal; Vanderhobli, Ram; Pelzner, Roger; Fried, Daniel

2015-02-24

Secondary caries stands as the leading reason for the failure of composite restorations and dentists spend more time replacing existing restorations than placing new ones. Current clinical strategies, and even modern visible light methods designed to detect decay, lack the sensitivity to distinguish incipient lesions, are confounded by staining on the surface and within the tooth, or are limited to detecting decay on the tooth surface. Near-IR (NIR) imaging methods, such as NIR reflectance and transillumination imaging, and optical coherence tomography are promising strategies for imaging secondary caries. Wavelengths longer than 1300-nm avoid interference from stain and exploit the greater transparency of sound enamel and dental composites, to provide increased contrast with demineralized tissues and improved imaging depth. The purpose of this study was to determine whether NIR transillumination (λ=1300-nm) and NIR cross-polarized reflectance (λ=1500-1700-nm) images can serve as reliable indicators of demineralization surrounding composite restorations. Twelve composite margins (n=12) consisting of class I, II & V restorations were chosen from ten extracted teeth. The samples were imaged in vitro using NIR transillumination and reflectance, polarization sensitive optical coherence tomography (PS-OCT) and a high-magnification digital visible light microscope. Samples were serially sectioned into 200- μ m slices for histological analysis using polarized light microscopy (PLM) and transverse microradiography (TMR). The results presented demonstrate the utility of NIR light for detecting recurrent decay and suggest that NIR images could be a reliable screening tool used in conjunction with PS-OCT for the detection and diagnosis of secondary caries.

In-vitro near-infrared imaging of natural secondary caries

NASA Astrophysics Data System (ADS)

Simon, Jacob C.; Lucas, Seth; Lee, Robert; Darling, Cynthia L.; Staninec, Michal; Vanderhobli, Ram; Pelzner, Roger; Fried, Daniel

2015-02-01

Secondary caries stands as the leading reason for the failure of composite restorations and dentists spend more time replacing existing restorations than placing new ones. Current clinical strategies, and even modern visible light methods designed to detect decay, lack the sensitivity to distinguish incipient lesions, are confounded by staining on the surface and within the tooth, or are limited to detecting decay on the tooth surface. Near-IR (NIR) imaging methods, such as NIR reflectance and transillumination imaging, and optical coherence tomography are promising strategies for imaging secondary caries. Wavelengths longer than 1300-nm avoid interference from stain and exploit the greater transparency of sound enamel and dental composites, to provide increased contrast with demineralized tissues and improved imaging depth. The purpose of this study was to determine whether NIR transillumination (λ=1300-nm) and NIR crosspolarized reflectance (λ=1500-1700-nm) images can serve as reliable indicators of demineralization surrounding composite restorations. Twelve composite margins (n=12) consisting of class I, II and V restorations were chosen from ten extracted teeth. The samples were imaged in vitro using NIR transillumination and reflectance, polarization sensitive optical coherence tomography (PS-OCT) and a high-magnification digital visible light microscope. Samples were serially sectioned into 200-μm slices for histological analysis using polarized light microscopy (PLM) and transverse microradiography (TMR). The results presented demonstrate the utility of NIR light for detecting recurrent decay and suggest that NIR images could be a reliable screening tool used in conjunction with PS-OCT for the detection and diagnosis of secondary caries.

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

Laser shock wave assisted patterning on NiTi shape memory alloy surfaces

NASA Astrophysics Data System (ADS)

Seyitliyev, Dovletgeldi; Li, Peizhen; Kholikov, Khomidkhodza; Grant, Byron; Karaca, Haluk E.; Er, Ali O.

2017-02-01

An advanced direct imprinting method with low cost, quick, and less environmental impact to create thermally controllable surface pattern using the laser pulses is reported. Patterned micro indents were generated on Ni50Ti50 shape memory alloys (SMA) using an Nd:YAG laser operating at 1064 nm combined with suitable transparent overlay, a sacrificial layer of graphite, and copper grid. Laser pulses at different energy densities which generates pressure pulses up to 10 GPa on the surface was focused through the confinement medium, ablating the copper grid to create plasma and transferring the grid pattern onto the NiTi surface. Scanning electron microscope (SEM) and optical microscope images of square pattern with different sizes were studied. One dimensional profile analysis shows that the depth of the patterned sample initially increase linearly with the laser energy until 125 mJ/pulse where the plasma further absorbs and reflects the laser beam. In addition, light the microscope image show that the surface of NiTi alloy was damaged due to the high power laser energy which removes the graphite layer.

Characterization of process-induced damage in Cu/low-k interconnect structure by microscopic infrared spectroscopy with polarized infrared light

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

Seki, Hirofumi, E-mail: Hirofumi-Seki@trc.toray.co.jp; Hashimoto, Hideki; Ozaki, Yukihiro

Microscopic Fourier-transform infrared (FT-IR) spectra are measured for a Cu/low-k interconnect structure using polarized IR light for different widths of low-k spaces and Cu lines, and for different heights of Cu lines, on Si substrates. Although the widths of the Cu line and the low-k space are 70 nm each, considerably smaller than the wavelength of the IR light, the FT-IR spectra of the low-k film were obtained for the Cu/low-k interconnect structure. A suitable method was established for measuring the process-induced damage in a low-k film that was not detected by the TEM-EELS (Transmission Electron Microscope-Electron Energy-Loss Spectroscopy) using microscopicmore » IR polarized light. Based on the IR results, it was presumed that the FT-IR spectra mainly reflect the structural changes in the sidewalls of the low-k films for Cu/low-k interconnect structures, and the mechanism of generating process-induced damage involves the generation of Si-OH groups in the low-k film when the Si-CH{sub 3} bonds break during the fabrication processes. The Si-OH groups attract moisture and the OH peak intensity increases. It was concluded that the increase in the OH groups in the low-k film is a sensitive indicator of low-k damage. We achieved the characterization of the process-induced damage that was not detected by the TEM-EELS and speculated that the proposed method is applicable to interconnects with line and space widths of 70 nm/70 nm and on shorter scales of leading edge devices. The location of process-induced damage and its mechanism for the Cu/low-k interconnect structure were revealed via the measurement method.« less

Crystal Structure, Magnetic and Optical Properties of Mn-Doped BiFeO₃ by Hydrothermal Synthesis.

PubMed

Zhang, Ning; Wei, Qinhua; Qin, Laishun; Chen, Da; Chen, Zhi; Niu, Feng; Wang, Jiangying; Huanag, Yuexiang

2017-01-01

In this paper, Mn doped BiFeO₃ were firstly synthesized by hydrothermal process. The influence of Mn doping on structural, optical and magnetic properties of BiFeO₃ was studied. The different amounts of Mn doping in BiFeO₃ were characterized by X-ray diffraction, Scanning Electron Microscope, Energy Dispersive X-ray Spectroscope, UV-Vis diffuse reflectance spectroscopy and magnetic measurements. The X-ray diffraction (XRD) patterns confirmed the formation of pure phase rhombohedral structure in BiFe(1−x) Mn (x) O₃ (x = 0.01, 0.03, 0.05, 0.07) samples. The morphologies and chemical compositions of as-prepared samples could be observed by Scanning Electron Microscope (SEM) and Energy Dispersive X-ray Spectroscope (EDS). A relative large saturated magnetization (Ms) of 0.53 emu/g for x = 0.07 sample was obtained at room temperature, which is considered to be Mn ions doping. UV-Vis diffuse reflectance spectroscopy showed strong absorption of light in the range of 200–1000 nm, indicating the optical band gap in the visible region for these samples. This implied that BiFe(1−x) Mn(x)O₃ may be a potential photocatalyst for utilizing solar energy.

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.

Utility and safety of a novel surgical microscope laser light source

PubMed Central

Bakhit, Mudathir S.; Suzuki, Kyouichi; Sakuma, Jun; Fujii, Masazumi; Murakami, Yuta; Ito, Yuhei; Sugano, Tetsuo; Saito, Kiyoshi

2018-01-01

Objective Tissue injuries caused by the thermal effects of xenon light microscopes have previously been reported. Due to this, the development of a safe microscope light source became a necessity. A newly developed laser light source is evaluated regarding its effectiveness and safety as an alternative to conventional xenon light source. Methods We developed and tested a new laser light source for surgical microscopes. Four experiments were conducted to compare xenon and laser lights: 1) visual luminance comparison, 2) luminous and light chromaticity measurements, 3) examination and analysis of visual fatigue, and 4) comparison of focal temperature elevation due to light source illumination using porcine muscle samples. Results Results revealed that the laser light could be used at a lower illumination value than the xenon light (p < 0.01). There was no significant difference in visual fatigue status between the laser light and the xenon light. The laser light was superior to the xenon light regarding luminous intensity and color chromaticity. The focal temperature elevation of the muscle samples was significantly higher when irradiated with xenon light in vitro than with laser light (p < 0.01). Conclusion The newly developed laser light source is more efficient and safer than a conventional xenon light source. It lacks harmful ultraviolet waves, has a longer lifespan, a lower focal temperature than that of other light sources, a wide range of brightness and color production, and improved safety for the user’s vision. Further clinical trials are necessary to validate the impact of this new light source on the patient’s outcome and prognosis. PMID:29390016

AccessScope project: Accessible light microscope for users with upper limb mobility or visual impairments.

PubMed

Mansoor, Awais; Ahmed, Wamiq M; Samarapungavan, Ala; Cirillo, John; Schwarte, David; Robinson, J Paul; Duerstock, Bradley S

2010-01-01

A web-based application was developed to remotely view slide specimens and control all functions of a research-level light microscopy workstation, called AccessScope. Students and scientists with upper limb mobility and visual impairments are often unable to use a light microscope by themselves and must depend on others in its operation. Users with upper limb mobility impairments and low vision were recruited to assist in the design process of the AccessScope personal computer (PC) user interface. Participants with these disabilities were evaluated in their ability to use AccessScope to perform microscopical tasks. AccessScope usage was compared with inspecting prescanned slide images by grading participants' identification and understanding of histological features and knowledge of microscope operation. With AccessScope subjects were able to independently perform common light microscopy functions through an Internet browser by employing different PC pointing devices or accessibility software according to individual abilities. Subjects answered more histology and microscope usage questions correctly after first participating in an AccessScope test session. AccessScope allowed users with upper limb or visual impairments to successfully perform light microscopy without assistance. This unprecedented capability is crucial for students and scientists with disabilities to perform laboratory coursework or microscope-based research and pursue science, technology, engineering, and mathematics fields.

Cu2S-Cu-TiO2 mesoporous carbon composites for the degradation of high concentration of methyl orange under visible light

NASA Astrophysics Data System (ADS)

Zhang, Liang; Zhao, Yuan; Zhong, Lvling; Wang, Yang; Chai, Shouning; Yang, Tao; Han, Xuanli

2017-11-01

A Schiff base compound was used to prepare a Cu2S-Cu-TiO2 mesoporous carbon composite photocatalyst (Cu2S-Cu-TiO2/MC) by a simple precipitation-carbonization method with a carbonization temperature of 750 °C. X-ray diffraction and x-ray photoelectron spectroscopy studies show that Cu2S, Cu, and TiO2 exist in Cu2S-Cu-TiO2/MC in the form of nanometer-sized particles. Scanning electron microscope and transmission electron microscope images show that the composites form a spherical carbon structure inlaid with Cu2S and Cu and coated TiO2. The Brunauer-Emmett-Teller test shows that the material has a large specific surface area (76.14 m2/g) and mesoporous structure. UV-vis diffuse reflection spectroscopy and photoluminescence spectroscopy indicate that the recombination of photo-generated electrons and holes in the samples were inhibited. The composites show good degradation performance in a high concentration (300 mg/L) of methyl orange (MO) solution under visible light. The composites exhibit great potential in the treatment of dyes for wastewater treatment.

Sporadic-inclusion body myositis (s-IBM) is not so prevalent in Istanbul/Turkey: a muscle biopsy based survey.

PubMed

Oflazer, P Serdaroglu; Deymeer, F; Parman, Y

2011-06-01

In a muscle biopsy based study, only 9 out of 5450 biopsy samples, received from all parts of greater Istanbul area, had typical clinical and most suggestive light microscopic sporadic-inclusion body myositis (s-IBM) findings. Two other patients with and ten further patients without characteristic light microscopic findings had referring diagnosis of s-IBM. As the general and the age-adjusted populations of Istanbul in 2010 were 13.255.685 and 2.347.300 respectively, the calculated corresponding 'estimated prevalences' of most suggestive s-IBM in the Istanbul area were 0.679 X 10(-6) and 3.834 X 10(-6). Since Istanbul receives heavy migration from all regions of Turkey and ours is the only muscle pathology laboratory in Istanbul, projection of these figures to the Turkish population was considered to be reasonable and an estimate of the prevalence of s-IBM in Turkey was obtained. The calculated 'estimated prevalence' of s-IBM in Turkey is lower than the previously reported rates from other countries. The wide variation in the prevalence rates of s-IBM may reflect different genetic, immunogenetic or environmental factors in different populations.

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.

How the confocal laser scanning microscope entered biological research.

PubMed

Amos, W B; White, J G

2003-09-01

A history of the early development of the confocal laser scanning microscope in the MRC Laboratory of Molecular Biology in Cambridge is presented. The rapid uptake of this technology is explained by the wide use of fluorescence in the 80s. The key innovations were the scanning of the light beam over the specimen rather than vice-versa and a high magnification at the level of the detector, allowing the use of a macroscopic iris. These were followed by an achromatic all-reflective relay system, a non-confocal transmission detector and novel software for control and basic image processing. This design was commercialized successfully and has been produced and developed over 17 years, surviving challenges from alternative technologies, including solid-state scanning systems. Lessons are pointed out from the unusual nature of the original funding and research environment. Attention is drawn to the slow adoption of the instrument in diagnostic medicine, despite promising applications.

ELLIPSOMETRY OF ELECTROCHEMICAL SURFACE LAYERS

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

Muller, R.H.

1977-06-01

Ellipsometry is concerned with the analysis and interpretation of changes in the state of polarization caused by reflection. The technique has found increasing interest in recent years for the measurement of thin films because it is unusually sensitive, disturbs the object minimally and can be applied to surfaces contained in any optically transparent medium. Film thicknesses amenable to measurement range from fractional monoatomic coverage to microscopic thicknesses. The measurement of changes in the state of polarization of light due to reflection provides an unusually sensitive tool for observing surface layers in any optically transparent environment. A fast, self-compensating ellipsometer hasmore » been used to observe the electrochemical formation of reacted surface layers. The optical effect of mass-transport boundary layers and component imperfections have been taken into account in the interpretation of results.« less

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.

Autofluorescence-based diagnostic UV imaging of tissues and cells

NASA Astrophysics Data System (ADS)

Renkoski, Timothy E.

Cancer is the second leading cause of death in the United States, and its early diagnosis is critical to improving treatment options and patient outcomes. In autofluorescence (AF) imaging, light of controlled wavelengths is projected onto tissue, absorbed by specific molecules, and re-emitted at longer wavelengths. Images of re-emitted light are used together with spectral information to infer tissue functional information and diagnosis. This dissertation describes AF imaging studies of three different organs using data collected from fresh human surgical specimens. In the ovary study, illumination was at 365 nm, and images were captured at 8 emission wavelengths. Measurements from a multispectral imaging system and fiber optic probe were used to map tissue diagnosis at every image pixel. For the colon and pancreas studies, instrumentation was developed extending AF imaging capability to sub-300 nm excitation. Images excited in the deep UV revealed tryptophan and protein content which are believed to change with disease state. Several excitation wavelength bands from 280 nm to 440 nm were investigated. Microscopic AF images collected in the pancreas study included both cultured and primary cells. Several findings are reported. A method of transforming fiber optic probe spectra for direct comparison with imager spectra was devised. Normalization of AF data by green reflectance data was found useful in correcting hemoglobin absorption. Ratio images, both AF and reflectance, were formulated to highlight growths in the colon. Novel tryptophan AF images were found less useful for colon diagnostics than the new ratio techniques. Microscopic tryptophan AF images produce useful visualization of cellular protein content, but their diagnostic value requires further study.

Reflection soft X-ray microscope and method

DOEpatents

Suckewer, Szymon; Skinner, Charles H.; Rosser, Roy

1993-01-01

A reflection soft X-ray microscope is provided by generating soft X-ray beams, condensing the X-ray beams to strike a surface of an object at a predetermined angle, and focusing the X-ray beams reflected from the surface onto a detector, for recording an image of the surface or near surface features of the object under observation.

Reflection soft X-ray microscope and method

DOEpatents

Suckewer, S.; Skinner, C.H.; Rosser, R.

1993-01-05

A reflection soft X-ray microscope is provided by generating soft X-ray beams, condensing the X-ray beams to strike a surface of an object at a predetermined angle, and focusing the X-ray beams reflected from the surface onto a detector, for recording an image of the surface or near surface features of the object under observation.

Holographic microscopy for in situ studies of microorganism motility

NASA Astrophysics Data System (ADS)

Nadeau, J.; Hu, S.; Jericho, S.; Lindensmith, C.

2011-12-01

Robust technologies for the detection and identification of microorganisms at low concentrations in complex liquid media are needed for numerous applications: environmental and medical microbiology, food safety, and for the search for microbial life elsewhere in the Solar System. The best current method for microbial enumeration is specific labeling with fluorescent dyes followed by high-resolution light microscopy. However, fluorescent techniques are difficult to use in situ in extreme environments (such as the Arctic and Antarctic or the open ocean) due to the fragility of the instruments and their high power demands. In addition, light microscopic techniques rarely provide insight into microbial motility behaviors. Tracking single cells would provide important insight into the physics of micron-scale motility as well as into key microbial phenomena such as surface attachment and invasiveness. An alternative to traditional light microscopy that is attracting increasing attention is holographic microscopy. Holographic microscopy works by illuminating the object of interest with coherent light from a laser. The light reflected from (or transmitted through) the object is then combined with a coherent reference beam to create an interference pattern that contains the phase and intensity information required to reconstruct a three dimensional image of the object. The interference pattern is recorded on a high resolution detector and can be used to computationally reconstruct a 3D image of the object. The lateral resolution of the image depends upon the wavelength of the light used, the laser power, camera quality, and external noise sources (vibration, stray light, and so forth). Although the principle is simple, technological barriers have prevented wider use of holographic microscopy. Laser sources and CCD cameras with the appropriate properties have only very recently become affordable. In addition, holographic microscopy leads to large data sets that are computationally intensive to reconstruct images from, so the technology to store and process large amounts of data are required. We have successfully deployed a digital in-line holographic microscope in lakes of the Canadian High Arctic and the open ocean. We present characteristic data sets from these experiments, as well as discussing how data acquisition and instrumentation can be improved. A design for a new type of autonomous, submersible holographic microscope incorporating an off-axis reference beam is presented, and future plans for controlled microbe-polymer studies are detailed.

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.

Simultaneous measurements of absorption spectrum and refractive index in a microfluidic system.

PubMed

Helseth, Lars Egil

2012-02-13

The characterization of dyes in various solvents requires determination of the absorption spectrum of the dye as well as the refractive index of the solvent. Typically, the refractive index of the solvent and the absorption spectrum of the solute are measured using separate experimental setups where significant liquid volumes are required. In this work the first optical measurement system that is able to do simultaneous measurements of the refractive index of the solvent and the spectral properties of the solute in a microscopic volume is presented. The laser dye Rhodamine 6G in glycerol is investigated, and the refractive index of the solution is monitored using the interference pattern of the light scattered off the channel, while its spectral properties is found by monitoring reflected light from the channel.

X ray microscope assembly and alignment support and advanced x ray microscope design and analysis

NASA Technical Reports Server (NTRS)

Shealy, David L.

1991-01-01

Considerable efforts have been devoted recently to the design, analysis, fabrication, and testing of spherical Schwarzschild microscopes for soft x ray application in microscopy and projection lithography. The spherical Schwarzschild microscope consists of two concentric spherical mirrors configured such that the third order spherical aberration and coma are zero. Since multilayers are used on the mirror substrates for x ray applications, it is desirable to have only two reflecting surfaces in a microscope. In order to reduce microscope aberrations and increase the field of view, generalized mirror surface profiles have been considered in this investigation. Based on incoherent and sine wave modulation transfer function (MTF) calculations, the object plane resolution of a microscope has been analyzed as a function of the object height and numerical aperture (NA) of the primary for several spherical Schwarzschild, conic, and aspherical head reflecting two mirror microscope configurations.

Effects of compression on human skin optical properties

NASA Astrophysics Data System (ADS)

Chan, Eric K.; Sorg, Brian S.; Protsenko, Dmitry E.; O'Neil, Michael P.; Motamedi, Massoud; Welch, Ashley J.

1997-08-01

Tissue optical properties are necessary parameters for prescribing light dosimetry in photomedicine. In many diagnostic or therapeutic applications where optical fiber probes are used, pressure is often applied to the tissue to reduce index mismatch and increase light transmittance. In this study, we have measured in vitro optical properties as a function of pressure with a visible-IR spectrophotometer. A spectral range of 400 - 1800 nm with a spectral resolution of 5 nm was used for all measurements. Skin specimens of two Hispanic donors and three caucasian donors were obtained from the tissue bank. Each specimen, sandwiched between microscope slides, was compressed by a spring-loaded apparatus. Then diffuse reflectance and transmittance of each sample were measured at no load and at approximately 0.1 and 1 kgf/cm2. Under compression, tissue thicknesses were reduced up to 78%. Generally, reflectance decreased while the overall transmittance increased under compression. The absorption and reduced scattering coefficients were calculated using the inverse adding doubling method. Compared with the no-load controls, there was an increase in the absorption and scattering coefficients among most of the compressed specimens.

Gold nanorods based diffusion reflection measurements: current status and perspectives for clinical applications

NASA Astrophysics Data System (ADS)

Ankri, Rinat; Fixler, Dror

2017-07-01

Optical imaging is a powerful tool for investigating the structure and function of tissues. Tissue optical imaging technologies are generally discussed under two broad regimes: microscopic and macroscopic, while the latter is widely investigated in the field of light-tissue interaction. Among the developed optical technologies for tissue investigation, the diffusion reflectance (DR) method is a simple and safe technology. However, this method suffers from low specificity and low signal-to-noise ratio, so the extraction of the tissue properties is not an easy task. In this review, we describe the use of gold nanorods (GNRs) in DR spectroscopy. The GNRs present unique optical properties which enhance the scattering and absorption properties of a tissue. The GNRs can be easily targeted toward abnormal sites in order to improve the DR signal and to distinguish between the healthy and the abnormal sites in the tissue, with high specificity. This article describes the use of the DR-GNRs method for the detection of cancer and atherosclerosis, from light transfer theory, through the extraction of the tissue properties using the diffusion theory and up to DR in vivo measurements.

Extending Whole Slide Imaging: Color Darkfield Internal Reflection Illumination (DIRI) for Biological Applications

PubMed Central

Namiki, Kana; Miyawaki, Atsushi; Ishikawa, Takuji

2017-01-01

Whole slide imaging (WSI) is a useful tool for multi-modal imaging, and in our work, we have often combined WSI with darkfield microscopy. However, traditional darkfield microscopy cannot use a single condenser to support high- and low-numerical-aperture objectives, which limits the modality of WSI. To overcome this limitation, we previously developed a darkfield internal reflection illumination (DIRI) microscope using white light-emitting diodes (LEDs). Although the developed DIRI is useful for biological applications, substantial problems remain to be resolved. In this study, we propose a novel illumination technique called color DIRI. The use of three-color LEDs dramatically improves the capability of the system, such that color DIRI (1) enables optimization of the illumination color; (2) can be combined with an oil objective lens; (3) can produce fluorescence excitation illumination; (4) can adjust the wavelength of light to avoid cell damage or reactions; and (5) can be used as a photostimulator. These results clearly illustrate that the proposed color DIRI can significantly extend WSI modalities for biological applications. PMID:28085892

Magnetically tunable oil droplet lens of deep-sea shrimp

NASA Astrophysics Data System (ADS)

Iwasaka, M.; Hirota, N.; Oba, Y.

2018-05-01

In this study, the tunable properties of a bio-lens from a deep-sea shrimp were investigated for the first time using magnetic fields. The skin of the shrimp exhibited a brilliantly colored reflection of incident white light. The light reflecting parts and the oil droplets in the shrimp's skin were observed in a glass slide sample cell using a digital microscope that operated in the bore of two superconducting magnets (maximum strengths of 5 and 13 T). In the ventral skin of the shrimp, which contained many oil droplets, some comparatively large oil droplets (50 to 150 μm in diameter) were present. A distinct response to magnetic fields was found in these large oil droplets. Further, the application of the magnetic fields to the sample cell caused a change in the size of the oil droplets. The phenomena observed in this work indicate that the oil droplets of deep sea shrimp can act as lenses in which the optical focusing can be modified via the application of external magnetic fields. The results of this study will make it possible to fabricate bio-inspired soft optical devices in future.

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.

Optical anisotropy in micromechanically rolled carbon nanotube forest

NASA Astrophysics Data System (ADS)

Razib, Mohd Asyraf bin Mohd; Rana, Masud; Saleh, Tanveer; Fan, Harrison; Koch, Andrew; Nojeh, Alireza; Takahata, Kenichi; Muthalif, Asan Gani Bin Abdul

2017-09-01

The bulk appearance of arrays of vertically aligned carbon nanotubes (VACNT arrays or CNT forests) is dark as they absorb most of the incident light. In this paper, two postprocessing techniques have been described where the CNT forest can be patterned by selective bending of the tips of the nanotubes using a rigid cylindrical tool. A tungsten tool was used to bend the vertical structure of CNTs with predefined parameters in two different ways as stated above: bending using the bottom surface of the tool (micromechanical bending (M2B)) and rolling using the side of the tool (micromechanical rolling (M2R)). The processed zone was investigated using a Field Emission Scanning Electron Microscope (FESEM) and optical setup to reveal the surface morphology and optical characteristics of the patterned CNTs on the substrate. Interestingly, the polarized optical reflection from the micromechanical rolled (M2R) sample was found to be significantly influenced by the rotation of the sample. It was observed that, if the polarization of the light is parallel to the alignment of the CNTs, the reflectance is at least 2 x higher than for the perpendicular direction. Furthermore, the reflectance varied almost linearly with good repeatability ( 10%) as the processed CNT forest sample was rotated from 0° to 90°. [Figure not available: see fulltext.

Water window imaging x ray microscope

NASA Technical Reports Server (NTRS)

Hoover, Richard B. (Inventor)

1992-01-01

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

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.

Spectral and Geological Characterization of Beach Components in Northern Puerto Rico

NASA Astrophysics Data System (ADS)

Caraballo Álvarez, I. O.; Torres-Perez, J. L.; Barreto, M.

2015-12-01

Understanding how changes in beach components may reflect beach processes is essential since variations along beach profiles can shed light on river and ocean processes influencing beach sedimentation and beachrock formation. It is likely these influences are related to beach proximity within the Río Grande de Manatí river mouth. Therefore, this study focuses on characterizing beach components at two sites in Manatí, Puerto Rico. Playa Machuca and Playa Tombolo, which are separated by eolianites, differ greatly in sediment size, mineralogy, and beachrock morphology. Several approaches were taken to geologically and spectrally characterize main beach components at each site. These approaches included field and microscopic laboratory identification, granulometry, and a comparison between remote sensing reflectance (Rrs) obtained with a field spectroradiometer and pre-existing spectral library signatures. Preliminary results indicate a positive correlation between each method. This study may help explore the possibility of using only Rrs to characterize beach and shallow submarine components for detailed image analysis and management of coastal features.This study focuses on characterizing beach components at two sites in Manatí, Puerto Rico. Playa Machuca and Playa Tombolo, two beaches that are separated by eolianites, differ greatly in sediment size and mineralogy, as well as in beachrock morphology. Understanding how changes in beach components may reflect beach processes is essential, since it is likely that differences are mostly related to each beaches' proximity to the Río Grande de Manatí river mouth. Hence, changes in components along beach profiles can shed light on the river's and the ocean's influence on beach sedimentation and beachrock formation. Several approaches were taken to properly geologically and spectrally characterize the main beach components at each site. These approaches included field and microscopic laboratory identification, granulometry, and a comparison between remote sensing reflectance (Rrs) obtained with a field spectroradiometer and the ENVI spectral library. Preliminary results show a positive correlation between each method. This study may help explore the possibility of using only Rrs to characterize beach and shallow submarine components for detailed image analysis and management of coastal features.

Lobular and cellular patterns of early hepatic glycogen deposition in the rat as observed by light and electron microscopic radioautography after injection of /sup 3/H-galactose

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

Michaels, J.E.; Hung, J.T.; Garfield, S.A.

1984-05-01

Very low hepatic glycogen levels are achieved by overnight fasting of adrenalectomized (ADX) rats. Subsequent injection of dexamethasone (DEX), a synthetic glucocorticoid, stimulates marked increases in glycogen synthesis. Using this system and injecting /sup 3/H-galactose as a glycogen precursor 1 hr prior to sacrifice, the intralobular and intracellular patterns of labeled glycogen deposition were studied by light (LM) and electron (EM) microscopic radioautography. LM radioautography revealed that 1 hr after DEX treatment, labeling patterns for both periportal and centrilobular hepatocytes resembled those in rats with no DEX treatment: 18% of the hepatocytes were unlabeled, and 82% showed light labeling. Twomore » hours after treatment with DEX, 14% of the hepatocytes remained unlabeled, and 78% were lightly labeled; however, 8% of the cells, located randomly throughout the lobule, were intensely labeled. An increased number of heavily labeled cells (26%) appeared 3 hr after DEX treatment; and by 5 hr 91% of the hepatocytes were intensely labeled. Label over the periportal cells at this time was aggregated, whereas centrilobular cells displayed dispersed label. EM radioautographs showed that 2 to 3 hr after DEX injection initial labeling of hepatocytes, regardless of their intralobular location, occurred over foci of smooth endoplasmic reticulum (SER) and small electron-dense particles of presumptive glycogen, and in areas of SER and distinct glycogen particles. After 5 hrs of treatment with DEX, the intracellular distribution of label reflected the glycogen patterns characteristic of periportal or centrilobular regions.« less

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.

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.

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

PubMed

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

2008-11-01

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

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.

Imaging, scattering, and spectroscopic systems for biomedical optics: Tools for bench top and clinical applications

NASA Astrophysics Data System (ADS)

Cottrell, William J.

Optical advances have had a profound impact on biology and medicine. The capabilities range from sensing biological analytes to whole animal and subcellular imaging and clinical therapies. The work presented in this thesis describes three independent and multifunctional optical systems, which explore clinical therapy at the tissue level, biological structure at the cell/organelle level, and the function of underlying fundamental cellular processes. First, we present a portable clinical instrument for delivering delta-aminolevulinic acid photodynamic therapy (ALA-PDT) while performing noninvasive spectroscopic monitoring in vivo. Using an off-surface probe, the instrument delivered the treatment beam to a user-defined field on the skin and performed reflectance and fluorescence spectroscopies at two regions within this field. The instrument was used to monitor photosensitizer fluorescence photobleaching, fluorescent photoproduct kinetics, and blood oxygen saturation during a clinical ALA-PDT trial on superficial basal cell carcinoma (sBCC). Protoporphyrin IX and photoproduct fluorescence excited by the 632.8 nm PDT treatment laser was collected between 665 and 775 nm. During a series of brief treatment interruptions at programmable time points, white-light reflectance spectra between 475 and 775 nm were acquired. Fluorescence spectra were corrected for the effects of absorption and scattering, informed by the reflectance measurements, and then decomposed into known fluorophore contributions in real time using a robust singular-value decomposition fitting routine. Reflectance spectra additionally provided information on hemoglobin oxygen saturation. We next describe the incorporation of this instrument into clinical trials at Roswell Park Cancer Institute (Buffalo, NY). In this trial we examined the effects of light irradiance on photodynamic efficiency and pain. The rate of singlet-oxygen production depends on the product of irradiance and photosensitizer and oxygen concentrations. High irradiance and/or photosensitizer levels cause inefficient treatment from oxygen depletion in preclinical models. This trial established the irradiance-dependence of patient tolerability to ALA-PDT of sBCC and a pain-threshold irradiance, below which patients did not experience significant pain or require anesthetic. The irradiance-dependence of sensitizer photobleaching was also used to determine an optimal irradiance that maximized treatment efficiency. The optimal fluence at a single low irradiance is yet to be determined. We additionally report the design, construction, and initial characterization of two optical systems used for cellular scattering measurements: a forward scattering white-light spectroscopy system used to characterize lysosomal refractive index and a multifunctional scattering and fluorescence microscope that exploited an angle-resolved forward-scattering geometry. The multifunctional scattering and fluorescence microscope employed brightfield, Fourier-filtered darkfield, direct imaging of the Fourier plane, angle-resolved scattering, and white-light scattering spectroscopy while preserving a fluorescence imaging channel. Lastly, we report on the development of a microscope-based system used for high-powered, focal laser photolysis. This system was used with cell-permeable caged messenger molecules and analyte specific fluorophores to provide local stimulation of intact cells and subsequent analyte monitoring. This provided a high-precision, non-invasive means for studying Ca2+ dynamics between cell types and between sub-cellular regions within a single cell type. The resulting studies compared the mechanisms underlying the Ca2+ signal globalization in these individual exocrine cell types and under regional messenger release.

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.

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.

Comparison of Confocal and Super-Resolution Reflectance Imaging of Metal Oxide Nanoparticles

PubMed Central

Guggenheim, Emily J.; Khan, Abdullah; Pike, Jeremy; Chang, Lynne; Lynch, Iseult; Rappoport, Joshua Z.

2016-01-01

The potential for human exposure to manufactured nanoparticles (NPs) has increased in recent years, in part through the incorporation of engineered particles into a wide range of commercial goods and medical applications. NP are ideal candidates for use as therapeutic and diagnostic tools within biomedicine, however concern exists regarding their efficacy and safety. Thus, developing techniques for the investigation of NP uptake into cells is critically important. Current intracellular NP investigations rely on the use of either Transmission Electron Microscopy (TEM), which provides ultrahigh resolution, but involves cumbersome sample preparation rendering the technique incompatible with live cell imaging, or fluorescent labelling, which suffers from photobleaching, poor bioconjugation and, often, alteration of NP surface properties. Reflected light imaging provides an alternative non-destructive label free technique well suited, but not limited to, the visualisation of NP uptake within model systems, such as cells. Confocal reflectance microscopy provides optical sectioning and live imaging capabilities, with little sample preparation. However confocal microscopy is diffraction limited, thus the X-Y resolution is restricted to ~250 nm, substantially larger than the <100 nm size of NPs. Techniques such as super-resolution light microscopy overcome this fundamental limitation, providing increased X-Y resolution. The use of Reflectance SIM (R-SIM) for NP imaging has previously only been demonstrated on custom built microscopes, restricting the widespread use and limiting NP investigations. This paper demonstrates the use of a commercial SIM microscope for the acquisition of super-resolution reflectance data with X-Y resolution of 115 nm, a greater than two-fold increase compared to that attainable with RCM. This increase in resolution is advantageous for visualising small closely spaced structures, such as NP clusters, previously unresolvable by RCM. This is advantageous when investigating the subcellular trafficking of NP within fluorescently labelled cellular compartments. NP signal can be observed using RCM, R-SIM and TEM and a direct comparison is presented. Each of these techniques has its own benefits and limitations; RCM and R-SIM provide novel complementary information while the combination of modalities provides a unique opportunity to gain additional information regarding NP uptake. The use of multiple imaging methods therefore greatly enhances the range of NPs that can be studied under label-free conditions. PMID:27695038

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...

Synthesis and visible-light photocatalytic performance of flower-like porous Bi5O7I

NASA Astrophysics Data System (ADS)

Yao, Lizhu; Shi, Lei; Wang, Fangxiao

2018-04-01

Flower-like porous Bi5O7I was successfully synthesized through an easy thermal decomposition of flower-like BiOI. And its chemical structure, morphology and optical property were thoroughly analyzed by x-ray diffraction, x-ray photoelectron spectroscopy, scanning electron microscope, energydispersive spectrometry elements mapping, transmission electron microscopy, N2 adsorption-desorption isotherm, BET, and UV–vis diffuse reflectance spectra. The visible-light photocatalytic elimination of rhodamine B (RhB) was investigated. The experimental results indicated that flower-like porous Bi5O7I exhibited enhanced photocatalytic activity for degrading RhB in comparsion of flower-like BiOI, g-C3N4 and N-doped TiO2. Additionally, the as-prepared flower-like porous Bi5O7I possessed catalytic stability after recycles.

New light on ion channel imaging by total internal reflection fluorescence (TIRF) microscopy.

PubMed

Yamamura, Hisao; Suzuki, Yoshiaki; Imaizumi, Yuji

2015-05-01

Ion channels play pivotal roles in a wide variety of cellular functions; therefore, their physiological characteristics, pharmacological responses, and molecular structures have been extensively investigated. However, the mobility of an ion channel itself in the cell membrane has not been examined in as much detail. A total internal reflection fluorescence (TIRF) microscope allows fluorophores to be imaged in a restricted region within an evanescent field of less than 200 nm from the interface of the coverslip and plasma membrane in living cells. Thus the TIRF microscope is useful for selectively visualizing the plasmalemmal surface and subplasmalemmal zone. In this review, we focused on a single-molecule analysis of the dynamic movement of ion channels in the plasma membrane using TIRF microscopy. We also described two single-molecule imaging techniques under TIRF microscopy: fluorescence resonance energy transfer (FRET) for the identification of molecules that interact with ion channels, and subunit counting for the determination of subunit stoichiometry in a functional channel. TIRF imaging can also be used to analyze spatiotemporal Ca(2+) events in the subplasmalemma. Single-molecule analyses of ion channels and localized Ca(2+) signals based on TIRF imaging provide beneficial pharmacological and physiological information concerning the functions of ion channels. Copyright © 2015 The Authors. Production and hosting by Elsevier B.V. All rights reserved.

Fly artifact documentation of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) - a forensically important blowfly species in Malaysia.

PubMed

Zuha, R M; Supriyani, M; Omar, B

2008-04-01

Analysis on fly artifacts produced by forensically important blowfly, Chrysomya megacephala (Fabricius) (Diptera:Calliphoridae), revealed several unique patterns. They can be divided into fecal spots, regurgitation spots and swiping stains. The characteristics of fecal spots are round with three distinct levels of pigmentation; creamy, brownish and darkly pigmented. Matrix of the spots appears cloudy. The round spots are symmetrical and non-symmetrical, delineated by irregular and darker perimeter which only visible in fairly colored fecal spots. Diameter of these artifacts ranged from 0.5 mm to 4 mm. Vomit or regurgitation spots are determined by the presence of craters due to sucking activity of blowflies and surrounded by thickly raised and darker colored perimeter. The size of these specks ranged from 1 mm to 2 mm. Matrix of the spots displays irregular surface and reflective under auxiliary microscope light. Swiping stains due to defecation by flies consists of two distinguishable segments, the body and tail. It can be seen as a tear drop-like, sperm-like, snake-like and irregular tadpole-like stain. The direction of body and tail is inconsistent and length ranged between 4.8 mm to 9.2 mm. A finding that should be highlighted in this observation is the presence of crater on tadpole-like swiping stain which is apparent by its raised border characteristic and reflective under auxiliary microscope light. The directionality of this darkly brown stain is random. This unique mix of regurgitation and swiping stain has never been reported before. Highlighting the features of artifacts produced by flies would hopefully add our understanding in differentiating them from blood spatters produced from victims at crime scenes.

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.

Polarization Imaging Apparatus

NASA Technical Reports Server (NTRS)

Zou, Yingyin K.; Chen, Qiushui

2010-01-01

A polarization imaging apparatus has shown promise as a prototype of instruments for medical imaging with contrast greater than that achievable by use of non-polarized light. The underlying principles of design and operation are derived from observations that light interacts with tissue ultrastructures that affect reflectance, scattering, absorption, and polarization of light. The apparatus utilizes high-speed electro-optical components for generating light properties and acquiring polarization images through aligned polarizers. These components include phase retarders made of OptoCeramic (registered TradeMark) material - a ceramic that has a high electro-optical coefficient. The apparatus includes a computer running a program that implements a novel algorithm for controlling the phase retarders, capturing image data, and computing the Stokes polarization images. Potential applications include imaging of superficial cancers and other skin lesions, early detection of diseased cells, and microscopic analysis of tissues. The high imaging speed of this apparatus could be beneficial for observing live cells or tissues, and could enable rapid identification of moving targets in astronomy and national defense. The apparatus could also be used as an analysis tool in material research and industrial processing.

Design and demonstration of multimodal optical scanning microscopy for confocal and two-photon imaging

NASA Astrophysics Data System (ADS)

Chun, Wanhee; Do, Dukho; Gweon, Dae-Gab

2013-01-01

We developed a multimodal microscopy based on an optical scanning system in order to obtain diverse optical information of the same area of a sample. Multimodal imaging researches have mostly depended on a commercial microscope platform, easy to use but restrictive to extend imaging modalities. In this work, the beam scanning optics, especially including a relay lens, was customized to transfer broadband (400-1000 nm) lights to a sample without any optical error or loss. The customized scanning optics guarantees the best performances of imaging techniques utilizing the lights within the design wavelength. Confocal reflection, confocal fluorescence, and two-photon excitation fluorescence images were obtained, through respective implemented imaging channels, to demonstrate imaging feasibility for near-UV, visible, near-IR continuous light, and pulsed light in the scanning optics. The imaging performances for spatial resolution and image contrast were verified experimentally; the results were satisfactory in comparison with theoretical results. The advantages of customization, containing low cost, outstanding combining ability and diverse applications, will contribute to vitalize multimodal imaging researches.

Resin rodlets in shale and coal (Lower Cretaceous), Baltimore Canyon Trough

USGS Publications Warehouse

Lyons, P.C.; Hatcher, P.G.; Minkin, J.A.; Thompson, C.L.; Larson, R.R.; Brown, Z.A.; Pheifer, R.N.

1984-01-01

Rodlets, occurring in shale and coal (uppermost Berriasian to middle Aptian, Lower Cretaceous), were identified from drill cuttings taken from depths between 9330 ft (2844 m) and 11, 460 ft (3493 m) in the Texaco et al., Federal Block 598, No. 2 well, in the Baltimore Canyon Trough. Under the binocular microscope, most of the rodlets appear black, but a few are reddish brown, or brownish and translucent on thin edges. They range in diameter from about 0.4 to 1.7 mm and are commonly flattened. The rodlets break with a conchoidal fracture, and some show an apparent cellular cast on their longitudinal surfaces. When polished and viewed in reflected light, the rodlets appear dark gray and have an average random reflectance of less than 0.1% whereas mean maximum reflectances are 0.48-0.55% for vitrinite in the associated shale and coal. These vitrinite reflectances indicate either subbituminous A or high-volatile C bituminous coal. The rodlets fluoresce dull gray yellow to dull yellow. The scanning electron microscope (SEM) and light microscope reveal the presence of swirl-like features in the rodlet interiors. Minerals associated with the rodlets occur as sand-size grains attached to the outer surface, as finely disseminated interior grains, and as fracture fillings. Electron microprobe and SEM-energy-dispersive X-ray (EDX) anlayses indicate that the minerals are dominantly clays (probably illite and chlorite) and iron disulfide; calcium carbonate, silicon dioxide, potassium aluminum silicate (feldspar), titanium dioxide, zinc sulfide, and iron sulfate minerals have been also identified. The rodlets were analyzed directly for C, H, N, O, and total S and are interpreted as true resins on the basis of C and H contents that range from 75.6 to 80.3 and from 7.4 to 8.7 wt. % (dry, ash-free basis), respectively. Elemental and infrared data support a composition similar to that of resinite from bituminous coal. Elements determined to be organically associated in the rodlets include S (0.2-0.5 wt.%), Cl (0.03-0.1 wt.%), and Si (0.05-0.08 wt.%). The ash content of the resin rodlets ranges from 4 to 24 wt.% and averages 12 wt.%. Total sulfur contents range from 1.7 to 3.6 wt.%. Resins of fossil plants are known to have little or no sulfur and ash; therefore, these data and the presence of minerals in fractures indicate that most of the sulfur and mineral matter were introduced into the resin partly or wholly after the time of brittle fracture of the resin. The probable source of the resin rodlets is fossil pinaceous conifer cones, which are known to have resin canals as much as 2400 ??m in diameter. ?? 1984.

Design of a normal incidence multilayer imaging X-ray microscope

NASA Astrophysics Data System (ADS)

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

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

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...

Chemical analyses of fossil bone.

PubMed

Zheng, Wenxia; Schweitzer, Mary Higby

2012-01-01

The preservation of microstructures consistent with soft tissues, cells, and other biological components in demineralized fragments of dinosaur bone tens of millions of years old was unexpected, and counter to current hypotheses of tissue, cellular, and molecular degradation. Although the morphological similarity of these tissues to extant counterparts was unmistakable, after at least 80 million years exposed to geochemical influences, morphological similarity is insufficient to support an endogenous source. To test this hypothesis, and to characterize these materials at a molecular level, we applied multiple independent chemical, molecular, and microscopic analyses to identify the presence of original components produced by the extinct organisms. Microscopic techniques included field emission scanning electron microscopy, analytical transmission electron microscopy, transmitted light microscopy (LM), and fluorescence microscopy (FM). The chemical and molecular techniques include enzyme-linked immunosorbant assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis, western blot (immunoblot), and attenuated total reflectance infrared spectroscopy. In situ analyses performed directly on tissues included immunohistochemistry and time-of-flight secondary ion mass spectrometry. The details of sample preparation and methodology are described in detail herein.

HISTOLOGIC, IMMUNOHISTOCHEMICAL, AND ELECTRON MICROSCOPIC CHARACTERIZATION OF A MALIGNANT IRIDOPHOROMA IN A DWARF BEARDED DRAGON (POGONA HENRYLAWSONI).

PubMed

de Brot, Simone; Sydler, Titus; Nufer, Lisbeth; Ruetten, Maja

2015-09-01

A dwarf bearded dragon (Pogona henrylawsoni) was presented with a white subcutaneous mandibular mass and multiple nodules in the oral mucosa, heart, liver, kidney, intestine, and visceral fat. Histologically, the tumor consisted of densely packed spindle-shaped cells with brow intracytoplasmic pigment that exhibited white-blue birefringence with polarized light. Immunohistochemical staining was negative for S-100 and weakly positive with melan A. Electron microscopic examination revealed cytoplasmic irregular and oblong empty spaces, laminated and often arranged into short stacks, compatible with reflecting platelet profiles typically seen in iridophores. However, in unstained ultrathin sections, electron-dense crystalline material was present, which filled the empty spaces described for stained sections before. Based on histology, immunohistochemistry, and biologic behavior, a malignant iridophoroma was diagnosed. To the authors' knowledge, iridophoromas in lizards have rarely been characterized by using electronic microscopy. Moreover, this is the first description of an iridophoroma in a dwarf bearded dragon.

Modeling of ultrashort pulse generation in mode-locked VECSELs

NASA Astrophysics Data System (ADS)

Kilen, I.; Koch, S. W.; Hader, J.; Moloney, J. V.

2016-03-01

We present a study of various models for the mode-locked pulse dynamics in a vertical external-cavity surface emitting laser with a saturable absorber. The semiconductor Bloch equations are used to model microscopically the light-matter interaction and the carrier dynamics. Maxwell's equations describe the pulse propagation. Scattering contributions due to higher order correlation effects are approximated using effective rates that are found from a comparison to solving the microscopic scattering equations on the second Born-Markov level. It is shown that the simulations result in the same mode-locked final state whether the system is initialized with a test pulse close to the final mode-locked pulse or the full field build-up from statistical noise is considered. The influence of the cavity design is studied. The longest pulses are found for a standard V-cavity while a linear cavity and a V-cavity with an high reflectivity mirror in the middle are shown to produce similar, much shorter pulses.

A new approach to preparing Bi2Zr2O7 photocatalysts for dye degradation

NASA Astrophysics Data System (ADS)

Luo, Yijia; Cao, Liyun; Huang, Jianfeng; Feng, Liangliang; Yao, Chunyan

2018-01-01

A new synthetic route is presented to prepared pure Bi2Zr2O7 material, in which a NaNO3/KNO3 molten salt is used to obtain the resulting Bi2Zr2O7 at a relatively low temperature of 400 °C under atmospheric pressure. Powder x-ray diffraction confirmed the structure type and purity of the as-prepared sample, and further revealed that a single-source Bi(OH)3 · Zr(OH)4 · nH2O complex precursor plays a crucial role to synthesize Bi2Zr2O7 nanocrystals. Scanning electron microscope and transmission electron microscope show the morphologies and sizes of Bi2Zr2O7 crystal in detail, and UV-vis diffuse reflectance measurements evidenced the wide light absorption range. Furthermore, the as-synthesized Bi2Zr2O7 with smaller particle size and larger specific surface area exhibit superior photocatalytic activities compared with the sample obtained without adding molten salts.

Chip-based wide field-of-view nanoscopy

NASA Astrophysics Data System (ADS)

Diekmann, Robin; Helle, Øystein I.; Øie, Cristina I.; McCourt, Peter; Huser, Thomas R.; Schüttpelz, Mark; Ahluwalia, Balpreet S.

2017-04-01

Present optical nanoscopy techniques use a complex microscope for imaging and a simple glass slide to hold the sample. Here, we demonstrate the inverse: the use of a complex, but mass-producible optical chip, which hosts the sample and provides a waveguide for the illumination source, and a standard low-cost microscope to acquire super-resolved images via two different approaches. Waveguides composed of a material with high refractive-index contrast provide a strong evanescent field that is used for single-molecule switching and fluorescence excitation, thus enabling chip-based single-molecule localization microscopy. Additionally, multimode interference patterns induce spatial fluorescence intensity variations that enable fluctuation-based super-resolution imaging. As chip-based nanoscopy separates the illumination and detection light paths, total-internal-reflection fluorescence excitation is possible over a large field of view, with up to 0.5 mm × 0.5 mm being demonstrated. Using multicolour chip-based nanoscopy, we visualize fenestrations in liver sinusoidal endothelial cells.

Anisotropic excitation of surface plasmon polaritons on a metal film by a scattering-type scanning near-field microscope with a non-rotationally-symmetric probe tip

NASA Astrophysics Data System (ADS)

Walla, Frederik; Wiecha, Matthias M.; Mecklenbeck, Nicolas; Beldi, Sabri; Keilmann, Fritz; Thomson, Mark D.; Roskos, Hartmut G.

2018-01-01

We investigated the excitation of surface plasmon polaritons on gold films with the metallized probe tip of a scattering-type scanning near-field optical microscope (s-SNOM). The emission of the polaritons from the tip, illuminated by near-infrared laser radiation, was found to be anisotropic and not circularly symmetric as expected on the basis of literature data. We furthermore identified an additional excitation channel via light that was reflected off the tip and excited the plasmon polaritons at the edge of the metal film. Our results, while obtained for a non-rotationally-symmetric type of probe tip and thus specific for this situation, indicate that when an s-SNOM is employed for the investigation of plasmonic structures, the unintentional excitation of surface waves and anisotropic surface wave propagation must be considered in order to correctly interpret the signatures of plasmon polariton generation and propagation.

Effect of Organic Substrates on the Photocatalytic Reduction of Cr(VI) by Porous Hollow Ga2O3 Nanoparticles

PubMed Central

Liu, Jin; Gan, Huihui; Wu, Hongzhang; Zhang, Xinlei; Zhang, Jun; Li, Lili; Wang, Zhenling

2018-01-01

Porous hollow Ga2O3 nanoparticles were successfully synthesized by a hydrolysis method followed by calcination. The prepared samples were characterized by field emission scanning electron microscope, transmission electron microscope, thermogravimetry and differential scanning calorimetry, UV-vis diffuse reflectance spectra and Raman spectrum. The porous structure of Ga2O3 nanoparticles can enhance the light harvesting efficiency, and provide lots of channels for the diffusion of Cr(VI) and Cr(III). Photocatalytic reduction of Cr(VI), with different initial pH and degradation of several organic substrates by porous hollow Ga2O3 nanoparticles in single system and binary system, were investigated in detail. The reduction rate of Cr(VI) in the binary pollutant system is markedly faster than that in the single Cr(VI) system, because Cr(VI) mainly acts as photogenerated electron acceptor. In addition, the type and concentration of organic substrates have an important role in the photocatalytic reduction of Cr(VI). PMID:29690548

Particle Shape Characterization of Lunar Regolith using Reflected Light Microscopy

NASA Astrophysics Data System (ADS)

McCarty, C. B.; Garcia, G. C.; Rickman, D.

2014-12-01

Automated identification of particles in lunar thin sections is necessary for practical measurement of particle shape, void characterization, and quantitative characterization of sediment fabric. This may be done using image analysis, but several aspects of the lunar regolith make such automations difficult. For example, many of the particles are shattered; others are aggregates of smaller particles. Sieve sizes of the particles span 5 orders of magnitude. The physical thickness of a thin section, at a nominal 30 microns, is large compared to the size of many of the particles. Image acquisition modes, such as SEM and reflected light, while superior to transmitted light, still have significant ambiguity as to the volume being sampled. It is also desirable to have a technique that is inexpensive, not resource intensive, and analytically robust. To this end, we have developed an image acquisition and processing protocol that identifies and delineates resolvable particles on the front surface of a lunar thin section using a petrographic microscope in reflected light. For a polished thin section, a grid is defined covering the entire thin section. The grid defines discrete images taken with 20% overlap, minimizing the number of particles that intersect image boundaries. In reflected light mode, two images are acquired at each grid location, with a closed aperture diaphragm. One image, A, is focused precisely on the front surface of the thin section. The second image, B, is made after the stage is brought toward the objective lens just slightly. A bright fringe line, analogous to a Becke line, appears inside all transparent particles at the front surface of the section in the second image. The added light in the bright line corresponds to a deficit around the particles. Particle identification is done using ImageJ and uses multiple steps. A hybrid 5x5 median filter is used to make images Af and Bf. This primarily removes very small particles just below the front surface of the section. Bf - (Bf/Af) is then computed. The division strongly enhances the fringe and the deficit, while minimizing the correlated information in A and B. The subtraction emphasizes the particle-epoxy boundaries. The resulting image is converted to binary, and then holes are filled. Cracks are removed using a median-based operator.

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...

Total Internal Reflection Microscopy (TIRM) as a nondestructive surface damage assessment tool

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

Liao, Z.M.; Cohen, S.J.; Taylor, J.R.

1994-10-01

An easy to use, nondestructive, method for evaluating subsurface damage in polished substrates has been established at LLNL. Subsurface damage has been related to laser damage in coated optical components used in high power, high repetition rate laser systems. Total Internal Reflection Microscopy (TIRM) has been shown to be a viable nondestructive technique in analyzing subsurface damage in optical components. A successful TIRM system has been established for evaluating subsurface damage on fused silica components. Laser light scattering from subsurface damage sites is collected through a Nomarski microscope. These images are then captured by a CCD camera for analysis onmore » a computer. A variety of optics, including components with intentional subsurface damage due to grinding and polishing, have been analyzed and their TIRM images compared to an existing destructive etching method. Methods for quantitative measurement of subsurface damage are also discussed.« less

Development of and Clinical Experience with a Simple Device for Performing Intraoperative Fluorescein Fluorescence Cerebral Angiography: Technical Notes.

PubMed

Ichikawa, Tsuyoshi; Suzuki, Kyouichi; Watanabe, Yoichi; Sato, Taku; Sakuma, Jun; Saito, Kiyoshi

2016-01-01

To perform intraoperative fluorescence angiography (FAG) under a microscope without an integrated FAG function with reasonable cost and sufficient quality for evaluation, we made a small and easy to use device for fluorescein FAG (FAG filter). We investigated the practical use of this FAG filter during aneurysm surgery, revascularization surgery, and brain tumor surgery. The FAG filter consists of two types of filters: an excitatory filter and a barrier filter. The excitatory filter excludes all wavelengths except for blue light and the barrier filter passes long waves except for blue light. By adding this FAG filter to a microscope without an integrated FAG function, light from the microscope illuminating the surgical field becomes blue, which is blocked by the barrier filter. We put the FAG filter on the objective lens of the operating microscope correctly and fluorescein sodium was injected intravenously or intra-arterially. Fluorescence (green light) from vessels in the surgical field and the dyed tumor were clearly observed through the microscope and recorded by a memory device. This method was easy and could be performed in a short time (about 10 seconds). Blood flow of small vessels deep in the surgical field could be observed. Blood flow stagnation could be evaluated. However, images from this method were inferior to those obtained by currently commercially available microscopes with an integrated FAG function. In brain tumor surgery, a stained tumor on the brain surface could be observed using this method. FAG could be performed with a microscope without an integrated FAG function easily with only this FAG filter.

Development of and Clinical Experience with a Simple Device for Performing Intraoperative Fluorescein Fluorescence Cerebral Angiography: Technical Notes

PubMed Central

ICHIKAWA, Tsuyoshi; SUZUKI, Kyouichi; WATANABE, Yoichi; SATO, Taku; SAKUMA, Jun; SAITO, Kiyoshi

2016-01-01

To perform intraoperative fluorescence angiography (FAG) under a microscope without an integrated FAG function with reasonable cost and sufficient quality for evaluation, we made a small and easy to use device for fluorescein FAG (FAG filter). We investigated the practical use of this FAG filter during aneurysm surgery, revascularization surgery, and brain tumor surgery. The FAG filter consists of two types of filters: an excitatory filter and a barrier filter. The excitatory filter excludes all wavelengths except for blue light and the barrier filter passes long waves except for blue light. By adding this FAG filter to a microscope without an integrated FAG function, light from the microscope illuminating the surgical field becomes blue, which is blocked by the barrier filter. We put the FAG filter on the objective lens of the operating microscope correctly and fluorescein sodium was injected intravenously or intra-arterially. Fluorescence (green light) from vessels in the surgical field and the dyed tumor were clearly observed through the microscope and recorded by a memory device. This method was easy and could be performed in a short time (about 10 seconds). Blood flow of small vessels deep in the surgical field could be observed. Blood flow stagnation could be evaluated. However, images from this method were inferior to those obtained by currently commercially available microscopes with an integrated FAG function. In brain tumor surgery, a stained tumor on the brain surface could be observed using this method. FAG could be performed with a microscope without an integrated FAG function easily with only this FAG filter. PMID:26597335

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.

Perspectives: Nanofibers and nanowires for disordered photonics

NASA Astrophysics Data System (ADS)

Pisignano, Dario; Persano, Luana; Camposeo, Andrea

2017-03-01

As building blocks of microscopically non-homogeneous materials, semiconductor nanowires and polymer nanofibers are emerging component materials for disordered photonics, with unique properties of light emission and scattering. Effects found in assemblies of nanowires and nanofibers include broadband reflection, significant localization of light, strong and collective multiple scattering, enhanced absorption of incident photons, synergistic effects with plasmonic particles, and random lasing. We highlight recent related discoveries, with a focus on material aspects. The control of spatial correlations in complex assemblies during deposition, the coupling of modes with efficient transmission channels provided by nanofiber waveguides, and the embedment of random architectures into individually coded nanowires will allow the potential of these photonic materials to be fully exploited, unconventional physics to be highlighted, and next-generation optical devices to be achieved. The prospects opened by this technology include enhanced random lasing and mode-locking, multi-directionally guided coupling to sensors and receivers, and low-cost encrypting miniatures for encoders and labels.

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.

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.

Color digital lensless holographic microscopy: laser versus LED illumination.

PubMed

Garcia-Sucerquia, Jorge

2016-08-20

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

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.

Cornea and ocular lens visualized with three-dimensional confocal microscopy

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1992-08-01

This paper demonstrates the advantages of three-dimensional reconstruction of the cornea and the ocular crystalline lens by confocal microscopy and volume rendering computer techniques. The advantages of noninvasive observation of ocular structures in living, unstained, unfixed tissue include the following: the tissue is in a natural living state without the artifacts of fixation, mechanical sectioning, and staining; the three-dimensional structure can be observed from any view point and quantitatively analyzed; the dynamics of morphological changes can be studied; and the use of confocal microscopic observation results in a reduction of the number of animals required for ocular morphometric studies. The main advantage is that the dynamic morphology of ocular structures can be investigated in living ocular tissue. A laser scanning confocal microscope was used in the reflected light mode to obtain the two- dimensional images from the cornea and the ocular lens of a freshly enucleated rabbit eye. The light source was an argon ion laser with 488 nm wavelength. The microscope objective was a Leitz 25X, NA 0.6 water immersion lens. The 400 micron thick cornea was optically sectioned into 133, three micron sections. The semi-transparent cornea and the in-situ ocular lens was visualized as high resolution, high contrast two-dimensional images. The under sampling resulted in a three-dimensional visualization rendering in which the corneal thickness (z-axis) is compressed. The structures observed in the cornea include: superficial epithelial cells and their nuclei, basal epithelial cells and their `beaded' cell borders, basal lamina, nerve plexus, nerve fibers, free nerve endings in the basal epithelial cells, nuclei of stromal keratocytes, and endothelial cells. The structures observed in the in-situ ocular lens include: lens capsule, lens epithelial cells, and individual lens fibers.

Imaging the spectral reflectance properties of bipolar radiofrequency-fused bowel tissue

NASA Astrophysics Data System (ADS)

Clancy, Neil T.; Arya, Shobhit; Stoyanov, Danail; Du, Xiaofei; Hanna, George B.; Elson, Daniel S.

2015-07-01

Delivery of radiofrequency (RF) electrical energy is used during surgery to heat and seal tissue, such as vessels, allowing resection without blood loss. Recent work has suggested that this approach may be extended to allow surgical attachment of larger tissue segments for applications such as bowel anastomosis. In a large series of porcine surgical procedures bipolar RF energy was used to resect and re-seal the small bowel in vivo with a commercial tissue fusion device (Ligasure; Covidien PLC, USA). The tissue was then imaged with a multispectral imaging laparoscope to obtain a spectral datacube comprising both fused and healthy tissue. Maps of blood volume, oxygen saturation and scattering power were derived from the measured reflectance spectra using an optimised light-tissue interaction model. A 60% increase in reflectance of visible light (460-700 nm) was observed after fusion, with the tissue taking on a white appearance. Despite this the distinctive shape of the haemoglobin absorption spectrum was still noticeable in the 460-600 nm wavelength range. Scattering power increased in the fused region in comparison to normal serosa, while blood volume and oxygen saturation decreased. Observed fusion-induced changes in the reflectance spectrum are consistent with the biophysical changes induced through tissue denaturation and increased collagen cross-linking. The multispectral imager allows mapping of the spatial extent of these changes and classification of the zone of damaged tissue. Further analysis of the spectral data in parallel with histopathological examination of excised specimens will allow correlation of the optical property changes with microscopic alterations in tissue structure.

Blue Marble Eastern Hemisphere

NASA Technical Reports Server (NTRS)

2002-01-01

Drawing on data from multiple satellite missions (not all collected at the same time), a team of NASA scientists and graphic artists created layers of global data for everything from the land surface, to polar sea ice, to the light reflected by the chlorophyll in the billions of microscopic plants that grow in the ocean. They wrapped these layers around a globe, set it against a black background, and simulated the hazy edge of the Earth's atmosphere (the limb) that appears in astronaut photography of the Earth. The land surface layer is based on photo-like surface reflectance observations (reflected sunlight) measured by the Moderate Resolution Imaging Spectroradiometer (MODIS) on NASA's Terra satellite in July 2004. The sea ice layer near the poles comes from Terra MODIS observations of daytime sea ice observed between August 28 and September 6, 2001. The ocean layer is a composite. In shallow water areas, the layer shows surface reflectances observed by Terra MODIS in July 2004. In the open ocean, the photo-like layer is overlaid with observations of the average ocean chlorophyll content for 2004. NASA's Aqua MODIS collected the chlorophyll data. The cloud layer shows a single-day snapshot of clouds observed by Terra MODIS across the planet on July 29, 2001. City lights on Earth's night side are visualized from data collected by the Defense Meteorological Satellite Program mission between 1994-1995. The topography layer is based on radar data collected by the Space Shuttle Endeavour during an 11-day mission in February of 2000. Topography over Antarctica comes from the Radarsat Antarctic Mapping Project, version 2.

Blue Marble Western Hemisphere

NASA Technical Reports Server (NTRS)

2002-01-01

Drawing on data from multiple satellite missions (not all collected at the same time), a team of NASA scientists and graphic artists created layers of global data for everything from the land surface, to polar sea ice, to the light reflected by the chlorophyll in the billions of microscopic plants that grow in the ocean. They wrapped these layers around a globe, set it against a black background, and simulated the hazy edge of the Earth's atmosphere (the limb) that appears in astronaut photography of the Earth. The land surface layer is based on photo-like surface reflectance observations (reflected sunlight) measured by the Moderate Resolution Imaging Spectroradiometer (MODIS) on NASA's Terra satellite in July 2004. The sea ice layer near the poles comes from Terra MODIS observations of daytime sea ice observed between August 28 and September 6, 2001. The ocean layer is a composite. In shallow water areas, the layer shows surface reflectances observed by Terra MODIS in July 2004. In the open ocean, the photo-like layer is overlaid with observations of the average ocean chlorophyll content for 2004. NASA's Aqua MODIS collected the chlorophyll data. The cloud layer shows a single-day snapshot of clouds observed by Terra MODIS across the planet on July 29, 2001. City lights on Earth's night side are visualized from data collected by the Defense Meteorological Satellite Program mission between 1994-1995. The topography layer is based on radar data collected by the Space Shuttle Endeavour during an 11-day mission in February of 2000. Topography over Antarctica comes from the Radarsat Antarctic Mapping Project, version 2.

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.

Fabrication of low reflective nanopore-type black Si layer using one-step Ni-assisted chemical etching for Si solar cell application

NASA Astrophysics Data System (ADS)

Takaloo, AshkanVakilipour; Kolahdouz, Mohammadreza; Poursafar, Jafar; Es, Firat; Turan, Rasit; Ki-Joo, Seung

2018-03-01

Nanotextured Si fabricated through metal-assisted chemical etching (MACE) technique exhibits a promising potential for producing antireflective layer for photovoltaic (PV) application. In this study, a novel single-step nickel (Ni) assisted etching technique was applied to produce an antireflective, nonporous Si (black Si) in an aqueous solution containing hydrofluoric acid (HF), hydrogen peroxide (H2O2) and NiSO4 at 40 °C. Field emission scanning electron microscope was used to characterize different morphologies of the textured Si. Optical reflection measurements of samples were carried out to compare the reflectivity of different morphologies. Results indicated that vertical as well as horizontal pores with nanosized diameters were bored in the Si wafer after 1 h treatment in the etching solution containing different molar ratios of H2O2 to HF. Increasing H2O2 concentration in electrochemical etching solution had a considerable influence on the morphology due to higher injection of positive charges from Ni atoms onto the Si surface. Optimized concentration of H2O2 led to formation of an antireflective layer with 2.1% reflectance of incident light.

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.

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.

Is in vivo analysis of urinary stone composition feasible? Evaluation of an experimental setup of a Raman system coupled to commercial lithotripsy laser fibers.

PubMed

Miernik, Arkadiusz; Eilers, Yvan; Nuese, Christoph; Bolwien, Carsten; Lambrecht, Armin; Hesse, Albrecht; Rassweiler, Jens J; Schlager, Daniel; Wilhelm, Konrad; Wetterauer, Ulrich; Schoenthaler, Martin

2015-10-01

Raman spectroscopy allows immediate analysis of stone composition. In vivo stone analysis during endoscopic treatment may offer advantages concerning surgical strategy and metaphylaxis. Urinary stone components were evaluated utilizing an experimental setup of a Raman system coupled to commercial laser fibers. Samples of paracetamol (acetaminophen) and human urinary stones with known Raman spectra were analyzed using an experimental Raman system coupled to common commercial lithotripsy laser fibers (200 and 940 µm). Two different excitation lasers were used at wavelengths of 532 and 785 nm. Numerical aperture of the fibers, proportion of reflected light reaching the CCD chip, and integration times were calculated. Mathematical signal correction was performed. Both the laser beam profile and the quality of light reflected by the specimens were impaired significantly when used with commercial fibers. Acquired spectra could no longer be assigned to a specific stone composition. Subsequent measurements revealed a strong intrinsic fluorescence of the fibers and poor light acquisition properties leading to a significant decrease in the Raman signal in comparison with a free-beam setup. This was true for both investigated fiber diameters and both wavelengths. Microscopic examination showed highly irregular fiber tip surfaces (both new and used fibers). Our results propose that laser excitation and light acquisition properties of commercial lithotripsy fibers impair detectable Raman signals significantly in a fiber-coupled setting. This study provides essential physical and technological information for the development of an advanced fiber-coupled system able to be used for immediate stone analysis during endoscopic stone therapy.

Multilayered phantoms with tunable optical properties for a better understanding of light/tissue interactions

NASA Astrophysics Data System (ADS)

Roig, Blandine; Koenig, Anne; Perraut, François; Piot, Olivier; Vignoud, Séverine; Lavaud, Jonathan; Manfait, Michel; Dinten, Jean-Marc

2015-03-01

Light/tissue interactions, like diffuse reflectance, endogenous fluorescence and Raman scattering, are a powerful means for providing skin diagnosis. Instrument calibration is an important step. We thus developed multilayered phantoms for calibration of optical systems. These phantoms mimic the optical properties of biological tissues such as skin. Our final objective is to better understand light/tissue interactions especially in the case of confocal Raman spectroscopy. The phantom preparation procedure is described, including the employed method to obtain a stratified object. PDMS was chosen as the bulk material. TiO2 was used as light scattering agent. Dye and ink were adopted to mimic, respectively, oxy-hemoglobin and melanin absorption spectra. By varying the amount of the incorporated components, we created a material with tunable optical properties. Monolayer and multilayered phantoms were designed to allow several characterization methods. Among them, we can name: X-ray tomography for structural information; Diffuse Reflectance Spectroscopy (DRS) with a homemade fibered bundle system for optical characterization; and Raman depth profiling with a commercial confocal Raman microscope for structural information and for our final objective. For each technique, the obtained results are presented and correlated when possible. A few words are said on our final objective. Raman depth profiles of the multilayered phantoms are distorted by elastic scattering. The signal attenuation through each single layer is directly dependent on its own scattering property. Therefore, determining the optical properties, obtained here with DRS, is crucial to properly correct Raman depth profiles. Thus, it would be permitted to consider quantitative studies on skin for drug permeation follow-up or hydration assessment, for instance.

X ray imaging microscope for cancer research

NASA Technical Reports Server (NTRS)

Hoover, Richard B.; Shealy, David L.; Brinkley, B. R.; Baker, Phillip C.; Barbee, Troy W., Jr.; Walker, Arthur B. C., Jr.

1991-01-01

The NASA technology employed during the Stanford MSFC LLNL Rocket X Ray Spectroheliograph flight established that doubly reflecting, normal incidence multilayer optics can be designed, fabricated, and used for high resolution x ray imaging of the Sun. Technology developed as part of the MSFC X Ray Microscope program, showed that high quality, high resolution multilayer x ray imaging microscopes are feasible. Using technology developed at Stanford University and at the DOE Lawrence Livermore National Laboratory (LLNL), Troy W. Barbee, Jr. has fabricated multilayer coatings with near theoretical reflectivities and perfect bandpass matching for a new rocket borne solar observatory, the Multi-Spectral Solar Telescope Array (MSSTA). Advanced Flow Polishing has provided multilayer mirror substrates with sub-angstrom (rms) smoothnesss for the astronomical x ray telescopes and x ray microscopes. The combination of these important technological advancements has paved the way for the development of a Water Window Imaging X Ray Microscope for cancer research.

Exploring photoreceptor reflectivity via multimodal imaging of outer retinal tubulation in advanced age-related macular degeneration

PubMed Central

Litts, Katie M.; Wang, Xiaolin; Clark, Mark E.; Owsley, Cynthia; Freund, K. Bailey; Curcio, Christine A.; Zhang, Yuhua

2016-01-01

Purpose To investigate the microscopic structure of outer retinal tubulation (ORT) and optical properties of cone photoreceptors in vivo, we studied ORT appearance by multimodal imaging, including spectral domain optical coherence tomography (SD-OCT) and adaptive optics scanning laser ophthalmoscopy (AOSLO). Methods Four eyes of 4 subjects with advanced AMD underwent color fundus photography, infrared reflectance imaging, SD-OCT, and AOSLO with a high-resolution research instrument. ORT was identified in closely spaced (11 μm) SD-OCT volume scans. Results ORT in cross-sectional and en face SD-OCT was a hyporeflective area representing a lumen surrounded by a hyperreflective border consisting of cone photoreceptor mitochondria and external limiting membrane, per previous histology. In contrast, ORT by AOSLO was a hyporeflective structure of the same shape as in en face SD-OCT but lacking visualizable cone photoreceptors. Conclusion Lack of ORT cone reflectivity by AOSLO indicates that cones have lost their normal directionality and waveguiding property due to loss of outer segments and subsequent retinal remodeling. Reflective ORT cones by SD-OCT, in contrast, may depend partly on mitochondria as light scatterers within inner segments of these degenerating cells, a phenomenon enhanced by coherent imaging. Multimodal imaging of ORT provides insight into cone degeneration and reflectivity sources in OCT. PMID:27584549

Refractive Index Imaging of Cells with Variable-Angle Near-Total Internal Reflection (TIR) Microscopy.

PubMed

Bohannon, Kevin P; Holz, Ronald W; Axelrod, Daniel

2017-10-01

The refractive index in the interior of single cells affects the evanescent field depth in quantitative studies using total internal reflection (TIR) fluorescence, but often that index is not well known. We here present method to measure and spatially map the absolute index of refraction in a microscopic sample, by imaging a collimated light beam reflected from the substrate/buffer/cell interference at variable angles of incidence. Above the TIR critical angle (which is a strong function of refractive index), the reflection is 100%, but in the immediate sub-critical angle zone, the reflection intensity is a very strong ascending function of incidence angle. By analyzing the angular position of that edge at each location in the field of view, the local refractive index can be estimated. In addition, by analyzing the steepness of the edge, the distance-to-substrate can be determined. We apply the technique to liquid calibration samples, silica beads, cultured Chinese hamster ovary cells, and primary culture chromaffin cells. The optical technique suffers from decremented lateral resolution, scattering, and interference artifacts. However, it still provides reasonable results for both refractive index (~1.38) and for distance-to-substrate (~150 nm) for the cells, as well as a lateral resolution to about 1 µm.

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.

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)

Modification of Ag nanoparticles on the surface of SrTiO3 particles and resultant influence on photoreduction of CO2

NASA Astrophysics Data System (ADS)

Shao, Kunjuan; Wang, Yanjie; Iqbal, Muzaffar; Lin, Lin; Wang, Kai; Zhang, Xuehua; He, Meng; He, Tao

2018-03-01

Modification of a wide-bandgap semiconductor with noble metals that can exhibit surface plasmon effect is an effective approach to make it responsive to the visible light. In this work, a series of cubic and all-edge-truncated SrTiO3 with and without thermal pretreatment in air are modified by Ag nanoparticles via photodeposition method. The crystal structure, morphology, loading amount of Ag nanoparticles, and optical properties of the obtained Ag-SrTiO3 nanomaterials are well characterized by powder X-ray diffraction, scanning microscope, transmission electron microscope, energy disperse X-ray spectroscopy, ICP-MS and UV-vis diffuse-reflection spectroscopy. The loading amount and size of the Ag nanoparticles can be controlled to some extent by tuning the photodeposition time via growth-dissolution mechanism. The Ag nanoparticles are inclined to deposit on different locations on the surface of cubic and truncated SrTiO3 with and without thermal pretreatment. The resultant SrTiO3 modified by Ag nanoparticles exhibits visible light activity for photocatalytic reduction of CO2, which is closely related to the oxygen vacancy induced by thermal pretreatment, size and amount of Ag nanoparticles. Accordingly, there is an optimized photodeposition time for the synthesis of the photocatalyst that exhibits the highest photocatalytic activity.

Pulsed holographic system for imaging through spatially extended scattering media

NASA Astrophysics Data System (ADS)

Kanaev, A. V.; Judd, K. P.; Lebow, P.; Watnik, A. T.; Novak, K. M.; Lindle, J. R.

2017-10-01

Imaging through scattering media is a highly sought capability for military, industrial, and medical applications. Unfortunately, nearly all recent progress was achieved in microscopic light propagation and/or light propagation through thin or weak scatterers which is mostly pertinent to medical research field. Sensing at long ranges through extended scattering media, for example turbid water or dense fog, still represents significant challenge and the best results are demonstrated using conventional approaches of time- or range-gating. The imaging range of such systems is constrained by their ability to distinguish a few ballistic photons that reach the detector from the background, scattered, and ambient photons, as well as from detector noise. Holography can potentially enhance time-gating by taking advantage of extra signal filtering based on coherence properties of the ballistic photons as well as by employing coherent addition of multiple frames. In a holographic imaging scheme ballistic photons of the imaging pulse are reflected from a target and interfered with the reference pulse at the detector creating a hologram. Related approaches were demonstrated previously in one-way imaging through thin biological samples and other microscopic scale scatterers. In this work, we investigate performance of holographic imaging systems under conditions of extreme scattering (less than one signal photon per pixel signal), demonstrate advantages of coherent addition of images recovered from holograms, and discuss image quality dependence on the ratio of the signal and reference beam power.

Photocatalytic degradation of Orange G dye under solar light using nanocrystalline semiconductor metal oxide.

PubMed

Thennarasu, G; Kavithaa, S; Sivasamy, A

2011-08-01

The photocatalytic degradation of Orange G (OG) dye has been investigated using synthesised nanocrystalline ZnO as a photocatalyst and sunlight as the irradiation source. The formation of ZnO prepared from its precursor was confirmed through FT-IR and powder X-ray diffraction analyses. Surface morphology was characterised by scanning electron microscope and transmission electron microscope analysis. Band gap energy of synthesised nanocrystalline ZnO was calculated using diffuse reflectance spectroscopy (DRS). Different experimental parameters such as effects of pH, dye concentrations and mass of catalyst were standardised in order to achieve complete degradation of the dye molecules under solar light irradiation. The kinetics of oxidation of OG was also studied. The complete degradation of OG was evident after 90 min of irradiation at an initial pH of 6.86. The degradation of OG was confirmed by UV-Visible spectrophotometer, high-pressure liquid chromatography, ESI-Mass and chemical oxygen demand analyses. The adsorption of dye onto catalytic surface was analysed employing model equations such as Langmuir and Freundlich isotherms, and it was found that the Langmuir isotherm model best fitted the adsorption data. The solar photodegradation of OG followed pseudo-first-order kinetics. HPLC and ESI-Mass analyses of the degraded samples suggested that the dye molecules were readily degraded under solar irradiation with nanocrystalline ZnO.

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

A programmable light engine for quantitative single molecule TIRF and HILO imaging.

PubMed

van 't Hoff, Marcel; de Sars, Vincent; Oheim, Martin

2008-10-27

We report on a simple yet powerful implementation of objective-type total internal reflection fluorescence (TIRF) and highly inclined and laminated optical sheet (HILO, a type of dark-field) illumination. Instead of focusing the illuminating laser beam to a single spot close to the edge of the microscope objective, we are scanning during the acquisition of a fluorescence image the focused spot in a circular orbit, thereby illuminating the sample from various directions. We measure parameters relevant for quantitative image analysis during fluorescence image acquisition by capturing an image of the excitation light distribution in an equivalent objective backfocal plane (BFP). Operating at scan rates above 1 MHz, our programmable light engine allows directional averaging by circular spinning the spot even for sub-millisecond exposure times. We show that restoring the symmetry of TIRF/HILO illumination reduces scattering and produces an evenly lit field-of-view that affords on-line analysis of evanescnt-field excited fluorescence without pre-processing. Utilizing crossed acousto-optical deflectors, our device generates arbitrary intensity profiles in BFP, permitting variable-angle, multi-color illumination, or objective lenses to be rapidly exchanged.

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.

Optimal detection pinhole for lowering speckle noise while maintaining adequate optical sectioning in confocal reflectance microscopes

PubMed Central

Rajadhyaksha, Milind

2012-01-01

Abstract. Coherent speckle influences the resulting image when narrow spectral line-width and single spatial mode illumination are used, though these are the same light-source properties that provide the best radiance-to-cost ratio. However, a suitable size of the detection pinhole can be chosen to maintain adequate optical sectioning while making the probability density of the speckle noise more normal and reducing its effect. The result is a qualitatively better image with improved contrast, which is easier to read. With theoretical statistics and experimental results, we show that the detection pinhole size is a fundamental parameter for designing imaging systems for use in turbid media. PMID:23224184

Mechanisms of structural colour in the Morpho butterfly: cooperation of regularity and irregularity in an iridescent scale.

PubMed Central

Kinoshita, Shuichi; Yoshioka, Shinya; Kawagoe, Kenji

2002-01-01

Structural colour in the Morpho butterfly originates from submicron structure within a scale and, for over a century, its colour and reflectivity have been explained as interference of light due to the multilayer of cuticle and air. However, this model fails to explain the extraordinarily uniform colour of the wing with respect to the observation direction. We have performed microscopic, optical and theoretical investigations, and have found that the separate lamellar structure with irregular heights is extremely important. Using a simple model, we have shown that the combined action of interference and diffraction is essential for the structural colour of the Morpho butterfly. PMID:12137569

Quantitative surface topography determination by Nomarski reflection microscopy. 2: Microscope modification, calibration, and planar sample experiments

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

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

1980-09-01

The application of reflective Nomarski differential interference contrast microscopy for the determination of quantitative sample topography data is presented. The discussion includes a review of key theoretical results presented previously plus the experimental implementation of the concepts using a commercial Momarski microscope. The experimental work included the modification and characterization of a commercial microscope to allow its use for obtaining quantitative sample topography data. System usage for the measurement of slopes on flat planar samples is also discussed. The discussion has been designed to provide the theoretical basis, a physical insight, and a cookbook procedure for implementation to allow thesemore » results to be of value to both those interested in the microscope theory and its practical usage in the metallography laboratory.« less

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.

Miniature in vivo MEMS-based line-scanned dual-axis confocal microscope for point-of-care pathology

PubMed Central

Yin, C.; Glaser, A.K.; Leigh, S. Y.; Chen, Y.; Wei, L.; Pillai, P. C. S.; Rosenberg, M. C.; Abeytunge, S.; Peterson, G.; Glazowski, C.; Sanai, N.; Mandella, M. J.; Rajadhyaksha, M.; Liu, J. T. C.

2016-01-01

There is a need for miniature optical-sectioning microscopes to enable in vivo interrogation of tissues as a real-time and noninvasive alternative to gold-standard histopathology. Such devices could have a transformative impact for the early detection of cancer as well as for guiding tumor-resection procedures. Miniature confocal microscopes have been developed by various researchers and corporations to enable optical sectioning of highly scattering tissues, all of which have necessitated various trade-offs in size, speed, depth selectivity, field of view, resolution, image contrast, and sensitivity. In this study, a miniature line-scanned (LS) dual-axis confocal (DAC) microscope, with a 12-mm diameter distal tip, has been developed for clinical point-of-care pathology. The dual-axis architecture has demonstrated an advantage over the conventional single-axis confocal configuration for reducing background noise from out-of-focus and multiply scattered light. The use of line scanning enables fast frame rates (16 frames/sec is demonstrated here, but faster rates are possible), which mitigates motion artifacts of a hand-held device during clinical use. We have developed a method to actively align the illumination and collection beams in a DAC microscope through the use of a pair of rotatable alignment mirrors. Incorporation of a custom objective lens, with a small form factor for in vivo clinical use, enables our device to achieve an optical-sectioning thickness and lateral resolution of 2.0 and 1.1 microns respectively. Validation measurements with reflective targets, as well as in vivo and ex vivo images of tissues, demonstrate the clinical potential of this high-speed optical-sectioning microscopy device. PMID:26977337

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.

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.

Phototoxic effects of an operating microscope on the ocular surface and tear film.

PubMed

Hwang, Hyung Bin; Kim, Hyun Seung

2014-01-01

We evaluated light exposure-induced dry eye syndrome by investigating the phototoxic effects of an operating microscope on the ocular surface and tear film in rabbits. Sixty eyes of 30 rabbits were divided into 3 groups based on the intensity of light exposure received from an operating microscope: Control group, no exposure to light; group A, 40,000-lx intensity for 30 minutes; and group B, 100,000-lx intensity for 30 minutes. To evaluate the potential damage to the ocular surface and tear film, Schirmer tests, rose bengal staining, and conjunctival impression cytology were performed before the light exposure and at 1, 3, and 5 days afterward. In addition, the expression of interleukin 1-beta was analyzed in tear samples. The expression of mucin 5AC was evaluated using immunofluorescence staining, and periodic acid-Schiff staining was conducted on conjunctival tissues. Corneal and conjunctival tissues were observed by means of electron microscopy. Potential damage to the ocular surface and tear film was found in the light-exposed groups as evidenced by decreased aqueous tear production, devitalized corneal and conjunctival epithelial cells, squamous metaplasia of conjunctival epithelial cells, decreased conjunctival goblet cell density, decreased expression of mucin 5AC, ultrastructural cellular damage to corneal and conjunctival tissues, and increased interleukin 1-beta expression in tears. This damage was more noticeable in group B than in group A (P < 0.05). Light exposure from an operating microscope had phototoxic effects on the ocular surface and tear film in this in vivo experiment. These changes seemed to intensify as the intensity of the light increased. Therefore, excessive light exposure during ophthalmic procedures could be a pathogenic factor in dry eye syndrome after a surgery is performed.

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.

Quantification of Concentration of Microalgae Anabaena Cylindrica, Coal-bed Methane Water Isolates Nannochloropsis Gaditana and PW-95 in Aquatic Solutions through Hyperspectral Reflectance Measurement and Analytical Model Establishment

NASA Astrophysics Data System (ADS)

Zhou, Z.; Zhou, X.; Apple, M. E.; Spangler, L.

2017-12-01

Three species of microalgae, Anabaena cylindrica (UTEX # 1611), coal-bed methane water isolates Nannochloropsis gaditana and PW-95 were cultured for the measurements of their hyperspectral profiles in different concentrations. The hyperspectral data were measured by an Analytical Spectral Devices (ASD) spectroradiomter with the spectral resolution of 1 nanometer over the wavelength ranges from 350nm to 1050 nm for samples of microalgae of different concentration. Concentration of microalgae was measured using a Hemocytometer under microscope. The objective of this study is to establish the relation between spectral reflectance and micro-algal concentration so that microalgae concentration can be measured remotely by space- or airborne hyperspectral or multispectral sensors. Two types of analytical models, linear reflectance-concentration model and Lamber-Beer reflectance-concentration model, were established for each species. For linear modeling, the wavelength with the maximum correlation coefficient between the reflectance and concentrations of algae was located and then selected for each species of algae. The results of the linear models for each species are shown in Fig.1(a), in which Refl_1, Refl_2, and Refl_3 represent the reflectance of Anabaena, N. Gaditana, and PW-95 respectively. C1, C2, and C3 represent the Concentrations of Anabaena, N. Gaditana, and PW-95 respectively. The Lamber-Beer models were based on the Lambert-Beer Law, which states that the intensity of light propagating in a substance dissolved in a fully transmitting solvent is directly proportional to the concentration of the substance and the path length of the light through the solution. Thus, for the Lamber-Beer modeling, a wavelength with large absorption in red band was selected for each species. The results of Lambert-Beer models for each species are shown in Fig.1(b). Based on the Lamber-Beer models, the absorption coefficient for the three different species will be quantified.

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

PubMed

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

2005-04-01

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

Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues.

PubMed

Sung, Kung-Bin; Liang, Chen; Descour, Michael; Collier, Tom; Follen, Michele; Richards-Kortum, Rebecca

2002-10-01

We have built a fiber-optic confocal reflectance microscope capable of imaging human tissues in near real time. Miniaturization of the objective lens and the mechanical components for positioning and axially scanning the objective enables the device to be used in inner organs of the human body. The lateral resolution is 2 micrometers and axial resolution is 10 micrometers. Confocal images of fixed tissue biopsies and the human lip in vivo have been obtained at 15 frames/s without any fluorescent stains. Both cell morphology and tissue architecture can be appreciated from images obtained with this microscope.

Sub-diffusive scattering parameter maps recovered using wide-field high-frequency structured light imaging.

PubMed

Kanick, Stephen Chad; McClatchy, David M; Krishnaswamy, Venkataramanan; Elliott, Jonathan T; Paulsen, Keith D; Pogue, Brian W

2014-10-01

This study investigates the hypothesis that structured light reflectance imaging with high spatial frequency patterns [Formula: see text] can be used to quantitatively map the anisotropic scattering phase function distribution [Formula: see text] in turbid media. Monte Carlo simulations were used in part to establish a semi-empirical model of demodulated reflectance ([Formula: see text]) in terms of dimensionless scattering [Formula: see text] and [Formula: see text], a metric of the first two moments of the [Formula: see text] distribution. Experiments completed in tissue-simulating phantoms showed that simultaneous analysis of [Formula: see text] spectra sampled at multiple [Formula: see text] in the frequency range [0.05-0.5] [Formula: see text] allowed accurate estimation of both [Formula: see text] in the relevant tissue range [0.4-1.8] [Formula: see text], and [Formula: see text] in the range [1.4-1.75]. Pilot measurements of a healthy volunteer exhibited [Formula: see text]-based contrast between scar tissue and surrounding normal skin, which was not as apparent in wide field diffuse imaging. These results represent the first wide-field maps to quantify sub-diffuse scattering parameters, which are sensitive to sub-microscopic tissue structures and composition, and therefore, offer potential for fast diagnostic imaging of ultrastructure on a size scale that is relevant to surgical applications.

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

Photovoltaic characteristics of natural light harvesting dye sensitized solar cells

NASA Astrophysics Data System (ADS)

Hafez, H. S.; Shenouda, S. S.; Fadel, M.

2018-03-01

In this work of research, anthocyanin as a natural dye obtained from raspberry fruits, was used and tested as a photon harvesting/electron donating dye in titanium dioxide nanoparticle-based DSSCs. A working photoelectrode made from TiO2 nanoparticles with an average particle size (10-40 nm) that is coated on Florine doped tin-oxide substrate, was prepared via a simple and low cost hydrothermal method. A detailed structural and morphological analysis of the TiO2 photoactive electrode was investigated by X-ray diffraction (XRD), diffuse reflectance spectrometer, transmission electron microscope (TEM) and scanning electron microscope (SEM). Complete photovoltaic characteristics including (current, voltage, outpower, and responsivity) of the natural anthocyanin based dye sensitized solar cell have been investigated under different illumination intensity ranging from 10 to 100 mW.cm- 2. The cell responsivity and efficiency of the fabricated solar cell under different illumination intensity were found to be in the range (R = 15.6-23.8 mA.W- 1 and η = 0.13-0.25) at AM = 1.5 conditions. This study is important for enhancing the future applications of the promising DSSC technology.

Measurement and characterization of external oil in the fried waxy maize starch granules using ATR-FTIR and XRD.

PubMed

Chen, Long; Tian, Yaoqi; Sun, Binghua; Cai, Canxin; Ma, Rongrong; Jin, Zhengyu

2018-03-01

Concerns regarding increased dietary oil uptake have prompted efforts to investigate the oil absorption and distribution in fried starchy foods. In the present study, attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, together with a chloroform-methanol method, was used to analyze the external and internal oil contents in fried starchy samples. The micromorphology of fried starchy samples was further investigated using scanning electron microscope (SEM), polarized light microscope (PLM) and confocal laser scanning microscopy (CLSM). The results indicated that large amounts of oil were absorbed in or within waxy maize starch, but the majority of oil was located near the surface layer of the starch granules. After defatting, the internal oil was thoroughly removed, while a small amount of external oil remained. As evidenced by the changes of the crystalline characteristics with the help of X-ray diffraction (XRD), the interaction between starch and lipids on the surface was confirmed to form V-type complex compounds during frying at high moisture. Copyright © 2017 Elsevier Ltd. All rights reserved.

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.

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.

Effect of surface roughness and subsurface damage on grazing-incidence x-ray scattering and specular reflectance.

PubMed

Lodha, G S; Yamashita, K; Kunieda, H; Tawara, Y; Yu, J; Namba, Y; Bennett, J M

1998-08-01

Grazing-incidence specular reflectance and near-specular scattering were measured at Al-K(alpha) (1.486-keV, 8.34-?) radiation on uncoated dielectric substrates whose surface topography had been measured with a scanning probe microscope and a mechanical profiler. Grazing-incidence specular reflectance was also measured on selected substrates at the Cu-K(alpha) (8.047-keV, 1.54-?) wavelength. Substrates included superpolished and conventionally polished fused silica; SiO(2) wafers; superpolished and precision-ground Zerodur; conventionally polished, float-polished, and precision-ground BK-7 glass; and superpolished and precision-ground silicon carbide. Roughnesses derived from x-ray specular reflectance and scattering measurements were in good agreement with topographic roughness values measured with a scanning probe microscope (atomic force microscope) and a mechanical profiler that included similar ranges of surface spatial wavelengths. The specular reflectance was also found to be sensitive to the density of polished surface layers and subsurface damage down to the penetration depth of the x rays. Density gradients and subsurface damage were found in the superpolished fused-silica and precision-ground Zerodur samples. These results suggest that one can nondestructively evaluate subsurface damage in transparent materials using grazing-incidence x-ray specular reflectance in the 1.5-8-keV range.

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.

Snow crystal imaging using scanning electron microscopy: III. Glacier ice, snow and biota

USGS Publications Warehouse

Rango, A.; Wergin, W.P.; Erbe, E.F.; Josberger, E.G.

2000-01-01

Low-temperature scanning electron microscopy (SEM) was used to observe metamorphosed snow, glacial firn, and glacial ice obtained from South Cascade Glacier in Washington State, USA. Biotic samples consisting of algae (Chlamydomonas nivalis) and ice worms (a species of oligochaetes) were also collected and imaged. In the field, the snow and biological samples were mounted on copper plates, cooled in liquid nitrogen, and stored in dry shipping containers which maintain a temperature of -196??C. The firn and glacier ice samples were obtained by extracting horizontal ice cores, 8 mm in diameter, at different levels from larger standard glaciological (vertical) ice cores 7.5 cm in diameter. These samples were cooled in liquid nitrogen and placed in cryotubes, were stored in the same dry shipping container, and sent to the SEM facility. In the laboratory, the samples were sputter coated with platinum and imaged by a low-temperature SEM. To image the firn and glacier ice samples, the cores were fractured in liquid nitrogen, attached to a specimen holder, and then imaged. While light microscope images of snow and ice are difficult to interpret because of internal reflection and refraction, the SEM images provide a clear and unique view of the surface of the samples because they are generated from electrons emitted or reflected only from the surface of the sample. In addition, the SEM has a great depth of field with a wide range of magnifying capabilities. The resulting images clearly show the individual grains of the seasonal snowpack and the bonding between the snow grains. Images of firn show individual ice crystals, the bonding between the crystals, and connected air spaces. Images of glacier ice show a crystal structure on a scale of 1-2 mm which is considerably smaller than the expected crystal size. Microscopic air bubbles, less than 15 ??m in diameter, clearly marked the boundaries between these crystal-like features. The life forms associated with the glacier were easily imaged and studied. The low-temperature SEM sample collecting and handling methods proved to be operable in the field; the SEM analysis is applicable to glaciological studies and reveals details unattainable by conventional light microscopic methods.Low temperature scanning electron microscopy (SEM) was used to observe metamorphosed snow, glacial firn, and glacial ice obtained from South Cascade Glacier in Washington State, USA. Biotic samples consisting of algae and ice worms were also collected and imaged. The SEM images provide a clear and unique view of the surface of the samples because they are generated from electrons emitted or reflected only from the surface of the sample. The SEM has a great depth of field with a wide range of magnifying capabilities.

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

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.

Implementation of PSF engineering in high-resolution 3D microscopy imaging with a LCoS (reflective) SLM

NASA Astrophysics Data System (ADS)

King, Sharon V.; Doblas, Ana; Patwary, Nurmohammed; Saavedra, Genaro; Martínez-Corral, Manuel; Preza, Chrysanthe

2014-03-01

Wavefront coding techniques are currently used to engineer unique point spread functions (PSFs) that enhance existing microscope modalities or create new ones. Previous work in this field demonstrated that simulated intensity PSFs encoded with a generalized cubic phase mask (GCPM) are invariant to spherical aberration or misfocus; dependent on parameter selection. Additional work demonstrated that simulated PSFs encoded with a squared cubic phase mask (SQUBIC) produce a depth invariant focal spot for application in confocal scanning microscopy. Implementation of PSF engineering theory with a liquid crystal on silicon (LCoS) spatial light modulator (SLM) enables validation of WFC phase mask designs and parameters by manipulating optical wavefront properties with a programmable diffractive element. To validate and investigate parameters of the GCPM and SQUBIC WFC masks, we implemented PSF engineering in an upright microscope modified with a dual camera port and a LCoS SLM. We present measured WFC PSFs and compare them to simulated PSFs through analysis of their effect on the microscope imaging system properties. Experimentally acquired PSFs show the same intensity distribution as simulation for the GCPM phase mask, the SQUBIC-mask and the well-known and characterized cubic-phase mask (CPM), first applied to high NA microscopy by Arnison et al.10, for extending depth of field. These measurements provide experimental validation of new WFC masks and demonstrate the use of the LCoS SLM as a WFC design tool. Although efficiency improvements are needed, this application of LCoS technology renders the microscope capable of switching among multiple WFC modes.

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.

Fast, label-free super-resolution live-cell imaging using rotating coherent scattering (ROCS) microscopy

NASA Astrophysics Data System (ADS)

Jünger, Felix; Olshausen, Philipp V.; Rohrbach, Alexander

2016-07-01

Living cells are highly dynamic systems with cellular structures being often below the optical resolution limit. Super-resolution microscopes, usually based on fluorescence cell labelling, are usually too slow to resolve small, dynamic structures. We present a label-free microscopy technique, which can generate thousands of super-resolved, high contrast images at a frame rate of 100 Hertz and without any post-processing. The technique is based on oblique sample illumination with coherent light, an approach believed to be not applicable in life sciences because of too many interference artefacts. However, by circulating an incident laser beam by 360° during one image acquisition, relevant image information is amplified. By combining total internal reflection illumination with dark-field detection, structures as small as 150 nm become separable through local destructive interferences. The technique images local changes in refractive index through scattered laser light and is applied to living mouse macrophages and helical bacteria revealing unexpected dynamic processes.

Chromatic confocal microscopy for multi-depth imaging of epithelial tissue

PubMed Central

Olsovsky, Cory; Shelton, Ryan; Carrasco-Zevallos, Oscar; Applegate, Brian E.; Maitland, Kristen C.

2013-01-01

We present a novel chromatic confocal microscope capable of volumetric reflectance imaging of microstructure in non-transparent tissue. Our design takes advantage of the chromatic aberration of aspheric lenses that are otherwise well corrected. Strong chromatic aberration, generated by multiple aspheres, longitudinally disperses supercontinuum light onto the sample. The backscattered light detected with a spectrometer is therefore wavelength encoded and each spectrum corresponds to a line image. This approach obviates the need for traditional axial mechanical scanning techniques that are difficult to implement for endoscopy and susceptible to motion artifact. A wavelength range of 590-775 nm yielded a >150 µm imaging depth with ~3 µm axial resolution. The system was further demonstrated by capturing volumetric images of buccal mucosa. We believe these represent the first microstructural images in non-transparent biological tissue using chromatic confocal microscopy that exhibit long imaging depth while maintaining acceptable resolution for resolving cell morphology. Miniaturization of this optical system could bring enhanced speed and accuracy to endomicroscopic in vivo volumetric imaging of epithelial tissue. PMID:23667789

Fast, label-free super-resolution live-cell imaging using rotating coherent scattering (ROCS) microscopy

PubMed Central

Jünger, Felix; Olshausen, Philipp v.; Rohrbach, Alexander

2016-01-01

Living cells are highly dynamic systems with cellular structures being often below the optical resolution limit. Super-resolution microscopes, usually based on fluorescence cell labelling, are usually too slow to resolve small, dynamic structures. We present a label-free microscopy technique, which can generate thousands of super-resolved, high contrast images at a frame rate of 100 Hertz and without any post-processing. The technique is based on oblique sample illumination with coherent light, an approach believed to be not applicable in life sciences because of too many interference artefacts. However, by circulating an incident laser beam by 360° during one image acquisition, relevant image information is amplified. By combining total internal reflection illumination with dark-field detection, structures as small as 150 nm become separable through local destructive interferences. The technique images local changes in refractive index through scattered laser light and is applied to living mouse macrophages and helical bacteria revealing unexpected dynamic processes. PMID:27465033

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

NASA Astrophysics Data System (ADS)

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

2014-11-01

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

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

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

NASA Astrophysics Data System (ADS)

Chandra, Subhash; Morrison, George H.

1995-05-01

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

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.

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.

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

PubMed

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

1989-01-01

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

Detection of nerve agent stimulants based on photoluminescent porous silicon interferometer

NASA Astrophysics Data System (ADS)

Kim, Seongwoong; Cho, Bomin; Sohn, Honglae

2012-09-01

Porous silicon (PSi) exhibiting dual optical properties, both Fabry-Pérot fringe and photolumincence, was developed and used as chemical sensors. PSi samples were prepared by an electrochemical etch of p-type silicon under the illumination of 300-W tungsten lamp during the etch process. The surface of PSi was characterized by cold field-emission scanning electron microscope. PSi samples exhibited a strong visible orange photoluminescence at 610 nm with an excitation wavelength of 460 nm as well as Fabry-Pérot fringe with a tungsten light source. Both reflectivity and photoluminescence were simultaneously measured under the exposure of organophosphate vapors. An increase of optical thickness and quenching photoluminescences under the exposure of various organophosphate vapors were observed.

Modeling the unidentified infrared emission with combinations of polycyclic aromatic hydrocarbons

NASA Technical Reports Server (NTRS)

Allamandola, L. J.; Hudgins, D. M.; Sandford, S. A.

1999-01-01

The infrared emission band spectrum associated with many different interstellar objects can be modeled successfully by using combined laboratory spectra of neutral and positively charged polycyclic aromatic hydrocarbons (PAHs). These model spectra, shown here for the first time, alleviate the principal spectroscopic criticisms previously leveled at the PAH hypothesis and demonstrate that mixtures of free molecular PAHs can indeed account for the overall appearance of the widespread interstellar infrared emission spectrum. Furthermore, these models give us insight into the structures, stabilities, abundances, and ionization balance of the interstellar PAH population. These, in turn, reflect conditions in the emission zones and shed light on the microscopic processes involved in the carbon nucleation, growth, and evolution in circumstellar shells and the interstellar medium.

Lucien J. Rubinstein: enduring contributions to neuro-oncology.

PubMed

Mut, Melike; Lopes, M Beatriz S; Shaffrey, Mark

2005-04-15

Dr. Lucien Rubinstein is best remembered for his significant contributions to the field of neuropathology, particularly in the classification of nervous system tumors. His accomplishments in basic neuro-oncology and in the formulation of diagnostic principles reflected a unique talent for synthesizing fundamental clinicopathological concepts based on skillful diagnostic investigation and a thorough understanding of neurobiology. Dr. Rubinstein was the leader in the establishment of cell cultures from central nervous system (CNS) tumors. He meticulously analyzed both light and electron microscopic features of CNS tumors, recorded his findings, and patiently drew sketches to be shared generously with his colleagues and students. As a pioneer in neuropathology, in his work Dr. Rubinstein set the foundation for many enduring concepts in neurosurgery, neuro-oncology, neurology, and basic tumor biology.

Fractography of a bis-GMA resin.

PubMed

Davis, D M; Waters, N E

1989-07-01

The fracture behavior of a bis-GMA resin was studied by means of the double-torsion test. The fracture parameter measured was the stress-intensity factor. Fracture occurred in either a stick-slip (unstable) or continuous (stable) manner, depending upon the test conditions. When stick-slip propagation occurred, the fracture surfaces showed characteristic crack-arrest lines. The fracture surfaces were examined by use of a reflected-light optical microscope. The stress-intensity factor for crack initiation was found to be related to the size of the crack-arrest line which, in turn, could be related to the Dugdale model for plastic zone size. The evidence supported the concept that the behavior of the crack during propagation was controlled by the amount of plastic deformation occurring at the crack tip.

Living Matter Observations with a Novel Hyperspectral Supercontinuum Confocal Microscope for VIS to Near-IR Reflectance Spectroscopy

PubMed Central

Bertani, Francesca R.; Ferrari, Luisa; Mussi, Valentina; Botti, Elisabetta; Costanzo, Antonio; Selci, Stefano

2013-01-01

A broad range hyper-spectroscopic microscope fed by a supercontinuum laser source and equipped with an almost achromatic optical layout is illustrated with detailed explanations of the design, implementation and data. The real novelty of this instrument, a confocal spectroscopic microscope capable of recording high resolution reflectance data in the VIS-IR spectral range from about 500 nm to 2.5 μm wavelengths, is the possibility of acquiring spectral data at every physical point as defined by lateral coordinates, X and Y, as well as at a depth coordinate, Z, as obtained by the confocal optical sectioning advantage. With this apparatus we collect each single scanning point as a whole spectrum by combining two linear spectral detector arrays, one CCD for the visible range, and one InGaAs infrared array, simultaneously available at the sensor output channel of the home made instrument. This microscope has been developed for biomedical analysis of human skin and other similar applications. Results are shown illustrating the technical performances of the instrument and the capability in extracting information about the composition and the structure of different parts or compartments in biological samples as well as in solid statematter. A complete spectroscopic fingerprinting of samples at microscopic level is shown possible by using statistical analysis on raw data or analytical reflectance models based on Abelés matrix transfer methods. PMID:24233077

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.

Simulation and Implementation of Moth-eye Structures as a Broadband Anti-Reflective Layer

NASA Astrophysics Data System (ADS)

Deshpande, Ketan S.

Conventional single layer thin anti-reflective coatings (ARCs) are only suitable for narrowband applications. A multilayer film stack is often employed for broadband applications. A coating of multiple layers with alternating low and high refractive index materials increases the overall cost of the system. This makes multilayer ARCs unsuitable for low-cost broadband applications. Since the discovery of moth-eye corneal nipple patterns and their potential applicability in the field of broadband ARCs, many studies have been carried out to fabricate these bio-inspired nanostructures with available manufacturing processes. Plasma etching processes used in microelectronic manufacturing are applied for creating these nanostructures at the Rochester Institute of Technology's Semiconductor & Microsystems Fabrication Laboratory (SMFL). Atomic Force Microscope (AFM) scanned surfaces of the nanostructure layer are simulated and characterized for their optical properties using a Finite-Difference Time Domain (FDTD) simulator from Lumerical Solutions, Inc. known as FDTD Solutions. Simulation results show that the layer is anti-reflective over 50 to 350 nm broadband of wavelengths at 0° angle of incidence. These simulation results were supported by ellipsometer reflection measurements off the actual samples at multiple angles of light incidence, which show a 10% to 15% decrease in reflection for 240 to 400 nm wavelengths. Further improvements in the optical efficiency of these structures can be achieved through simulation-fabrication-characterization cycles performed for this project. The optimized nanostructures can then serve the purpose of low-cost anti-reflective coatings for solar cells and similar applications.

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.

The reflectivity, wettability and scratch durability of microsurface features molded in the injection molding process using a dynamic tool tempering system

NASA Astrophysics Data System (ADS)

Kuhn, Sascha; Burr, August; Kübler, Michael; Deckert, Matthias; Bleesen, Christoph

2011-02-01

In this paper the replication qualities of periodically and randomly arranged micro-features molded in the injection molding process and their effects on surface properties are studied. The features are molded in PC, PMMA and PP at different mold wall temperatures in order to point out the necessity and profitability of a variotherm mold wall temperature control system. A one-dimensional heat conduction model is proposed to predict the cycle times of the variotherm injection molding processes. With regard to these processes, the molding results are compared to the molded surface feature heights using an atomic force microscope. In addition, the effects of the molded surface features on macroscopic surfaces are characterized in terms of light reflection using a spectrometer and in terms of water wettability by measuring the static contact angle. Furthermore, due to the sensitivity of the surface features on the molded parts, their durability is compared in a scratch test with a diamond tip. This leads to successful implementation in applications in which the optical appearance, in terms of gloss and reflection, and the water repellence, in terms of drag flow and adhesion, are of importance.

Nanoscale cellular imaging with scanning angle interference microscopy.

PubMed

DuFort, Christopher; Paszek, Matthew

2014-01-01

Fluorescence microscopy is among the most widely utilized tools in cell and molecular biology due to its ability to noninvasively obtain time-resolved images of live cells with molecule-specific contrast. In this chapter, we describe a simple high-resolution technique, scanning angle interference microscopy (SAIM), for the imaging and localization of fluorescent molecules with nanometer precision along the optical axis. In SAIM, samples above a reflective surface are sequentially scanned with an excitation laser at varying angles of incidence. Interference patterns generated between the incident and reflected lights result in an emission intensity that depends on the height of a fluorophore above the silicon surface and the angle of the incident radiation. The measured fluorescence intensities are then fit to an optical model to localize the labeled molecules along the z-axis with 5-10 nm precision and diffraction-limited lateral resolution. SAIM is easily implemented on widely available commercial total internal reflection fluorescence microscopes, offering potential for widespread use in cell biology. Here, we describe the setup of SAIM and its application for imaging cellular structures near (<1 μm) the sample substrate. © 2014 Elsevier Inc. All rights reserved.

Enhanced selective photocatalytic reduction of CO2 to CH4 over plasmonic Au modified g-C3N4 photocatalyst under UV-vis light irradiation

NASA Astrophysics Data System (ADS)

Li, Hailong; Gao, Yan; Xiong, Zhuo; Liao, Chen; Shih, Kaimin

2018-05-01

A series of Au-g-C3N4 (Au-CN) catalysts were prepared through a NaBH4-reduction method using g-C3N4 (CN) from pyrolysis of urea as precursor. The catalysts' surface area, crystal structure, surface morphology, chemical state, functional group composition and optical properties were characterized by X-ray diffraction, transmission electron microscope, X-ray photoelectron spectroscopy, ultraviolet visible (UV-vis) diffuse reflectance spectra, fourier transform infrared, photoluminescence and transient photocurrent analysis. The carbon dioxide (CO2) photoreduction activities under ultraviolet visible (UV-vis) light irradiation were significantly enhanced when gold (Au) was loaded on the surface of CN. 2Au-CN catalyst with Au to CN mole ratio of 2% showed the best catalytic activity. After 2 h UV-vis light irradiation, the methane (CH4) yield over the 2Au-CN catalyst was 9.1 times higher than that over the pure CN. The CH4 selectivity also greatly improved for the 2Au-CN compared to the CN. The deposited Au nanoparticles facilitated the separation of electron-hole pairs on the CN surface. Moreover, the surface plasmon resonance effect of Au further promoted the generation of hot electrons and visible light absorption. Therefore, Au loading significantly improved CO2 photoreduction performance of CN under UV-vis light irradiation.

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).

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.

Multispectral digital lensless holographic microscopy: from femtosecond laser to white light LED

NASA Astrophysics Data System (ADS)

Garcia-Sucerquia, J.

2015-04-01

The use of femtosecond laser radiation and super bright white LED in digital lensless holographic microscopy is presented. For the ultrafast laser radiation two different configurations of operation of the microscope are presented and the dissimilar performance of each one analyzed. The microscope operating with a super bright white light LED in combination with optical filters shows very competitive performance as it is compared with more expensive optical sources. The broadband emission of both radiation sources allows the multispectral imaging of biological samples to obtain spectral responses and/or full color images of the microscopic specimens; sections of the head of a Drosophila melanogaster fly are imaged in this contribution. The simple, solid, compact, lightweight, and reliable architecture of digital lensless holographic microscopy operating with broadband light sources to image biological specimens exhibiting micrometer-sized details is evaluated in the present contribution.

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

Ultrafast dark-field surface inspection with hybrid-dispersion laser scanning

NASA Astrophysics Data System (ADS)

Yazaki, Akio; Kim, Chanju; Chan, Jacky; Mahjoubfar, Ata; Goda, Keisuke; Watanabe, Masahiro; Jalali, Bahram

2014-06-01

High-speed surface inspection plays an important role in industrial manufacturing, safety monitoring, and quality control. It is desirable to go beyond the speed limitation of current technologies for reducing manufacturing costs and opening a new window onto a class of applications that require high-throughput sensing. Here, we report a high-speed dark-field surface inspector for detection of micrometer-sized surface defects that can travel at a record high speed as high as a few kilometers per second. This method is based on a modified time-stretch microscope that illuminates temporally and spatially dispersed laser pulses on the surface of a fast-moving object and detects scattered light from defects on the surface with a sensitive photodetector in a dark-field configuration. The inspector's ability to perform ultrafast dark-field surface inspection enables real-time identification of difficult-to-detect features on weakly reflecting surfaces and hence renders the method much more practical than in the previously demonstrated bright-field configuration. Consequently, our inspector provides nearly 1000 times higher scanning speed than conventional inspectors. To show our method's broad utility, we demonstrate real-time inspection of the surface of various objects (a non-reflective black film, transparent flexible film, and reflective hard disk) for detection of 10 μm or smaller defects on a moving target at 20 m/s within a scan width of 25 mm at a scan rate of 90.9 MHz. Our method holds promise for improving the cost and performance of organic light-emitting diode displays for next-generation smart phones, lithium-ion batteries for green electronics, and high-efficiency solar cells.

Wakata performs microscopic analysis of the NanoRacks Module-38 Petri Dishes

NASA Image and Video Library

2014-01-13

ISS038-E-029082 (12 Jan. 2014) --- Japan Aerospace Exploration Agency astronaut Koichi Wakata, Expedition 38 flight engineer, performs microscopic analysis of the NanoRacks Module-38 Petri Dishes, using Celestron Reflective Microscope, in the Kibo laboratory of the International Space Station. These Module-38 experiments are designed by students as part of a competition sponsored by the International Space School Educational Trust (ISSET). This experiment examines three-dimensional growth of slime mold in petri dishes utilizing the NanoRacks Microscopes Facility.

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.

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.

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.

Value of Reflected Light Microscopy in Teaching.

ERIC Educational Resources Information Center

Pasteris, Jill Dill

1983-01-01

Briefly reviews some optical and other physical properties of minerals that can be determined in reflected/incident light. Topics include optical properties of minerals, reflectance, internal reflections, color, bireflectance and reflection pleochroism, anisotropism, zonation, and reflected light microscopy as a teaching tool in undergraduate…

Design and analysis of multilayer x ray/XUV microscope

NASA Technical Reports Server (NTRS)

Shealy, David L.

1990-01-01

The design and analysis of a large number of normal incidence multilayer x ray microscopes based on the spherical mirror Schwarzschild configuration is examined. Design equations for the spherical mirror Schwarzschild microscopes are summarized and used to evaluate mirror parameters for microscopes with magnifications ranging from 2 to 50x. Ray tracing and diffraction analyses are carried out for many microscope configurations to determine image resolution as a function of system parameters. The results are summarized in three publication included herein. A preliminary study of advanced reflecting microscope configurations, where aspherics are used in place of the spherical microscope mirror elements, has indicated that the aspherical elements will improve off-axis image resolution and increase the effective field of view.

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

Nonimaging Optical Illumination System

DOEpatents

Winston, Roland

1994-08-02

A nonimaging illumination optical device for producing selected intensity output over an angular range. The device includes a light reflecting surface (24, 26) around a light source (22) which is disposed opposite the aperture opening of the light reflecting surface (24, 26). The light source (22) has a characteristic dimension which is small relative to one or more of the distance from the light source (22) to the light reflecting surface (24, 26) or the angle subtended by the light source (22) at the light reflecting surface (24, 26).

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.

[Phenotype-based primary screening for drugs promoting neuronal subtype differentiation in embryonic stem cells with light microscope].

PubMed

Gao, Yi-ning; Wang, Dan-ying; Pan, Zong-fu; Mei, Yu-qin; Wang, Zhi-qiang; Zhu, Dan-yan; Lou, Yi-jia

2012-07-01

To set up a platform for phenotype-based primary screening of drug candidates promoting neuronal subtype differentiation in embryonic stem cells (ES) with light microscope. Hanging drop culture 4-/4+ method was employed to harvest the cells around embryoid body (EB) at differentiation endpoint. Morphological evaluation for neuron-like cells was performed with light microscope. Axons for more than three times of the length of the cell body were considered as neuron-like cells. The compound(s) that promote neuron-like cells was further evaluated. Icariin (ICA, 10(-6)mol/L) and Isobavachin (IBA, 10(-7)mol/L) were selected to screen the differentiation-promoting activity on ES cells. Immunofluorescence staining with specific antibodies (ChAT, GABA) was used to evaluate the neuron subtypes. The cells treated with IBA showed neuron-like phenotype, but the cells treated with ICA did not exhibit the morphological changes. ES cells treated with IBA was further confirmed to be cholinergic and GABAergic neurons. Phenotypic screening with light microscope for molecules promoting neuronal differentiation is an effective method with advantages of less labor and material consuming and time saving, and false-positive results derived from immunofluorescence can be avoided. The method confirms that IBA is able to facilitate ES cells differentiating into neuronal cells, including cholinergic neurons and GABAergic neurons.

Photovoltaic module with light reflecting backskin

DOEpatents

Gonsiorawski, Ronald C [Danvers, MA

2007-07-03

A photovoltaic module comprises electrically interconnected and mutually spaced photovoltaic cells that are encapsulated by a light-transmitting encapsulant between a light-transparent front cover and a back cover, with the back cover sheet being an ionomer/nylon alloy embossed with V-shaped grooves running in at least two directions and coated with a light reflecting medium so as to provide light-reflecting facets that are aligned with the spaces between adjacent cells and oriented so as to reflect light falling in those spaces back toward said transparent front cover for further internal reflection onto the solar cells, whereby substantially all of the reflected light will be internally reflected from said cover sheet back to the photovoltaic cells, thereby increasing the current output of the module. The internal reflector improves power output by as much as 67%.

Müller glial cells contribute to dim light vision in the spectacled caiman (Caiman crocodilus fuscus): Analysis of retinal light transmission.

PubMed

Agte, Silke; Savvinov, Alexey; Karl, Anett; Zayas-Santiago, Astrid; Ulbricht, Elke; Makarov, Vladimir I; Reichenbach, Andreas; Bringmann, Andreas; Skatchkov, Serguei N

2018-05-16

In this study, we show the capability of Müller glial cells to transport light through the inverted retina of reptiles, specifically the retina of the spectacled caimans. Thus, confirming that Müller cells of lower vertebrates also improve retinal light transmission. Confocal imaging of freshly isolated retinal wholemounts, that preserved the refractive index landscape of the tissue, indicated that the retina of the spectacled caiman is adapted for vision under dim light conditions. For light transmission experiments, we used a setup with two axially aligned objectives imaging the retina from both sides to project the light onto the inner (vitreal) surface and to detect the transmitted light behind the retina at the receptor layer. Simultaneously, a confocal microscope obtained images of the Müller cells embedded within the vital tissue. Projections of light onto several representative Müller cell trunks within the inner plexiform layer, i.e. (i) trunks with a straight orientation, (ii) trunks which are formed by the inner processes and (iii) trunks which get split into inner processes, were associated with increases in the intensity of the transmitted light. Projections of light onto the periphery of the Müller cell endfeet resulted in a lower intensity of transmitted light. In this way, retinal glial (Müller) cells support dim light vision by improving the signal-to-noise ratio which increases the sensitivity to light. The field of illuminated photoreceptors mainly include rods reflecting the rod dominance of the of tissue. A subpopulation of Müller cells with downstreaming cone cells led to a high-intensity illumination of the cones, while the surrounding rods were illuminated by light of lower intensity. Therefore, Müller cells that lie in front of cones may adapt the intensity of the transmitted light to the different sensitivities of cones and rods, presumably allowing a simultaneous vision with both receptor types under dim light conditions. Copyright © 2018 Elsevier Ltd. 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

The CLAS12-RICH hybrid geometry

NASA Astrophysics Data System (ADS)

Angelini, Giovanni; CLAS12-RICH Collaboration

2017-01-01

A Ring-imaging Cherenkov detector (RICH) has been designed for the CLAS12 spectrometer (JLAB, Hall B) in order to increase the particle identification. Among the approved physics program focused upon 3D imaging of the nucleon, some Semi Inclusive Deep Inelastic Scattering experiments (E12-09-007, E12-09-008, E12-09-009) demand an efficient kaon identification across the momentum range from 3 to 8 GeV/c. The detector exploits a novel elaborated hybrid geometry based on a complex focusing mirror system that will reduce the area instrumented with photon detectors. For forward scattered particles (θ <12°) with momenta p = 3-8 GeV/c, a proximity imaging method with direct Cherenkov light detection will be used. For larger angles of 12° < θ <35° and momenta of p = 3-6 GeV/c, the Cherenkov light will be focused by a spherical mirror, undergo two further passes through the aerogel radiator and will be reflected from planar mirrors before detection. A carefully study on reflections has been performed considering microscopic and macroscopic effects. In addition, a new feature has been introduced in the CLAS12 simulation software in order to generate the geometry of the detector by using a computer-aided design (CAD) file for an accurate geometrical description. U.S. Department of Energy, GWU Columbian College Art and Science Facilitating Fund Award (CCAS CCFF).

Dynamic quantitative phase images of pond life, insect wings, and in vitro cell cultures

NASA Astrophysics Data System (ADS)

Creath, Katherine

2010-08-01

This paper presents images and data of live biological samples taken with a novel Linnik interference microscope. The specially designed optical system enables instantaneous and 3D video measurements of dynamic motions within and among live cells without the need for contrast agents. This "label-free", vibration insensitive imaging system enables measurement of biological objects in reflection using harmless light levels with current magnifications of 10X (NA 0.3) and 20X (NA 0.5) and wavelengths of 660 nm and 785 nm over fields of view from several hundred microns up to a millimeter. At the core of the instrument is a phasemeasurement camera (PMC) enabling simultaneous measurement of multiple interference patterns utilizing a pixelated phase mask taking advantage of the polarization properties of light. Utilizing this technology enables the creation of phase image movies in real time at video rates so that dynamic motions and volumetric changes can be tracked. Objects are placed on a reflective surface in liquid under a coverslip. Phase values are converted to optical thickness data enabling volumetric, motion and morphological studies. Data from a number of different mud puddle organisms such as paramecium, flagellates and rotifers will be presented, as will measurements of flying ant wings and cultures of human breast cancer cells. These data highlight examples of monitoring different biological processes and motions. The live presentation features 4D phase movies of these examples.

Optical based tactile shear and normal load sensor

DOEpatents

Salisbury, Curt Michael

2015-06-09

Various technologies described herein pertain to a tactile sensor that senses normal load and/or shear load. The tactile sensor includes a first layer and an optically transparent layer bonded together. At least a portion of the first layer is made of optically reflective material. The optically transparent layer is made of resilient material (e.g., clear silicone rubber). The tactile sensor includes light emitter/light detector pair(s), which respectively detect either normal load or shear load. Light emitter(s) emit light that traverses through the optically transparent layer and reflects off optically reflective material of the first layer, and light detector(s) detect and measure intensity of reflected light. When a normal load is applied, the optically transparent layer compresses, causing a change in reflected light intensity. When shear load is applied, a boundary between optically reflective material and optically absorptive material is laterally displaced, causing a change in reflected light intensity.

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

PubMed

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

2013-05-20

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

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.

Ellipsoidal cell flow system

DOEpatents

Salzman, Gary C.; Mullaney, Paul F.

1976-01-01

The disclosure relates to a system incorporating an ellipsoidal flow chamber having light reflective walls for low level light detection in practicing cellular analysis. The system increases signal-to-noise ratio by a factor of ten over prior art systems. In operation, laser light passes through the primary focus of the ellipsoid. A controlled flow of cells simultaneously passes through this focus so that the laser light impinges on the cells and is modulated by the cells. The reflective walls of the ellipsoid reflect the cell-modulated light to the secondary focus of the ellipsoid. A tapered light guide at the secondary focus picks up a substantial portion of modulated reflective light and directs it onto a light detector to produce a signal. The signal is processed to obtain the intensity distribution of the modulated light and hence sought after characteristics of the cells. In addition, cells may be dyed so as to fluoresce in response to the laser light and their fluorescence may be processed as cell-modulated light above described. A light discriminating filter would be used to distinguish reflected modulated laser light from reflected fluorescent light.

Solar module having reflector between cells

DOEpatents

Kardauskas, Michael J.

1999-01-01

A photovoltaic module comprising an array of electrically interconnected photovoltaic cells disposed in a planar and mutually spaced relationship between a light-transparent front cover member in sheet form and a back sheet structure is provided with a novel light-reflecting means disposed between adjacent cells for reflecting light falling in the areas between cells back toward said transparent cover member for further internal reflection onto the solar cells. The light-reflecting comprises a flexible plastic film that has been embossed so as to have a plurality of small V-shaped grooves in its front surface, and a thin light-reflecting coating on said front surface, the portions of said coating along the sides of said grooves forming light-reflecting facets, said grooves being formed so that said facets will reflect light impinging thereon back into said transparent cover sheet with an angle of incidence greater than the critical angle, whereby substantially all of the reflected light will be internally reflected from said cover sheet back to said solar modules, thereby increasing the current output of the module.

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.

Apparatus for and method of correcting for astigmatism in a light beam reflected off of a light reflecting surface

DOEpatents

Sawicki, R.H.; Sweatt, W.

1985-11-21

A technique for adjustably correcting for astigmatism in a light beam is disclosed herein. This technique defines a flat, rectangular light reflecting surface having opposite reinforced side edges and which is resiliently bendable, to a limited extent, into different concave and/or convex cylindrical curvatures about a particular axis and provides for adjustably bending the light reflecting surface into one of different curvatures depending upon the astigmatism to be corrected and for fixedly maintaining the curvature selected. In the embodiment disclosed, the light reflecting surface is adjustably bendable into the selected cylindrical curvature by application of a particular bending moment to the reinforced side edges of the light reflecting surface.

Apparatus for and method of correcting for astigmatism in a light beam reflected off of a light reflecting surface

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

Sawicki, R.H.; Sweatt, W.

1985-11-21

A technique for adjustably correcting for astigmatism in a light beam is disclosed herein. This technique defines a flat, rectangular light reflecting surface having opposite reinforced side edges and which is resiliently bendable, to a limited extent, into different concave and/or convex cylindrical curvatures about a particular axis and provides for adjustably bending the light reflecting surface into one of different curvatures depending upon the astigmatism to be corrected and for fixedly maintaining the curvature selected. In the embodiment disclosed, the light reflecting surface is adjustably bendable into the selected cylindrical curvature by application of a particular bending moment tomore » the reinforced side edges of the light reflecting surface.« less

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.

Characterization and Beneficiation Studies of a Low Grade Bauxite Ore

NASA Astrophysics Data System (ADS)

Rao, D. S.; Das, B.

2014-10-01

A low grade bauxite sample of central India was thoroughly characterized with the help of stereomicroscope, reflected light microscope and electron microscope using QEMSCAN. A few hand picked samples were collected from different places of the mine and were subjected to geochemical characterization studies. The geochemical studies indicated that most of the samples contain high silica and low alumina, except a few which are high grade. Mineralogically the samples consist of bauxite (gibbsite and boehmite), ferruginous mineral phases (goethite and hematite), clay and silicate (quartz), and titanium bearing minerals like rutile and ilmenite. Majority of the gibbsite, boehmite and gibbsitic oolites contain clay, quartz and iron and titanium mineral phases within the sample as inclusions. The sample on an average contains 39.1 % Al2O3 and 12.3 % SiO2, and 20.08 % of Fe2O3. Beneficiation techniques like size classification, sorting, scrubbing, hydrocyclone and magnetic separation were employed to reduce the silica content suitable for Bayer process. The studies indicated that, 50 % by weight with 41 % Al2O3 containing less than 5 % SiO2 could be achieved. The finer sized sample after physical beneficiation still contains high silica due to complex mineralogical associations.

Reconstruction of explicit structural properties at the nanoscale via spectroscopic microscopy

NASA Astrophysics Data System (ADS)

Cherkezyan, Lusik; Zhang, Di; Subramanian, Hariharan; Taflove, Allen; Backman, Vadim

2016-02-01

The spectrum registered by a reflected-light bright-field spectroscopic microscope (SM) can quantify the microscopically indiscernible, deeply subdiffractional length scales within samples such as biological cells and tissues. Nevertheless, quantification of biological specimens via any optical measures most often reveals ambiguous information about the specific structural properties within the studied samples. Thus, optical quantification remains nonintuitive to users from the diverse fields of technique application. In this work, we demonstrate that the SM signal can be analyzed to reconstruct explicit physical measures of internal structure within label-free, weakly scattering samples: characteristic length scale and the amplitude of spatial refractive-index (RI) fluctuations. We present and validate the reconstruction algorithm via finite-difference time-domain solutions of Maxwell's equations on an example of exponential spatial correlation of RI. We apply the validated algorithm to experimentally measure structural properties within isolated cells from two genetic variants of HT29 colon cancer cell line as well as within a prostate tissue biopsy section. The presented methodology can lead to the development of novel biophotonics techniques that create two-dimensional maps of explicit structural properties within biomaterials: the characteristic size of macromolecular complexes and the variance of local mass density.

Photovoltaic characteristics of natural light harvesting dye sensitized solar cells.

PubMed

Hafez, H S; Shenouda, S S; Fadel, M

2018-03-05

In this work of research, anthocyanin as a natural dye obtained from raspberry fruits, was used and tested as a photon harvesting/electron donating dye in titanium dioxide nanoparticle-based DSSCs. A working photoelectrode made from TiO 2 nanoparticles with an average particle size (10-40nm) that is coated on Florine doped tin-oxide substrate, was prepared via a simple and low cost hydrothermal method. A detailed structural and morphological analysis of the TiO 2 photoactive electrode was investigated by X-ray diffraction (XRD), diffuse reflectance spectrometer, transmission electron microscope (TEM) and scanning electron microscope (SEM). Complete photovoltaic characteristics including (current, voltage, outpower, and responsivity) of the natural anthocyanin based dye sensitized solar cell have been investigated under different illumination intensity ranging from 10 to 100mW.cm -2 . The cell responsivity and efficiency of the fabricated solar cell under different illumination intensity were found to be in the range (R=15.6-23.8mA.W -1 and η=0.13-0.25) at AM=1.5 conditions. This study is important for enhancing the future applications of the promising DSSC technology. Copyright © 2017 Elsevier B.V. All rights reserved.

Microscopic image processing systems for measuring nonuniform film thickness profiles

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

Liu, A.H.; Plawsky, J.L.; DasGupta, S.

1994-01-01

In very thin liquid films. transport processes are controlled by the temperature and the interfacial intermolecular force field which is a function of the film thickness profile and interfacial properties. The film thickness profile and interfacial properties can be measured most efficiently using a microscopic image processing system. IPS, to record the intensity pattern of the reflected light from the film. There are two types of IPS: an image analyzing interferometer (IAI) and/or an image scanning ellipsometer (ISE). The ISE is a novel technique to measure the two dimensional thickness profile of a nonuniform, thin film, from 1 nm upmore » to several {mu}m, in a steady state as well as in a transient state. It is a full field imaging technique which can study every point on the surface simultaneously with high spatial resolution and thickness sensitivity, i.e., it can measure and map the 2-D film thickness profile. Using the ISE, the transient thickness profile of a draining thin liquid film was measured and modeled. The interfacial conditions were determined in situ by measuring the Hamaker constant. The ISE and IAI systems are compared.« less

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.

Optical nulling apparatus and method for testing an optical surface

NASA Technical Reports Server (NTRS)

Olczak, Eugene (Inventor); Hannon, John J. (Inventor); Dey, Thomas W. (Inventor); Jensen, Arthur E. (Inventor)

2008-01-01

An optical nulling apparatus for testing an optical surface includes an aspheric mirror having a reflecting surface for imaging light near or onto the optical surface under test, where the aspheric mirror is configured to reduce spherical aberration of the optical surface under test. The apparatus includes a light source for emitting light toward the aspheric mirror, the light source longitudinally aligned with the aspheric mirror and the optical surface under test. The aspheric mirror is disposed between the light source and the optical surface under test, and the emitted light is reflected off the reflecting surface of the aspheric mirror and imaged near or onto the optical surface under test. An optical measuring device is disposed between the light source and the aspheric mirror, where light reflected from the optical surface under test enters the optical measuring device. An imaging mirror is disposed longitudinally between the light source and the aspheric mirror, and the imaging mirror is configured to again reflect light, which is first reflected from the reflecting surface of the aspheric mirror, onto the optical surface under test.

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

Improved Oxygen-Beam Texturing of Glucose-Monitoring Optics

NASA Technical Reports Server (NTRS)

Banks, Bruce A.

2006-01-01

An improved method has been devised for using directed, hyperthermal beams of oxygen atoms and ions to impart desired textures to the tips of polymethylmethacrylate [PMMA] optical fibers to be used in monitoring the glucose content of blood. The improved method incorporates, but goes beyond, the method described in Texturing Blood-Glucose- Monitoring Optics Using Oxygen Beams (LEW-17642-1), NASA Tech Briefs, Vol. 29, No. 4 (April 2005), page 11a. The basic principle of operation of such a glucose-monitoring sensor is as follows: The textured surface of the optical fiber is coated with chemicals that interact with glucose in such a manner as to change the reflectance of the surface. Light is sent down the optical fiber and is reflected from, the textured surface. The resulting change in reflectance of the light is measured as an indication of the concentration of glucose. The required texture on the ends of the optical fibers is a landscape of microscopic cones or pillars having high aspect ratios (microscopic structures being taller than they are wide). The average distance between hills must be no more than about 5 mso that blood cells (which are wider) cannot enter the valleys between the hills, where they would interfere with optical sensing of glucose in the blood plasma. On the other hand, the plasma is required to enter the valleys, and high aspect ratio structures are needed to maximize the surface area in contact with the plasma, thereby making it possible to obtain a given level of optical glucose-measurement sensitivity with a relatively small volume of blood. There is an additional requirement that the hills be wide enough that a sufficient amount of light can propagate into them and, after reflection, can propagate out of them. The method described in the cited prior article produces a texture comprising cones and pillars that conform to the average-distance and aspect-ratio requirements. However, a significant fraction of the cones and pillars are so narrow that not enough light can propagate along them. The improved method makes it possible to form wider cones and pillars while still satisfying the average-distance and aspect-ratio requirements. In the improved method, as in the previously reported method, multiple optical fibers are first bundled together for simultaneous texturing of their distal tips. However, prior to texturing by exposure to an oxygen beam, the tips are first coated by vapor deposition of a thin, sparse layer of aluminum: The exposure to the aluminum vapor must be short enough (typically of the order of seconds) so that the aluminum nucleates into islands separated by uncoated areas. The coated tips are textured by exposure to a directed beam of hyperthermal (kinetic energy >1 eV) oxygen atoms and/or ions in a vacuum chamber, as in the previously reported method. The aluminum islands partially shield the underlying PMMA from oxidation and erosion by the beam, so that the cones or pillars remaining after texturing are wider than they would otherwise be. To some extent, the dimensions of the hills and the distances between them can be tailored through choice of the thickness of the aluminum coat and/or the oxygen-beam fluence. The figure illustrates an example of texturing of the tip of a PMMA optical fiber without and with prior aluminum coating.

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

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

PubMed Central

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

2015-01-01

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

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

PubMed

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

2014-10-01

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

Analysis of biological time-lapse microscopic experiment from the point of view of the information theory.

PubMed

Štys, Dalibor; Urban, Jan; Vaněk, Jan; Císař, Petr

2011-06-01

We report objective analysis of information in the microscopic image of the cell monolayer. The process of transfer of information about the cell by the microscope is analyzed in terms of the classical Shannon information transfer scheme. The information source is the biological object, the information transfer channel is the whole microscope including the camera chip. The destination is the model of biological system. The information contribution is analyzed as information carried by a point to overall information in the image. Subsequently we obtain information reflection of the biological object. This is transformed in the biological model which, in information terminology, is the destination. This, we propose, should be constructed as state transitions in individual cells modulated by information bonds between the cells. We show examples of detected cell states in multidimensional state space. This space is reflected as colour channel intensity phenomenological state space. We have also observed information bonds and show examples of them.

Analysis of biological time-lapse microscopic experiment from the point of view of the information theory.

PubMed

Stys, Dalibor; Urban, Jan; Vanek, Jan; Císar, Petr

2010-07-01

We report objective analysis of information in the microscopic image of the cell monolayer. The process of transfer of information about the cell by the microscope is analyzed in terms of the classical Shannon information transfer scheme. The information source is the biological object, the information transfer channel is the whole microscope including the camera chip. The destination is the model of biological system. The information contribution is analyzed as information carried by a point to overall information in the image. Subsequently we obtain information reflection of the biological object. This is transformed in the biological model which, in information terminology, is the destination. This, we propose, should be constructed as state transitions in individual cells modulated by information bonds between the cells. We show examples of detected cell states in multidimensional state space reflected in space an colour channel intensity phenomenological state space. We have also observed information bonds and show examples of them. Copyright 2010 Elsevier Ltd. All rights reserved.

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

Cytochemical features common to nucleoli and cytoplasmic nucleoloids of Olea europaea meiocytes: detection of rRNA by in situ hybridization.

PubMed

Alché, J D; Fernández, M C; Rodríguez-García, M I

1994-02-01

We used light and electron microscopic techniques to study the composition of cytoplasmic nucleoloids during meiotic division in Olea europaea. Nucleoloids were found in two clearly distinguishable morphological varieties: one similar in morphology to the nucleolus, and composed mainly of dense fibrillar component, and another surrounded by many ribosome-like particles. Cytochemical and immunocytochemical techniques showed similar reactivities in nucleoloids and the nucleolus: both are ribonucleoproteic in nature, and possess argyrophillic, argentaffinic and highly phosphorylated proteins. Immunohistochemical techniques failed to detect DNA in either structure. In situ hybridization to a 18 S rRNA probe demonstrated the presence of ribosomal transcripts in both the nucleolus and nucleoloids. These similarities in morphology and composition may reflect similar functionalities.

Using an atom interferometer to take the Gedanken out of Feynman's Gedankenexperiment

NASA Astrophysics Data System (ADS)

Pritchard, David E.; Hammond, Troy D.; Lenef, Alan; Rubenstein, Richard A.; Smith, Edward T.; Chapman, Michael S.; Schmiedmayer, Jörg

1997-01-01

We give a description of two experiments performed in an atom interferometer at MIT. By scattering a single photon off of the atom as it passes through the interferometer, we perform a version of a classic gedankenexperiment, a demonstration of a Feynman light microscope. As path information about the atom is gained, contrast in the atom fringes (coherence) is lost. The lost coherence is then recovered by observing only atoms which scatter photons into a particular final direction. This paper reflects the main emphasis of D. E. Pritchard's talk at the RIS meeting. Information about other topics covered in that talk, as well as a review of all of the published work performed with the MIT atom/molecule interferometer, is available on the world wide web at http://coffee.mit.edu/.

Estimated Mid-Infrared (200-2000 cm-1) Optical Constants of Some Silica Polymorphs

NASA Astrophysics Data System (ADS)

Glotch, Timothy; Rossman, G. R.; Michalski, J. R.

2006-09-01

We use Lorentz-Lorenz dispersion analysis to model the mid-infrared (200-2000 cm-1) optical constants, of opal-A, opal-CT, and tridymite. These minerals, which are all polymorphs of silica (SiO2), are potentially important in the analysis of thermal emission spectra acquired by the Mars Global Surveyor Thermal Emission Spectrometer (MGS-TES) and Mars Exploration Rover Mini-TES instruments in orbit and on the surface of Mars as well as emission spectra acquired by telescopes of planetary disks and dust and debris clouds in young solar systems. Mineral samples were crushed, washed, and sieved and emissivity spectra of the >100; μm size fraction were acquired at Arizona State University's emissivity spectroscopy laboratory. Therefore, the spectra and optical constants are representative of all crystal orientations. Ideally, emissivity or reflectance measurements of single polished crystals or fine powders pressed to compact disks are used for the determination of mid-infrared optical constants. Measurements of these types of surfaces eliminate or minimize multiple reflections, providing a specular surface. Our measurements, however, likely produce a reasonable approximation of specular emissivity or reflectance, as the minimum particle size is greater than the maximum wavelength of light measured. Future work will include measurement of pressed disks of powdered samples in emission and reflection, and when possible, small single crystals under an IR reflectance microscope, which will allow us to assess the variability of spectra and optical constants under different sample preparation and measurement conditions.

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.

Front lighted optical tooling method and apparatus

DOEpatents

Stone, W.J.

1983-06-30

An optical tooling method and apparatus uses a front lighted shadowgraphic technique to enhance visual contrast of reflected light. The apparatus includes an optical assembly including a fiducial mark, such as cross hairs, reflecting polarized light with a first polarization, a polarizing element backing the fiducial mark and a reflective surface backing the polarizing element for reflecting polarized light bypassing the fiducial mark and traveling through the polarizing element. The light reflected by the reflecting surface is directed through a second pass of the polarizing element toward the frontal direction with a polarization differing from the polarization of the light reflected by the fiducial mark. When used as a tooling target, the optical assembly may be mounted directly to a reference surface or may be secured in a mounting, such as a magnetic mounting. The optical assembly may also be mounted in a plane defining structure and used as a spherometer in conjunction with an optical depth measuring instrument.

Apparatus for and method of correcting for astigmatism in a light beam reflected off of a light reflecting surface

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

Sawicki, R.H.; Sweatt, W.

1987-03-03

An apparatus is described for correcting for astigmatism in a light beam reflected off of a light reflecting surface, comprising: (a) a first means defining a flat, rectangular light reflecting surface which is resiliently bendable, to a limited extent, into different concave and/or convex cylindrical curvatures about a particular axis. The first means is configured so that the light reflecting surface can be adjustably bent into the selected cylindrical curvature by applying a particular bending moment to the first means with respect to the surface, depending upon the curvature desired. The first means includes an integrally formed body member havingmore » a main plate-like segment including a front fact defining the light reflecting surface and a pair of spaced-apart flange segments extending rearwardly of the main segment; and (b) second means acting on the first means for adjustably bending the light reflecting surface into a particular selected one of the different cylindrical curvatures, depending upon the astigmatism to be corrected for.« less

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.

Beagle I and II Voyages: Charles Darwin's rocks and the quest for Mars rock; the Open University's virtual microscope has both

NASA Astrophysics Data System (ADS)

Schwenzer, S. P.; Tindle, A. G.; Anand, M.; Gibson, E. K.; Pearson, V. K.; Pemberton, D.; Pillinger, C.; Smith, C. L.; Whalley, P.; Kelley, S. P.

2011-12-01

Exploration is in itself a fascinating subject, and a strong draw to engaging the public in understanding science. Nearly two hundred years ago Charles Darwin took part in an exploration of the Earth, and more recently we have begun to explore the solar system and in particular the surface of Mars. The engagement is made easier if an element of exploration is involved in the public engagement, using modern internet and even mobile technologies. The Open University combines all those aspects in a series of virtual microscopes for Earth science that are freely available on the web, installed in museums, or built into its teaching material. The basis of the virtual microscope is a mosaic of several hundred microscopic images of each thin section taken in plane polarised light, between crossed polars and in reflected light, which are then assembled into three high resolution images. Rotation movies for selected points in the thin section illustrate changing optical properties such as birefringence. The user is able to pan and zoom around to explore the section, studying the mineralogy and rock texture, and view the rotation movies linked to points in the section to see the changing birefringence colours. We have created several collections of terrestrial rocks, mainly for teaching purposes, and outreach directly linked to exploration: Charles Darwin returned from the Voyage of the Beagle with a large variety of rock samples, and although thin sections were not being made at that time, they were created from his rocks in the late 19th century. The historic material is part of the "Darwin the Geologist" exhibition at the Sedgwick Museum in Cambridge. Our Darwin virtual microscope includes hand specimen illustrations and thin sections together with documentation and an interactive map allow internet users and museum visitors alike to have a close look at Darwin's rocks and study the petrology of them. Charles Darwin explored distant horizons on Earth in the 19th century; in the 20th century the Apollo astronauts set foot on the Moon, returning valuable rock samples to Earth. Through collaboration between NASA and the OU it became possible to show lunar samples as virtual thin sections. The Beagle II mission represented a new voyage, following Charles Darwin's footsteps, to horizons well beyond the Earth - on a journey to investigate the planet Mars. Although no samples have yet been returned from the red planet, we do have access to Martian meteorites. Like Moon rock samples, these meteorites are rare and very valuable. So, one way to make them accessible to the general public is via the internet using our virtual microscope technology. Within the framework of the EUROPLANET project, and in collaboration with the Natural History Museum in London we are making such meteorites freely available to all. We plan to extend this collection and make it openly accessible for teaching and outreach activities anywhere and any time. Our current microscopes are located at http://microscope.open.ac.uk.

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).

Light microscopical structure of the excurrent ducts and distribution of spermatozoa in the Australian rodents Pseudomys australis and Notomys alexis.

PubMed Central

Peirce, E J; Breed, W G

1989-01-01

The light microscopical structure of the male excurrent ducts and the distribution of spermatozoa were examined in two species of Australian rodents, the plains rat, Pseudomys australis, and the hopping mouse, Notomys alexis. In plains rats the microstructure of the ductus epididymidis and ductus deferens was similar to that of the common laboratory rodents, with the majority of the spermatozoa being found in the cauda epididymides. By contrast, in the hopping mouse, the structure of the cauda epididymidis differed significantly as the height of the epithelium and stereocilia did not decrease from the distal caput to the cauda region, and luminal diameter did not increase markedly along its length. In addition, few spermatozoa were stored in the cauda region of the tract, and as many as 60% were located in the ductus deferens, the distal portion of which displayed a highly infolded epithelium and underlying lamina propria. These differences in histological structure of the hopping mouse excurrent ducts presumably reflect divergence in function of the various regions of the tract. Although the functional implications of the present findings remain to be determined, this study demonstrates the considerable plasticity in the male excurrent ducts amongst the hydromyine rodents of Australia. Images Figs. 1-2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 10 Fig. 11 Fig. 12 Fig. 13 Fig. 14 Fig. 15 Fig. 16 Fig. 17 Fig. 18 Fig. 19 Fig. 20 Fig. 21 Figs. 22-23 Fig. 24 Fig. 25 Fig. 26 PMID:2808117

Fabrication of magnetic Fe@ZnO0.6S0.4 nanocomposite for visible-light-driven photocatalytic inactivation of Escherichia coli

NASA Astrophysics Data System (ADS)

Peng, Ziling; Wu, Dan; Wang, Wei; Tan, Fatang; Ng, Tsz Wai; Chen, Jianguo; Qiao, Xueliang; Wong, Po Keung

2017-02-01

Bacterial inactivation by magnetic photocatalysts has now received growing interests due to the easy separation for recycle and reuse of photocatalysts. In this study, magnetic Fe@ZnO0.6S0.4 photocatalyst was prepared by a facile two-step precipitation method. Multiple techniques such as X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), UV-vis diffused reflectance spectra (UV-vis DRS) and vibrating sample magnetometer (VSM) were employed to characterize the structure, morphology and physicochemical properties of the photocatalyst. The as-obtained Fe@ZnO0.6S0.4 possessing magnetic property was easily collected from the reaction system by a magnet. Under white light-emitting-diode (LED) lamp irradiation, Fe@ZnO0.6S0.4 nanocomposite could completely inactivate 7-log of Escherichia coli K-12 within 5 h. More importantly, almost no decrease of photocatalytic efficiency in bacterial inactivation was observed even after five consecutive cycles, demonstrating Fe@ZnO0.6S0.4 exhibited good stability for reuse. The low released rate of Fe2+/Fe3+ and Zn2+ from Fe@ZnO0.6S0.4 composite further indicated the photocatalyst showed low cytotoxicity to bacterium and high stability under LED lamp irradiation. Facile preparation, high photocatalytic efficiency, good stability and reusability, and magnetic recovery property endow Fe@ZnO0.6S0.4 nanocomposite to be a promising photocatalytic material for bacterial inactivation.

Three-dimensional confocal microscopy of the living cornea and ocular lens

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1991-07-01

The three-dimensional reconstruction of the optic zone of the cornea and the ocular crystalline lens has been accomplished using confocal microscopy and volume rendering computer techniques. A laser scanning confocal microscope was used in the reflected light mode to obtain the two-dimensional images from the cornea and the ocular lens of a freshly enucleated rabbit eye. The light source was an argon ion laser with a 488 nm wavelength. The microscope objective was a Leitz X25, NA 0.6 water immersion lens. The 400 micron thick cornea was optically sectioned into 133 three micron sections. The semi-transparent cornea and the in-situ ocular lens was visualized as high resolution, high contrast two-dimensional images. The structures observed in the cornea include: superficial epithelial cells and their nuclei, basal epithelial cells and their 'beaded' cell borders, basal lamina, nerve plexus, nerve fibers, nuclei of stromal keratocytes, and endothelial cells. The structures observed in the in- situ ocular lens include: lens capsule, lens epithelial cells, and individual lens fibers. The three-dimensional data sets of the cornea and the ocular lens were reconstructed in the computer using volume rendering techniques. Stereo pairs were also created of the two- dimensional ocular images for visualization. The stack of two-dimensional images was reconstructed into a three-dimensional object using volume rendering techniques. This demonstration of the three-dimensional visualization of the intact, enucleated eye provides an important step toward quantitative three-dimensional morphometry of the eye. The important aspects of three-dimensional reconstruction are discussed.

A comparison of antibacterial and antibiofilm efficacy of phenothiazinium dyes between Gram positive and Gram negative bacterial biofilm.

PubMed

Misba, Lama; Zaidi, Sahar; Khan, Asad U

2017-06-01

Antimicrobial photodynamic therapy (APDT) is a process that generates reactive oxygen species (ROS) in presence of photosensitizer, visible light and oxygen which destroys the bacterial cells. We investigated the photoinactivation efficiency of phenothiazinium dyes and the effect of ROS generation on Gram positive and Gram negative bacterial cell as well as on biofilm. Enterococcus faecalis and Klebsiella pneumonia were incubated with all the three phenothiazinium dyes and exposed to 630nm of light. After PDT, colony forming unit (CFU) were performed to estimate the cell survival fraction. Intracellular reactive oxygen species (ROS) was detected by DCFH-DA. Crystal violet (CV) assay and extracellular polysaccharides (EPS) reduction assay were performed to analyze antibiofilm effect. Confocal laser electron microscope (CLSM) scanning electron microscope (SEM) was performed to assess the disruption of biofilm. 8log 10 reduction in bacterial count was observed in Enterococcus faecalis while 3log 10 in Klebsiella pneumoniae. CV and EPS reduction assay revealed that photodynamic inhibition was more pronounced in Enterococcus faecalis. In addition to this CLSM and SEM study showed an increase in cell permeability of propidium iodide and leakage of cellular constituents in treated preformed biofilm which reflects the antibiofilm action of photodynamic therapy. We conclude that Gram-positive bacteria (Enterococcus faecalis) are more susceptible to APDT due to increased level of ROS generation inside the cell, higher photosensitizer binding efficiency and DNA degradation. Phenothiazinium dyes are proved to be highly efficient against both planktonic and biofilm state of cells. Copyright © 2017 Elsevier B.V. All rights reserved.

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.

Infrared/microwave (IR/MW) micromirror array beam combiner design and analysis.

PubMed

Tian, Yi; Lv, Lijun; Jiang, Liwei; Wang, Xin; Li, Yanhong; Yu, Haiming; Feng, Xiaochen; Li, Qi; Zhang, Li; Li, Zhuo

2013-08-01

We investigated the design method of an infrared (IR)/microwave (MW) micromirror array type of beam combiner. The size of micromirror is in microscopic levels and comparable to MW wavelengths, so that the MW will not react in these dimensions, whereas the much shorter optical wavelengths will be reflected by them. Hence, the MW multilayered substrate was simplified and designed using transmission line theory. The beam combiner used an IR wavefront-division imaging technique to reflect the IR radiation image to the unit under test (UUT)'s pupil in a parallel light path. In addition, the boresight error detected by phase monopulse radar was analyzed using a moment-of method (MoM) and multilevel fast multipole method (MLFMM) acceleration technique. The boresight error introduced by the finite size of the beam combiner was less than 1°. Finally, in order to verify the wavefront-division imaging technique, a prototype of a micromirror array was fabricated, and IR images were tested. The IR images obtained by the thermal imager verified the correctness of the wavefront-division imaging technique.

Group-III nitride VCSEL structures grown by molecular beam epitaxy

NASA Astrophysics Data System (ADS)

Ng, HockMin; Moustakas, Theodore D.

2000-07-01

III-nitride VCSEL structures designed for electron-beam pumping have been grown by molecular beam epitaxy (MBE). The structures consist of a sapphire substrate on which an AlN/GaN distributed Bragg reflector (DBR) with peak reflectance >99% at 402 nm is deposited. The active region consists of a 2-(lambda) cavity with 25 In0.1Ga0.9N/GaN multiquantum wells (MQWs) whose emission coincides with the high reflectance region of the DBR. The thicknesses of the InGaN wells and the GaN barriers are 35 angstrom and 75 angstrom respectively. The top reflector consists of a silver metallic mirror which prevents charging effects during electron-beam pumping. The structure was pumped from the top- side with a cw electron-beam using a modified cathodoluminescence (CL) system mounted on a scanning electron microscope chamber. Light output was collected from the polished sapphire substrate side. Measurements performed at 100 K showed intense emission at 407 nm with narrowing of the linewidth with increasing beam current. A narrow emission linewidth of 0.7 nm was observed indicating the onset of stimulated emission.

Nonimaging Optical Illumination System

DOEpatents

Winston, Roland

1994-02-22

A nonimaging illumination or concentration optical device. An optical device is provided having a light source, a light reflecting surface with an opening and positioned partially around the light source which is opposite the opening of the light reflecting surface. The light reflecting surface is disposed to produce a substantially uniform intensity output with the reflecting surface defined in terms of a radius vector R.sub.i in conjunction with an angle .phi..sub.i between R.sub.i, a direction from the source and an angle .theta..sub.i between direct forward illumination and the light ray reflected once from the reflecting surface. R.sub.i varies as the exponential of tan (.phi..sub.i -.theta..sub.i)/2 integrated over .phi..sub.i.

Photoelectrochemical Properties of CuS-GeO2-TiO2 Composite Coating Electrode

PubMed Central

Wen, Xinyu; Zhang, Huawei

2016-01-01

The ITO (indium tin oxide) conductive glass-matrix CuS-GeO2-TiO2 composite coating was generated via EPD (electrophoretic deposition) and followed by a sintering treatment at 450°C for 40 minutes. Characterizations of the CuS-GeO2-TiO2 composite coating were taken by SEM (scanning electron microscope), XRD (X-ray diffraction), EDX (energy dispersive X-ray), UV-Vis DRS (ultraviolet-visible diffuse reflection spectrum), and FT-IR (Fourier transform infrared spectroscopy). Results showed that CuS and GeO2 had dispersed in this CuS-GeO2-TiO2 composite coating (mass percentages for CuS and GeO2 were 1.23% and 2.79%, respectively). The electrochemical studies (cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and Tafel polarization) of this CuS-GeO2-TiO2 composite coating electrode were performed in pH = 9.51 Na2CO3-NaHCO3 buffer solution containing 0.50 mol/L CH3OH under the conditions of visible light, ultraviolet light (λ = 365 nm), and dark (without light irradiation as control), respectively. Electrochemical studies indicated that this CuS-GeO2-TiO2 composite coating electrode had better photoelectrocatalytic activity than the pure TiO2 electrode in the electrocatalysis of methanol under visible light. PMID:27055277

Photoelectrochemical Properties of CuS-GeO2-TiO2 Composite Coating Electrode.

PubMed

Wen, Xinyu; Zhang, Huawei

2016-01-01

The ITO (indium tin oxide) conductive glass-matrix CuS-GeO2-TiO2 composite coating was generated via EPD (electrophoretic deposition) and followed by a sintering treatment at 450°C for 40 minutes. Characterizations of the CuS-GeO2-TiO2 composite coating were taken by SEM (scanning electron microscope), XRD (X-ray diffraction), EDX (energy dispersive X-ray), UV-Vis DRS (ultraviolet-visible diffuse reflection spectrum), and FT-IR (Fourier transform infrared spectroscopy). Results showed that CuS and GeO2 had dispersed in this CuS-GeO2-TiO2 composite coating (mass percentages for CuS and GeO2 were 1.23% and 2.79%, respectively). The electrochemical studies (cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and Tafel polarization) of this CuS-GeO2-TiO2 composite coating electrode were performed in pH = 9.51 Na2CO3-NaHCO3 buffer solution containing 0.50 mol/L CH3OH under the conditions of visible light, ultraviolet light (λ = 365 nm), and dark (without light irradiation as control), respectively. Electrochemical studies indicated that this CuS-GeO2-TiO2 composite coating electrode had better photoelectrocatalytic activity than the pure TiO2 electrode in the electrocatalysis of methanol under visible light.

Comparative ultrastructure of vallate, foliate and fungiform taste buds of golden Syrian hamster.

PubMed

Miller, R L; Chaudhry, A P

1976-01-01

A fine-structure study of the hamster fungiform, foliate and vallate taste buds was undertaken for comparative purposes. All three taste bud types shared in common composition of the dark cells, light cells, basal cells, nerve fibers and nerve endings and undifferentiated peripheral cells, but morphological difference existed among them. The foliate and vallate taste buds were quite similar in their ultrastructural morphology. Their dark cells displayed long apical necks, long apical microvilli, apical osmiophilic secretory granules and an abundant rough endoplasmic reticulum. The dark cells of the fungiform taste buds, however, showed no neck formation and lacked apical osmiophilic granules. They had short apical microvilli and relatively scant rough endoplasmic reticulum. There was no difference in the fine structure features of the light cells, basal cells and neural elements of different types of taste buds. Both light and dark cells were much more readily distinguishable in foliate and vallate buds than in fungiform buds at both light-and electron-microscopic levels. Foliate and vallate buds demonstrated homogeneous dense substance within the taste pores while fungiform pores were frequently empty. It is speculated that the differences in taste bud morphology may be due to their different lingual locations and/or may be a reflection of the differences in the inductive influences from different nerves. Furthermore, structural differences may be responsible for varying thresholds to different taste modalities.

Microscopic analysis of "iron spot" on blue-and-white porcelain from Jingdezhen imperial kiln in early Ming dynasty (14th-15th century).

PubMed

Wang, Wenxuan; Zhu, Jian; Jiang, Jianxin; Xu, Changqing; Wu, Shurong; Guan, Li; Zhang, Zhaoxia; Wu, Menglei; Du, Jingnan

2016-11-01

"Sumali," as an imported cobalt ore from overseas, was a sort of precious and valuable pigment used for imperial kilns only, which produces characteristic "iron spot" to blue-and-white porcelain in early Ming Dynasty (A.D. 14th-15th century). Although there were some old studies on it, the morphology and formation of iron spot has not been fully investigated and understood. Therefore, five selected samples with typical spot from Jingdezhen imperial kiln in Ming Yongle periods (A.D. 1403-1424) were analyzed by various microscopic analysis including 3D digital microscope, SEM-EDS and EPMA. According to SEM images, samples can be divided into three groups: un-reflected "iron spot" without crystals, un-reflected "iron spot" with crystals and reflected "iron spot" with crystals. Furthermore, 3D micro-images revealed that "iron spots" separate out dendritic or snow-shaped crystals of iron only on and parallel to the surface of glaze for which "iron spot" show strong metallic luster. Combining with microscopic observation and microanalysis on crystallization and non-crystallization areas, it indicates that firing oxygen concentration is the ultimate causation of forming reflective iron spot which has a shallower distribution below the surface and limits crystals growing down. More details about characters of "iron spot" used "Sumali" were found and provided new clues to coloration, formation mechanism and porcelain producing technology of imperial kiln from 14th to 15th centuries of China. © 2016 Wiley Periodicals, Inc.

Apparatus for and method of correcting for astigmatism in a light beam reflected off of a light reflecting surface

DOEpatents

Sawicki, Richard H.; Sweatt, William

1987-01-01

A technique for adjustably correcting for astigmatism in a light beam is disclosed herein. This technique utilizes first means which defines a flat, rectangular light reflecting surface having opposite reinforced side edges and which is resiliently bendable, to a limited extent, into different concave and/or convex cylindrical curvatures about a particular axis and second means acting on the first means for adjustably bending the light reflecting surface into a particular selected one of the different curvatures depending upon the astigmatism to be corrected for and for fixedly maintaining the curvature selected. In the embodiment disclosed, the light reflecting surface is adjustably bendable into the selected cylindrical curvature by application of a particular bending moment to the reinforced side edges of the light reflecting surface.

Design of a dynamic biofilm imaging cell for white-light interferometric microscopy

DOE PAGES

Larimer, Curtis; Brann, Michelle; Suter, Jonathan D.; ...

2017-05-10

In microbiology research there is a strong need for next generation imaging and sensing instrumentation that will enable minimally invasive and label-free investigation of soft, hydrated structures such as in bacterial biofilms. White light interferometry (WLI) can provide high resolution images of surface topology without the use of fluorescent labels but is not typically used to image biofilms because there is insufficient refractive index contrast to induce reflection from the biofilm’s interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ, especially in a non-destructive manner. In this report, we build onmore » our prior description of static biofilm imaging and describe the design of a dynamic imaging flow cell that enables monitoring the thickness and topology of live biofilms over time using a WLI microscope. The microfluidic system is specifically designed to create a reflective interface on the surface of biofilms while minimizing disruption of fragile structures. The imaging cell was also designed to accommodate limitations imposed by the depth of focus of the microscope’s objective lens. Example images of live biofilm samples are shown in order to illustrate the ability of the flow cell and WLI instrument to 1) support bacterial growth and biofilm development, 2) image biofilm structure that reflects growth in flow conditions, and 3) monitor biofilm development over time non-destructively. In future work, the apparatus described here will enable surface metrology measurements (roughness, surface area, etc.) of biofilms and may be used to observe changes in biofilm structure in response to changes in environmental conditions (e.g., flow velocity, availability of nutrients, and presence of biocides). Furthermore, this development will open new opportunities for the use of WLI in bioimaging.« less

Design of a dynamic biofilm imaging cell for white-light interferometric microscopy

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

Larimer, Curtis; Brann, Michelle; Suter, Jonathan D.

In microbiology research there is a strong need for next generation imaging and sensing instrumentation that will enable minimally invasive and label-free investigation of soft, hydrated structures such as in bacterial biofilms. White light interferometry (WLI) can provide high resolution images of surface topology without the use of fluorescent labels but is not typically used to image biofilms because there is insufficient refractive index contrast to induce reflection from the biofilm’s interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ, especially in a non-destructive manner. In this report, we build onmore » our prior description of static biofilm imaging and describe the design of a dynamic imaging flow cell that enables monitoring the thickness and topology of live biofilms over time using a WLI microscope. The microfluidic system is specifically designed to create a reflective interface on the surface of biofilms while minimizing disruption of fragile structures. The imaging cell was also designed to accommodate limitations imposed by the depth of focus of the microscope’s objective lens. Example images of live biofilm samples are shown in order to illustrate the ability of the flow cell and WLI instrument to 1) support bacterial growth and biofilm development, 2) image biofilm structure that reflects growth in flow conditions, and 3) monitor biofilm development over time non-destructively. In future work, the apparatus described here will enable surface metrology measurements (roughness, surface area, etc.) of biofilms and may be used to observe changes in biofilm structure in response to changes in environmental conditions (e.g., flow velocity, availability of nutrients, and presence of biocides). Furthermore, this development will open new opportunities for the use of WLI in bioimaging.« less

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.

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.

Ultraviolet micro-Raman spectrograph for the detection of small numbers of bacterial cells

NASA Astrophysics Data System (ADS)

Chadha, S.; Nelson, W. H.; Sperry, J. F.

1993-11-01

The construction of a practical UV micro-Raman spectrograph capable of selective excitation of bacterial cells and other microscopic samples has been described. A reflective objective is used to focus cw laser light on a sample and at the same time collect the scattered light at 180°. With the aid of a quartz lens the image produced is focused on the slits of a spectrograph equipped with a single 2400 grooves/mm grating optimized for 250 nm. Spectra were detected by means of a blue-intensified diode array detector. Resonance Raman spectra of Bacillus subtilis and Flavobacterium capsulatum excited by the 257.2 nm output of a cw laser were recorded in the 900-1800 cm-1 region. Bacterial cells were immobilized on a quartz plate by means of polylysine and were counted visually. Cooling was required to retard sample degradation. Sample sizes ranged from 1 to 50 cells with excitation times varying from 15 to 180 s. Excellent spectra have been obtained from 20 cells in 15 s using a spectrograph having only 3% throughput.

Synergistic effect on the visible light activity of Ti3+ doped TiO2 nanorods/boron doped graphene composite

PubMed Central

Xing, Mingyang; Li, Xiao; Zhang, Jinlong

2014-01-01

TiO2/graphene (TiO2-x/GR) composites, which are Ti3+ self-doped TiO2 nanorods decorated on boron doped graphene sheets, were synthesized via a simple one-step hydrothermal method using low-cost NaBH4 as both a reducing agent and a boron dopant on graphene. The resulting TiO2 nanorods were about 200 nm in length with exposed (100) and (010) facets. The samples were characterized by X-ray diffraction (XRD), UV-visible diffuse reflectance spectroscopy, X-band electron paramagnetic resonance (EPR), X-ray photoelectron spectra (XPS), transmission electron microscope (TEM), Raman, and Fourier-transform infrared spectroscopy (FTIR). The XRD results suggest that the prepared samples have an anatase crystalline structure. All of the composites tested exhibited improved photocatalytic activities as measured by the degradation of methylene blue and phenol under visible light irradiation. This improvement was attributed to the synergistic effect of Ti3+ self-doping on TiO2 nanorods and boron doping on graphene. PMID:24974890

Nondestructive and in situ determination of graphene layers using optical fiber Fabry-Perot interference

NASA Astrophysics Data System (ADS)

Li, Cheng; Peng, Xiaobin; Liu, Qianwen; Gan, Xin; Lv, Ruitao; Fan, Shangchun

2017-02-01

Thickness measurement plays an important role for characterizing optomechanical behaviors of graphene. From the view of graphene-based Fabry-Perot (F-P) sensors, a simple, nondestructive and in situ method of determining the thickness of nanothick graphene membranes was demonstrated by using optical fiber F-P interference. Few-layer/multilayer graphene sheets were suspendedly adhered onto the endface of a ferrule with a 125 µm inner diameter by van der Waals interactions to construct micro F-P cavities. Along with the Fresnel’s law and complex index of refraction of the membrane working as a light reflector of an F-P interferometer, the optical reflectivity of graphene was modeled to investigate the effects of light wavelength and temperature. Then the average thickness of graphene membranes were extracted by F-P interference demodulation, and yielded a very strong cross-correlation coefficient of 99.95% with the experimental results observed by Raman spectrum and atomic force microscope. The method could be further extended for determining the number of layers of other 2D materials.

Two-Solvent Method Synthesis of NiO/ZnO Nanoparticles Embedded in Mesoporous SBA-15: Photocatalytic Properties Study.

PubMed

Dai, Peng; Yan, Tao-Tao; Yu, Xin-Xin; Bai, Zhi-Man; Wu, Ming-Zai

2016-12-01

Different loadings of NiO/ZnO nanoparticles embedded in mesoporous silica (SBA-15) were prepared via a two-solvent method with the ordered hexagonal mesoporous structure of SBA-15 kept. X-ray diffraction, transmission electron microscope, X-ray photoelectron spectroscopy, diffusive reflective UV-vis spectroscopy, and N2 adsorption porosimetry were employed to characterize the nanocomposites. The results indicate that the ordered hexagonal mesoporous structure of SBA-15 is kept and the absorption band edges of the nanocomposites shift into the ultraviolet light regime. The photocatalytic activity of our samples for degradation of methylene orange was investigated under UV light irradiation, and the results show that the nanocomposites have higher photodegradation ability toward methylene orange than commercial pure P-25. The photocatalytic activity of the nanocomposites was found to be dependent on both the adsorption ability of the SBA-15 and the photocatalytic activity of NiO-ZnO nanoparticles encapsulated in SBA-15. In addition, there is an optimal loading of NiO-ZnO nanoparticles. Too high or low loading will lower the photodegradation ability of the nanocomposites.

High resolution surface plasmon microscopy for cell imaging

NASA Astrophysics Data System (ADS)

Argoul, F.; Monier, K.; Roland, T.; Elezgaray, J.; Berguiga, L.

2010-04-01

We introduce a new non-labeling high resolution microscopy method for cellular imaging. This method called SSPM (Scanning Surface Plasmon Microscopy) pushes down the resolution limit of surface plasmon resonance imaging (SPRi) to sub-micronic scales. High resolution SPRi is obtained by the surface plasmon lauching with a high numerical aperture objective lens. The advantages of SPPM compared to other high resolution SPRi's rely on three aspects; (i) the interferometric detection of the back reflected light after plasmon excitation, (ii) the twodimensional scanning of the sample for image reconstruction, (iii) the radial polarization of light, enhancing both resolution and sensitivity. This microscope can afford a lateral resolution of - 150 nm in liquid environment and - 200 nm in air. We present in this paper images of IMR90 fibroblasts obtained with SSPM in dried environment. Internal compartments such as nucleus, nucleolus, mitochondria, cellular and nuclear membrane can be recognized without labelling. We propose an interpretation of the ability of SSPM to reveal high index contrast zones by a local decomposition of the V (Z) function describing the response of the SSPM.

Superluminescence from an optically pumped molecular tunneling junction by injection of plasmon induced hot electrons

PubMed Central

Braun, Kai; Wang, Xiao; Kern, Andreas M; Adler, Hilmar; Peisert, Heiko; Chassé, Thomas; Zhang, Dai

2015-01-01

Summary Here, we demonstrate a bias-driven superluminescent point light-source based on an optically pumped molecular junction (gold substrate/self-assembled molecular monolayer/gold tip) of a scanning tunneling microscope, operating at ambient conditions and providing almost three orders of magnitude higher electron-to-photon conversion efficiency than electroluminescence induced by inelastic tunneling without optical pumping. A positive, steadily increasing bias voltage induces a step-like rise of the Stokes shifted optical signal emitted from the junction. This emission is strongly attenuated by reversing the applied bias voltage. At high bias voltage, the emission intensity depends non-linearly on the optical pump power. The enhanced emission can be modelled by rate equations taking into account hole injection from the tip (anode) into the highest occupied orbital of the closest substrate-bound molecule (lower level) and radiative recombination with an electron from above the Fermi level (upper level), hence feeding photons back by stimulated emission resonant with the gap mode. The system reflects many essential features of a superluminescent light emitting diode. PMID:26171286

Synthesis of N-doped potassium tantalate perovskite material for environmental applications

NASA Astrophysics Data System (ADS)

Rao, Martha Purnachander; Nandhini, Vellangattupalayam Ponnusamy; Wu, Jerry J.; Syed, Asad; Ameen, Fuad; Anandan, Sambandam

2018-02-01

Nitrogen containing potassium tantalate perovskite material has been synthesized by the solvothermal method using urea (CH4N2O) as a nitrogen source. The as-prepared sample was characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), diffuse reflectance spectroscopy (DRS), scanning electron microscope (SEM), and energy-dispersive X-ray spectroscopy (EDX) and X-ray photoelectron spectroscopy (XPS). The particle size of nitrogen containing KTaO3 observed from SEM images was found to be 100-150 nm. Doping KTaO3 with nitrogen causes reduction of band gap from 3.5 to 2.54 eV. The incorporation of Nitrogen into the crystal lattice of KTaO3 not only extended the absorption of light from UV (ultraviolet) region to visible region and also enhanced the photocatalytic activity. As prepared nitrogen containing KTaO3 samples exhibit cubic-like morphology and noticed efficient photocatalytic activity towards methylene blue dye degradation under visible light illumination. The intermediates formed during photodegradation were identified by mass spectrometry (GC-MS) and proposed suitable degradation pathway.

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.

Comparison between reflectance confocal microscopy and two-photon microscopy in early detection of cutaneous radiation injury in a mouse model in-vivo.

PubMed

Jang, Won Hyuk; Kwon, Soonjae; Shim, Sehwan; Jang, Won-Suk; Myung, Jae Kyung; Yang, Sejung; Park, Sunhoo; Kim, Ki Hean

2018-05-12

Cutaneous radiation injury (CRI) is a skin injury caused by high dose exposure of ionizing radiation (IR). For proper treatment, early detection of CRI before clinical symptoms is important. Optical microscopic techniques such as reflectance confocal microscopy (RCM) and two-photon microscopy (TPM) have been tested as the early diagnosis method by detecting cellular changes. In this study, RCM and TPM were compared in the detection of cellular changes caused by CRI in an in-vivo mouse model. CRI was induced on the mouse hindlimb skin with various IR doses and the injured skin regions were imaged longitudinally by both modalities until the onset of clinical symptoms. Both RCM and TPM detected the changes of epidermal cells and sebaceous glands before clinical symptoms in different optical contrasts. RCM detected changes of cell morphology and scattering property based on light reflection. TPM detected detail changes of cellular structures based on autofluorescence of cells. Since both RCM and TPM were sensitive to the early-stage CRI by using different contrasts, the optimal method for clinical CRI diagnosis could be either individual methods or their combination. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Chemical imaging of structured SAMs with a novel SFG microscope

NASA Astrophysics Data System (ADS)

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

2002-11-01

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

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.

Real-time, non-invasive microscopic confirmation of clinical diagnosis of bullous pemphigoid using in vivo reflectance confocal microscopy.

PubMed

Ardigò, M; Agozzino, M; Amorosi, B; Moscarella, E; Cota, C; de Abreu, L; Berardesca, E

2014-05-01

Bullous pemphigoid is an autoimmune disease affecting prevalently the elder. In vivo reflectance confocal microscopy is a non-invasive technique for real-time imaging of the skin with cellular-level resolution. No previous data has been reported about confocal microscopy of bullous pemphigoid. Aim of this preliminary study is the evaluation of the potential of in vivo reflectance confocal microscopy for real-time, microscopical confirmation of clinical bullous pemphigoid diagnosis. A total of nine lesions from patients affected by pemphigoid underwent in vivo reflectance confocal microscopy before histological examination. In our preliminary study, confocal microscopy showed high grade of correspondence to histopathology. In particular, presence of sub-epidermal cleft and variable amount of oedema of the upper dermis associated with inflammatory cells infiltration were seen as prevalent confocal features in the bullous lesions considered. Differently, in urticarial lesions, no specific features could be appreciated at confocal analysis beside the presence of signs of spongiosis and perivascular inflammation. Confocal microscopy seems to be useful for in vivo, microscopical confirmation of the clinical suspect of bullous pemphigoid and for biopsy site selection in urticarial lesions to obtain a more significant specimen for histopathological examination. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

The approach to reflection x-ray microscopy below the critical angles

NASA Astrophysics Data System (ADS)

Artyukov, Igor A.; Busarov, Alexander; Popov, Nikolay L.; Vinogradov, Alexander V.

2017-05-01

There is a quest for new knowledge and methods to study various materials and processes on surfaces and interfaces at the nanoscale. It concerns ablation, phase transitions, physical and chemical transformations, dissolution, selforganization etc. Obviously, to achieve an appropriate resolution it is necessary to use a corresponding wavelength . Higher resolution can be obtained with shorter wavelengths. On the other hand, in surface modification, ablation, study of buried interfaces etc. the penetration length of radiation into the materials, which depends on the wavelength and angle of incidence, plays important role... Considering these factors the experimental studies in nano-physics and nanotechnology are usually carried out using X-ray radiation with a photon energy of 0.1-10 keV. As far as surfaces and films are investigated, it is reasonable to use an X-ray microscope operating in the reflection mode. However, in this spectral range a substantial portion of the radiation is reflected only at small grazing angles (e.g. <= 10°). Thus, the idea of grazing incidence reflection-mode X-ray microscope has been developed. In this paper, we consider one of possible schemes of such an X-ray microscope. Our analysis and simulation is based on the extension of the Fresnel propagation theory to tilted object problems.

Relationship between light scattering and absorption due to cytochrome c oxidase reduction during loss of tissue viability in brains of rats

NASA Astrophysics Data System (ADS)

Kawauchi, Satoko; Sato, Shunichi; Ooigawa, Hidetoshi; Nawashiro, Hiroshi; Ishihara, Miya; Kikuchi, Makoto

2008-02-01

We performed simultaneous measurement of light scattering and absorption due to reduction of cytochrome c oxidase as intrinsic optical signals that are related to morphological characteristics and energy metabolism, respectively, for rat brains after oxygen/glucose deprivation by saline infusion. To detect change in light scattering, we determined the wavelength that was the most insensitive to change in light absorption due to the reduction of cytochrome c oxidase on the basis of multiwavelength analysis of diffuse reflectance data set for each rat. Then the relationships between scattering signal and absorption signals related to the reductions of heme aa 3 (605 nm) and CuA (830 nm) in cytochrome c oxidase were examined. Measurements showed that after starting saline infusion, the reduction of heme aa 3 started first; thereafter triphasic, large scattering change occurred (200-300 s), during which the reduction of CuA started. Despite such complex behaviors of IOSs, almost linear correlations were seen between the scattering signal and the heme aa 3-related absorption signal, while a relatively large animal-to-animal variation was observed in the correlation between the scattering signal and CuA-related absorption signal. Transmission electron microscopic observation revealed that dendritic swelling and mitochondrial deformation occurred in the cortical surface tissue after the triphasic scattering change. These results suggest that mitochondrial energy failure accompanies morphological alteration in the brain tissue and results in change in light scattering; light scattering will become an important indicator of tissue viability in brain.

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

Gofron, K. J., E-mail: kgofron@bnl.gov; Cai, Y. Q.; Coburn, D. S.

A novel on-axis X-ray microscope with 3 µm resolution, 3x magnification, and a working distance of 600 mm for in-situ sample alignment and X-ray beam visualization for the Inelastic X-ray Scattering (IXS) beamline at NSLS-II is presented. The microscope uses reflective optics, which minimizes dispersion, and allows imaging from Ultraviolet (UV) to Infrared (IR) with specifically chosen objective components (coatings, etc.). Additionally, a portable high resolution X-ray microscope for KB mirror alignment and X-ray beam characterization was developed.

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.

Phytochrome-Mediated Detection of Changes in Reflected Light

PubMed Central

Mancinelli, Alberto L.

1991-01-01

Measurements of phytochrome photoequilibria and photoconversion rates in vivo, in seedlings of Cucurbita pepo L. exposed to light in growth chambers, indicate that significant changes in the state of phytochrome can be brought about by changes in the quality and quantity of the light reflected from the walls of the growth chambers. The changes in reflected light, although large, were small in terms of the total radiation (direct light from the lamps plus wall-reflected light) to which the seedlings were exposed. The conditions used were approximate simulations of direct and reflected sunlight conditions in the natural environment. Keeping in mind the limitations imposed by the approximation of the simulations, the results from this study are consistent with the hypothesis that, in the natural environment, a plant might be capable of detecting the presence of nearby plants, before being shaded by them, through the phytochrome-mediated perception of changes in reflected light. PMID:16667942

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.

Enhanced photocatalytic efficiency of NiS/TiO{sub 2} composite catalysts using sunset yellow, an azo dye under day light illumination

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

Rajamanickam, D.; Dhatshanamurthi, P.; Shanthi, M., E-mail: shanthimsm@gmail.com

2015-01-15

Highlights: • NiS/TiO{sub 2} was successfully synthesized by sol–gel method. • This new method of preparation gives a homogeneous dispersion of NiS on TiO{sub 2}. • Degradation activity of NiS/TiO{sub 2} is found to be more efficient than other catalysts. • Addition of oxidants enhances the degradation efficiency significantly. • COD measurements reveal the complete mineralization of dye molecules. • The catalyst is found to be reusable. - Abstract: To improve the solar light induced photocatalytic application performances of TiO{sub 2}, in this study, the NiS modified TiO{sub 2} composite photocatalysts with various ratios of NiS to TiO{sub 2} weremore » prepared by sol–gel method. The catalyst was characterized by X-ray diffraction (XRD), high resolution scanning electron microscope (HR-SEM), high resolution transmission electron microscope (HR-TEM), energy dispersive spectra (EDS), diffuse reflectance spectra (DRS), photoluminescence spectra (PL), X-ray photoelectron spectroscopy (XPS), and Brunauer–Emmett–Teller (B–E–T) surface area measurement methods. The photocatalytic activity of NiS/TiO{sub 2} was investigated for the degradation of sunset yellow (SY) in aqueous solution using solar light. The NiS/TiO{sub 2} is found to be more efficient than prepared TiO{sub 2} and TiO{sub 2}–P25 at pH 7 for the mineralization of SY. The effects of operational parameters such as the amount of photocatalyst, dye concentration and initial pH on photo mineralization of SY have been analyzed. The degradation was strongly enhanced in the presence of oxidants such as H{sub 3}K{sub 5}O{sub 18}S{sub 4} (Oxone), KIO{sub 4}, and KBrO{sub 3}. The mineralization of SY has been identified by COD measurements. The catalyst is found to be reusable.« less

Low frequency acoustic microscope

DOEpatents

Khuri-Yakub, Butrus T.

1986-11-04

A scanning acoustic microscope is disclosed for the detection and location of near surface flaws, inclusions or voids in a solid sample material. A focused beam of acoustic energy is directed at the sample with its focal plane at the subsurface flaw, inclusion or void location. The sample is scanned with the beam. Detected acoustic energy specularly reflected and mode converted at the surface of the sample and acoustic energy reflected by subsurface flaws, inclusions or voids at the focal plane are used for generating an interference signal which is processed and forms a signal indicative of the subsurface flaws, inclusions or voids.

Comparative Photoluminescence Properties and Judd-Ofelt Analysis of Eu3+ Ion-Activated Metal Molybdate Phosphors A2MoO6:Eu3+ (A = La, Y, Gd and Bi)

NASA Astrophysics Data System (ADS)

Han, Bing; Liu, Bingkun; Zhang, Jie; Li, Pengju; Shi, Hengzhen

2017-07-01

A class of red-emitting Eu3+ ion-activated metal molybdate A2MoO6:Eu3+ (A = La, Y, Gd and Bi) phosphors were synthesized by a conventional high-temperature solid-state reaction method. The x-ray diffraction patterns, scanning electron microscope images, Fourier transform infrared spectra, ultraviolet-visible diffuse reflection spectra as well as photoluminescence properties were measured to characterize the as-prepared samples. The photoluminescence properties including excitation/emission spectra, decay curves, Commission Internationale de L'Eclairage chromaticity coordinates and quantum efficiency were comparatively investigated in detail. The Judd-Ofelt theory was also applied to understand the radiative properties of f-f transitions of Eu3+ ions in this system for the first time. The as-prepared phosphors can be effectively excited with near-ultraviolet and/or blue light, and exhibit red emission belonging to the prevailing 5D0 → 7F2 transitions of Eu3+ with short decay time (millisecond level). The results demonstrated that A2MoO6:Eu3+ (A = La, Y, Gd and Bi) phosphors could have potential application as red-emitting phosphors in white light-emitting diodes based on near-ultraviolet and/or blue light-emitting diode chips.

A Comparative Study of Microscopic Images Captured by a Box Type Digital Camera Versus a Standard Microscopic Photography Camera Unit

PubMed Central

Desai, Nandini J.; Gupta, B. D.; Patel, Pratik Narendrabhai

2014-01-01

Introduction: Obtaining images of slides viewed by a microscope can be invaluable for both diagnosis and teaching.They can be transferred among technologically-advanced hospitals for further consultation and evaluation. But a standard microscopic photography camera unit (MPCU)(MIPS-Microscopic Image projection System) is costly and not available in resource poor settings. The aim of our endeavour was to find a comparable and cheaper alternative method for photomicrography. Materials and Methods: We used a NIKON Coolpix S6150 camera (box type digital camera) with Olympus CH20i microscope and a fluorescent microscope for the purpose of this study. Results: We got comparable results for capturing images of light microscopy, but the results were not as satisfactory for fluorescent microscopy. Conclusion: A box type digital camera is a comparable, less expensive and convenient alternative to microscopic photography camera unit. PMID:25478350

Five years of experience teaching pathology to dental students using the WebMicroscope

PubMed Central

2011-01-01

Background We describe development and evaluation of the user-friendly web based virtual microscopy - WebMicroscope for teaching and learning dental students basic and oral pathology. Traditional students microscopes were replaced by computer workstations. Methods The transition of the basic and oral pathology courses from light to virtual microscopy has been completed gradually over a five-year period. A pilot study was conducted in academic year 2005/2006 to estimate the feasibility of integrating virtual microscopy into a traditional light microscopy-based pathology course. The entire training set of glass slides was subsequently converted to virtual slides and placed on the WebMicroscope server. Giving access to fully digitized slides on the web with a browser and a viewer plug-in, the computer has become a perfect companion of the student. Results The study material consists now of over 400 fully digitized slides which covering 15 entities in basic and systemic pathology and 15 entities in oral pathology. Digitized slides are linked with still macro- and microscopic images, organized with clinical information into virtual cases and supplemented with text files, syllabus, PowerPoint presentations and animations on the web, serving additionally as material for individual studies. After their examinations, the students rated the use of the software, quality of the images, the ease of handling the images, and the effective use of virtual slides during the laboratory practicals. Responses were evaluated on a standardized scale. Because of the positive opinions and support from the students, the satisfaction surveys had shown a progressive improvement over the past 5 years. The WebMicroscope as a didactic tool for laboratory practicals was rated over 8 on a 1-10 scale for basic and systemic pathology and 9/10 for oral pathology especially as various students’ suggestions were implemented. Overall, the quality of the images was rated as very good. Conclusions An overwhelming majority of our students regarded a possibility of using virtual slides at their convenience as highly desirable. Our students and faculty consider the use of the virtual microscope for the study of basic as well as oral pathology as a significant improvement over the light microscope. PMID:21489183

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.

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.

Transmitting and reflecting diffuser. [for ultraviolet light

NASA Technical Reports Server (NTRS)

Keafer, L. S., Jr.; Burcher, E. E.; Kopia, L. P. (Inventor)

1973-01-01

A near-Lambertian diffuser is described which transmits and reflects ultraviolet light. An ultraviolet grade fused silica substrate is coated with vaporized fuse silica. The coating thickness is controlled, one thickness causing ultraviolet light to diffuse and another thickness causing ultraviolet light to reflect a near Lambertian pattern.

Application of a reflective microscope objective for multiphoton microscopy.

PubMed

Kabir, Mohammad M; Choubal, Aakash M; Toussaint, Kimani C

2018-04-20

Reflective objectives (ROs) mitigate chromatic aberration across a broad wavelength range. Yet, a systematic performance characterisation of ROs has not been done. In this paper, we compare the performance of a 0.5 numerical-aperture (NA) reflective objective (RO) with a 0.55 NA standard glass objective (SO), using two-photon fluorescence (TPF) and second-harmonic generation (SHG). For experiments spanning ∼1 octave in the visible and NIR wavelengths, the SO leads to defocusing errors of 25-40% for TPF images of subdiffraction fluorescent beads and 10-12% for SHG images of collagen fibres. The corresponding error for the RO is ∼4% for both imaging modalities. This work emphasises the potential utility of ROs for multimodal multiphoton microscopy applications. © 2018 The Authors Journal of Microscopy © 2018 Royal Microscopical Society.

Light reflecting apparatus including a multi-aberration light reflecting surface

DOEpatents

Sawicki, Richard H.; Sweatt, William

1987-01-01

A light reflecting apparatus including a multi-aberration bendable light reflecting surface is disclosed herein. This apparatus includes a structural assembly comprised of a rectangular plate which is resiliently bendable, to a limited extent, and which has a front side defining the multi-aberration light reflecting surface and an opposite back side, and a plurality of straight leg members rigidly connected with the back side of the plate and extending rearwardly therefrom. The apparatus also includes a number of different adjustment mechanisms, each of which is connected with specific ones of the leg members. These mechanisms are adjustably movable in different ways for applying corresponding forces to the leg members in order to bend the rectangular plate and light reflecting surface into different predetermined curvatures and which specifically include quadratic and cubic curvatures corresponding to different optical aberrations.

A light reflecting apparatus including a multi-aberration light reflecting surface

DOEpatents

Sawicki, R.H.; Sweatt, W.

1985-11-21

A light reflecting apparatus including a multi-aberration bendable light reflecting surface is disclosed herein. This apparatus includes a structural assembly comprised of a rectangular plate which is resiliently bendable, to a limited extent, and which has a front side defining the multi-aberration light reflecting surface and an opposite back side, and a plurality of straight leg members rigidly connected with the back side of the plate and extending rearwardly therefrom. The apparatus also includes a number of different adjustment mechanisms, each of which is connected with specific ones of the leg members. These mechanisms are adjustably movable in different ways for applying corresponding forces to the leg members in order to bend the rectangular plate and light reflecting surface into different predetermined curvatures and which specifically include quadratic and cubic curvatures corresponding to different optical aberrations.

Electron and Light Microscopy Techniques Suitable for Studying Fatigue Damage in a Crystallized Glass Ceramic

NASA Technical Reports Server (NTRS)

Harrell, Shelley; Zaretsky, Erwin V.

1961-01-01

The crystals of Pyroceram are randomly oriented and highly reflective so that standard microscopy techniques are not satisfactory for studying this material. Standard replicating procedures proved difficult to use. New microscopy techniques and procedures have therefore been developed. A method for locating, orienting, and identifying specific areas to be viewed with an electron microscope is described. This method not require any special equipment. Plastic replicas were found to be unsatisfactory because of their tendency to adhere to Pryoceram. This caused them to tear when released or resulted in artifacts. Preshadowed silicon monoxide replicas were satisfactory but required a releasing agent. A method of depositing the releasing agent is described. To polish specimens without evidence of fire-polishing, it was found necessary to use a vibratory polishing technique. Chrome oxide was used as the abrasive and either water or kerosene as the lubricant. Vibratory polishing is extremely slow, but surfaces so polished show no evidence of fire polishing, even when examined by electron microscopy. The most satisfactory etching process used for Pyroceram 9608 consisted of a primary etch of 5 milliliters of hydrochloric acid (concentrated), 5 milliliters of hydrogen fluoride (45 percent), and 45 milliliters of water, and a secondary etch with methyl alcohol replacing the water. Best results were obtained with total etching times from 25 to 30 seconds. Staining of the Pyroceram surface with a Sanford's marker was found to be an expedient way to reduce the glare of reflected light.

Polymer-cholesteric liquid-crystalline composites with a broad light reflection band

NASA Astrophysics Data System (ADS)

Mitov, Michel

2016-05-01

Cholesteric liquid crystals selectively reflect the light. The reflection bandgap is typically limited to 100 nm in the visible spectrum and, at the best, 50% of the unpolarized incident light is reflected. Solutions are found in biopolymers and polymer-liquid crystal composite materials to go beyond these limits.

Optics: Light, Color, and Their Uses. An Educator's Guide with Activities in Science and Mathematics.

ERIC Educational Resources Information Center

National Aeronautics and Space Administration, Huntsville, AL. George C. Marshall Space Flight Center.

This educator's guide from discusses optics, light, color and their uses. Activities include: (1) "Reflection of Light with a Plane (Flat) Mirror--Trace a Star"; (2) "Reflection of Light with Two Plane Mirrors--Double Mirrors Placed at a 90-Degree Angle"; (3) "Reflection of Light with Two Plane Mirrors--Double Mirrors Placed at a Number of…

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…

Penetration of immunoreagents in Vibratome-sectioned brain: a light and electron microscopic study.

PubMed

Piekut, D T; Casey, S M

1983-05-01

Immunocytochemical studies on the localization of peptides at the ultrastructural level have most frequently involved the application of the peroxidase--antiperoxidase (PAP) method of immunocytochemistry and the use of the preembedding or postembedding staining procedures. The present study was designed to determine the depth of penetration of Vibratome tissue sections by immunoreagents used in the preembedding method in which immunostaining of unembedded fixed tissue sections is accomplished prior to tissue dehydration and embedment. Our data indicate that penetration of immunoreagents is restricted to the superficial 8-9 micrometers of a 80-micrometers thick Vibratome tissue section of hypothalamus of brain using antisera generated against arginine vasopressin. The final immunoreaction product visualized in a Vibratome tissue section may reflect only a fraction of the amount of hormone contained within the thickness of the tissue section.

Detection of Human Ig G Using Photoluminescent Porous Silicon Interferometer.

PubMed

Cho, Bomin; Kim, Seongwoong; Woo, Hee-Gweon; Kim, Sungsoo; Sohn, Honglae

2015-02-01

Photoluminescent porous silicon (PSi) interferometers having dual optical properties, both Fabry-Pérot fringe and photolumincence (PL), have been developed and used as biosensors for detection of Human Immunoglobin G (Ig G). PSi samples were prepared by electrochemical etching of p-type silicon under white light exposure. The surface of PSi was characterized using a cold field emission scanning electron microscope. The sensor system studied consisted of a single layer of porous silicon modified with Protein A. The system was probed with various fragments of aqueous human immunoglobin G (Ig G) analyte. Both reflectivity and PL were simultaneously measured under the exposure of human Ig G. An increase of optical thickness and decrease of PL were obtained under the exposure of human Ig G. Detection limit of 500 fM was observed for the human Ig G.

Label-free and live cell imaging by interferometric scattering microscopy.

PubMed

Park, Jin-Sung; Lee, Il-Buem; Moon, Hyeon-Min; Joo, Jong-Hyeon; Kim, Kyoung-Hoon; Hong, Seok-Cheol; Cho, Minhaeng

2018-03-14

Despite recent remarkable advances in microscopic techniques, it still remains very challenging to directly observe the complex structure of cytoplasmic organelles in live cells without a fluorescent label. Here we report label-free and live-cell imaging of mammalian cell, Escherischia coli , and yeast, using interferometric scattering microscopy, which reveals the underlying structures of a variety of cytoplasmic organelles as well as the underside structure of the cells. The contact areas of the cells attached onto a glass substrate, e.g. , focal adhesions and filopodia, are clearly discernible. We also found a variety of fringe-like features in the cytoplasmic area, which may reflect the folded structures of cytoplasmic organelles. We thus anticipate that the label-free interferometric scattering microscopy can be used as a powerful tool to shed interferometric light on in vivo structures and dynamics of various intracellular phenomena.

Fourier Transform Infrared Spectroscopy Part III. Applications.

ERIC Educational Resources Information Center

Perkins, W. D.

1987-01-01

Discusses the use of the FT-IR spectrometer in analyses that were previously avoided. Examines some of the applications of this spectroscopy with aqueous solutions, circular internal reflection, samples with low transmission, diffuse reflectance, infrared emission, and the infrared microscope. (TW)

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.

Analytic reflected light curves for exoplanets

NASA Astrophysics Data System (ADS)

Haggard, Hal M.; Cowan, Nicolas B.

2018-07-01

The disc-integrated reflected brightness of an exoplanet changes as a function of time due to orbital and rotational motions coupled with an inhomogeneous albedo map. We have previously derived analytic reflected light curves for spherical harmonic albedo maps in the special case of a synchronously rotating planet on an edge-on orbit (Cowan, Fuentes & Haggard). In this paper, we present analytic reflected light curves for the general case of a planet on an inclined orbit, with arbitrary spin period and non-zero obliquity. We do so for two different albedo basis maps: bright points (δ-maps), and spherical harmonics (Y_ l^m-maps). In particular, we use Wigner D-matrices to express an harmonic light curve for an arbitrary viewing geometry as a non-linear combination of harmonic light curves for the simpler edge-on, synchronously rotating geometry. These solutions will enable future exploration of the degeneracies and information content of reflected light curves, as well as fast calculation of light curves for mapping exoplanets based on time-resolved photometry. To these ends, we make available Exoplanet Analytic Reflected Lightcurves, a simple open-source code that allows rapid computation of reflected light curves.

Spectroscopy of reflection-asymmetric nuclei with relativistic energy density functionals

NASA Astrophysics Data System (ADS)

Xia, S. Y.; Tao, H.; Lu, Y.; Li, Z. P.; Nikšić, T.; Vretenar, D.

2017-11-01

Quadrupole and octupole deformation energy surfaces, low-energy excitation spectra, and transition rates in 14 isotopic chains: Xe, Ba, Ce, Nd, Sm, Gd, Rn, Ra, Th, U, Pu, Cm, Cf, and Fm, are systematically analyzed using a theoretical framework based on a quadrupole-octupole collective Hamiltonian (QOCH), with parameters determined by constrained reflection-asymmetric and axially symmetric relativistic mean-field calculations. The microscopic QOCH model based on the PC-PK1 energy density functional and δ -interaction pairing is shown to accurately describe the empirical trend of low-energy quadrupole and octupole collective states, and predicted spectroscopic properties are consistent with recent microscopic calculations based on both relativistic and nonrelativistic energy density functionals. Low-energy negative-parity bands, average octupole deformations, and transition rates show evidence for octupole collectivity in both mass regions, for which a microscopic mechanism is discussed in terms of evolution of single-nucleon orbitals with deformation.

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.

Scanning computed confocal imager

DOEpatents

George, John S.

2000-03-14

There is provided a confocal imager comprising a light source emitting a light, with a light modulator in optical communication with the light source for varying the spatial and temporal pattern of the light. A beam splitter receives the scanned light and direct the scanned light onto a target and pass light reflected from the target to a video capturing device for receiving the reflected light and transferring a digital image of the reflected light to a computer for creating a virtual aperture and outputting the digital image. In a transmissive mode of operation the invention omits the beam splitter means and captures light passed through the target.

Comparisons of Modeled and Observed Reflectivities and Fall Speeds for Snowfall of Varied Riming Degree During Winter Storms on Long Island, New York

NASA Technical Reports Server (NTRS)

Molthan, Andrew L.; Colle, Brian A.; Yuter, Sandra E.; Stark, David

2016-01-01

Derived radar reflectivity and fall speed for four Weather Research and Forecasting model bulk microphysical parameterizations (BMPs) run at 1.33 km grid spacing are compared with ground-based, vertically-pointing Ku-band radar, scanning S- band radar, and in situ measurements at Stony Brook, NY. Simulations were partitioned into periods of observed riming degree as determined manually using a stereo microscope and camera during nine winter storms. Simulations were examined to determine whether the selected BMPs captured the effects of varying riming intensities, provided a reasonable match to the vertical structure of radar reflectivity or fall speed, and whether they produced reasonable surface fall speed distributions. Schemes assuming non spherical mass-diameter relationships yielded reflectivity distributions closer to observed values. All four schemes examined in this study provided a better match to the observed, vertical structure of reflectivity during moderate riming than light riming periods. The comparison of observed and simulated snow fall speeds had mixed results. One BMP produced episodes of excessive cloud water at times, resulting in fall speeds that were too large. However, most schemes had frequent periods of little or no cloud water during moderate riming periods and thus underpredicted the snow fall speeds at lower levels. Short, 1-4 hour periods with relatively steady snow conditions were used to compare BMP and observed size and fall speed distributions. These limited data suggest the examined BMPs underpredict fall speeds of cold-type snow habits and underrepresent aggregates larger than 4 mm diameter.

Tandem resonator reflectance modulator

DOEpatents

Fritz, I.J.; Wendt, J.R.

1994-09-06

A wide band optical modulator is grown on a substrate as tandem Fabry-Perot resonators including three mirrors spaced by two cavities. The absorption of one cavity is changed relative to the absorption of the other cavity by an applied electric field, to cause a change in total reflected light, as light reflecting from the outer mirrors is in phase and light reflecting from the inner mirror is out of phase with light from the outer mirrors. 8 figs.

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

Effective light coupling in reflective fiber optic distance sensors using a double-clad fiber

NASA Astrophysics Data System (ADS)

Werzinger, Stefan; Härteis, Lisa; Köhler, Aaron; Engelbrecht, Rainer; Schmauss, Bernhard

2017-04-01

Many fiber optic distance sensors use a reflective configuration, where a light beam is launched from an optical fiber, reflected from a target and coupled back into the fiber. While singlemode fibers (SMF) provide low-loss, high-performance components and a well-defined output beam, the coupling of the reflected light into the SMF is very sensitive to mechanical misalignments and scattering at the reflecting target. In this paper we use a double-clad fiber (DCF) and a DCF coupler to obtain an enhanced multimodal coupling of reflected light into the fiber. Increased power levels and robustness are achieved compared to a pure SMF configuration.

Two-Dimensional Standing Wave Total Internal Reflection Fluorescence Microscopy: Superresolution Imaging of Single Molecular and Biological Specimens

PubMed Central

Chung, Euiheon; Kim, Daekeun; Cui, Yan; Kim, Yang-Hyo; So, Peter T. C.

2007-01-01

The development of high resolution, high speed imaging techniques allows the study of dynamical processes in biological systems. Lateral resolution improvement of up to a factor of 2 has been achieved using structured illumination. In a total internal reflection fluorescence microscope, an evanescence excitation field is formed as light is total internally reflected at an interface between a high and a low index medium. The <100 nm penetration depth of evanescence field ensures a thin excitation region resulting in low background fluorescence. We present even higher resolution wide-field biological imaging by use of standing wave total internal reflection fluorescence (SW-TIRF). Evanescent standing wave (SW) illumination is used to generate a sinusoidal high spatial frequency fringe pattern on specimen for lateral resolution enhancement. To prevent thermal drift of the SW, novel detection and estimation of the SW phase with real-time feedback control is devised for the stabilization and control of the fringe phase. SW-TIRF is a wide-field superresolution technique with resolution better than a fifth of emission wavelength or ∼100 nm lateral resolution. We demonstrate the performance of the SW-TIRF microscopy using one- and two-directional SW illumination with a biological sample of cellular actin cytoskeleton of mouse fibroblast cells as well as single semiconductor nanocrystal molecules. The results confirm the superior resolution of SW-TIRF in addition to the merit of a high signal/background ratio from TIRF microscopy. PMID:17483188

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.

Smart Optical Material Characterization System and Method

NASA Technical Reports Server (NTRS)

Choi, Sang Hyouk (Inventor); Park, Yeonjoon (Inventor)

2015-01-01

Disclosed is a system and method for characterizing optical materials, using steps and equipment for generating a coherent laser light, filtering the light to remove high order spatial components, collecting the filtered light and forming a parallel light beam, splitting the parallel beam into a first direction and a second direction wherein the parallel beam travelling in the second direction travels toward the material sample so that the parallel beam passes through the sample, applying various physical quantities to the sample, reflecting the beam travelling in the first direction to produce a first reflected beam, reflecting the beam that passes through the sample to produce a second reflected beam that travels back through the sample, combining the second reflected beam after it travels back though the sample with the first reflected beam, sensing the light beam produced by combining the first and second reflected beams, and processing the sensed beam to determine sample characteristics and properties.

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.

Reflecting microscope system with a 0.99 numerical aperture designed for three-dimensional fluorescence imaging of individual molecules at cryogenic temperatures

PubMed Central

Inagawa, H.; Toratani, Y.; Motohashi, K.; Nakamura, I.; Matsushita, M.; Fujiyoshi, S.

2015-01-01

We have developed a cryogenic fluorescence microscope system, the core of which is a reflecting objective that consists of spherical and aspherical mirrors. The use of an aspherical mirror allows the reflecting objective to have a numerical aperture (NA) of up to 0.99, which is close to the maximum possible NA of 1.03 in superfluid helium. The performance of the system at a temperature of 1.7 K was tested by recording a three-dimensional fluorescence image of individual quantum dots using excitation wavelengths (λex) of 532 nm and 635 nm. At 1.7 K, the microscope worked with achromatic and nearly diffraction-limited performance. The 1/e2 radius (Γ) of the point spread function of the reflecting objective in the lateral (xy) direction was 0.212 ± 0.008 μm at λex = 532 nm and was less than 1.2 times the simulated value for a perfectly polished objective. The radius Γ in the axial (z) direction was 0.91 ± 0.04 μm at λex = 532 nm and was less than 1.4 times the simulated value of Γ. The chromatic aberrations between the two wavelengths were one order of magnitude smaller than Γ in each direction. PMID:26239746

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.

Fabrication, characterization, and photocatalytic property of α-Fe2O3/graphene oxide composite

NASA Astrophysics Data System (ADS)

Li, Hong; Zhao, Qidong; Li, Xinyong; Zhu, Zhengru; Tade, Moses; Liu, Shaomin

2013-06-01

Spindle-shaped microstructure of α-Fe2O3 was successfully synthesized by a simple hydrothermal method. The α-Fe2O3/graphene oxide (GO) composites was prepared using a modified Hummers' strategy. The properties of the samples were systematically investigated by X-ray powder diffraction (XRD), UV-Vis diffuse reflectance spectrophotometer, transmission electron microscope, atomic force microscope, X-ray photoelectron spectroscopy, and Raman spectroscopy (Raman) techniques. GO nanosheets act as supporting materials for anchoring the α-Fe2O3 particles. The average crystallite sizes of the α-Fe2O3 and α-Fe2O3/GO samples are ca. 27 and 24 nm, respectively. The possible growth of α-Fe2O3 onto GO layers led to a higher absorbance capacity for visible light by α-Fe2O3/GO than α-Fe2O3 composite. The photocatalytic degradation of toluene over the α-Fe2O3 and α-Fe2O3/GO samples under xenon-lamp irradiation was comparatively studied by in situ FTIR technique. The results indicate that the α-Fe2O3/GO sample synthesized exhibited a higher capacity for the degradation of toluene. The composite of α-Fe2O3/GO could be promisingly applied in photo-driven air purification.

A near-field scanning microwave microscope for characterization of inhomogeneous photovoltaics.

PubMed

Weber, J C; Schlager, J B; Sanford, N A; Imtiaz, A; Wallis, T M; Mansfield, L M; Coakley, K J; Bertness, K A; Kabos, P; Bright, V M

2012-08-01

We present a near-field scanning microwave microscope (NSMM) that has been configured for imaging photovoltaic samples. Our system incorporates a Pt-Ir tip inserted into an open-ended coaxial cable to form a weakly coupled resonator, allowing the microwave reflection S(11) signal to be measured across a sample over a frequency range of 1 GHz - 5 GHz. A phase-tuning circuit increased impedance-measurement sensitivity by allowing for tuning of the S(11) minimum down to -78 dBm. A bias-T and preamplifier enabled simultaneous, non-contact measurement of the DC tip-sample current, and a tuning fork feedback system provided simultaneous topographic data. Light-free tuning fork feedback provided characterization of photovoltaic samples both in the dark and under illumination at 405 nm. NSMM measurements were obtained on an inhomogeneous, third-generation Cu(In,Ga)Se(2) (CIGS) sample. The S(11) and DC current features were found to spatially broaden around grain boundaries with the sample under illumination. The broadening is attributed to optically generated charge that becomes trapped and changes the local depletion of the grain boundaries, thereby modifying the local capacitance. Imaging provided by the NSMM offers a new RF methodology to resolve and characterize nanoscale electrical features in photovoltaic materials and devices.

Web-based virtual microscopy at the RWTH Aachen University: didactic concept, methods and analysis of acceptance by the students.

PubMed

Merk, Magdalene; Knuechel, Ruth; Perez-Bouza, Alberto

2010-12-20

Fundamental knowledge of microscopic anatomy and pathology has always been an essential part in medical education. The traditional didactic concept comprises theoretical and practical lessons using a light microscope and glass slides. High-speed Internet connections and technical improvement in whole-slide digital microscopy (commonly termed "virtual microscopy") provide a new and attractive approach for both teachers and students. High picture quality and unlimited temporal and spatial availability of histology samples from different fields are key advantages of web-based digital microscopy. In this report we discuss the technical requirements, system efficiency, optical resolution and didactic concept. Furthermore, we present a review of the experience gained in the course of one year based on an analysis of student acceptance. Three groups with a total of 192 students between the 3rd and 5th year of medical studies attending the practical courses of general and advanced histopathology had access to both glass-mounted and digitalized slides. Prior to exams, students were asked to answer an anonymous questionnaire. The results of the study reflect the high acceptance and intensive use of the web-based digital histology by students, thus encouraging the development of further Web-based learning strategies for the teaching of histology and pathology. 2010 Elsevier GmbH. All rights reserved.

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 pragmatic guide to multiphoton microscope design

PubMed Central

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

2016-01-01

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

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

3D interferometric microscope: color visualization of engineered surfaces for industrial applications

NASA Astrophysics Data System (ADS)

Schmit, Joanna; Novak, Matt; Bui, Son

2015-09-01

3D microscopes based on white light interference (WLI) provide precise measurement for the topography of engineering surfaces. However, the display of an object in its true colors as observed under white illumination is often desired; this traditionally has presented a challenge for WLI-based microscopes. Such 3D color display is appealing to the eye and great for presentations, and also provides fast evaluation of certain characteristics like defects, delamination, or deposition of different materials. Determination of color as observed by interferometric objectives is not straightforward; we will present how color imaging capabilities similar to an ordinary microscope can be obtained in interference microscopes based on WLI and we will give measurement and imaging examples of a few industrial samples.

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.

Metal-free inactivation of E. coli O157:H7 by fullerene/C3N4 hybrid under visible light irradiation.

PubMed

Ouyang, Kai; Dai, Ke; Chen, Hao; Huang, Qiaoyun; Gao, Chunhui; Cai, Peng

2017-02-01

Interest has grown in developing safe and high-performance photocatalysts based on metal-free materials for disinfection of bacterial pathogens under visible light irradiation. In this paper, the C 60 /C 3 N 4 and C 70 /C 3 N 4 hybrids were synthesized by a hydrothermal method, and characterized by X-ray diffraction (XRD), UV-vis diffuse reflection spectroscopy (UV-vis DRS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and high revolution transmission electron microscope (HRTEM). The performance of photocatalytic disinfection was investigated by the inactivation of Escherichia coli O157:H7. Both C 60 /C 3 N 4 and C 70 /C 3 N 4 hybrids showed similar crystalline structure and morphology with C 3 N 4 ; however, the two composites exhibited stronger bacterial inactivation than C 3 N 4 . In particular, C 70 /C 3 N 4 showed the highest bactericidal efficiency and was detrimental to all E. coli O157:H7 in 4h irradiation. Compared to C 3 N 4 , the enhancement of photocatalytic activity of composites could be attributed to the effective transfer of the photoinduced electrons under visible light irradiation. Owing to the excellent performance of fullerenes (C 60 , C 70 )/C 3 N 4 composites, a visible light response and environmental friendly photocatalysts for disinfection were achieved. Copyright © 2016. Published by Elsevier Inc.

High-resolution light microscopy of nanoforms

NASA Astrophysics Data System (ADS)

Vodyanoy, Vitaly; Pustovyy, Oleg; Vainrub, Arnold

2007-09-01

We developed a high resolution light imaging system. Diffraction gratings with 100 nm width lines as well as less than 100 nm size features of different-shaped objects are clearly visible on a calibrated microscope test slide (Vainrub et al., Optics Letters, 2006, 31, 2855). The two-point resolution increase results from a known narrowing of the central diffraction peak for the annular aperture. Better visibility and advanced contrast of the smallest features in the image are due to enhancement of high spatial frequencies in the optical transfer function. The imaging system is portable, low energy, and battery operated. It has been adapted to use in both transmitting and reflecting light. It is particularly applicable for motile nanoform systems where structure and functions can be depicted in real time. We have isolated micrometer and submicrometer particles, termed proteons, from human and animal blood. Proteons form by reversible seeded aggregation of proteins around proteon nucleating centers (PNCs). PNCs are comprised of 1-2nm metallic nanoclusters containing 40-300 atoms. Proteons are capable of spontaneous assembling into higher nanoform systems assuming structure of complicated topology. The arrangement of complex proteon system mimics the structure of a small biological cell. It has structures that imitate membrane and nucleolus or nuclei. Some of these nanoforms are motile. They interact and divide. Complex nanoform systems can spontaneously reduce to simple proteons. The physical properties of these nanoforms could shed some light on the properties of early life forms or forms at extreme conditions.

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.

Bidirectional reflectance distribution function of Spectralon white reflectance standard illuminated by incoherent unpolarized and plane-polarized light.

PubMed

Bhandari, Anak; Hamre, Børge; Frette, Øvynd; Zhao, Lu; Stamnes, Jakob J; Kildemo, Morten

2011-06-01

A Lambert surface would appear equally bright from all observation directions regardless of the illumination direction. However, the reflection from a randomly scattering object generally has directional variation, which can be described in terms of the bidirectional reflectance distribution function (BRDF). We measured the BRDF of a Spectralon white reflectance standard for incoherent illumination at 405 and 680 nm with unpolarized and plane-polarized light from different directions of incidence. Our measurements show deviations of the BRDF for the Spectralon white reflectance standard from that of a Lambertian reflector that depend both on the angle of incidence and the polarization states of the incident light and detected light. The non-Lambertian reflection characteristics were found to increase more toward the direction of specular reflection as the angle of incidence gets larger.

Light Reflector

NASA Astrophysics Data System (ADS)

1988-01-01

Ultra Sales, Inc.'s fluorescent lighting fixture gets a boost in reflectivity through installation of Lightdriver, a thin tough thermoplastic film plated with aluminum, capable of reflecting 95 percent of visible light striking it. Lightdriver increases brightness without adding bulbs, and allows energy savings by removing some bulbs because the mirrorlike surface cuts light loss generally occasioned by conventional low reflectivity white painted surface above the bulbs in many fluorescent fixtures. Forty-five percent reduction in lighting electricity is attainable.

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

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.

Integrated Emissivity And Temperature Measurement

DOEpatents

Poulsen, Peter

2005-11-08

A multi-channel spectrometer and a light source are used to measure both the emitted and the reflected light from a surface which is at an elevated temperature relative to its environment. In a first method, the temperature of the surface and emissivity in each wavelength is calculated from a knowledge of the spectrum and the measurement of the incident and reflected light. In the second method, the reflected light is measured from a reference surface having a known reflectivity and the same geometry as the surface of interest and the emitted and the reflected light are measured for the surface of interest. These measurements permit the computation of the emissivity in each channel of the spectrometer and the temperature of the surface of interest.

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.

Wide acceptance angle, high concentration ratio, optical collector

NASA Technical Reports Server (NTRS)

Kruer, Mark A. (Inventor)

1991-01-01

A cassegrain optical system provides improved collection of off-axis light yet is still characterized by a high concentration ratio. The optical system includes a primary mirror for collecting incoming light and reflecting the light to a secondary mirror which, in turn, reflects the light to a solar cell or other radiation collection device. The primary mirror reflects incoming on-axis light onto an annular section of the secondary mirror and results in the reflection of a substantial amount of incoming off-axis light onto the remainder of the secondary mirror. Thus light which would otherwise be lost to the system will be captured by the collector. Furthermore, the off-axis sections of the secondary mirror may be of a different geometrical shape than the on-axis annular section so as to optimize the amount of off-axis light collected.

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.

Electrowetting-actuated optical switch based on total internal reflection.

PubMed

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

2015-04-01

In this paper we demonstrate a liquid optical switch based on total internal reflection. Two indium tin oxide electrodes are fabricated on the bottom substrate. A conductive liquid (Liquid 1) is placed on one side of the chamber and surrounded by a density-matched silicone oil (Liquid 2). In initial state, when the light beam illuminates the interface of the two liquids, it just meets the conditions of total internal reflection. The light is totally reflected by Liquid 2, and the device shows light-off state. When we apply a voltage to the other side of the indium tin oxide electrode, Liquid 1 stretched towards this side of the substrate and the curvature of the liquid-liquid interface changes. The light beam is refracted by Liquid 1 and the device shows light-on state. So the device can achieve the functions of an optical switch. Because the light beam can be totally reflected by the liquid, the device can attain 100% light intensity attenuation. Our experiments show that the response time from light-on (off) to light-off (on) are 130 and 132 ms, respectively. The proposed optical switch has potential applications in variable optical attenuators, information displays, and light shutters.

Light reflection models for computer graphics.

PubMed

Greenberg, D P

1989-04-14

During the past 20 years, computer graphic techniques for simulating the reflection of light have progressed so that today images of photorealistic quality can be produced. Early algorithms considered direct lighting only, but global illumination phenomena with indirect lighting, surface interreflections, and shadows can now be modeled with ray tracing, radiosity, and Monte Carlo simulations. This article describes the historical development of computer graphic algorithms for light reflection and pictorially illustrates what will be commonly available in the near future.

Silica-gold nanoshells as potential intraoperative molecular probes for HER2-overexpression in ex vivo breast tissue using near-infrared reflectance confocal microscopy.

PubMed

Bickford, Lissett R; Agollah, Germaine; Drezek, Rebekah; Yu, Tse-Kuan

2010-04-01

Obtaining negative margins is critical for breast cancer patients undergoing conservation therapy in order to reduce the reemergence of the original cancer. Currently, breast cancer tumor margins are examined in a pathology lab either while the patient is anesthetized or after the surgical procedure has been terminated. These current methods often result in cancer cells present at the surgical resection margin due to inadequate margin assessment at the point of care. Due to such limitations evident in current diagnoses, tools for increasing the accuracy and speed of tumor margin detection directly in the operating room are still needed. We are exploring the potential of using a nano-biophotonics system to facilitate intraoperative tumor margin assessment ex vivo at the cellular level. By combining bioconjugated silica-based gold nanoshells, which scatter light in the near-infrared, with a portable FDA-approved reflectance confocal microscope, we first validate the use of gold nanoshells as effective reflectance-based imaging probes by evaluating the contrast enhancement of three different HER2-overexpressing cell lines. Additionally, we demonstrate the ability to detect HER2-overexpressing cells in human tissue sections within 5 min of incubation time. This work supports the use of targeted silica-based gold nanoshells as potential real-time molecular probes for HER2-overexpression in human tissue.

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.

Catheter based mid-infrared reflectance and reflectance generated absorption spectroscopy

DOEpatents

Holman, Hoi-Ying N

2013-10-29

A method of characterizing conditions in a tissue, by (a) providing a catheter that has a light source that emits light in selected wavenumbers within the range of mid-IR spectrum; (b) directing the light from the catheter to an area of tissue at a location inside a blood vessel of a subject; (c) collecting light reflected from the location and generating a reflectance spectra; and (d) comparing the reflectance spectra to a reference spectra of normal tissue, whereby a location having an increased number of absorbance peaks at said selected wavenumbers indicates a tissue inside the blood vessel containing a physiological marker for atherosclerosis.

Single-lens computed tomography imaging spectrometer and method of capturing spatial and spectral information

NASA Technical Reports Server (NTRS)

Wilson, Daniel W. (Inventor); Johnson, William R. (Inventor); Bearman, Gregory H. (Inventor)

2011-01-01

Computed tomography imaging spectrometers ("CTISs") employing a single lens are provided. The CTISs may be either transmissive or reflective, and the single lens is either configured to transmit and receive uncollimated light (in transmissive systems), or is configured to reflect and receive uncollimated light (in reflective systems). An exemplary transmissive CTIS includes a focal plane array detector, a single lens configured to transmit and receive uncollimated light, a two-dimensional grating, and a field stop aperture. An exemplary reflective CTIS includes a focal plane array detector, a single mirror configured to reflect and receive uncollimated light, a two-dimensional grating, and a field stop aperture.

Apparatus for and method of correcting for aberrations in a light beam

DOEpatents

Sawicki, Richard H.

1996-01-01

A technique for adjustably correcting for aberrations in a light beam is disclosed herein. This technique utilizes first means which defines a flat, circular light reflecting surface having opposite reinforced circumferential edges and a central post and which is resiliently distortable, to a limited extent, into different concave and/or convex curvatures, which may be Gaussian-like, about the central axis, and second means acting on the first means for adjustably distorting the light reflecting surface into a particular selected one of the different curvatures depending upon the aberrations to be corrected for and for fixedly maintaining the curvature selected. In the embodiment disclosed, the light reflecting surface is adjustably distorted into the selected curvature by application of particular axial moments to the central post on the opposite side from the light reflecting surface and lateral moments to the circumference of the reflecting surface.

Apparatus for and method of correcting for aberrations in a light beam

DOEpatents

Sawicki, R.H.

1996-09-17

A technique for adjustably correcting for aberrations in a light beam is disclosed herein. This technique utilizes first means which defines a flat, circular light reflecting surface having opposite reinforced circumferential edges and a central post and which is resiliently distortable, to a limited extent, into different concave and/or convex curvatures, which may be Gaussian-like, about the central axis, and second means acting on the first means for adjustably distorting the light reflecting surface into a particular selected one of the different curvatures depending upon the aberrations to be corrected for and for fixedly maintaining the curvature selected. In the embodiment disclosed, the light reflecting surface is adjustably distorted into the selected curvature by application of particular axial moments to the central post on the opposite side from the light reflecting surface and lateral moments to the circumference of the reflecting surface. 8 figs.

Lamp method and apparatus using multiple reflections

DOEpatents

MacLennan, Donald A.; Turner, Brian; Kipling, Kent

1999-01-01

A method wherein the light in a sulfur or selenium lamp is reflected through the fill a multiplicity of times to convert ultraviolet radiation to visible. A light emitting device comprised of an electrodeless envelope which bears a light reflecting covering around a first portion which does not crack due to differential thermal expansion and which has a second portion which comprises a light transmissive aperture.

Changes in functional metabolism in the rat central nervous system following spaceflight

NASA Technical Reports Server (NTRS)

Murakami, D. M.; Miller, J. D.; Fuller, C. A.

1985-01-01

The neuronal metabolism and soma size of neurons within the paraventricular nucleus (PVN) and the supraoptic nucleus of rats are analyzed. Five male Sprague-Dawley rats were flown on Spacelab 3 for 7 days under a 12:12 light/dark cycle and unlimited food and water, and a control group was kept on the ground under similar conditions. The preparation of the hypothalamus of the rats for microscopic examination using thionin or the cytochrome oxidase (CYOX) technique is described. CYOX activity and soma size within the PVN are evaluated. The effects of water drinking pattern and space flight on CYOX activity and soma size are investigated. The data reveal that the flight rats with normal drinking patterns display a decrease in neuronal metabolism within the vasopressin-containing neurons of the hypothalamus and this metabolic change may reflect fluid shifts caused by microgravity.

Freshwater Fossil Pearls from the Nihewan Basin, Early Early Pleistocene.

PubMed

Li, Su-Ping; Yao, Pei-Yi; Li, Jin-Feng; Ferguson, David Kay; Min, Long-Rui; Chi, Zhen-Qing; Wang, Yong; Yao, Jian-Xin; Sha, Jin-Geng

2016-01-01

Fossil blister pearls attached to the shells of an Anodonta mollusk from China, early Early Pleistocene, are reported here for the first time. The pearls were investigated in detail using a variety of methods. Micro-CT scanning of the fossil pearls was carried out to discover the inner structure and the pearl nucleus. Using CTAn software, changes in the gray levels of the biggest pearl, which reflect the changing density of the material, were investigated. The results provide us with some clues on how these pearls were formed. Sand grains, shell debris or material with a similar density could have stimulated the development of these pearls. X-ray diffraction analysis of one fossil pearl and the shell to which it was attached reveals that only aragonite exists in both samples. The internal structures of our fossil shells and pearls were investigated using a Scanning Electron Microscope. These investigations throw some light on pearl development in the past.

Studies of Islands on Freely Suspended Bubbles of Smectic Liquid Crystal

NASA Technical Reports Server (NTRS)

Pattanaporkratana, A.; Mavel, B.; Park, C. S.; Maclennan, J. E.; Clark, N. A.

2002-01-01

We have constructed an optical system for observing the internal structure of freely suspended smectic liquid crystal bubbles using a reflected light microscope. Liquid crystal bubbles can have thicker circular regions (islands) which can easily be generated by shrinking the bubble diameter. The diameter of these islands is approximately 10 microns and they are typically up to five times thicker than the surrounding liquid crystal film (500 angstroms). In the Laboratory, the location of the islands is strongly influenced by gravity, which causes the majority of islands to migrate to the bottom half of the bubble. We will describe the size and thickness distributions of islands and their time evolution, and also discuss two-dimensional hydrodynamics and turbulence of smectic bubbles, the shapes of islands and holes affected by bubble vibrations, and the interactions between islands, which we have probed using optical tweezers.

Acousto-optical tunable filter for combined wideband, spectral, and optical coherence microscopy.

PubMed

Machikhin, Alexander S; Pozhar, Vitold E; Viskovatykh, Alexander V; Burmak, Ludmila I

2015-09-01

A multimodal technique for inspection of microscopic objects by means of wideband optical microscopy, spectral microscopy, and optical coherence microscopy is described, implemented, and tested. The key feature is the spectral selection of light in the output arm of an interferometer with use of the specialized imaging acousto-optical tunable filter. In this filter, two interfering optical beams are diffracted via the same ultrasound wave without destruction of interference image structure. The basic requirements for the acousto-optical tunable filter are defined, and mathematical formulas for calculation of its parameters are derived. Theoretical estimation of the achievable accuracy of the 3D image reconstruction is presented and experimental proofs are given. It is demonstrated that spectral imaging can also be accompanied by measurement of the quantitative reflectance spectra. Examples of inspection of optically transparent and nontransparent samples demonstrate the applicability of the technique.

Qualitative investigation of fresh human scalp hair with full-field optical coherence tomography

NASA Astrophysics Data System (ADS)

Choi, Woo June; Pi, Long-Quan; Min, Gihyeon; Lee, Won-Soo; Lee, Byeong Ha

2012-03-01

We have investigated depth-resolved cellular structures of unmodified fresh human scalp hairs with ultrahigh-resolution full-field optical coherence tomography (FF-OCT). The Linnik-type white light interference microscope has been home-implemented to observe the micro-internal layers of human hairs in their natural environment. In hair shafts, FF-OCT has qualitatively revealed the cellular hair compartments of cuticle and cortex layers involved in keratin filaments and melanin granules. No significant difference between black and white hair shafts was observed except for absence of only the melanin granules in the white hair, reflecting that the density of the melanin granules directly affects the hair color. Anatomical description of plucked hair bulbs was also obtained with the FF-OCT in three-dimensions. We expect this approach will be useful for evaluating cellular alteration of natural hairs on cosmetic assessment or diagnosis of hair diseases.

High throughput, parallel imaging and biomarker quantification of human spermatozoa by ImageStream flow cytometry.

PubMed

Buckman, Clayton; George, Thaddeus C; Friend, Sherree; Sutovsky, Miriam; Miranda-Vizuete, Antonio; Ozanon, Christophe; Morrissey, Phil; Sutovsky, Peter

2009-12-01

Spermatid specific thioredoxin-3 protein (SPTRX-3) accumulates in the superfluous cytoplasm of defective human spermatozoa. Novel ImageStream technology combining flow cytometry with cell imaging was used for parallel quantification and visualization of SPTRX-3 protein in defective spermatozoa of five men from infertile couples. The majority of the SPTRX-3 containing cells were overwhelmingly spermatozoa with a variety of morphological defects, detectable in the ImageStream recorded images. Quantitative parameters of relative SPTRX-3 induced fluorescence measured by ImageStream correlated closely with conventional flow cytometric measurements of the same sample set and reflected the results of clinical semen evaluation. Image Stream quantification of SPTRX-3 combines and surpasses the informative value of both conventional flow cytometry and light microscopic semen evaluation. The observed patterns of the retention of SPTRX-3 in the sperm samples from infertility patients support the view that SPTRX3 is a biomarker of male infertility.

Determining biological tissue optical properties via integrating sphere spatial measurements

DOEpatents

Baba, Justin S [Knoxville, TN; Letzen, Brian S [Coral Springs, FL

2011-01-11

An optical sample is mounted on a spatial-acquisition apparatus that is placed in or on an enclosure. An incident beam is irradiated on a surface of the sample and the specular reflection is allowed to escape from the enclosure through an opening. The spatial-acquisition apparatus is provided with a light-occluding slider that moves in front of the sample to block portions of diffuse scattering from the sample. As the light-occluding slider moves across the front of the sample, diffuse light scattered into the area of the backside of the light-occluding slider is absorbed by back side surface of the light-occluding slider. By measuring a baseline diffuse reflectance without a light-occluding slider and subtracting measured diffuse reflectance with a light-occluding slider therefrom, diffuse reflectance for the area blocked by the light-occluding slider can be calculated.

Method of Detecting Coliform Bacteria and Escherichia Coli Bacteria from Reflected Light

NASA Technical Reports Server (NTRS)

Vincent, Robert (Inventor)

2013-01-01

The present invention relates to a method of detecting coliform bacteria in water from reflected light and a method of detecting Eschericha Coli bacteria in water from reflected light, and also includes devices for the measurement, calculation and transmission of data relating to that method.

Systems and Methods for Integrated Emissivity and Temperature Measurement of a Surface

DOEpatents

Poulsen, Peter

2005-11-08

A multi-channel spectrometer and a light source are used to measure both the emitted and the reflected light from a surface which is at an elevated temperature relative to its environment. In a first method, the temperature of the surface and emissivity in each wavelength is calculated from a knowledge of the spectrum and the measurement of the incident and reflected light. In the second method, the reflected light is measured from a reference surface having a known reflectivity and the same geometry as the surface of interest and the emitted and the reflected light are measured for the surface of interest. These measurements permit the computation of the emissivity in each channel of the spectrometer and the temperature of the surface of interest.