Transmissive Nanohole Arrays for Massively-Parallel Optical Biosensing

PubMed Central

2015-01-01

A high-throughput optical biosensing technique is proposed and demonstrated. This hybrid technique combines optical transmission of nanoholes with colorimetric silver staining. The size and spacing of the nanoholes are chosen so that individual nanoholes can be independently resolved in massive parallel using an ordinary transmission optical microscope, and, in place of determining a spectral shift, the brightness of each nanohole is recorded to greatly simplify the readout. Each nanohole then acts as an independent sensor, and the blocking of nanohole optical transmission by enzymatic silver staining defines the specific detection of a biological agent. Nearly 10000 nanoholes can be simultaneously monitored under the field of view of a typical microscope. As an initial proof of concept, biotinylated lysozyme (biotin-HEL) was used as a model analyte, giving a detection limit as low as 0.1 ng/mL. PMID:25530982

Augmented microscopy with near-infrared fluorescence detection

NASA Astrophysics Data System (ADS)

Watson, Jeffrey R.; Martirosyan, Nikolay; Skoch, Jesse; Lemole, G. Michael; Anton, Rein; Romanowski, Marek

2015-03-01

Near-infrared (NIR) fluorescence has become a frequently used intraoperative technique for image-guided surgical interventions. In procedures such as cerebral angiography, surgeons use the optical surgical microscope for the color view of the surgical field, and then switch to an electronic display for the NIR fluorescence images. However, the lack of stereoscopic, real-time, and on-site coregistration adds time and uncertainty to image-guided surgical procedures. To address these limitations, we developed the augmented microscope, whereby the electronically processed NIR fluorescence image is overlaid with the anatomical optical image in real-time within the optical path of the microscope. In vitro, the augmented microscope can detect and display indocyanine green (ICG) concentrations down to 94.5 nM, overlaid with the anatomical color image. We prepared polyacrylamide tissue phantoms with embedded polystyrene beads, yielding scattering properties similar to brain matter. In this model, 194 μM solution of ICG was detectable up to depths of 5 mm. ICG angiography was then performed in anesthetized rats. A dynamic process of ICG distribution in the vascular system overlaid with anatomical color images was observed and recorded. In summary, the augmented microscope demonstrates NIR fluorescence detection with superior real-time coregistration displayed within the ocular of the stereomicroscope. In comparison to other techniques, the augmented microscope retains full stereoscopic vision and optical controls including magnification and focus, camera capture, and multiuser access. Augmented microscopy may find application in surgeries where the use of traditional microscopes can be enhanced by contrast agents and image guided delivery of therapeutics, including oncology, neurosurgery, and ophthalmology.

Shape oscillations of microparticles on an optical microscope stage.

PubMed

Zhu, Z M; Apfel, R E

1985-11-01

A modulated acoustic radiation pressure technique to produce quadrupole shape oscillations of drops ranging in diameter from 50-220 micron has been used by us. These drops have been suspended by acoustic levitation in a small chamber mounted on a stage of an optical microscope, which allowed easy viewing. The fission of drops and the deformation of sea urchin eggs were also observed.

Joint Services Electronics Program Annual Progress Report.

DTIC Science & Technology

1987-10-15

polarizability of free carriers in the semiconductor perturb the index of refraction which can be detected in a Nomarski -type optical interferometer. For...interferomters. However, the charge probe relies on a different physical effect and operates by interferometrically detecting the phase change induced in an... Nomarski microscope systems. These techniques will be applied, eventually, in our real-time V.. scanning optical microscope described below. Recently

Resolution enhancement techniques in microscopy

NASA Astrophysics Data System (ADS)

Cremer, Christoph; Masters, Barry R.

2013-05-01

We survey the history of resolution enhancement techniques in microscopy and their impact on current research in biomedicine. Often these techniques are labeled superresolution, or enhanced resolution microscopy, or light-optical nanoscopy. First, we introduce the development of diffraction theory in its relation to enhanced resolution; then we explore the foundations of resolution as expounded by the astronomers and the physicists and describe the conditions for which they apply. Then we elucidate Ernst Abbe's theory of optical formation in the microscope, and its experimental verification and dissemination to the world wide microscope communities. Second, we describe and compare the early techniques that can enhance the resolution of the microscope. Third, we present the historical development of various techniques that substantially enhance the optical resolution of the light microscope. These enhanced resolution techniques in their modern form constitute an active area of research with seminal applications in biology and medicine. Our historical survey of the field of resolution enhancement uncovers many examples of reinvention, rediscovery, and independent invention and development of similar proposals, concepts, techniques, and instruments. Attribution of credit is therefore confounded by the fact that for understandable reasons authors stress the achievements from their own research groups and sometimes obfuscate their contributions and the prior art of others. In some cases, attribution of credit is also made more complex by the fact that long term developments are difficult to allocate to a specific individual because of the many mutual connections often existing between sometimes fiercely competing, sometimes strongly collaborating groups. Since applications in biology and medicine have been a major driving force in the development of resolution enhancing approaches, we focus on the contribution of enhanced resolution to these fields.

SIL-STED microscopy technique enhancing super-resolution of fluorescence microscopy

NASA Astrophysics Data System (ADS)

Park, No-Cheol; Lim, Geon; Lee, Won-sup; Moon, Hyungbae; Choi, Guk-Jong; Park, Young-Pil

2017-08-01

We have characterized a new type STED microscope which combines a high numerical aperture (NA) optical head with a solid immersion lens (SIL), and we call it as SIL-STED microscope. The advantage of a SIL-STED microscope is that its high NA of the SIL makes it superior to a general STED microscope in lateral resolution, thus overcoming the optical diffraction limit at the macromolecular level and enabling advanced super-resolution imaging of cell surface or cell membrane structure and function Do. This study presents the first implementation of higher NA illumination in a STED microscope limiting the fluorescence lateral resolution to about 40 nm. The refractive index of the SIL which is made of material KTaO3 is about 2.23 and 2.20 at a wavelength of 633 nm and 780 nm which are used for excitation and depletion in STED imaging, respectively. Based on the vector diffraction theory, the electric field focused by the SILSTED microscope is numerically calculated so that the numerical results of the point dispersion function of the microscope and the expected resolution could be analyzed. For further investigation, fluorescence imaging of nano size fluorescent beads is fulfilled to show improved performance of the technique.

Optical method for distance and displacement measurements of the probe-sample separation in a scanning near-field optical microscope

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

Santamaria, L.; Siller, H. R.; Garcia-Ortiz, C. E., E-mail: cegarcia@cicese.mx

In this work, we present an alternative optical method to determine the probe-sample separation distance in a scanning near-field optical microscope. The experimental method is based in a Lloyd’s mirror interferometer and offers a measurement precision deviation of ∼100 nm using digital image processing and numerical analysis. The technique can also be strategically combined with the characterization of piezoelectric actuators and stability evaluation of the optical system. It also opens the possibility for the development of an automatic approximation control system valid for probe-sample distances from 5 to 500 μm.

Skin suturing and cortical surface viral infusion improves imaging of neuronal ensemble activity with head-mounted miniature microscopes.

PubMed

Li, Xinjian; Cao, Vania Y; Zhang, Wenyu; Mastwal, Surjeet S; Liu, Qing; Otte, Stephani; Wang, Kuan Hong

2017-11-01

In vivo optical imaging of neural activity provides important insights into brain functions at the single-cell level. Cranial windows and virally delivered calcium indicators are commonly used for imaging cortical activity through two-photon microscopes in head-fixed animals. Recently, head-mounted one-photon microscopes have been developed for freely behaving animals. However, minimizing tissue damage from the virus injection procedure and maintaining window clarity for imaging can be technically challenging. We used a wide-diameter glass pipette at the cortical surface for infusing the viral calcium reporter AAV-GCaMP6 into the cortex. After infusion, the scalp skin over the implanted optical window was sutured to facilitate postoperative recovery. The sutured scalp was removed approximately two weeks later and a miniature microscope was attached above the window to image neuronal activity in freely moving mice. We found that cortical surface virus infusion efficiently labeled neurons in superficial layers, and scalp skin suturing helped to maintain the long-term clarity of optical windows. As a result, several hundred neurons could be recorded in freely moving animals. Compared to intracortical virus injection and open-scalp postoperative recovery, our methods minimized tissue damage and dura overgrowth underneath the optical window, and significantly increased the experimental success rate and the yield of identified neurons. Our improved cranial surgery technique allows for high-yield calcium imaging of cortical neurons with head-mounted microscopes in freely behaving animals. This technique may be beneficial for other optical applications such as two-photon microscopy, multi-site imaging, and optogenetic modulation. Published by Elsevier B.V.

Advantages of microscope-integrated intraoperative online optical coherence tomography: usage in Boston keratoprosthesis type I surgery

NASA Astrophysics Data System (ADS)

Siebelmann, Sebastian; Steven, Philipp; Hos, Deniz; Hüttmann, Gereon; Lankenau, Eva; Bachmann, Björn; Cursiefen, Claus

2016-01-01

Boston keratoprosthesis (KPro) type I is a technique to treat patients with corneal diseases that are not amenable to conventional keratoplasty. Correct assembly and central implantation of the prosthesis are crucial for postoperative visual recovery. This study investigates the potential benefit of intraoperative optical coherence tomography (OCT) to monitor KPro surgery. Retrospective case series are presented for two patients who underwent Boston KPro type I implantation. The surgery in both patients was monitored intraoperatively using a commercially available intraoperative OCT (iOCT) device mounted on a surgical microscope. Microscope-integrated intraoperative OCT was able to evaluate the correct assembly and implantation of the KPro. All parts of the prosthesis were visible, and interfaces between the corneal graft and titanium backplate or anterior optics were clearly depictable. Moreover, iOCT visualized a gap between the backplate and graft in one case, and in the other case, a gap between the anterior optic and graft. Neither gap was visible with a conventional surgical microscope. The gap between the anterior optic and the graft could easily be corrected. Microscope-integrated iOCT delivers enhanced information, adding to the normal surgical microscope view during KPro surgery. Correct assembly can be controlled as well as the correct placement of the Boston KPro into the anterior chamber.

Evaluation of a completely robotized neurosurgical operating microscope.

PubMed

Kantelhardt, Sven R; Finke, Markus; Schweikard, Achim; Giese, Alf

2013-01-01

Operating microscopes are essential for most neurosurgical procedures. Modern robot-assisted controls offer new possibilities, combining the advantages of conventional and automated systems. We evaluated the prototype of a completely robotized operating microscope with an integrated optical coherence tomography module. A standard operating microscope was fitted with motors and control instruments, with the manual control mode and balance preserved. In the robot mode, the microscope was steered by a remote control that could be fixed to a surgical instrument. External encoders and accelerometers tracked microscope movements. The microscope was additionally fitted with an optical coherence tomography-scanning module. The robotized microscope was tested on model systems. It could be freely positioned, without forcing the surgeon to take the hands from the instruments or avert the eyes from the oculars. Positioning error was about 1 mm, and vibration faded in 1 second. Tracking of microscope movements, combined with an autofocus function, allowed determination of the focus position within the 3-dimensional space. This constituted a second loop of navigation independent from conventional infrared reflector-based techniques. In the robot mode, automated optical coherence tomography scanning of large surface areas was feasible. The prototype of a robotized optical coherence tomography-integrated operating microscope combines the advantages of a conventional manually controlled operating microscope with a remote-controlled positioning aid and a self-navigating microscope system that performs automated positioning tasks such as surface scans. This demonstrates that, in the future, operating microscopes may be used to acquire intraoperative spatial data, volume changes, and structural data of brain or brain tumor tissue.

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.

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

NASA Astrophysics Data System (ADS)

Romo, Jaime E., Jr.

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

Laser beam shaping for biomedical microscopy techniques

NASA Astrophysics Data System (ADS)

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

2016-04-01

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

Applied physics: Optical trapping for space mirrors.

PubMed

McGloin, David

2014-02-27

Might it be possible to create mirrors for space telescopes, using nothing but microscopic particles held in place by light? A study that exploits a technique called optical binding provides a step towards this goal.

Environmental Integrity of Coating/Metal Interface

DTIC Science & Technology

1989-03-01

occurred. For this explanation to be correct, microblisters would be optically detectable using a microscope with Nomarsky contrast or by examining the...polymer surface with an interferometric microscope. Both of these techniques have been tried, but neither of them was able to detect any surface

Review on Microstructure Analysis of Metals and Alloys Using Image Analysis Techniques

NASA Astrophysics Data System (ADS)

Rekha, Suganthini; Bupesh Raja, V. K.

2017-05-01

The metals and alloys find vast application in engineering and domestic sectors. The mechanical properties of the metals and alloys are influenced by their microstructure. Hence the microstructural investigation is very critical. Traditionally the microstructure is studied using optical microscope with suitable metallurgical preparation. The past few decades the computers are applied in the capture and analysis of the optical micrographs. The advent of computer softwares like digital image processing and computer vision technologies are a boon to the analysis of the microstructure. In this paper the literature study of the various developments in the microstructural analysis, is done. The conventional optical microscope is complemented by the use of Scanning Electron Microscope (SEM) and other high end equipments.

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.

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

NASA Technical Reports Server (NTRS)

Choudry, A.; Dupuis, N.

1988-01-01

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

Dynamic-focusing microscope objective for optical coherence tomography

NASA Astrophysics Data System (ADS)

Murali, Supraja; Rolland, Jannick

2007-01-01

Optical Coherence Tomography (OCT) is a novel optical imaging technique that has assumed significant importance in bio-medical imaging in the last two decades because it is non-invasive and provides accurate, high resolution images of three dimensional cross-sections of body tissue, exceeding the capabilities of the current predominant imaging technique - ultrasound. In this paper, the application of high resolution OCT, known as optical coherence microscopy (OCM) is investigated for in vivo detection of abnormal skin pathology for the early diagnosis of cancer. A main challenge in OCM is maintaining invariant resolution throughout the sample. The technology presented is based on a dynamic focusing microscope imaging probe conceived for skin imaging and the detection of abnormalities in the epithelium. A novel method for dynamic focusing in the biological sample is presented using variable-focus lens technology to obtain three dimensional images with invariant resolution throughout the cross-section and depth of the sample is presented and discussed. A low coherence broadband source centered at near IR wavelengths is used to illuminate the sample. The design, analysis and predicted performance of the dynamic focusing microscope objective designed for dynamic three dimensional imaging at 5μm resolution for the chosen broadband spectrum is presented.

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

NASA Astrophysics Data System (ADS)

Patel, Shreyas J.

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

A wide field-of-view microscope based on holographic focus grid

NASA Astrophysics Data System (ADS)

Wu, Jigang; Cui, Xiquan; Zheng, Guoan; Lee, Lap Man; Yang, Changhuei

2010-02-01

We have developed a novel microscope technique that can achieve wide field-of-view (FOV) imaging and yet possess resolution that is comparable to conventional microscope. The principle of wide FOV microscope system breaks the link between resolution and FOV magnitude of traditional microscopes. Furthermore, by eliminating bulky optical elements from its design and utilizing holographic optical elements, the wide FOV microscope system is more cost-effective. In our system, a hologram was made to focus incoming collimated beam into a focus grid. The sample is put in the focal plane and the transmissions of the focuses are detected by an imaging sensor. By scanning the incident angle of the incoming beam, the focus grid will scan across the sample and the time-varying transmission can be detected. We can then reconstruct the transmission image of the sample. The resolution of microscopic image is limited by the size of the focus formed by the hologram. The scanning area of each focus spot is determined by the separation of the focus spots and can be made small for fast imaging speed. We have fabricated a prototype system with a 2.4-mm FOV and 1-μm resolution. The prototype system was used to image onion skin cells for a demonstration. The preliminary experiments prove the feasibility of the wide FOV microscope technique, and the possibility of a wider FOV system with better resolution.

Embedded Fiber Optic Sensors for Measuring Transient Detonation/Shock Behavior;Time-of-Arrival Detection and Waveform Determination.

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

Chavez, Marcus Alexander; Willis, Michael David; Covert, Timothy Todd

2014-09-01

The miniaturization of explosive components has driven the need for a corresponding miniaturization of the current diagnostic techniques available to measure the explosive phenomena. Laser interferometry and the use of spectrally coated optical windows have proven to be an essential interrogation technique to acquire particle velocity time history data in one- dimensional gas gun and relatively large-scale explosive experiments. A new diagnostic technique described herein allows for experimental measurement of apparent particle velocity time histories in microscale explosive configurations and can be applied to shocks/non-shocks in inert materials. The diagnostic, Embedded Fiber Optic Sensors (EFOS), has been tested in challengingmore » microscopic experimental configurations that give confidence in the technique's ability to measure the apparent particle velocity time histories of an explosive with pressure outputs in the tenths of kilobars to several kilobars. Embedded Fiber Optic Sensors also allow for several measurements to be acquired in a single experiment because they are microscopic, thus reducing the number of experiments necessary. The future of EFOS technology will focus on further miniaturization, material selection appropriate for the operating pressure regime, and extensive hydrocode and optical analysis to transform apparent particle velocity time histories into true particle velocity time histories as well as the more meaningful pressure time histories.« less

Closed loop adaptive optics for microscopy without a wavefront sensor.

PubMed

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

2010-02-24

A three-dimensional wide-field image of a small fluorescent bead contains more than enough information to accurately calculate the wavefront in the microscope objective back pupil plane using the phase retrieval technique. The phase-retrieved wavefront can then be used to set a deformable mirror to correct the point-spread function (PSF) of the microscope without the use of a wavefront sensor. This technique will be useful for aligning the deformable mirror in a widefield microscope with adaptive optics and could potentially be used to correct aberrations in samples where small fluorescent beads or other point sources are used as reference beacons. Another advantage is the high resolution of the retrieved wavefont as compared with current Shack-Hartmann wavefront sensors. Here we demonstrate effective correction of the PSF in 3 iterations. Starting from a severely aberrated system, we achieve a Strehl ratio of 0.78 and a greater than 10-fold increase in maximum intensity.

Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.

PubMed

Hayashi, Shinichi; Okada, Yasushi

2015-05-01

Most current superresolution (SR) microscope techniques surpass the diffraction limit at the expense of temporal resolution, compromising their applications to live-cell imaging. Here we describe a new SR fluorescence microscope based on confocal microscope optics, which we name the spinning disk superresolution microscope (SDSRM). Theoretically, the SDSRM is equivalent to a structured illumination microscope (SIM) and achieves a spatial resolution of 120 nm, double that of the diffraction limit of wide-field fluorescence microscopy. However, the SDSRM is 10 times faster than a conventional SIM because SR signals are recovered by optical demodulation through the stripe pattern of the disk. Therefore a single SR image requires only a single averaged image through the rotating disk. On the basis of this theory, we modified a commercial spinning disk confocal microscope. The improved resolution around 120 nm was confirmed with biological samples. The rapid dynamics of micro-tubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30-100 frames/s. Because our method requires only small optical modifications, it will enable an easy upgrade from an existing spinning disk confocal to a SR microscope for live-cell imaging. © 2015 Hayashi and Okada. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy

PubMed Central

Lim, Daryl; Ford, Tim N.; Chu, Kengyeh K.; Mertz, Jerome

2011-01-01

We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish. PMID:21280920

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy.

PubMed

Lim, Daryl; Ford, Tim N; Chu, Kengyeh K; Mertz, Jerome

2011-01-01

We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish.

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy

NASA Astrophysics Data System (ADS)

Lim, Daryl; Ford, Tim N.; Chu, Kengyeh K.; Mertz, Jerome

2011-01-01

We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish.

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.

Simple fiber-optic confocal microscopy with nanoscale depth resolution beyond the diffraction barrier.

PubMed

Ilev, Ilko; Waynant, Ronald; Gannot, Israel; Gandjbakhche, Amir

2007-09-01

A novel fiber-optic confocal approach for ultrahigh depth-resolution (

Hybrid Microscopy: Enabling Inexpensive High-Performance Imaging through Combined Physical and Optical Magnifications.

PubMed

Zhang, Yu Shrike; Chang, Jae-Byum; Alvarez, Mario Moisés; Trujillo-de Santiago, Grissel; Aleman, Julio; Batzaya, Byambaa; Krishnadoss, Vaishali; Ramanujam, Aishwarya Aravamudhan; Kazemzadeh-Narbat, Mehdi; Chen, Fei; Tillberg, Paul W; Dokmeci, Mehmet Remzi; Boyden, Edward S; Khademhosseini, Ali

2016-03-15

To date, much effort has been expended on making high-performance microscopes through better instrumentation. Recently, it was discovered that physical magnification of specimens was possible, through a technique called expansion microscopy (ExM), raising the question of whether physical magnification, coupled to inexpensive optics, could together match the performance of high-end optical equipment, at a tiny fraction of the price. Here we show that such "hybrid microscopy" methods--combining physical and optical magnifications--can indeed achieve high performance at low cost. By physically magnifying objects, then imaging them on cheap miniature fluorescence microscopes ("mini-microscopes"), it is possible to image at a resolution comparable to that previously attainable only with benchtop microscopes that present costs orders of magnitude higher. We believe that this unprecedented hybrid technology that combines expansion microscopy, based on physical magnification, and mini-microscopy, relying on conventional optics--a process we refer to as Expansion Mini-Microscopy (ExMM)--is a highly promising alternative method for performing cost-effective, high-resolution imaging of biological samples. With further advancement of the technology, we believe that ExMM will find widespread applications for high-resolution imaging particularly in research and healthcare scenarios in undeveloped countries or remote places.

Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry.

PubMed

Schwertner, M; Booth, M J; Neil, M A A; Wilson, T

2004-01-01

Confocal or multiphoton microscopes, which deliver optical sections and three-dimensional (3D) images of thick specimens, are widely used in biology. These techniques, however, are sensitive to aberrations that may originate from the refractive index structure of the specimen itself. The aberrations cause reduced signal intensity and the 3D resolution of the instrument is compromised. It has been suggested to correct for aberrations in confocal microscopes using adaptive optics. In order to define the design specifications for such adaptive optics systems, one has to know the amount of aberrations present for typical applications such as with biological samples. We have built a phase stepping interferometer microscope that directly measures the aberration of the wavefront. The modal content of the wavefront is extracted by employing Zernike mode decomposition. Results for typical biological specimens are presented. It was found for all samples investigated that higher order Zernike modes give only a small contribution to the overall aberration. Therefore, these higher order modes can be neglected in future adaptive optics sensing and correction schemes implemented into confocal or multiphoton microscopes, leading to more efficient designs.

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

NASA Astrophysics Data System (ADS)

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

2011-09-01

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

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.

Optical fabrication and testing; Proceedings of the Meeting, Singapore, Oct. 22-27, 1990

NASA Astrophysics Data System (ADS)

Lorenzen, Manfred; Campbell, Duncan R.; Johnson, Craig W.

1991-03-01

Various papers on optical fabrication and testing are presented. Individual topics addressed include: interferometry with laser diodes, new methods for economic production of prisms and lenses, interferometer accuracy and precision, optical testing with wavelength scanning interferometer, digital Talbot interferometer, high-sensitivity interferometric technique for strain measurements, absolute interferometric testing of spherical surfaces, contouring using gratings created on an LCD panel, three-dimensional inspection using laser-based dynamic fringe projection, noncontact optical microtopography, laser scan microscope and infrared laser scan microscope, photon scanning tunneling microscopy. Also discussed are: combination-matching problems in the layout design of minilaser rangefinder, design and testing of a cube-corner array for laser ranging, mode and far-field pattern of diode laser-phased arrays, new glasses for optics and optoelectronics, optical properties of Li-doped ZnO films, application and machining of Zerodur for optical purposes, finish machining of optical components in mass production.

Experimental stress–strain analysis of tapered silica optical fibers with nanofiber waist

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

Holleis, S.; Hoinkes, T.; Wuttke, C.

2014-04-21

We experimentally determine tensile force–elongation diagrams of tapered optical fibers with a nanofiber waist. The tapered optical fibers are produced from standard silica optical fibers using a heat and pull process. Both, the force–elongation data and scanning electron microscope images of the rupture points indicate a brittle material. Despite the small waist radii of only a few hundred nanometers, our experimental data can be fully explained by a nonlinear stress–strain model that relies on material properties of macroscopic silica optical fibers. This is an important asset when it comes to designing miniaturized optical elements as one can rely on themore » well-founded material characteristics of standard optical fibers. Based on this understanding, we demonstrate a simple and non-destructive technique that allows us to determine the waist radius of the tapered optical fiber. We find excellent agreement with independent scanning electron microscope measurements of the waist radius.« less

Optical fabrication and testing; Proceedings of the Meeting, Singapore, Oct. 22-27, 1990

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

Lorenzen, M.; Campbell, D.R.; Johnson, C.W.

1991-01-01

Various papers on optical fabrication and testing are presented. Individual topics addressed include: interferometry with laser diodes, new methods for economic production of prisms and lenses, interferometer accuracy and precision, optical testing with wavelength scanning interferometer, digital Talbot interferometer, high-sensitivity interferometric technique for strain measurements, absolute interferometric testing of spherical surfaces, contouring using gratings created on an LCD panel, three-dimensional inspection using laser-based dynamic fringe projection, noncontact optical microtopography, laser scan microscope and infrared laser scan microscope, photon scanning tunneling microscopy. Also discussed are: combination-matching problems in the layout design of minilaser rangefinder, design and testing of a cube-corner arraymore » for laser ranging, mode and far-field pattern of diode laser-phased arrays, new glasses for optics and optoelectronics, optical properties of Li-doped ZnO films, application and machining of Zerodur for optical purposes, finish machining of optical components in mass production.« less

Review of advanced imaging techniques

PubMed Central

Chen, Yu; Liang, Chia-Pin; Liu, Yang; Fischer, Andrew H.; Parwani, Anil V.; Pantanowitz, Liron

2012-01-01

Pathology informatics encompasses digital imaging and related applications. Several specialized microscopy techniques have emerged which permit the acquisition of digital images (“optical biopsies”) at high resolution. Coupled with fiber-optic and micro-optic components, some of these imaging techniques (e.g., optical coherence tomography) are now integrated with a wide range of imaging devices such as endoscopes, laparoscopes, catheters, and needles that enable imaging inside the body. These advanced imaging modalities have exciting diagnostic potential and introduce new opportunities in pathology. Therefore, it is important that pathology informaticists understand these advanced imaging techniques and the impact they have on pathology. This paper reviews several recently developed microscopic techniques, including diffraction-limited methods (e.g., confocal microscopy, 2-photon microscopy, 4Pi microscopy, and spatially modulated illumination microscopy) and subdiffraction techniques (e.g., photoactivated localization microscopy, stochastic optical reconstruction microscopy, and stimulated emission depletion microscopy). This article serves as a primer for pathology informaticists, highlighting the fundamentals and applications of advanced optical imaging techniques. PMID:22754737

Optical and Acoustical Techniques for Non-viral Gene Delivery to Mammalian Cells and In-situ Study of Cytoskeletal Mechanics

NASA Astrophysics Data System (ADS)

Ma, Zili

Since the first optical microscope invented by Anton van Leeuwenhoek in 1674, the great development of laser technique and its applications in biophotonics have helped us reveal the mechanisms underlying numerous biological activities gradually. The introduction of fs lasers to the studies of biology has emerged as a fast developing area calling for the efforts and skills both from optics and electric engineering and biology and medicine. Due to the fast update of laser source techniques, there has been an increasing number of commercialized fs lasers available for this growing market of biophotonics. To better utilize the potential offered by fs lasers, we studied the technique of optical gene delivery and tried to narrow the gap between laboratorial research and industrial/clinical applications, in that the strict experimental conditions of specific optical laboratorial studies are generally not appropriate for the practical biological applications. To carry out our experiments, we built a two-stage amplifier fs laser system to generate the desired pulse train. The laser pulse train was coupled into an invert fluorescence microscope for the imaging and manipulation of each cell. To overcome limitations brought by the tight focus of laser beam due to high NA objective, we introduced gold nanorods (GNRs), a metallic nanomaterial, with tunable optical property. With these additional membrane for membrane permeabilization, which could significantly improve the manipulation speed than that based on the tightly focused laser. We used GFP plasmid to demonstrate the applications of this technique in gene delivery, and successfully transfected and GFP-expressed cells were observed one day after the optical transfection. Additionally, as an important trend of biophotonics, the integration of optics with microfluidic chips has become the new frontier of both biology and engineering. Here we firstly demonstrated a technique of gene delivery by an on-chip device generating surface acoustic waves, which not only achieved a high efficiency of cells permeabilization in a quick speed, but also allowed us to observe the permeabilization process in real time by microscope. This device is also compatible with biophotonics studies based on fs laser, which can be further developed as a powerful tool for optical gene delivery with the capability of precisely controlling the fluid on-chip by SAW. SAW devices could also achieve exogenous gene delivery through the cell membrane without the need of adding chemical agents. Our results showed that the membrane of mammalian adherent cells could be effectively perforated transiently by applying a SAW. The transfection of pEGFP plasmids into endothelial cells was carried out successfully via this SAW-induced cell perforation. The expression of GFP was observed after 24-hour incubation subsequent to the SAW treatment. In regard to the application of fs lasers in cellular and subcellular level studies, we applied the optical nanoscissoring technique based on fs lasers in biomechanical studies to study the mechanical properties of single SF in-situ. Integrated into a confocal microscope, the fs laser showed great power in manipulating targeted in-situ subcellular structures under real-time imaging without damaging nearby regions. Here, how oxidative challenges would alter the mechanical properties of SFs in myoblasts was firstly investigated using the optical nanoscissoring technique to comprehend the whole picture of muscle tissue injury and repair from the basics. The prestress of stress fibers after the oxidative challenges was found through our modified viscoelastic retraction model and experiment result.

Optically Sectioned Imaging of Microvasculature of In-Vivo and Ex-Vivo Thick Tissue Models with Speckle-illumination HiLo Microscopy and HiLo Image Processing Implementation in MATLAB Architecture

NASA Astrophysics Data System (ADS)

Suen, Ricky Wai

The work described in this thesis covers the conversion of HiLo image processing into MATLAB architecture and the use of speckle-illumination HiLo microscopy for use of ex-vivo and in-vivo imaging of thick tissue models. HiLo microscopy is a wide-field fluorescence imaging technique and has been demonstrated to produce optically sectioned images comparable to confocal in thin samples. The imaging technique was developed by Jerome Mertz and the Boston University Biomicroscopy Lab and has been implemented in our lab as a stand-alone optical setup and a modification to a conventional fluorescence microscope. Speckle-illumination HiLo microscopy combines two images taken under speckle-illumination and standard uniform-illumination to generate an optically sectioned image that reject out-of-focus fluorescence. The evaluated speckle contrast in the images is used as a weighting function where elements that move out-of-focus have a speckle contrast that decays to zero. The experiments shown here demonstrate the capability of our HiLo microscopes to produce optically-sectioned images of the microvasculature of ex-vivo and in-vivo thick tissue models. The HiLo microscope were used to image the microvasculature of ex-vivo mouse heart sections prepared for optical histology and the microvasculature of in-vivo rodent dorsal window chamber models. Studies in label-free surface profiling with HiLo microscopy is also presented.

Quantitative high dynamic range beam profiling for fluorescence microscopy

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

Mitchell, T. J., E-mail: t.j.mitchell@dur.ac.uk; Saunter, C. D.; O’Nions, W.

2014-10-15

Modern developmental biology relies on optically sectioning fluorescence microscope techniques to produce non-destructive in vivo images of developing specimens at high resolution in three dimensions. As optimal performance of these techniques is reliant on the three-dimensional (3D) intensity profile of the illumination employed, the ability to directly record and analyze these profiles is of great use to the fluorescence microscopist or instrument builder. Though excitation beam profiles can be measured indirectly using a sample of fluorescent beads and recording the emission along the microscope detection path, we demonstrate an alternative approach where a miniature camera sensor is used directly withinmore » the illumination beam. Measurements taken using our approach are solely concerned with the illumination optics as the detection optics are not involved. We present a miniature beam profiling device and high dynamic range flux reconstruction algorithm that together are capable of accurately reproducing quantitative 3D flux maps over a large focal volume. Performance of this beam profiling system is verified within an optical test bench and demonstrated for fluorescence microscopy by profiling the low NA illumination beam of a single plane illumination microscope. The generality and success of this approach showcases a widely flexible beam amplitude diagnostic tool for use within the life sciences.« less

The influence of voltage applied between the electrodes on optical and morphological properties of the InGaN thin films grown by thermionic vacuum arc.

PubMed

Özen, Soner; Şenay, Volkan; Pat, Suat; Korkmaz, Şadan

2016-01-01

The aim of this research is to investigate the optical and morphological properties of the InGaN thin films deposited onto amorphous glass substrates in two separate experiments with two different voltages applied between the electrodes, i.e. 500 and 600 V by means of the thermionic vacuum arc technique. This technique is original for thin film deposition and it enables thin film production in a very short period of time. The optical and morphological properties of the films were investigated by using field emission scanning electron microscope, atomic force microscope, spectroscopic ellipsometer, reflectometer, spectrophotometer, and optical tensiometer. Optical properties were also supported by empirical relations. The deposition rates were calculated as 3 and 3.3 nm/sec for 500 and 600 V, respectively. The increase in the voltage also increased the refractive index, grain size, root mean square roughness and surface free energy. According to the results of the wetting experiments, InGaN samples were low-wettable, also known as hydrophobic. © Wiley Periodicals, Inc.

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.

A combined confocal and magnetic resonance microscope for biological studies

NASA Astrophysics Data System (ADS)

Majors, Paul D.; Minard, Kevin R.; Ackerman, Eric J.; Holtom, Gary R.; Hopkins, Derek F.; Parkinson, Christopher I.; Weber, Thomas J.; Wind, Robert A.

2002-12-01

Complementary data acquired with different microscopy techniques provide a basis for establishing a more comprehensive understanding of cell function in health and disease, particularly when results acquired with different methodologies can be correlated in time and space. In this article, a novel microscope is described for studying live cells simultaneously with both confocal scanning laser fluorescence optical microscopy and magnetic resonance microscopy. The various design considerations necessary for integrating these two complementary techniques are discussed, the layout and specifications of the instrument are given, and examples of confocal and magnetic resonance images of large frog cells and model tumor spheroids obtained with the compound microscope are presented.

Electron microscopic and optical studies of prism faces of synthetic quartz

NASA Technical Reports Server (NTRS)

Buzek, B. C.; Vagh, A. S.

1977-01-01

Application of electron and optical microscopic techniques to the study of growth spirals on quartz crystal faces is described. Attention is centered on the centers of the spirals and on screw ledges; overhanging kinks are revealed on one side of the spiral centers. The possibility that these special features may have developed after growth of the crystals went to completion is explored. The conjecture is raised that such structures might result from adsorption of growth-inhibiting impurities at the center of the growth spiral on the quartz habit faces.

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

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

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

2016-01-28

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

Fast and Accurate Cell Tracking by a Novel Optical-Digital Hybrid Method

NASA Astrophysics Data System (ADS)

Torres-Cisneros, M.; Aviña-Cervantes, J. G.; Pérez-Careta, E.; Ambriz-Colín, F.; Tinoco, Verónica; Ibarra-Manzano, O. G.; Plascencia-Mora, H.; Aguilera-Gómez, E.; Ibarra-Manzano, M. A.; Guzman-Cabrera, R.; Debeir, Olivier; Sánchez-Mondragón, J. J.

2013-09-01

An innovative methodology to detect and track cells using microscope images enhanced by optical cross-correlation techniques is proposed in this paper. In order to increase the tracking sensibility, image pre-processing has been implemented as a morphological operator on the microscope image. Results show that the pre-processing process allows for additional frames of cell tracking, therefore increasing its robustness. The proposed methodology can be used in analyzing different problems such as mitosis, cell collisions, and cell overlapping, ultimately designed to identify and treat illnesses and malignancies.

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

PubMed Central

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-01-01

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications. PMID:26525841

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

PubMed

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-11-03

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.

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

NASA Astrophysics Data System (ADS)

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-11-01

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.

Assessing the Penetrating Abilities of Experimental Preparation with Dental Infiltrant Features Using Optical Microscope: Preliminary Study.

PubMed

Skucha-Nowak, Małgorzata; Machorowska-Pieniążek, Agnieszka; Tanasiewicz, Marta

2016-01-01

The aim of the infiltration technique is to penetrate demineralized enamel with a low viscosity resin. Icon® (DMG) is the first ever and so far the only dental infiltrant. Bacteriostaticity is one of the properties that should be inherent in dental infiltrants, but Icon lacks this feature. The aim of the preliminary study was to properly choose a dye which would allow us to assess the penetrating abilities of our own, experimental preparation with features of a dental infiltrant with bacteriostatic properties and to compare using an optical microscope the depth of infiltration of the designed experimental preparation with the infiltrant available on the market. The preparation is supposed to infiltrate decalcified human enamel and be assessed with an optical microscope. Eosin, neutral fuchsine and methylene blue were added to experimental preparation with dental infiltrant features and to Icon® (DMG) in order to assess the depth of penetration of the experimental solution into the decalcified layers of enamel. The experimental solution mixes well with eosin, neutral fuchsine, and methylene blue. During the preliminary study, the authors concluded that the experimental solution mixes well with methylene blue, neutral fuchsine, and eosin. An addition of eosin to a preparation which infiltrates inner, demineralized enamel layers, facilitates the assessment of such a preparation with an optical microscope. A designed experimental solution with the main ingredients, i.e., 2-hydroxyethyl methacrylate (HEMA) and tetraethylene glycol dimethacrylate (TEGDMA) with a ratio of 75% to 25% penetrates the demineralized (decalcified) inner parts of the enamel and polymerizes when exposed to light. In order to assess the infiltration of the experimental solution into the demineralized enamel layers, it is required to improve the measurement techniques that utilize optical microscopy.

Refractive-index profiling of embedded microstructures in optical materials

NASA Astrophysics Data System (ADS)

Dave, Digant P.; Milner, Thomas E.

2002-04-01

We describe use of a phase-sensitive low-coherence reflectometer to measure spatial variation of refractive index in optical materials. The described interferometric technique is demonstrated to be a valuable tool to profile the refractive index of optical elements such as integrated waveguides and photowritten optical microstructures. As an example, a refractive-index profile is mapped of a microstructure written in a microscope glass slide with an ultrashort-pulse laser.

Structured illumination 3D microscopy using adaptive lenses and multimode fibers

NASA Astrophysics Data System (ADS)

Czarske, Jürgen; Philipp, Katrin; Koukourakis, Nektarios

2017-06-01

Microscopic techniques with high spatial and temporal resolution are required for in vivo studying biological cells and tissues. Adaptive lenses exhibit strong potential for fast motion-free axial scanning. However, they also lead to a degradation of the achievable resolution because of aberrations. This hurdle can be overcome by digital optical technologies. We present a novel High-and-Low-frequency (HiLo) 3D-microscope using structured illumination and an adaptive lens. Uniform illumination is used to obtain optical sectioning for the high-frequency (Hi) components of the image, and nonuniform illumination is needed to obtain optical sectioning for the low-frequency (Lo) components of the image. Nonuniform illumination is provided by a multimode fiber. It ensures robustness against optical aberrations of the adaptive lens. The depth-of-field of our microscope can be adjusted a-posteriori by computational optics. It enables to create flexible scans, which compensate for irregular axial measurement positions. The adaptive HiLo 3D-microscope provides an axial scanning range of 1 mm with an axial resolution of about 4 microns and sub-micron lateral resolution over the full scanning range. In result, volumetric measurements with high temporal and spatial resolution are provided. Demonstration measurements of zebrafish embryos with reporter gene-driven fluorescence in the thyroid gland are presented.

Closed loop adaptive optics for microscopy without a wavefront sensor

PubMed Central

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

2013-01-01

A three-dimensional wide-field image of a small fluorescent bead contains more than enough information to accurately calculate the wavefront in the microscope objective back pupil plane using the phase retrieval technique. The phase-retrieved wavefront can then be used to set a deformable mirror to correct the point-spread function (PSF) of the microscope without the use of a wavefront sensor. This technique will be useful for aligning the deformable mirror in a widefield microscope with adaptive optics and could potentially be used to correct aberrations in samples where small fluorescent beads or other point sources are used as reference beacons. Another advantage is the high resolution of the retrieved wavefont as compared with current Shack-Hartmann wavefront sensors. Here we demonstrate effective correction of the PSF in 3 iterations. Starting from a severely aberrated system, we achieve a Strehl ratio of 0.78 and a greater than 10-fold increase in maximum intensity. PMID:24392198

Single-Shot Optical Sectioning Using Two-Color Probes in HiLo Fluorescence Microscopy

PubMed Central

Muro, Eleonora; Vermeulen, Pierre; Ioannou, Andriani; Skourides, Paris; Dubertret, Benoit; Fragola, Alexandra; Loriette, Vincent

2011-01-01

We describe a wide-field fluorescence microscope setup which combines HiLo microscopy technique with the use of a two-color fluorescent probe. It allows one-shot fluorescence optical sectioning of thick biological moving sample which is illuminated simultaneously with a flat and a structured pattern at two different wavelengths. Both homogenous and structured fluorescence images are spectrally separated at detection and combined similarly with the HiLo microscopy technique. We present optically sectioned full-field images of Xenopus laevis embryos acquired at 25 images/s frame rate. PMID:21641327

Suspension and simple optical characterization of two-dimensional membranes

NASA Astrophysics Data System (ADS)

Northeast, David B.; Knobel, Robert G.

2018-03-01

We report on a method for suspending two-dimensional crystal materials in an electronic circuit using an only photoresists and solvents. Graphene and NbSe2 are suspended tens of nanometers above metal electrodes with clamping diameters of several microns. The optical cavity formed from the membrane/air/metal structures enables a quick method to measure the number of layers and the gap separation using comparisons between the expected colour and optical microscope images. This characterization technique can be used with just an illuminated microscope with a digital camera which makes it adaptable to environments where other means of characterization are not possible, such as inside nitrogen glove boxes used in handling oxygen-sensitive materials.

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

NASA Astrophysics Data System (ADS)

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

2014-06-01

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

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

PubMed

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

2014-06-01

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

Multimodal full-field optical coherence tomography on biological tissue: toward all optical digital pathology

NASA Astrophysics Data System (ADS)

Harms, F.; Dalimier, E.; Vermeulen, P.; Fragola, A.; Boccara, A. C.

2012-03-01

Optical Coherence Tomography (OCT) is an efficient technique for in-depth optical biopsy of biological tissues, relying on interferometric selection of ballistic photons. Full-Field Optical Coherence Tomography (FF-OCT) is an alternative approach to Fourier-domain OCT (spectral or swept-source), allowing parallel acquisition of en-face optical sections. Using medium numerical aperture objective, it is possible to reach an isotropic resolution of about 1x1x1 ìm. After stitching a grid of acquired images, FF-OCT gives access to the architecture of the tissue, for both macroscopic and microscopic structures, in a non-invasive process, which makes the technique particularly suitable for applications in pathology. Here we report a multimodal approach to FF-OCT, combining two Full-Field techniques for collecting a backscattered endogeneous OCT image and a fluorescence exogeneous image in parallel. Considering pathological diagnosis of cancer, visualization of cell nuclei is of paramount importance. OCT images, even for the highest resolution, usually fail to identify individual nuclei due to the nature of the optical contrast used. We have built a multimodal optical microscope based on the combination of FF-OCT and Structured Illumination Microscopy (SIM). We used x30 immersion objectives, with a numerical aperture of 1.05, allowing for sub-micron transverse resolution. Fluorescent staining of nuclei was obtained using specific fluorescent dyes such as acridine orange. We present multimodal images of healthy and pathological skin tissue at various scales. This instrumental development paves the way for improvements of standard pathology procedures, as a faster, non sacrificial, operator independent digital optical method compared to frozen sections.

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

NASA Astrophysics Data System (ADS)

Upputuri, Paul Kumar; Pramanik, Manojit

2017-12-01

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

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

NASA Astrophysics Data System (ADS)

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

2018-04-01

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

Nanoscale characterization of local structures and defects in photonic crystals using synchrotron-based transmission soft X-ray microscopy

PubMed Central

Nho, Hyun Woo; Kalegowda, Yogesh; Shin, Hyun-Joon; Yoon, Tae Hyun

2016-01-01

For the structural characterization of the polystyrene (PS)-based photonic crystals (PCs), fast and direct imaging capabilities of full field transmission X-ray microscopy (TXM) were demonstrated at soft X-ray energy. PS-based PCs were prepared on an O2-plasma treated Si3N4 window and their local structures and defects were investigated using this label-free TXM technique with an image acquisition speed of ~10 sec/frame and marginal radiation damage. Micro-domains of face-centered cubic (FCC (111)) and hexagonal close-packed (HCP (0001)) structures were dominantly found in PS-based PCs, while point and line defects, FCC (100), and 12-fold symmetry structures were also identified as minor components. Additionally, in situ observation capability for hydrated samples and 3D tomographic reconstruction of TXM images were also demonstrated. This soft X-ray full field TXM technique with faster image acquisition speed, in situ observation, and 3D tomography capability can be complementally used with the other X-ray microscopic techniques (i.e., scanning transmission X-ray microscopy, STXM) as well as conventional characterization methods (e.g., electron microscopic and optical/fluorescence microscopic techniques) for clearer structure identification of self-assembled PCs and better understanding of the relationship between their structures and resultant optical properties. PMID:27087141

Neutron-induced defects in optical fibers

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

Rizzolo, S., E-mail: serena.rizzolo@univ-st-etienne.fr; Dipartimento di Fisica e Chimica, Università di Palermo, Palermo; and Areva Centre Technique, Le Creusot

2014-10-21

We present a study on 0.8 MeV neutron-induced defects up to fluences of 10{sup 17} n/cm{sup 2} in fluorine doped optical fibers by using electron paramagnetic resonance, optical absorption and confocal micro-luminescence techniques. Our results allow to address the microscopic mechanisms leading to the generation of Silica-related point-defects such as E', H(I), POR and NBOH Centers.

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

PubMed

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

2018-01-01

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

Mathematical model of a DIC position sensing system within an optical trap

NASA Astrophysics Data System (ADS)

Wulff, Kurt D.; Cole, Daniel G.; Clark, Robert L.

2005-08-01

The quantitative study of displacements and forces of motor proteins and processes that occur at the microscopic level and below require a high level of sensitivity. For optical traps, two techniques for position sensing have been accepted and used quite extensively: quadrant photodiodes and an interferometric position sensing technique based on DIC imaging. While quadrant photodiodes have been studied in depth and mathematically characterized, a mathematical characterization of the interferometric position sensor has not been presented to the authors' knowledge. The interferometric position sensing method works off of the DIC imaging capabilities of a microscope. Circularly polarized light is sent into the microscope and the Wollaston prism used for DIC imaging splits the beam into its orthogonal components, displacing them by a set distance determined by the user. The distance between the axes of the beams is set so the beams overlap at the specimen plane and effectively share the trapped microsphere. A second prism then recombines the light beams and the exiting laser light's polarization is measured and related to position. In this paper we outline the mathematical characterization of a microsphere suspended in an optical trap using a DIC position sensing method. The sensitivity of this mathematical model is then compared to the QPD model. The mathematical model of a microsphere in an optical trap can serve as a calibration curve for an experimental setup.

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

PubMed

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

2017-11-01

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

Subsurface Growth Of Silicide Structures In Silicon

NASA Technical Reports Server (NTRS)

Fathauer, Robert W.; George, Thomas; Pike, William T.; Schowalter, Leo

1993-01-01

Technique shows promise for fabrication of novel electronic, optoelectronic, and electro-optical devices. Experiments demonstrated feasibility of growing microscopic single-crystal CoSi2 structures beneath surfaces of Si substrates.

Ultrafast chirped optical waveform recorder using a time microscope

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

Bennett, Corey Vincent

2015-04-21

A new technique for capturing both the amplitude and phase of an optical waveform is presented. This technique can capture signals with many THz of bandwidths in a single shot (e.g., temporal resolution of about 44 fs), or be operated repetitively at a high rate. That is, each temporal window (or frame) is captured single shot, in real time, but the process may be run repeatedly or single-shot. By also including a variety of possible demultiplexing techniques, this process is scalable to recoding continuous signals.

Optimal model-based sensorless adaptive optics for epifluorescence microscopy.

PubMed

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

2018-01-01

We report on a universal sample-independent sensorless adaptive optics method, based on modal optimization of the second moment of the fluorescence emission from a point-like excitation. Our method employs a sample-independent precalibration, performed only once for the particular system, to establish the direct relation between the image quality and the aberration. The method is potentially applicable to any form of microscopy with epifluorescence detection, including the practically important case of incoherent fluorescence emission from a three dimensional object, through minor hardware modifications. We have applied the technique successfully to a widefield epifluorescence microscope and to a multiaperture confocal microscope.

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.

Microstructural study of the nickel-base alloy WAZ-20 using qualitative and quantitative electron optical techniques

NASA Technical Reports Server (NTRS)

Young, S. G.

1973-01-01

The NASA nickel-base alloy WAZ-20 was analyzed by advanced metallographic techniques to qualitatively and quantitatively characterize its phases and stability. The as-cast alloy contained primary gamma-prime, a coarse gamma-gamma prime eutectic, a gamma-fine gamma prime matrix, and MC carbides. A specimen aged at 870 C for 1000 hours contained these same constituents and a few widely scattered high W particles. No detrimental phases (such as sigma or mu) were observed. Scanning electron microscope, light metallography, and replica electron microscope methods are compared. The value of quantitative electron microprobe techniques such as spot and area analysis is demonstrated.

Plant Histology: Clearing and the Optical Section.

ERIC Educational Resources Information Center

Freeman, H. E.

1985-01-01

Clearing is a simple and rapid technique in which 75 percent lactic acid is used to remove pigments and cytoplasmic contents of fresh leaves, enabling microscopic view of various internal leaf layers. Procedures for using the technique (which helps students gain a more thorough understanding of plant anatomy) are given. (DH)

Differential polarization nonlinear optical microscopy with adaptive optics controlled multiplexed beams.

PubMed

Samim, Masood; Sandkuijl, Daaf; Tretyakov, Ian; Cisek, Richard; Barzda, Virginijus

2013-09-09

Differential polarization nonlinear optical microscopy has the potential to become an indispensable tool for structural investigations of ordered biological assemblies and microcrystalline aggregates. Their microscopic organization can be probed through fast and sensitive measurements of nonlinear optical signal anisotropy, which can be achieved with microscopic spatial resolution by using time-multiplexed pulsed laser beams with perpendicular polarization orientations and photon-counting detection electronics for signal demultiplexing. In addition, deformable membrane mirrors can be used to correct for optical aberrations in the microscope and simultaneously optimize beam overlap using a genetic algorithm. The beam overlap can be achieved with better accuracy than diffraction limited point-spread function, which allows to perform polarization-resolved measurements on the pixel-by-pixel basis. We describe a newly developed differential polarization microscope and present applications of the differential microscopy technique for structural studies of collagen and cellulose. Both, second harmonic generation, and fluorescence-detected nonlinear absorption anisotropy are used in these investigations. It is shown that the orientation and structural properties of the fibers in biological tissue can be deduced and that the orientation of fluorescent molecules (Congo Red), which label the fibers, can be determined. Differential polarization microscopy sidesteps common issues such as photobleaching and sample movement. Due to tens of megahertz alternating polarization of excitation pulses fast data acquisition can be conveniently applied to measure changes in the nonlinear signal anisotropy in dynamically changing in vivo structures.

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 Review of Adaptive Optics Optical Coherence Tomography: Technical Advances, Scientific Applications, and the Future.

PubMed

Jonnal, Ravi S; Kocaoglu, Omer P; Zawadzki, Robert J; Liu, Zhuolin; Miller, Donald T; Werner, John S

2016-07-01

Optical coherence tomography (OCT) has enabled "virtual biopsy" of the living human retina, revolutionizing both basic retina research and clinical practice over the past 25 years. For most of those years, in parallel, adaptive optics (AO) has been used to improve the transverse resolution of ophthalmoscopes to foster in vivo study of the retina at the microscopic level. Here, we review work done over the last 15 years to combine the microscopic transverse resolution of AO with the microscopic axial resolution of OCT, building AO-OCT systems with the highest three-dimensional resolution of any existing retinal imaging modality. We surveyed the literature to identify the most influential antecedent work, important milestones in the development of AO-OCT technology, its applications that have yielded new knowledge, research areas into which it may productively expand, and nascent applications that have the potential to grow. Initial efforts focused on demonstrating three-dimensional resolution. Since then, many improvements have been made in resolution and speed, as well as other enhancements of acquisition and postprocessing techniques. Progress on these fronts has produced numerous discoveries about the anatomy, function, and optical properties of the retina. Adaptive optics OCT continues to evolve technically and to contribute to our basic and clinical knowledge of the retina. Due to its capacity to reveal cellular and microscopic detail invisible to clinical OCT systems, it is an ideal companion to those instruments and has the demonstrable potential to produce images that can guide the interpretation of clinical findings.

An application of computer image-processing and filmy replica technique to the copper electroplating method of stress analysis

NASA Astrophysics Data System (ADS)

Sugiura, M.; Seika, M.

1994-02-01

In this study, a new technique to measure the density of slip-bands automatically is developed, namely, a TV image of the slip-bands observed through a microscope is directly processed by an image-processing system using a personal computer and an accurate value of the density of slip-bands is measured quickly. In the case of measuring the local stresses in machine parts of large size with the copper plating foil, the direct observation of slip-bands through an optical microscope is difficult. In this study, to facilitate a technique close to the direct microscopic observation of slip-bands in the foil attached to a large-sized specimen, the replica method using a platic film of acetyl cellulose is applied to replicate the slip-bands in the attached foil.

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

NASA Astrophysics Data System (ADS)

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

2018-02-01

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

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.

New clinical opportunities for retinal vascular imaging: adaptive optics to OCT angiography

NASA Astrophysics Data System (ADS)

Rosen, Richard; Chui, Toco; Weitz, Rishard; Dubra, Alfredo; Carroll, Joseph; Garcia, Patricia; Pinhas, Alexander; Scripsema, Nicole; Mo, Shelley; Agemy, Steven; Krawitz, Brian

2018-03-01

As techniques of retinal imaging have evolved, anatomic features that were only assessable in the laboratory have become available in the clinic for patient care. The retinal capillaries were initially described on microscope sections in the pathology laboratory. As optical methods have advanced these features have become part of the routine clinical landscape inspected daily by physicians. This paper briefly traces the evolution of these techniques and shows how they fit into the modern diagnostic armamentarium of ophthalmic retinal care.

Detecting Phase Boundaries in Hard-Sphere Suspensions

NASA Technical Reports Server (NTRS)

McDowell, Mark; Rogers, Richard B.; Gray, Elizabeth

2009-01-01

A special image-data-processing technique has been developed for use in experiments that involve observation, via optical microscopes equipped with electronic cameras, of moving boundaries between the colloidal-solid and colloidal-liquid phases of colloidal suspensions of monodisperse hard spheres. During an experiment, it is necessary to adjust the position of a microscope to keep the phase boundary within view. A boundary typically moves at a speed of the order of microns per hour. Because an experiment can last days or even weeks, it is impractical to require human intervention to keep the phase boundary in view. The present image-data-processing technique yields results within a computation time short enough to enable generation of automated-microscope-positioning commands to track the moving phase boundary

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.

Photon theory hypothesis about photon tunneling microscope's subwavelength resolution

NASA Astrophysics Data System (ADS)

Zhu, Yanbin; Ma, Junfu

1995-09-01

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

Hyper-spectral imaging in scanning-confocal-fluorescence microscopy using a novel broadband diffractive optic

NASA Astrophysics Data System (ADS)

Wang, Peng; Ebeling, Carl G.; Gerton, Jordan; Menon, Rajesh

In this paper, we demonstrate hyper-spectral imaging of fluorescent microspheres in a scanning-confocal-fluorescence microscope by spatially dispersing the spectra using a novel broadband diffractive optic, and applying a nonlinear optimization technique to extract the full-incident spectra. This broadband diffractive optic has a designed optical efficiency of over 90% across the entire visible spectrum. We used this technique to create two-color images of two fluorophores and also extracted their emission spectra with good fidelity. This method can be extended to image both spatially and spectrally overlapping fluorescent samples. Full control in the number of emission spectra and the feasibility of enhanced imaging speed are demonstrated as well.

Fabrication and characterization of optical super-smooth surfaces

NASA Astrophysics Data System (ADS)

Schmitt, Dirk-Roger; Kratz, Frank; Ringel, Gabriele A.; Mangelsdorf, Juergen; Creuzet, Francois; Garratt, John D.

1995-08-01

Intercomparison roughness measurements have been carried out at supersmooth artefacts fabricated from BK7, fused silica, and Zerodur. The surface parameters were determined using a special prototype of the mechanical profiler Nanostep (Rank Taylor Hobson), the Optical Heterodyne Profiler Z5500 (Zygo), and an Atomic Force Microscope (Park Scientific) with an improved acquisition technique. The intercomparison was performed after the range of collected spatial wavelength for each instrument was adjusted using digital filtering techniques. It is demonstrated for different roughness ranges that are applied superpolishing techniques yield supersmooth artefacts which can be used for more intercomparisons.

Manipulation of mammalian cells using a single-fiber optical microbeam

PubMed Central

Mohanty, Samarendra K.; Mohanty, Khyati S.; Berns, Michael W.

2014-01-01

The short working distance of microscope objectives has severely restricted the application of optical micromanipulation techniques at larger depths. We show the first use of fiber-optic tweezers toward controlled guidance of neuronal growth cones and stretching of neurons. Further, by mode locking, the fiber-optic tweezers beam was converted to fiber-optic scissors, enabling dissection of neuronal processes and thus allowing study of the subsequent response of neurons to localized injury. At high average powers, lysis of a three-dimensionally trapped cell was accomplished. PMID:19021429

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

NASA Astrophysics Data System (ADS)

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

2014-11-01

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

Dual-mode optical microscope based on single-pixel imaging

NASA Astrophysics Data System (ADS)

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

2016-07-01

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

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

PubMed

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

2016-02-08

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

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

NASA Astrophysics Data System (ADS)

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

2016-02-01

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

Characterization of a subwavelength-scale 3D void structure using the FDTD-based confocal laser scanning microscopic image mapping technique.

PubMed

Choi, Kyongsik; Chon, James W; Gu, Min; Lee, Byoungho

2007-08-20

In this paper, a simple confocal laser scanning microscopic (CLSM) image mapping technique based on the finite-difference time domain (FDTD) calculation has been proposed and evaluated for characterization of a subwavelength-scale three-dimensional (3D) void structure fabricated inside polymer matrix. The FDTD simulation method adopts a focused Gaussian beam incident wave, Berenger's perfectly matched layer absorbing boundary condition, and the angular spectrum analysis method. Through the well matched simulation and experimental results of the xz-scanned 3D void structure, we first characterize the exact position and the topological shape factor of the subwavelength-scale void structure, which was fabricated by a tightly focused ultrashort pulse laser. The proposed CLSM image mapping technique based on the FDTD can be widely applied from the 3D near-field microscopic imaging, optical trapping, and evanescent wave phenomenon to the state-of-the-art bio- and nanophotonics.

Interferometric scanning optical microscope for surface characterization.

PubMed

Offside, M J; Somekh, M G

1992-11-01

A phase-sensitive scanning optical microscope is described that can measure surface height changes down to 0.1 nm. This is achieved by using two heterodyne Michelson interferometers in parallel. One interferometer probes the sample with a tightly focused beam, and the second has a collimated beam that illuminates a large area of the surface, providing a large area on sample reference. This is facilitated by using a specially constructed objective lens that permits the relative areas illuminated by the two probe beams to be varied both arbitrarily and independently, thus ensuring an accurate absolute phase measurement. We subtracted the phase outputs from each interferometer to provide the sample phase information, canceling the phase noise resulting from microphonics in the process. Results from a prototype version of the microscope are presented that demonstrate the advantages of the system over existing techniques.

Vapor-condensation-assisted optical microscopy for ultralong carbon nanotubes and other nanostructures.

PubMed

Wang, Jiangtao; Li, Tianyi; Xia, Bingyu; Jin, Xiang; Wei, Haoming; Wu, Wenyun; Wei, Yang; Wang, Jiaping; Liu, Peng; Zhang, Lina; Li, Qunqing; Fan, Shoushan; Jiang, Kaili

2014-06-11

Here we present a simple yet powerful approach for the imaging of nanostructures under an optical microscope with the help of vapor condensation on their surfaces. Supersaturated water vapor will first form a nanometer-sized water droplet on the condensation nuclei on the surface of nanostructures, and then the water droplet will grow bigger and scatter more light to make the outline of the nanostructure be visible under dark-field optical microscope. This vapor-condensation-assisted (VCA) optical microscopy is applicable to a variety of nanostructures from ultralong carbon nanotubes to functional groups, generating images with contrast coming from the difference in density of the condensation sites, and does not induce any impurities to the specimens. Moreover, this low-cost and efficient technique can be conveniently integrated with other facilities, such as Raman spectroscope and so forth, which will pave the way for widespread applications.

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.

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.

A Review of Adaptive Optics Optical Coherence Tomography: Technical Advances, Scientific Applications, and the Future

PubMed Central

Jonnal, Ravi S.; Kocaoglu, Omer P.; Zawadzki, Robert J.; Liu, Zhuolin; Miller, Donald T.; Werner, John S.

2016-01-01

Purpose Optical coherence tomography (OCT) has enabled “virtual biopsy” of the living human retina, revolutionizing both basic retina research and clinical practice over the past 25 years. For most of those years, in parallel, adaptive optics (AO) has been used to improve the transverse resolution of ophthalmoscopes to foster in vivo study of the retina at the microscopic level. Here, we review work done over the last 15 years to combine the microscopic transverse resolution of AO with the microscopic axial resolution of OCT, building AO-OCT systems with the highest three-dimensional resolution of any existing retinal imaging modality. Methods We surveyed the literature to identify the most influential antecedent work, important milestones in the development of AO-OCT technology, its applications that have yielded new knowledge, research areas into which it may productively expand, and nascent applications that have the potential to grow. Results Initial efforts focused on demonstrating three-dimensional resolution. Since then, many improvements have been made in resolution and speed, as well as other enhancements of acquisition and postprocessing techniques. Progress on these fronts has produced numerous discoveries about the anatomy, function, and optical properties of the retina. Conclusions Adaptive optics OCT continues to evolve technically and to contribute to our basic and clinical knowledge of the retina. Due to its capacity to reveal cellular and microscopic detail invisible to clinical OCT systems, it is an ideal companion to those instruments and has the demonstrable potential to produce images that can guide the interpretation of clinical findings. PMID:27409507

Single-shot optical sectioning using two-color probes in HiLo fluorescence microscopy.

PubMed

Muro, Eleonora; Vermeulen, Pierre; Ioannou, Andriani; Skourides, Paris; Dubertret, Benoit; Fragola, Alexandra; Loriette, Vincent

2011-06-08

We describe a wide-field fluorescence microscope setup which combines HiLo microscopy technique with the use of a two-color fluorescent probe. It allows one-shot fluorescence optical sectioning of thick biological moving sample which is illuminated simultaneously with a flat and a structured pattern at two different wavelengths. Both homogenous and structured fluorescence images are spectrally separated at detection and combined similarly with the HiLo microscopy technique. We present optically sectioned full-field images of Xenopus laevis embryos acquired at 25 images/s frame rate. Copyright © 2011 Biophysical Society. Published by Elsevier Inc. All rights reserved.

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.

Precise measurement of surface plasmon forces at a metal-dielectric interface using a calibrated evanescent wave

NASA Astrophysics Data System (ADS)

Liu, Lulu; Woolf, Alex

2015-03-01

By observing the motion of an optically trapped microscopic colloid, sub-piconewton static and dynamical forces have been measured using a technique called photonic force microscopy. This technique, though potentially powerful, has in the past struggled to make precise measurements in the vicinity of a reflective or metallic interface, due to distortions of the optical field. We introduce a new in-situ, contact-free calibration method for particle tracking using an evanescent wave, and demonstrate its expanded capability by the precise measurement of forces of interaction between a single colloid and the optical field generated by a propagating surface plasmon polariton on gold.

Microscopic Studies of Quantum Phase Transitions in Optical Lattices

NASA Astrophysics Data System (ADS)

Bakr, Waseem S.

2011-12-01

In this thesis, I report on experiments that microscopically probe quantum phase transitions of ultracold atoms in optical lattices. We have developed a "quantum gas microscope" that allowed, for the first time, optical imaging and manipulation of single atoms in a quantum-degenerate gas on individual sites of an optical lattice. This system acts as a quantum simulator of strongly correlated materials, which are currently the subject of intense research because of the technological potential of high--T c superconductors and spintronic materials. We have used our microscope to study the superfluid to Mott insulator transition in bosons and a magnetic quantum phase transition in a spin system. In our microscopic study of the superfluid-insulator transition, we have characterized the on-site number statistics in a space- and time-resolved manner. We observed Mott insulators with fidelities as high as 99%, corresponding to entropies of 0.06kB per particle. We also measured local quantum dynamics and directly imaged the shell structure of the Mott insulator. I report on the first quantum magnetism experiments in optical lattices. We have realized a quantum Ising chain in a magnetic field, and observed a quantum phase transition between a paramagnet and antiferromagnet. We achieved strong spin interactions by encoding spins in excitations of a Mott insulator in a tilted lattice. We detected the transition by measuring the total magnetization of the system across the transition using in-situ measurements as well as the Neel ordering in the antiferromagnetic state using noise-correlation techniques. We characterized the dynamics of domain formation in the system. The spin mapping introduced opens up a new path to realizing more exotic states in optical lattices including spin liquids and quantum valence bond solids. As our system sizes become larger, simulating their physics on classical computers will require exponentially larger resources because of entanglement build-up near a quantum phase transition. We have demonstrated a quantum simulator in which all degrees of freedom can be read out microscopically, allowing the simulation of quantum many-body systems with manageable resources. More generally, the ability to image and manipulate individual atoms in optical lattices opens an avenue towards scalable quantum computation.

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

Photothermal camera port accessory for microscopic thermal diffusivity imaging

NASA Astrophysics Data System (ADS)

Escola, Facundo Zaldívar; Kunik, Darío; Mingolo, Nelly; Martínez, Oscar Eduardo

2016-06-01

The design of a scanning photothermal accessory is presented, which can be attached to the camera port of commercial microscopes to measure thermal diffusivity maps with micrometer resolution. The device is based on the thermal expansion recovery technique, which measures the defocusing of a probe beam due to the curvature induced by the local heat delivered by a focused pump beam. The beam delivery and collecting optics are built using optical fiber technology, resulting in a robust optical system that provides collinear pump and probe beams without any alignment adjustment necessary. The quasiconfocal configuration for the signal collection using the same optical fiber sets very restrictive conditions on the positioning and alignment of the optical components of the scanning unit, and a detailed discussion of the design equations is presented. The alignment procedure is carefully described, resulting in a system so robust and stable that no further alignment is necessary for the day-to-day use, becoming a tool that can be used for routine quality control, operated by a trained technician.

Non linear optical investigations of silver nanoparticles synthesised by curcumin reduction

NASA Astrophysics Data System (ADS)

Dhanya, N. P.

2017-11-01

Metal nanoparticles have considerable applications in assorted fields like medicine, biology, photonics, metallurgy etc. Optical applications of Silver nanoparticles are of significant interest among researchers nowadays. In this paper, we report a single step chemical reduction of silver nanoparticles with Curcumin both as a reducing and stabilising agent at room temperature. Structural, plasmonic and non linear optical properties of the prepared nanoparticles are explored using Scanning Electron Microscope, Transmission Electron Microscope, UV absorption spectrometry, Spectroflurometry and Z scan. UV-Vis absorption studies affirm the Surface Plasmon Resonance (SPR) absorption and spectroflurometric studies announce the emission spectrum of the prepared silvernanoparticles at 520 nm. SEM and TEM images uphold the existence of uniform sized, spherical silvernanoparticles. Nonlinear optical studies are accomplished with the open aperture z scan technique in the nanosecond regime. The nonlinearity is in virtue of saturable absorption, two-photon absorption and excited state absorption. The marked nonlinearity and optical limiting of the Curcumin reduced silvernanoparticles enhances its photonic applications.

Re-scan confocal microscopy: scanning twice for better resolution.

PubMed

De Luca, Giulia M R; Breedijk, Ronald M P; Brandt, Rick A J; Zeelenberg, Christiaan H C; de Jong, Babette E; Timmermans, Wendy; Azar, Leila Nahidi; Hoebe, Ron A; Stallinga, Sjoerd; Manders, Erik M M

2013-01-01

We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required.

Interactions of Small-Scale Physical Mixing Processes with the Structure, Morphology and Bloom Dynamics and Optics of Non-Spheroid Phytoplankton

DTIC Science & Technology

2001-09-30

microscopic imaging techniques, and microscopic video- cinematography protocols for both phytoplankton and zooplankton for use in current laboratory...phytoplankton, zooplankton and bioluminescence papers, and examined data/figures for layered structures. Imaging and Cinematography : Off-the-shelf...to preview it as a work-in-progress, email me (jrines@gso.uri.edu), and I will provide you with a temporary URL. Imaging and Cinematography

Atomic force-multi-optical imaging integrated microscope for monitoring molecular dynamics in live cells.

PubMed

Trache, Andreea; Meininger, Gerald A

2005-01-01

A novel hybrid imaging system is constructed integrating atomic force microscopy (AFM) with a combination of optical imaging techniques that offer high spatial resolution. The main application of this instrument (the NanoFluor microscope) is the study of mechanotransduction with an emphasis on extracellular matrix-integrin-cytoskeletal interactions and their role in the cellular responses to changes in external chemical and mechanical factors. The AFM allows the quantitative assessment of cytoskeletal changes, binding probability, adhesion forces, and micromechanical properties of the cells, while the optical imaging applications allow thin sectioning of the cell body at the coverslip-cell interface, permitting the study of focal adhesions using total internal reflection fluorescence (TIRF) and internal reflection microscopy (IRM). Combined AFM-optical imaging experiments show that mechanical stimulation at the apical surface of cells induces a force-generating cytoskeletal response, resulting in focal contact reorganization on the basal surface that can be monitored in real time. The NanoFluor system is also equipped with a novel mechanically aligned dual camera acquisition system for synthesized Forster resonance energy transfer (FRET). The integrated NanoFluor microscope system is described, including its characteristics, applications, and limitations.

Fractal propagation method enables realistic optical microscopy simulations in biological tissues

PubMed Central

Glaser, Adam K.; Chen, Ye; Liu, Jonathan T.C.

2017-01-01

Current simulation methods for light transport in biological media have limited efficiency and realism when applied to three-dimensional microscopic light transport in biological tissues with refractive heterogeneities. We describe here a technique which combines a beam propagation method valid for modeling light transport in media with weak variations in refractive index, with a fractal model of refractive index turbulence. In contrast to standard simulation methods, this fractal propagation method (FPM) is able to accurately and efficiently simulate the diffraction effects of focused beams, as well as the microscopic heterogeneities present in tissue that result in scattering, refractive beam steering, and the aberration of beam foci. We validate the technique and the relationship between the FPM model parameters and conventional optical parameters used to describe tissues, and also demonstrate the method’s flexibility and robustness by examining the steering and distortion of Gaussian and Bessel beams in tissue with comparison to experimental data. We show that the FPM has utility for the accurate investigation and optimization of optical microscopy methods such as light-sheet, confocal, and nonlinear microscopy. PMID:28983499

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

PubMed

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

2014-12-11

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

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

PubMed Central

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

2017-01-01

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

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

NASA Astrophysics Data System (ADS)

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

2017-02-01

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

EDITORIAL: Recent developments in biomedical optics

NASA Astrophysics Data System (ADS)

Wang, Ruikang K.; Hebden, Jeremy C.; Tuchin, Valery V.

2004-04-01

The rapid growth in laser and photonic technology has resulted in new tools being proposed and developed for use in the medical and biological sciences. Specifically, a discipline known as biomedical optics has emerged which is providing a broad variety of optical techniques and instruments for diagnostic, therapeutic and basic science applications. New laser sources, detectors and measurement techniques are yielding powerful new methods for the study of diseases on all scales, from single molecules, to specific tissues and whole organs. For example, novel laser microscopes permit spectroscopic and force measurements to be performed on single protein molecules; new optical devices provide information on molecular dynamics and structure to perform `optical biopsy' non-invasively and almost instantaneously; and optical coherence tomography and diffuse optical tomography allow visualization of specific tissues and organs. Using genetic promoters to derive luciferase expression, bioluminescence methods can generate molecular light switches, which serve as functional indicator lights reporting cellular conditions and responses in living animals. This technique could allow rapid assessment of and response to the effects of anti-tumour drugs, antibiotics, or antiviral drugs. This issue of Physics in Medicine and Biology highlights recent research in biomedical optics, and is based on invited contributions to the International Conference on Advanced Laser Technology (Focused on Biomedical Optics) held at Cranfield University at Silsoe on 19--23 September 2003. This meeting included sessions devoted to: diffuse optical imaging and spectroscopy; optical coherence tomography and coherent domain techniques; optical sensing and applications in life science; microscopic, spectroscopic and opto-acoustic imaging; therapeutic and diagnostic applications; and laser interaction with organic and inorganic materials. Twenty-one papers are included in this special issue. The first paper gives an overview on the current status of scanning laser ophthalmoscopy and its role in bioscience and medicine, while the second paper describes the current problems in tissue engineering and the potential role for optical coherence tomography. The following seven papers present and discuss latest developments in infrared spectroscopy and diffuse optical tomography for medical diagnostics. Eight further papers report recent advances in optical coherence tomography, covering new and evolving methods and instrumentation, theoretical and numerical modelling, and its clinical applications. The remaining papers cover miscellaneous topics in biomedical optics, including new developments in opto-acoustic imaging techniques, laser speckle imaging of blood flow in microcirculations, and potential of hollow-core photonic-crystal fibres for laser dentistry. We thank all the authors for their valuable contributions and their prompt responses to reviewers' comments. We are also very grateful to the reviewers for their hard work and their considerable efforts to meet tight deadlines.

Real-time terahertz near-field microscope.

PubMed

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

2011-04-25

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

High Speed Photomicrography

NASA Astrophysics Data System (ADS)

Hyzer, William G.

1983-03-01

One of the most challenging areas in applying high-speed photography and videography in the plant and laboratory is in the recording of rapid events at macro and microscopic scales. Depth of field, exposure efficiency, working distance, and required exposure time are all reduced as optical magnification is increased, which severely taxes the skill and ingenuity of workers interested in recording any fast moving phenomena through the microscope or with magnifying lenses. This paper defines the problems inherent in photographing within macro and microscopic ranges and offers a systematic approach to optimizing the selection of equipment and choice of applicable techniques.

The Scanning Optical Microscope: An Overview

NASA Astrophysics Data System (ADS)

Kino, G. S.; Corte, T. R.; Xiao, G. Q.

1988-07-01

In the last few years there has been a resurgence in research on optical microscopes. One reason stems from the invention of the acoustic microscope by Quate and Lemons,1 and the realization that some of the same principles could be applied to the optical microscope. The acoustic microscope has better transverse definition for the same wavelength than the standard optical microscope and at the same time has far better range definition. Consequently, Kompfner, who was involved with the work on the early acoustic microscope, decided to try out similar scanning microscope principles with optics, and started a group with Wilson and Sheppard to carry out such research at Oxford.2 Sometime earlier, Petran et a13 had invented the tandem scanning microscope which used many of the same principles. Now, in our laboratory at Stanford, these ideas on the tandem scanning microscope and the scanning optical microscope are converging. Another aspect of this work, which stems from the earlier experience with the acoustic microscope, involves measurement of both phase and amplitude of the optical beam. It is also possible to use scanned optical microscopy for other purposes. For instance, an optical beam can be used to excite electrons and holes in semiconductors, and the generated current can be measured. By scanning the optical beam over the semiconductor, an image can be obtained of the regions where there is strong or weak electron hole generation. This type of microscope is called OBIC (Optical Beam Induced Current). A second application involves fluorescent imaging of biological materials. Here we have the excellent range definition of a scanning optical microscope which eliminates unwanted glare from regions of the material where the beam is unfocused.3 A third application is focused on the heating effect of the light beam. With such a system, images can be obtained which are associated with changes in the thermal properties of a material, changes in recombination rates in semiconductors, and differences in material properties associated with either acoustic or thermal effects.4,5 Thus, the range of scanning optical microscopy applications is very large. In the main, the most important applications have been to semiconductors and to biology.

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.

Controlling dispersion of graphene nanoplatelets in aqueous solution by ultrasonic technique

NASA Astrophysics Data System (ADS)

Wang, Baomin; Jiang, Ruishuang; Song, Wanzeng; Liu, Hui

2017-08-01

The homogenous graphene nanoplatelets (GNP) suspension had been prepared through ultrasonic exfoliation in the presence of methylcellulose (MC) as dispersant. The influence of different sonication times on dispersing of aqueous GNP suspension was monitored by UV-Vis absorbance, sedimentation test, optical microscope and transmission electron microscope (TEM). The study of UV-Vis absorbance verifies that the minimum sonication time to break the 0.1 g/L concentration of bundled GNPs is 20 min; furthermore, the GNP suspension achieved the best dispersion, when sonication time increased up to 80 min. From optical microscope images of GNPs, the agglomeration of GNPs was broken by enough sonication energy, and the distribution of GNPs particles became more uniform. The dispersing mechanism had been discussed and simulated by HRTEM image. The bundled GNPs were exfoliated by cavitation effect of ultrasonic irradiation, meanwhile, the dispersant adsorbed on the surface of GNPs prevented re-entanglement by forming steric hindrance.

Practical guidelines for implementing adaptive optics in fluorescence microscopy

NASA Astrophysics Data System (ADS)

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

2018-02-01

In life sciences, interest in the microscopic imaging of increasingly complex three dimensional samples, such as cell spheroids, zebrafish embryos, and in vivo applications in small animals, is growing quickly. Due to the increasing complexity of samples, more and more life scientists are considering the implementation of adaptive optics in their experimental setups. While several approaches to adaptive optics in microscopy have been reported, it is often difficult and confusing for the microscopist to choose from the array of techniques and equipment. In this poster presentation we offer a small guide to adaptive optics providing general guidelines for successful adaptive optics implementation.

Two-Photon Imaging with Diffractive Optical Elements

PubMed Central

Watson, Brendon O.; Nikolenko, Volodymyr; Yuste, Rafael

2009-01-01

Two-photon imaging has become a useful tool for optical monitoring of neural circuits, but it requires high laser power and serial scanning of each pixel in a sample. This results in slow imaging rates, limiting the measurements of fast signals such as neuronal activity. To improve the speed and signal-to-noise ratio of two-photon imaging, we introduce a simple modification of a two-photon microscope, using a diffractive optical element (DOE) which splits the laser beam into several beamlets that can simultaneously scan the sample. We demonstrate the advantages of DOE scanning by enhancing the speed and sensitivity of two-photon calcium imaging of action potentials in neurons from neocortical brain slices. DOE scanning can easily improve the detection of time-varying signals in two-photon and other non-linear microscopic techniques. PMID:19636390

RMB identification based on polarization parameters inversion imaging

NASA Astrophysics Data System (ADS)

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

2016-10-01

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

Combinatorial materials synthesis and high-throughput screening: an integrated materials chip approach to mapping phase diagrams and discovery and optimization of functional materials.

PubMed

Xiang, X D

Combinatorial materials synthesis methods and high-throughput evaluation techniques have been developed to accelerate the process of materials discovery and optimization and phase-diagram mapping. Analogous to integrated circuit chips, integrated materials chips containing thousands of discrete different compositions or continuous phase diagrams, often in the form of high-quality epitaxial thin films, can be fabricated and screened for interesting properties. Microspot x-ray method, various optical measurement techniques, and a novel evanescent microwave microscope have been used to characterize the structural, optical, magnetic, and electrical properties of samples on the materials chips. These techniques are routinely used to discover/optimize and map phase diagrams of ferroelectric, dielectric, optical, magnetic, and superconducting materials.

Production and characterization of pure cryogenic inertial fusion targets

NASA Astrophysics Data System (ADS)

Boyd, B. A.; Kamerman, G. W.

An experimental cryogenic inertial fusion target generator and two optical techniques for automated target inspection are described. The generator produces 100 microns diameter solid hydrogen spheres at a rate compatible with fueling requirements of conceptual inertial fusion power plants. A jet of liquified hydrogen is disrupted into droplets by an ultrasonically excited nozzle. The droplets solidify into microspheres while falling through a chamber maintained below the hydrogen triple point pressure. Stable operation of the generator has been demonstrated for up to three hours. The optical inspection techniques are computer aided photomicrography and coarse diffraction pattern analysis (CDPA). The photomicrography system uses a conventional microscope coupled to a computer by a solid state camera and digital image memory. The computer enhances the stored image and performs feature extraction to determine pellet parameters. The CDPA technique uses Fourier transform optics and a special detector array to perform optical processing of a target image.

Thermal radiation scanning tunnelling microscopy

NASA Astrophysics Data System (ADS)

de Wilde, Yannick; Formanek, Florian; Carminati, Rémi; Gralak, Boris; Lemoine, Paul-Arthur; Joulain, Karl; Mulet, Jean-Philippe; Chen, Yong; Greffet, Jean-Jacques

2006-12-01

In standard near-field scanning optical microscopy (NSOM), a subwavelength probe acts as an optical `stethoscope' to map the near field produced at the sample surface by external illumination. This technique has been applied using visible, infrared, terahertz and gigahertz radiation to illuminate the sample, providing a resolution well beyond the diffraction limit. NSOM is well suited to study surface waves such as surface plasmons or surface-phonon polaritons. Using an aperture NSOM with visible laser illumination, a near-field interference pattern around a corral structure has been observed, whose features were similar to the scanning tunnelling microscope image of the electronic waves in a quantum corral. Here we describe an infrared NSOM that operates without any external illumination: it is a near-field analogue of a night-vision camera, making use of the thermal infrared evanescent fields emitted by the surface, and behaves as an optical scanning tunnelling microscope. We therefore term this instrument a `thermal radiation scanning tunnelling microscope' (TRSTM). We show the first TRSTM images of thermally excited surface plasmons, and demonstrate spatial coherence effects in near-field thermal emission.

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

Miranda, Adelaide; De Beule, Pieter A. A., E-mail: pieter.de-beule@inl.int; Martins, Marco

Combined microscopy techniques offer the life science research community a powerful tool to investigate complex biological systems and their interactions. Here, we present a new combined microscopy platform based on fluorescence optical sectioning microscopy through aperture correlation microscopy with a Differential Spinning Disk (DSD) and nanomechanical mapping with an Atomic Force Microscope (AFM). The illumination scheme of the DSD microscope unit, contrary to standard single or multi-point confocal microscopes, provides a time-independent illumination of the AFM cantilever. This enables a distortion-free simultaneous operation of fluorescence optical sectioning microscopy and atomic force microscopy with standard probes. In this context, we discussmore » sample heating due to AFM cantilever illumination with fluorescence excitation light. Integration of a DSD fluorescence optical sectioning unit with an AFM platform requires mitigation of mechanical noise transfer of the spinning disk. We identify and present two solutions to almost annul this noise in the AFM measurement process. The new combined microscopy platform is applied to the characterization of a DOPC/DOPS (4:1) lipid structures labelled with a lipophilic cationic indocarbocyanine dye deposited on a mica substrate.« less

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

NASA Astrophysics Data System (ADS)

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

2009-07-01

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

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

NASA Astrophysics Data System (ADS)

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

2017-01-01

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

Re-scan confocal microscopy: scanning twice for better resolution

PubMed Central

De Luca, Giulia M.R.; Breedijk, Ronald M.P.; Brandt, Rick A.J.; Zeelenberg, Christiaan H.C.; de Jong, Babette E.; Timmermans, Wendy; Azar, Leila Nahidi; Hoebe, Ron A.; Stallinga, Sjoerd; Manders, Erik M.M.

2013-01-01

We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required. PMID:24298422

Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections

NASA Astrophysics Data System (ADS)

DeArmond, Fredrick Michael

As optical microscopy techniques continue to improve, most notably the development of super-resolution optical microscopy which garnered the Nobel Prize in Chemistry in 2014, renewed emphasis has been placed on the development and use of fluorescence microscopy techniques. Of particular note is a renewed interest in multiphoton excitation due to a number of inherent properties of the technique including simplified optical filtering, increased sample penetration, and inherently confocal operation. With this renewed interest in multiphoton fluorescence microscopy, comes an increased demand for robust non-linear fluorescent markers, and characterization of the associated tool set. These factors have led to an experimental setup to allow a systematized approach for identifying and characterizing properties of fluorescent probes in the hopes that the tool set will provide researchers with additional information to guide their efforts in developing novel fluorophores suitable for use in advanced optical microscopy techniques as well as identifying trends for their synthesis. Hardware was setup around a software control system previously developed. Three experimental tool sets were set up, characterized, and applied over the course of this work. These tools include scanning multiphoton fluorescence microscope with single molecule sensitivity, an interferometric autocorrelator for precise determination of the bandwidth and pulse width of the ultrafast Titanium Sapphire excitation source, and a simplified fluorescence microscope for the measurement of two-photon absorption cross sections. Resulting values for two-photon absorption cross sections and two-photon absorption action cross sections for two standardized fluorophores, four commercially available fluorophores, and ten novel fluorophores are presented as well as absorption and emission spectra.

Introduction to the virtual special issue on super-resolution imaging techniques

NASA Astrophysics Data System (ADS)

Cao, Liangcai; Liu, Zhengjun

2017-12-01

Until quite recently, the resolution of optical imaging instruments, including telescopes, cameras and microscopes, was considered to be limited by the diffraction of light and by image sensors. In the past few years, many exciting super-resolution approaches have emerged that demonstrate intriguing ways to bypass the classical limit in optics and detectors. More and more research groups are engaged in the study of advanced super-resolution schemes, devices, algorithms, systems, and applications [1-6]. Super-resolution techniques involve new methods in science and engineering of optics [7,8], measurements [9,10], chemistry [11,12] and information [13,14]. Promising applications, particularly in biomedical research and semiconductor industry, have been successfully demonstrated.

Method to deterministically study photonic nanostructures in different experimental instruments.

PubMed

Husken, B H; Woldering, L A; Blum, C; Vos, W L

2009-01-01

We describe an experimental method to recover a single, deterministically fabricated nanostructure in various experimental instruments without the use of artificially fabricated markers, with the aim to study photonic structures. Therefore, a detailed map of the spatial surroundings of the nanostructure is made during the fabrication of the structure. These maps are made using a series of micrographs with successively decreasing magnifications. The graphs reveal intrinsic and characteristic geometric features that can subsequently be used in different setups to act as markers. As an illustration, we probe surface cavities with radii of 65 nm on a silica opal photonic crystal with various setups: a focused ion beam workstation; a scanning electron microscope (SEM); a wide field optical microscope and a confocal microscope. We use cross-correlation techniques to recover a small area imaged with the SEM in a large area photographed with the optical microscope, which provides a possible avenue to automatic searching. We show how both structural and optical reflectivity data can be obtained from one and the same nanostructure. Since our approach does not use artificial grids or markers, it is of particular interest for samples whose structure is not known a priori, like samples created solely by self-assembly. In addition, our method is not restricted to conducting samples.

[Intraoperative Optical Coherence Tomography (MI-OCT) for the Treatment of Corneal Dystrophies].

PubMed

Siebelmann, Sebastian; Matthaei, Mario; Heindl, Ludwig M; Bachmann, Björn O; Cursiefen, Claus

2018-06-01

The surgical treatment of corneal dystrophies develops rapidly as the use of lamellar corneal grafting techniques continue. While penetrating keratoplasty was the gold standard for treating a variety of dystrophies a few years ago, the affected layers of the cornea can, nowadays, be selectively replaced or ablated using laser technology. Of particular importance for these methods is optical coherence tomography, which has recently been integrated into surgical microscopes (MI-OCT). Literature overview from PubMed and Google.scholar.de supplemented with own imaging data. The MI-OCT enables the intraoperative real-time monitoring of different ophthalmic surgical procedures, such as deep anterior lamellar keratoplasty, Descemet's membrane endothelial keratoplasty, as well as minimally-invasive procedures, such as phototherapeutic keratectomy. In addition, it enables an evaluation of the cornea, but also of structures of the anterior chamber, in situations in which the cornea, for example, is clouded by an edema. Microscope-integrated, intraoperative optical coherence tomography (MI-OCT) represents a useful supplement to the normal surgical microscope. It is superior to the sole surgical microscope, especially in already severely clouded corneas, and represents a sensible supplement, especially for novel lamellar transplantation procedures. Prospective randomized trials are necessary to increase safety and efficacy when using MI-OCT for different indications. Georg Thieme Verlag KG Stuttgart · New York.

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection.

PubMed

Mertz, Jerome; Kim, Jinhyun

2010-01-01

It is well known that light-sheet illumination can enable optically sectioned wide-field imaging of macroscopic samples. However, the optical sectioning capacity of a light-sheet macroscope is undermined by sample-induced scattering or aberrations that broaden the thickness of the sheet illumination. We present a technique to enhance the optical sectioning capacity of a scanning light-sheet microscope by out-of-focus background rejection. The technique, called HiLo microscopy, makes use of two images sequentially acquired with uniform and structured sheet illumination. An optically sectioned image is then synthesized by fusing high and low spatial frequency information from both images. The benefits of combining light-sheet macroscopy and HiLo background rejection are demonstrated in optically cleared whole mouse brain samples, using both green fluorescent protein (GFP)-fluorescence and dark-field scattered light contrast.

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection

NASA Astrophysics Data System (ADS)

Mertz, Jerome; Kim, Jinhyun

2010-01-01

It is well known that light-sheet illumination can enable optically sectioned wide-field imaging of macroscopic samples. However, the optical sectioning capacity of a light-sheet macroscope is undermined by sample-induced scattering or aberrations that broaden the thickness of the sheet illumination. We present a technique to enhance the optical sectioning capacity of a scanning light-sheet microscope by out-of-focus background rejection. The technique, called HiLo microscopy, makes use of two images sequentially acquired with uniform and structured sheet illumination. An optically sectioned image is then synthesized by fusing high and low spatial frequency information from both images. The benefits of combining light-sheet macroscopy and HiLo background rejection are demonstrated in optically cleared whole mouse brain samples, using both green fluorescent protein (GFP)-fluorescence and dark-field scattered light contrast.

Seamless stitching of tile scan microscope images.

PubMed

Legesse, F B; Chernavskaia, O; Heuke, S; Bocklitz, T; Meyer, T; Popp, J; Heintzmann, R

2015-06-01

For diagnostic purposes, optical imaging techniques need to obtain high-resolution images of extended biological specimens in reasonable time. The field of view of an objective lens, however, is often smaller than the sample size. To image the whole sample, laser scanning microscopes acquire tile scans that are stitched into larger mosaics. The appearance of such image mosaics is affected by visible edge artefacts that arise from various optical aberrations which manifest in grey level jumps across tile boundaries. In this contribution, a technique for stitching tiles into a seamless mosaic is presented. The stitching algorithm operates by equilibrating neighbouring edges and forcing the brightness at corners to a common value. The corrected image mosaics appear to be free from stitching artefacts and are, therefore, suited for further image analysis procedures. The contribution presents a novel method to seamlessly stitch tiles captured by a laser scanning microscope into a large mosaic. The motivation for the work is the failure of currently existing methods for stitching nonlinear, multimodal images captured by our microscopic setups. Our method eliminates the visible edge artefacts that appear between neighbouring tiles by taking into account the overall illumination differences among tiles in such mosaics. The algorithm first corrects the nonuniform brightness that exists within each of the tiles. It then compensates for grey level differences across tile boundaries by equilibrating neighbouring edges and forcing the brightness at the corners to a common value. After these artefacts have been removed further image analysis procedures can be applied on the microscopic images. Even though the solution presented here is tailored for the aforementioned specific case, it could be easily adapted to other contexts where image tiles are assembled into mosaics such as in astronomical or satellite photos. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Development of x-ray laminography under an x-ray microscopic condition

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

Hoshino, Masato; Uesugi, Kentaro; Takeuchi, Akihisa

2011-07-15

An x-ray laminography system under an x-ray microscopic condition was developed to obtain a three-dimensional structure of laterally-extended planar objects which were difficult to observe by x-ray tomography. An x-ray laminography technique was introduced to an x-ray transmission microscope with zone plate optics. Three prototype sample holders were evaluated for x-ray imaging laminography. Layered copper grid sheets were imaged as a laminated sample. Diatomite powder on a silicon nitride membrane was measured to confirm the applicability of this method to non-planar micro-specimens placed on the membrane. The three-dimensional information of diatom shells on the membrane was obtained at a spatialmore » resolution of sub-micron. Images of biological cells on the membrane were also obtained by using a Zernike phase contrast technique.« less

3-D optical profilometry at micron scale with multi-frequency fringe projection using modified fibre optic Lloyd's mirror technique

NASA Astrophysics Data System (ADS)

Inanç, Arda; Kösoğlu, Gülşen; Yüksel, Heba; Naci Inci, Mehmet

2018-06-01

A new fibre optic Lloyd's mirror method is developed for extracting 3-D height distribution of various objects at the micron scale with a resolution of 4 μm. The fibre optic assembly is elegantly integrated to an optical microscope and a CCD camera. It is demonstrated that the proposed technique is quite suitable and practical to produce an interference pattern with an adjustable frequency. By increasing the distance between the fibre and the mirror with a micrometre stage in the Lloyd's mirror assembly, the separation between the two bright fringes is lowered down to the micron scale without using any additional elements as part of the optical projection unit. A fibre optic cable, whose polymer jacket is partially stripped, and a microfluidic channel are used as test objects to extract their surface topographies. Point by point sensitivity of the method is found to be around 8 μm, changing a couple of microns depending on the fringe frequency and the measured height. A straightforward calibration procedure for the phase to height conversion is also introduced by making use of the vertical moving stage of the optical microscope. The phase analysis of the acquired image is carried out by One Dimensional Continuous Wavelet Transform for which the chosen wavelet is the Morlet wavelet and the carrier removal of the projected fringe patterns is achieved by reference subtraction. Furthermore, flexible multi-frequency property of the proposed method allows measuring discontinuous heights where there are phase ambiguities like 2π by lowering the fringe frequency and eliminating the phase ambiguity.

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

PubMed

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

2017-12-01

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

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

NASA Astrophysics Data System (ADS)

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

2017-12-01

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

Diffraction Pattern Analysis as an Optical Inspection Technique

DTIC Science & Technology

1991-08-01

BACKGROUND Diameters of fiber samples have commonly been measured manually with an optical microscope. Marcuse and Presby developed an automatic...by analyzing the back-scattered light when a beam of laser light impinged upon the fiber [2]. Presby and Marcuse extended this back-scattering tech...be im- proved further in order to become a feasible method for detecting a small number of blocked openings in CRT screens. 20 REFERENCES 1. Marcuse

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

PubMed

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

2018-05-16

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

Monitoring damage growth in titanium matrix composites using acoustic emission

NASA Technical Reports Server (NTRS)

Bakuckas, J. G., Jr.; Prosser, W. H.; Johnson, W. S.

1993-01-01

The application of the acoustic emission (AE) technique to locate and monitor damage growth in titanium matrix composites (TMC) was investigated. Damage growth was studied using several optical techniques including a long focal length, high magnification microscope system with image acquisition capabilities. Fracture surface examinations were conducted using a scanning electron microscope (SEM). The AE technique was used to locate damage based on the arrival times of AE events between two sensors. Using model specimens exhibiting a dominant failure mechanism, correlations were established between the observed damage growth mechanisms and the AE results in terms of the events amplitude. These correlations were used to monitor the damage growth process in laminates exhibiting multiple modes of damage. Results revealed that the AE technique is a viable and effective tool to monitor damage growth in TMC.

Measurements and properties of ice particles and carbon dioxide bubbles in aqueous mixture utilizing optical techniques

NASA Astrophysics Data System (ADS)

Diallo, Amadou O.

Optical techniques are used to determine the size, shape and many other properties of particles ranging from the micro to a nano-level. These techniques have endless applications. This research is based on a project assigned by a "Vendor" that wants anonymity. The Leica optical microscope and the Dark Field Polarizing Metallurgical Microscope is used to determine the size and count of ice crystals (Vendors products) in multiple time frames. Since the ice temperature influences, its symmetry and the shape is subject to changes at room temperature (300 K) and the atmospheric pressure that is exerted on the ice crystals varies. The ice crystals are in a mixture of water, electrolytes and carbon dioxide with the optical spectroscopy (Qpod2) and Spectra suite, the optical density of the ice crystals is established from the absorbance and transmission measurements. The optical density in this case is also referred to as absorption; it is plotted with respect to a frequency (GHz), wavelength (nm) or Raman shift (1/cm) which shows the light colliding with the ice particles and CO2. Depending on the peaks positions, it is possible to profile the ice crystal sizes using a mean distribution plots. The region of absorbency wavelength expected for the ice is in the visible range; the water molecules in the (UV) Ultra-violet range and the CO2 in the (IR) infrared region. It is also possible to obtain the reflection and transmission output as a percentage change with the wavelengths ranging from 200 to 1100 nm. The refractive index of the ice can be correlated to the density based on the optical acoustic theorem, or Mie Scattering Theory. The viscosity of the ice crystals and the solutions from which the ice crystals are made of as well are recorded with the SV-10 viscometer. The baseline viscosity is used as reference and set lower than that of the ice crystals. The Zeta potential of the particles present in the mixture are approximated by first finding the viscosity of the solution where the pH level contribute to the surface charges, afterward use Stoke's diameter to compute the settling velocity of the bubbles, or alternatively record it under the microscope. With those parameters in hand the surface charge of the bubble (zeta potential) is approximated.

Nanopatterning of optical surfaces during low-energy ion beam sputtering

NASA Astrophysics Data System (ADS)

Liao, Wenlin; Dai, Yifan; Xie, Xuhui

2014-06-01

Ion beam figuring (IBF) provides a highly deterministic method for high-precision optical surface fabrication, whereas ion-induced microscopic morphology evolution would occur on surfaces. Consequently, the fabrication specification for surface smoothness must be seriously considered during the IBF process. In this work, low-energy ion nanopatterning of our frequently used optical material surfaces is investigated to discuss the manufacturability of an ultrasmooth surface. The research results indicate that ion beam sputtering (IBS) can directly smooth some amorphous or amorphizable material surfaces, such as fused silica, Si, and ULE under appropriate processing conditions. However, for IBS of a Zerodur surface, preferential sputtering together with curvature-dependent sputtering overcome ion-induced smoothing mechanisms, leading to the granular nanopatterns' formation and the coarsening of the surface. Furthermore, the material property difference at microscopic scales and the continuous impurity incorporation would affect the ion beam smoothing of optical surfaces. Overall, IBS can be used as a promising technique for ultrasmooth surface fabrication, which strongly depends on processing conditions and material characters.

Multifocal Fluorescence Microscope for Fast Optical Recordings of Neuronal Action Potentials

PubMed Central

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

2015-01-01

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

L-Phenylalanine functionalized silver nanoparticles: Photocatalytic and nonlinear optical applications

NASA Astrophysics Data System (ADS)

Nidya, M.; Umadevi, M.; Sankar, Pranitha; Philip, Reji; Rajkumar, Beulah J. M.

2015-04-01

An extensive study on the behavior of L-Phenylalanine capped silver nanoparticles (Phe-Ag NPs) in the aqueous phase and in a sol-gel thin film showed different UV/Vis, Transmission Electron Microscope (TEM), Dynamic Light Scattering and Zeta potential profiles. Scanning Electron Microscope (SEM) images of the samples in the sol gel film showed Ag embedded in the SiO2 matrix. Surface Enhanced Raman Spectra (SERS) confirmed that both in the aqueous media and in the sol gel film, the attachment of Phe to the Ag NP surface was through the benzene ring, with the sol-gel film showing a better enhancement. Photocatalytic degradation of crystal violet was measured spectrophotometrically using Phe-Ag NPs as a nanocatalyst under visible light illumination. Intensity-dependent nonlinear optical absorption of Phe-Ag measured using the open aperture Z-scan technique revealed that the material is an efficient optical limiter with potential applications.

Positron studies in catalysis research

NASA Astrophysics Data System (ADS)

During the past eight months, the authors have made progress in several areas relevant to the eventual use of positron techniques in catalysis research. They have come closer to the completion of their positron microscope, and at the same time have performed several studies in their non-microscopic positron spectrometer which should ultimately be applicable to catalysis. The current status of the efforts in each of these areas is summarized in the following sections: Construction of the positron microscope (optical element construction, data collection software, and electronic sub-assemblies); Doppler broadening spectroscopy of metal silicide; Positron lifetime spectroscopy of glassy polymers; and Positron lifetime measurements of pore-sizes in zeolites.

Development of 1500mm Wide Wrought Magnesium Alloys by Twin Roll Casting Technique in Turkey

NASA Astrophysics Data System (ADS)

Duygulu, Ozgur; Ucuncuoglu, Selda; Oktay, Gizem; Temur, Deniz Sultan; Yucel, Onuralp; Kaya, Ali Arslan

Magnesium alloy AZ31, AZ61, AZ91, AM50 and AM60 sheets were produced by twin roll casting first time in Turkey. Sheets of 4.5-6.5mm thick and 1500mm width were successfully achieved. Microstructure of the sheet was analyzed by optical microscope, scanning electron microscope (SEM) and transmission electron microscope (TEM). Semi-quantitative analyses were performed by SEM-EDS. In addition, X-ray studies were performed for both characterization and texture purposes. Mechanical properties were investigated by tensile tests and also hardness measurements. Homogenization and annealing heat treatments were performed on the produced sheets.

Near-field microscopy and fluorescence spectroscopy: application to chromosomes labelled with different fluorophores.

PubMed

Mahieu-Williame, L; Falgayrettes, P; Nativel, L; Gall-Borrut, P; Costa, L; Salehzada, T; Bisbal, C

2010-04-01

We have coupled a spectrophotometer with a scanning near-field optical microscope to obtain, with a single scan, simultaneously scanning near-field optical microscope fluorescence images at different wavelengths as well as topography and transmission images. Extraction of the fluorescence spectra enabled us to decompose the different wavelengths of the fluorescence signals which normally overlap. We thus obtained images of the different fluorescence emissions of acridine orange bound to single or double stranded nucleic acids in human metaphase chromosomes before and after DNAse I or RNAse A treatment. The analysis of these images allowed us to visualize some specific chromatin areas where RNA is associated with DNA showing that such a technique could be used to identify multiple components within a cell.

On-Chip Biomedical Imaging

PubMed Central

Göröcs, Zoltán; Ozcan, Aydogan

2012-01-01

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

Manufacture and calibration of optical supersmooth roughness artifacts for intercomparisons

NASA Astrophysics Data System (ADS)

Ringel, Gabriele A.; Kratz, Frank; Schmitt, Dirk-Roger; Mangelsdorf, Juergen; Creuzet, Francois; Garratt, John D.

1995-09-01

Intercomparison roughness measurements have been carried out on supersmooth artifacts fabricated from BK7, fused silica, and Zerodur. The surface parameters were determined using the optical heterodyne profiler Z5500 (Zygo), a special prototype of the mechanical profiler Nanostep (Rank Taylor Hobson), and an Atomic Force Microscope (Park Scientific Instruments) with an improved acquisition technique. The intercomparison was performed after the range of collected spatial wavelengths for each instrument was adjusted using digital filtering techniques. It is demonstrated for different roughness ranges that the applied superpolishing techniques yield supersmooth artifacts which can be used for more intercomparisons. More than 100 samples were investigated. Criteria were developed to select artifacts from the sample stock.

Hybrid Microscopy: Enabling Inexpensive High-Performance Imaging through Combined Physical and Optical Magnifications

NASA Astrophysics Data System (ADS)

Zhang, Yu Shrike; Chang, Jae-Byum; Alvarez, Mario Moisés; Trujillo-de Santiago, Grissel; Aleman, Julio; Batzaya, Byambaa; Krishnadoss, Vaishali; Ramanujam, Aishwarya Aravamudhan; Kazemzadeh-Narbat, Mehdi; Chen, Fei; Tillberg, Paul W.; Dokmeci, Mehmet Remzi; Boyden, Edward S.; Khademhosseini, Ali

2016-03-01

To date, much effort has been expended on making high-performance microscopes through better instrumentation. Recently, it was discovered that physical magnification of specimens was possible, through a technique called expansion microscopy (ExM), raising the question of whether physical magnification, coupled to inexpensive optics, could together match the performance of high-end optical equipment, at a tiny fraction of the price. Here we show that such “hybrid microscopy” methods—combining physical and optical magnifications—can indeed achieve high performance at low cost. By physically magnifying objects, then imaging them on cheap miniature fluorescence microscopes (“mini-microscopes”), it is possible to image at a resolution comparable to that previously attainable only with benchtop microscopes that present costs orders of magnitude higher. We believe that this unprecedented hybrid technology that combines expansion microscopy, based on physical magnification, and mini-microscopy, relying on conventional optics—a process we refer to as Expansion Mini-Microscopy (ExMM)—is a highly promising alternative method for performing cost-effective, high-resolution imaging of biological samples. With further advancement of the technology, we believe that ExMM will find widespread applications for high-resolution imaging particularly in research and healthcare scenarios in undeveloped countries or remote places.

Real-time restoration of white-light confocal microscope optical sections

PubMed Central

Balasubramanian, Madhusudhanan; Iyengar, S. Sitharama; Beuerman, Roger W.; Reynaud, Juan; Wolenski, Peter

2009-01-01

Confocal microscopes (CM) are routinely used for building 3-D images of microscopic structures. Nonideal imaging conditions in a white-light CM introduce additive noise and blur. The optical section images need to be restored prior to quantitative analysis. We present an adaptive noise filtering technique using Karhunen–Loéve expansion (KLE) by the method of snapshots and a ringing metric to quantify the ringing artifacts introduced in the images restored at various iterations of iterative Lucy–Richardson deconvolution algorithm. The KLE provides a set of basis functions that comprise the optimal linear basis for an ensemble of empirical observations. We show that most of the noise in the scene can be removed by reconstructing the images using the KLE basis vector with the largest eigenvalue. The prefiltering scheme presented is faster and does not require prior knowledge about image noise. Optical sections processed using the KLE prefilter can be restored using a simple inverse restoration algorithm; thus, the methodology is suitable for real-time image restoration applications. The KLE image prefilter outperforms the temporal-average prefilter in restoring CM optical sections. The ringing metric developed uses simple binary morphological operations to quantify the ringing artifacts and confirms with the visual observation of ringing artifacts in the restored images. PMID:20186290

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

PubMed

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

2008-07-01

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

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.

Laser scanning confocal microscopy: history, applications, and related optical sectioning techniques.

PubMed

Paddock, Stephen W; Eliceiri, Kevin W

2014-01-01

Confocal microscopy is an established light microscopical technique for imaging fluorescently labeled specimens with significant three-dimensional structure. Applications of confocal microscopy in the biomedical sciences include the imaging of the spatial distribution of macromolecules in either fixed or living cells, the automated collection of 3D data, the imaging of multiple labeled specimens and the measurement of physiological events in living cells. The laser scanning confocal microscope continues to be chosen for most routine work although a number of instruments have been developed for more specific applications. Significant improvements have been made to all areas of the confocal approach, not only to the instruments themselves, but also to the protocols of specimen preparation, to the analysis, the display, the reproduction, sharing and management of confocal images using bioinformatics techniques.

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

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

Spangenberg, Thomas; Goettlicher, Joerg; Steininger, Ralph

2009-01-29

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

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection

PubMed Central

Mertz, Jerome; Kim, Jinhyun

2010-01-01

It is well known that light-sheet illumination can enable optically sectioned wide-field imaging of macroscopic samples. However, the optical sectioning capacity of a light-sheet macroscope is undermined by sample-induced scattering or aberrations that broaden the thickness of the sheet illumination. We present a technique to enhance the optical sectioning capacity of a scanning light-sheet microscope by out-of-focus background rejection. The technique, called HiLo microscopy, makes use of two images sequentially acquired with uniform and structured sheet illumination. An optically sectioned image is then synthesized by fusing high and low spatial frequency information from both images. The benefits of combining light-sheet macroscopy and HiLo background rejection are demonstrated in optically cleared whole mouse brain samples, using both green fluorescent protein (GFP)-fluorescence and dark-field scattered light contrast. PMID:20210471

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2010-06-29

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P.; Chernobrod, Boris M.

2009-11-10

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of impaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P.; Chernobrod, Boris M.

2007-12-11

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2010-07-13

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2009-10-27

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Microstructures and properties of rapidly solidified alloys

NASA Technical Reports Server (NTRS)

Shechtman, D.; Horowitz, E.

1984-01-01

The microstructure and properties of rapidly solidified aluminum alloys were researched. The effects of powder and flake chemistry and morphology and alternative consolidation processing parameters are being conducted. Samples of the powders being utilized were obtained for comprehensive metallurgical characterization. Seven aluminum alloys in the form of thin foils were studied by a variety of techniques including optical metallography, scanning electron microscope, and transmission electron microscope. Details of the microstructural characteristics are presented along with a discussion of the solidification process. A better understanding of the microstructure of the rapidly solidified aluminum alloys prepared by a variety of techniques such as roller quenching, the vacuum atomized procedure, ultrasonically atomized in inert atmospheres, and atomized in flue gas was provided.

In situ focus characterization by ablation technique to enable optics alignment at an XUV FEL source

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

Gerasimova, N.; Dziarzhytski, S.; Weigelt, H.

2013-06-15

In situ focus characterization is demonstrated by working at an extreme ultraviolet (XUV) free-electron laser source using ablation technique. Design of the instrument reported here allows reaching a few micrometres resolution along with keeping the ultrahigh vacuum conditions and ensures high-contrast visibility of ablative imprints on optically transparent samples, e.g., PMMA. This enables on-line monitoring of the beam profile changes and thus makes possible in situ alignment of the XUV focusing optics. A good agreement between focal characterizations retrieved from in situ inspection of ablative imprints contours and from well-established accurate ex situ analysis with Nomarski microscope has been observedmore » for a typical micro-focus experiment.« less

Sub-nanosecond time-resolved near-field scanning magneto-optical microscope.

PubMed

Rudge, J; Xu, H; Kolthammer, J; Hong, Y K; Choi, B C

2015-02-01

We report on the development of a new magnetic microscope, time-resolved near-field scanning magneto-optical microscope, which combines a near-field scanning optical microscope and magneto-optical contrast. By taking advantage of the high temporal resolution of time-resolved Kerr microscope and the sub-wavelength spatial resolution of a near-field microscope, we achieved a temporal resolution of ∼50 ps and a spatial resolution of <100 nm. In order to demonstrate the spatiotemporal magnetic imaging capability of this microscope, the magnetic field pulse induced gyrotropic vortex dynamics occurring in 1 μm diameter, 20 nm thick CoFeB circular disks has been investigated. The microscope provides sub-wavelength resolution magnetic images of the gyrotropic motion of the vortex core at a resonance frequency of ∼240 MHz.

The Scanning Optical Microscope.

ERIC Educational Resources Information Center

Sheppard, C. J. R.

1978-01-01

Describes the principle of the scanning optical microscope and explains its advantages over the conventional microscope in the improvement of resolution and contrast, as well as the possibility of producing a picture from optical harmonies generated within the specimen.

Brain plasticity and functionality explored by nonlinear optical microscopy

NASA Astrophysics Data System (ADS)

Sacconi, L.; Allegra, L.; Buffelli, M.; Cesare, P.; D'Angelo, E.; Gandolfi, D.; Grasselli, G.; Lotti, J.; Mapelli, J.; Strata, P.; Pavone, F. S.

2010-02-01

In combination with fluorescent protein (XFP) expression techniques, two-photon microscopy has become an indispensable tool to image cortical plasticity in living mice. In parallel to its application in imaging, multi-photon absorption has also been used as a tool for the dissection of single neurites with submicrometric precision without causing any visible collateral damage to the surrounding neuronal structures. In this work, multi-photon nanosurgery is applied to dissect single climbing fibers expressing GFP in the cerebellar cortex. The morphological consequences are then characterized with time lapse 3-dimensional two-photon imaging over a period of minutes to days after the procedure. Preliminary investigations show that the laser induced fiber dissection recalls a regenerative process in the fiber itself over a period of days. These results show the possibility of this innovative technique to investigate regenerative processes in adult brain. In parallel with imaging and manipulation technique, non-linear microscopy offers the opportunity to optically record electrical activity in intact neuronal networks. In this work, we combined the advantages of second-harmonic generation (SHG) with a random access (RA) excitation scheme to realize a new microscope (RASH) capable of optically recording fast membrane potential events occurring in a wide-field of view. The RASH microscope, in combination with bulk loading of tissue with FM4-64 dye, was used to simultaneously record electrical activity from clusters of Purkinje cells in acute cerebellar slices. Complex spikes, both synchronous and asynchronous, were optically recorded simultaneously across a given population of neurons. Spontaneous electrical activity was also monitored simultaneously in pairs of neurons, where action potentials were recorded without averaging across trials. These results show the strength of this technique in describing the temporal dynamics of neuronal assemblies, opening promising perspectives in understanding the computations of neuronal networks.

Optical coherence tomography - principles and applications

NASA Astrophysics Data System (ADS)

Fercher, A. F.; Drexler, W.; Hitzenberger, C. K.; Lasser, T.

2003-02-01

There have been three basic approaches to optical tomography since the early 1980s: diffraction tomography, diffuse optical tomography and optical coherence tomography (OCT). Optical techniques are of particular importance in the medical field, because these techniques promise to be safe and cheap and, in addition, offer a therapeutic potential. Advances in OCT technology have made it possible to apply OCT in a wide variety of applications but medical applications are still dominating. Specific advantages of OCT are its high depth and transversal resolution, the fact, that its depth resolution is decoupled from transverse resolution, high probing depth in scattering media, contact-free and non-invasive operation, and the possibility to create various function dependent image contrasting methods. This report presents the principles of OCT and the state of important OCT applications. OCT synthesises cross-sectional images from a series of laterally adjacent depth-scans. At present OCT is used in three different fields of optical imaging, in macroscopic imaging of structures which can be seen by the naked eye or using weak magnifications, in microscopic imaging using magnifications up to the classical limit of microscopic resolution and in endoscopic imaging, using low and medium magnification. First, OCT techniques, like the reflectometry technique and the dual beam technique were based on time-domain low coherence interferometry depth-scans. Later, Fourier-domain techniques have been developed and led to new imaging schemes. Recently developed parallel OCT schemes eliminate the need for lateral scanning and, therefore, dramatically increase the imaging rate. These schemes use CCD cameras and CMOS detector arrays as photodetectors. Video-rate three-dimensional OCT pictures have been obtained. Modifying interference microscopy techniques has led to high-resolution optical coherence microscopy that achieved sub-micrometre resolution. This report is concluded with a short presentation of important OCT applications. Ophthalmology is, due to the transparent ocular structures, still the main field of OCT application. The first commercial instrument too has been introduced for ophthalmic diagnostics (Carl Zeiss Meditec AG). Advances in using near-infrared light, however, opened the path for OCT imaging in strongly scattering tissues. Today, optical in vivo biopsy is one of the most challenging fields of OCT application. High resolution, high penetration depth, and its potential for functional imaging attribute to OCT an optical biopsy quality, which can be used to assess tissue and cell function and morphology in situ. OCT can already clarify the relevant architectural tissue morphology. For many diseases, however, including cancer in its early stages, higher resolution is necessary. New broad-bandwidth light sources, like photonic crystal fibres and superfluorescent fibre sources, and new contrasting techniques, give access to new sample properties and unmatched sensitivity and resolution.

Assessment of nerve ultrastructure by fibre-optic confocal microscopy.

PubMed

Cushway, T R; Lanzetta, M; Cox, G; Trickett, R; Owen, E R

1996-01-01

Fibre-optic technology combined with confocality produces a microscope capable of optical thin sectioning. In this original study, tibial nerves have been stained in a rat model with a vital dye, 4-(4-diethylaminostyryl)-N-methylpyridinium iodide, and analysed by fibre-optic confocal microscopy to produce detailed images of nerve ultrastructure. Schwann cells, nodes of Ranvier and longitudinal myelinated sheaths enclosing axons were clearly visible. Single axons appeared as brightly staining longitudinal structures. This allowed easy tracing of multiple signal axons within the nerve tissue. An accurate measurement of internodal lengths was easily accomplished. This technique is comparable to current histological techniques, but does not require biopsy, thin sectioning or tissue fixing. This study offers a standard for further in vivo microscopy, including the possibility of monitoring the progression of nerve regeneration following microsurgical neurorraphy.

Optical, dielectric and morphological studies of sol-gel derived nanocrystalline TiO2 films.

PubMed

Vishwas, M; Sharma, Sudhir Kumar; Narasimha Rao, K; Mohan, S; Gowda, K V Arjuna; Chakradhar, R P S

2009-10-15

Nanocrystalline TiO(2) films have been synthesized on glass and silicon substrates by sol-gel technique. The films have been characterized with optical reflectance/transmittance in the wavelength range 300-1000 nm and the optical constants (n, k) were estimated by using envelope technique as well as spectroscopic ellipsometry. Morphological studies have been carried out using atomic force microscope (AFM). Metal-Oxide-Silicon (MOS) capacitor was fabricated using conducting coating on TiO(2) film deposited on silicon. The C-V measurements show that the film annealed at 300 degrees C has a dielectric constant of 19.80. The high percentage of transmittance, low surface roughness and high dielectric constant suggests that it can be used as an efficient anti-reflection coating on silicon and other optical coating applications and also as a MOS capacitor.

Formation of thin film like assembly of exfoliated C3N4 nanoflakes by solvent non-evaporative method using centrifuge

NASA Astrophysics Data System (ADS)

Tejasvi, Ravi; Basu, Suddhasatwa

2017-12-01

A simple method for depositing a thin film of nanomaterial on a substrate using centrifugation technique has been developed, whereby solvent evaporation is prevented and solvent reuse is possible. The centrifuge technique of deposition yields uniform, smooth thin film irrespective of substrate surface texture. The deposited TiO2/eC3N4 film studied, through field emission scanning electron microscope, atomic force microscope, and optical surface profilometer, shows variation in surface roughness on the basis of centrifugation speeds. Initially film coverage improves and surface roughness decreases with the increase in rpm of the centrifuge and the surface roughness slightly increases with further increase in rpm. The photoelectrochemical studies of TiO2/eC3N4 films suggest that the centrifuge technique forms better heterojunctions compared to that by spin coating technique leading to enhanced photoelectrochemical water splitting.

Photonic Microhand with Autonomous Action.

PubMed

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

2017-11-01

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

Optical monitoring of neuronal activity at high frame rate with a digital random-access multiphoton (RAMP) microscope.

PubMed

Otsu, Yo; Bormuth, Volker; Wong, Jerome; Mathieu, Benjamin; Dugué, Guillaume P; Feltz, Anne; Dieudonné, Stéphane

2008-08-30

Two-photon microscopy offers the promise of monitoring brain activity at multiple locations within intact tissue. However, serial sampling of voxels has been difficult to reconcile with millisecond timescales characteristic of neuronal activity. This is due to the conflicting constraints of scanning speed and signal amplitude. The recent use of acousto-optic deflector scanning to implement random-access multiphoton microscopy (RAMP) potentially allows to preserve long illumination dwell times while sampling multiple points-of-interest at high rates. However, the real-life abilities of RAMP microscopy regarding sensitivity and phototoxicity issues, which have so far impeded prolonged optical recordings at high frame rates, have not been assessed. Here, we describe the design, implementation and characterisation of an optimised RAMP microscope. We demonstrate the application of the microscope by monitoring calcium transients in Purkinje cells and cortical pyramidal cell dendrites and spines. We quantify the illumination constraints imposed by phototoxicity and show that stable continuous high-rate recordings can be obtained. During these recordings the fluorescence signal is large enough to detect spikes with a temporal resolution limited only by the calcium dye dynamics, improving upon previous techniques by at least an order of magnitude.

Mie scattering off coated microbubbles

NASA Astrophysics Data System (ADS)

Nelissen, Radboud; Koene, Elmer; Hilgenfeldt, Sascha; Versluis, Michel

2002-11-01

The acoustic behavior of coated microbubbles depends on parameters of the shell coating, which are in turn dependent on bubble size. More intimate knowledge of this size dependence is required for an improved modeling of a distribution of coated microbubbles such as found in ultrasound contrast agents (UCA). Here a setup is designed to simultaneously measure the optical and acoustic response of an ultrasound-driven single bubble contained in a capillary or levitated by the pressure field of a focused transducer. Optical detection is done by Mie scattering through an inverted microscope. Acoustical detection of the single bubble by a receiving transducer is made possible because of the large working distance of the microscope. For Mie scattering investigation of excited bubbles, two regimes can be distinguished, which require different detection techniques: Conventional wide-angle detection through the microscope objective is sufficient for bubbles of radius exceeding 10 mum. For smaller bubbles, two narrow-aperture detectors are used to reconstruct the bubble dynamics from the complex angle-dependence of the scattered light.

Scanning laser microscope for imaging nanostructured superconductors

NASA Astrophysics Data System (ADS)

Ishida, Takekazu; Arai, Kohei; Akita, Yukio; Miyanari, Mitsunori; Minami, Yusuke; Yotsuya, Tsutomu; Kato, Masaru; Satoh, Kazuo; Uno, Mayumi; Shimakage, Hisashi; Miki, Shigehito; Wang, Zhen

2010-10-01

The nanofabrication of superconductors yields various interesting features in superconducting properties. A variety of different imaging techniques have been developed for probing the local superconducting profiles. A scanning pulsed laser microscope has been developed by the combination of the XYZ piezo-driven stages and an optical fiber with an aspheric focusing lens. The scanning laser microscope is used to understand the position-dependent properties of a superconducting MgB 2 stripline of length 100 μm and width of 3 μm under constant bias current. Our results show that the superconducting stripline can clearly be seen in the contour image of the scanning laser microscope on the signal voltage. It is suggested from the observed image that the inhomogeneity is relevant in specifying the operating conditions such as detection efficiency of the sensor.

Scanning Tunneling Optical Resonance Microscopy Developed

NASA Technical Reports Server (NTRS)

Bailey, Sheila G.; Raffaelle, Ryne P.; Lau, Janis E.; Jenkins, Phillip P.; Castro, Stephanie L.; Tin, Padetha; Wilt, David M.; Pal, Anna Maria; Fahey, Stephen D.

2004-01-01

The ability to determine the in situ optoelectronic properties of semiconductor materials has become especially important as the size of device architectures has decreased and the development of complex microsystems has increased. Scanning Tunneling Optical Resonance Microscopy, or STORM, can interrogate the optical bandgap as a function of its position within a semiconductor micro-structure. This technique uses a tunable solidstate titanium-sapphire laser whose output is "chopped" using a spatial light modulator and is coupled by a fiber-optic connector to a scanning tunneling microscope in order to illuminate the tip-sample junction. The photoenhanced portion of the tunneling current is spectroscopically measured using a lock-in technique. The capabilities of this technique were verified using semiconductor microstructure calibration standards that were grown by organometallic vapor-phase epitaxy. Bandgaps characterized by STORM measurements were found to be in good agreement with the bulk values determined by transmission spectroscopy and photoluminescence and with the theoretical values that were based on x-ray diffraction results.

Wide field of view common-path lateral-shearing digital holographic interference microscope.

PubMed

Vora, Priyanka; Trivedi, Vismay; Mahajan, Swapnil; Patel, Nimit; Joglekar, Mugdha; Chhaniwal, Vani; Moradi, Ali-Reza; Javidi, Bahram; Anand, Arun

2017-12-01

Quantitative three-dimensional (3-D) imaging of living cells provides important information about the cell morphology and its time variation. Off-axis, digital holographic interference microscopy is an ideal tool for 3-D imaging, parameter extraction, and classification of living cells. Two-beam digital holographic microscopes, which are usually employed, provide high-quality 3-D images of micro-objects, albeit with lower temporal stability. Common-path digital holographic geometries, in which the reference beam is derived from the object beam, provide higher temporal stability along with high-quality 3-D images. Self-referencing geometry is the simplest of the common-path techniques, in which a portion of the object beam itself acts as the reference, leading to compact setups using fewer optical elements. However, it has reduced field of view, and the reference may contain object information. Here, we describe the development of a common-path digital holographic microscope, employing a shearing plate and converting one of the beams into a separate reference by employing a pin-hole. The setup is as compact as self-referencing geometry, while providing field of view as wide as that of a two-beam microscope. The microscope is tested by imaging and quantifying the morphology and dynamics of human erythrocytes. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

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

NASA Astrophysics Data System (ADS)

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

2016-03-01

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

Microscope collision protection apparatus

DOEpatents

DeNure, Charles R.

2001-10-23

A microscope collision protection apparatus for a remote control microscope which protects the optical and associated components from damage in the event of an uncontrolled collision with a specimen, regardless of the specimen size or shape. In a preferred embodiment, the apparatus includes a counterbalanced slide for mounting the microscope's optical components. This slide replaces the rigid mounts on conventional upright microscopes with a precision ball bearing slide. As the specimen contacts an optical component, the contacting force will move the slide and the optical components mounted thereon. This movement will protect the optical and associated components from damage as the movement causes a limit switch to be actuated, thereby stopping all motors responsible for the collision.

Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo: A Model of Single Vessel Stroke.

PubMed

Talley Watts, Lora; Zheng, Wei; Garling, R Justin; Frohlich, Victoria C; Lechleiter, James Donald

2015-06-23

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to image cellular events in an intact animal. Additionally, the ability to induce physiological disease states such as stroke in vivo increases its utility. The technique described herein allows for physiological assessment of cellular responses within the CNS following a stroke and can be adapted for other pathological conditions being studied. The technique presented uses laser excitation of the photosensitive dye Rose Bengal in vivo to induce a focal ischemic event in a single blood vessel. The video protocol demonstrates the preparation of a thin-skulled cranial window over the somatosensory cortex in a mouse for the induction of a Rose Bengal photothrombotic event keeping injury to the underlying dura matter and brain at a minimum. Surgical preparation is initially performed under a dissecting microscope with a custom-made surgical/imaging platform, which is then transferred to a confocal microscope equipped with an inverted objective adaptor. Representative images acquired utilizing this protocol are presented as well as time-lapse sequences of stroke induction. This technique is powerful in that the same area can be imaged repeatedly on subsequent days facilitating longitudinal in vivo studies of pathological processes following stroke.

LIPS database with LIPService: a microscopic image database of intracellular structures in Arabidopsis guard cells.

PubMed

Higaki, Takumi; Kutsuna, Natsumaro; Hasezawa, Seiichiro

2013-05-16

Intracellular configuration is an important feature of cell status. Recent advances in microscopic imaging techniques allow us to easily obtain a large number of microscopic images of intracellular structures. In this circumstance, automated microscopic image recognition techniques are of extreme importance to future phenomics/visible screening approaches. However, there was no benchmark microscopic image dataset for intracellular organelles in a specified plant cell type. We previously established the Live Images of Plant Stomata (LIPS) database, a publicly available collection of optical-section images of various intracellular structures of plant guard cells, as a model system of environmental signal perception and transduction. Here we report recent updates to the LIPS database and the establishment of a database table, LIPService. We updated the LIPS dataset and established a new interface named LIPService to promote efficient inspection of intracellular structure configurations. Cell nuclei, microtubules, actin microfilaments, mitochondria, chloroplasts, endoplasmic reticulum, peroxisomes, endosomes, Golgi bodies, and vacuoles can be filtered using probe names or morphometric parameters such as stomatal aperture. In addition to the serial optical sectional images of the original LIPS database, new volume-rendering data for easy web browsing of three-dimensional intracellular structures have been released to allow easy inspection of their configurations or relationships with cell status/morphology. We also demonstrated the utility of the new LIPS image database for automated organelle recognition of images from another plant cell image database with image clustering analyses. The updated LIPS database provides a benchmark image dataset for representative intracellular structures in Arabidopsis guard cells. The newly released LIPService allows users to inspect the relationship between organellar three-dimensional configurations and morphometrical parameters.

Optical second harmonic images of 90 deg domain structure in BaTiO3 and periodically inverted antiparallel domains in LiTaO3

NASA Astrophysics Data System (ADS)

Uesu, Y.; Kurimura, S.; Yamamoto, Y.

1995-04-01

Applied is a microscope to observations of 90 deg ferroelectric domain structure in BaTiO3 and inverted periodically are ferroelectric domains in LiTaO3. It is founded that the second harmonic generation microscope gives information which cannot be obtained by ordinary optical microscopes. The developed nonlinear optical microscope builds two dimensional second harmonic image of a specimen with inhomogenous distribution of d(sub ijk) and applied the microscope to observations of inhomogeneity in some nonlinear-optical organic microcrystals.

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

PubMed Central

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

2008-01-01

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

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

NASA Astrophysics Data System (ADS)

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

2003-01-01

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

Analysis of the chicken retina with an adaptive optics multiphoton microscope.

PubMed

Bueno, Juan M; Giakoumaki, Anastasia; Gualda, Emilio J; Schaeffel, Frank; Artal, Pablo

2011-06-01

The structure and organization of the chicken retina has been investigated with an adaptive optics multiphoton imaging microscope in a backward configuration. Non-stained flat-mounted retinal tissues were imaged at different depths, from the retinal nerve fiber layer to the outer segment, by detecting the intrinsic nonlinear fluorescent signal. From the stacks of images corresponding to the different retinal layers, volume renderings of the entire retina were reconstructed. The density of photoreceptors and ganglion cells layer were directly estimated from the images as a function of the retinal eccentricity. The maximum anatomical resolving power at different retinal eccentricities was also calculated. This technique could be used for a better characterization of retinal alterations during myopia development, and may be useful for visualization of retinal pathologies and intoxication during pharmacological studies.

Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells.

PubMed

Nygate, Yoav N; Singh, Gyanendra; Barnea, Itay; Shaked, Natan T

2018-06-01

We present a new technique for obtaining simultaneous multimodal quantitative phase and fluorescence microscopy of biological cells, providing both quantitative phase imaging and molecular specificity using a single camera. Our system is based on an interferometric multiplexing module, externally positioned at the exit of an optical microscope. In contrast to previous approaches, the presented technique allows conventional fluorescence imaging, rather than interferometric off-axis fluorescence imaging. We demonstrate the presented technique for imaging fluorescent beads and live biological cells.

Pump-probe optical microscopy for imaging nonfluorescent chromophores.

PubMed

Wei, Lu; Min, Wei

2012-06-01

Many chromophores absorb light intensely but have undetectable fluorescence. Hence microscopy techniques other than fluorescence are highly desirable for imaging these chromophores inside live cells, tissues, and organisms. The recently developed pump-probe optical microscopy techniques provide fluorescence-free contrast mechanisms by employing several fundamental light-molecule interactions including excited state absorption, stimulated emission, ground state depletion, and the photothermal effect. By using the pump pulse to excite molecules and the subsequent probe pulse to interrogate the created transient states on a laser scanning microscope, pump-probe microscopy offers imaging capability with high sensitivity and specificity toward nonfluorescent chromophores. Single-molecule sensitivity has even been demonstrated. Here we review and summarize the underlying principles of this emerging class of molecular imaging techniques.

Numerical dispersion compensation for Partial Coherence Interferometry and Optical Coherence Tomography.

PubMed

Fercher, A; Hitzenberger, C; Sticker, M; Zawadzki, R; Karamata, B; Lasser, T

2001-12-03

Dispersive samples introduce a wavelength dependent phase distortion to the probe beam. This leads to a noticeable loss of depth resolution in high resolution OCT using broadband light sources. The standard technique to avoid this consequence is to balance the dispersion of the sample byarrangingadispersive materialinthereference arm. However, the impact of dispersion is depth dependent. A corresponding depth dependent dispersion balancing technique is diffcult to implement. Here we present a numerical dispersion compensation technique for Partial Coherence Interferometry (PCI) and Optical Coherence Tomography (OCT) based on numerical correlation of the depth scan signal with a depth variant kernel. It can be used a posteriori and provides depth dependent dispersion compensation. Examples of dispersion compensated depth scan signals obtained from microscope cover glasses are presented.

Characterization of ZnO:SnO{sub 2} (50:50) thin film deposited by RF magnetron sputtering technique

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

Cynthia, S. R.; Sanjeeviraja, C.; Ponmudi, S.

2016-05-06

Zinc oxide (ZnO) and tin oxide (SnO{sub 2}) thin films have attracted significant interest recently for use in optoelectronic application such as solar cells, flat panel displays, photonic devices, laser diodes and gas sensors because of their desirable electrical and optical properties and wide band gap. In the present study, thin films of ZnO:SnO{sub 2} (50:50) were deposited on pre-cleaned microscopic glass substrate by RF magnetron sputtering technique. The substrate temperature and RF power induced changes in structural, surface morphological, compositional and optical properties of the films have been studied.

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

PubMed

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

2013-11-05

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

Optical Coherence Tomography for Retinal Surgery: Perioperative Analysis to Real-Time Four-Dimensional Image-Guided Surgery.

PubMed

Carrasco-Zevallos, Oscar M; Keller, Brenton; Viehland, Christian; Shen, Liangbo; Seider, Michael I; Izatt, Joseph A; Toth, Cynthia A

2016-07-01

Magnification of the surgical field using the operating microscope facilitated profound innovations in retinal surgery in the 1970s, such as pars plana vitrectomy. Although surgical instrumentation and illumination techniques are continually developing, the operating microscope for vitreoretinal procedures has remained essentially unchanged and currently limits the surgeon's depth perception and assessment of subtle microanatomy. Optical coherence tomography (OCT) has revolutionized clinical management of retinal pathology, and its introduction into the operating suite may have a similar impact on surgical visualization and treatment. In this article, we review the evolution of OCT for retinal surgery, from perioperative analysis to live volumetric (four-dimensional, 4D) image-guided surgery. We begin by briefly addressing the benefits and limitations of the operating microscope, the progression of OCT technology, and OCT applications in clinical/perioperative retinal imaging. Next, we review intraoperative OCT (iOCT) applications using handheld probes during surgical pauses, two-dimensional (2D) microscope-integrated OCT (MIOCT) of live surgery, and volumetric MIOCT of live surgery. The iOCT discussion focuses on technological advancements, applications during human retinal surgery, translational difficulties and limitations, and future directions.

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.

Optical Coherence Tomography for Retinal Surgery: Perioperative Analysis to Real-Time Four-Dimensional Image-Guided Surgery

PubMed Central

Carrasco-Zevallos, Oscar M.; Keller, Brenton; Viehland, Christian; Shen, Liangbo; Seider, Michael I.; Izatt, Joseph A.; Toth, Cynthia A.

2016-01-01

Magnification of the surgical field using the operating microscope facilitated profound innovations in retinal surgery in the 1970s, such as pars plana vitrectomy. Although surgical instrumentation and illumination techniques are continually developing, the operating microscope for vitreoretinal procedures has remained essentially unchanged and currently limits the surgeon's depth perception and assessment of subtle microanatomy. Optical coherence tomography (OCT) has revolutionized clinical management of retinal pathology, and its introduction into the operating suite may have a similar impact on surgical visualization and treatment. In this article, we review the evolution of OCT for retinal surgery, from perioperative analysis to live volumetric (four-dimensional, 4D) image-guided surgery. We begin by briefly addressing the benefits and limitations of the operating microscope, the progression of OCT technology, and OCT applications in clinical/perioperative retinal imaging. Next, we review intraoperative OCT (iOCT) applications using handheld probes during surgical pauses, two-dimensional (2D) microscope-integrated OCT (MIOCT) of live surgery, and volumetric MIOCT of live surgery. The iOCT discussion focuses on technological advancements, applications during human retinal surgery, translational difficulties and limitations, and future directions. PMID:27409495

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

NASA Astrophysics Data System (ADS)

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

2018-03-01

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

Correlative super-resolution fluorescence microscopy combined with optical coherence microscopy

NASA Astrophysics Data System (ADS)

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

2015-03-01

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

Mirrorlike pulsed laser deposited tungsten thin film.

PubMed

Mostako, A T T; Rao, C V S; Khare, Alika

2011-01-01

Mirrorlike tungsten thin films on stainless steel substrate deposited via pulsed laser deposition technique in vacuum (10(-5) Torr) is reported, which may find direct application as first mirror in fusion devices. The crystal structure of tungsten film is analyzed using x-ray diffraction pattern, surface morphology of the tungsten films is studied with scanning electron microscope and atomic force microscope. The film composition is identified using energy dispersive x-ray. The specular and diffuse reflectivities with respect to stainless steel substrate of the tungsten films are recorded with FTIR spectra. The thickness and the optical quality of pulsed laser deposition deposited films are tested via interferometric technique. The reflectivity is approaching about that of the bulk for the tungsten film of thickness ∼782 nm.

Effect of etching time on morphological, optical, and electronic properties of silicon nanowires.

PubMed

Nafie, Nesma; Lachiheb, Manel Abouda; Bouaicha, Mongi

2012-07-16

Owing to their interesting electronic, mechanical, optical, and transport properties, silicon nanowires (SiNWs) have attracted much attention, giving opportunities to several potential applications in nanoscale electronic, optoelectronic devices, and silicon solar cells. For photovoltaic application, a superficial film of SiNWs could be used as an efficient antireflection coating. In this work we investigate the morphological, optical, and electronic properties of SiNWs fabricated at different etching times. Characterizations of the formed SiNWs films were performed using a scanning electron microscope, ultraviolet-visible-near-infrared spectroscopy, and light-beam-induced-current technique. The latter technique was used to determine the effective diffusion length in SiNWs films. From these investigations, we deduce that the homogeneity of the SiNWs film plays a key role on the electronic properties.

Quantitative readout of optically encoded gold nanorods using an ordinary dark-field microscope.

PubMed

Mercatelli, Raffaella; Ratto, Fulvio; Centi, Sonia; Soria, Silvia; Romano, Giovanni; Matteini, Paolo; Quercioli, Franco; Pini, Roberto; Fusi, Franco

2013-10-21

In this paper we report on a new use for dark-field microscopy in order to retrieve two-dimensional maps of optical parameters of a thin sample such as a cryptograph, a histological section, or a cell monolayer. In particular, we discuss the construction of quantitative charts of light absorbance and scattering coefficients of a polyvinyl alcohol film that was embedded with gold nanorods and then etched using a focused mode-locked Ti:Sapphire oscillator. Individual pulses from this laser excite plasmonic oscillations of the gold nanorods, thus triggering plastic deformations of the particles and their environment, which are confined within a few hundred nm of the light focus. In turn, these deformations modify the light absorbance and scattering landscape, which can be measured with optical resolution in a dark-field microscope equipped with an objective of tuneable numerical aperture. This technique may prove to be valuable for various applications, such as the fast readout of optically encoded data or to model functional interactions between light and biological tissue at the level of cellular organelles, including the photothermolysis of cancer.

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

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

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

2015-01-19

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

Development of an ultrasound microscope combined with optical microscope for multiparametric characterization of a single cell.

PubMed

Arakawa, Mototaka; Shikama, Joe; Yoshida, Koki; Nagaoka, Ryo; Kobayashi, Kazuto; Saijo, Yoshifumi

2015-09-01

Biomechanics of the cell has been gathering much attention because it affects the pathological status in atherosclerosis and cancer. In the present study, an ultrasound microscope system combined with optical microscope for characterization of a single cell with multiple ultrasound parameters was developed. The central frequency of the transducer was 375 MHz and the scan area was 80 × 80 μm with up to 200 × 200 sampling points. An inverted optical microscope was incorporated in the design of the system, allowing for simultaneous optical observations of cultured cells. Two-dimensional mapping of multiple ultrasound parameters, such as sound speed, attenuation, and acoustic impedance, as well as the thickness, density, and bulk modulus of specimen/cell under investigation, etc., was realized by the system. Sound speed and thickness of a 3T3-L1 fibroblast cell were successfully obtained by the system. The ultrasound microscope system combined with optical microscope further enhances our understanding of cellular biomechanics.

Evanescent wave assisted nanomaterial coating.

PubMed

Mondal, Samir K; Pal, Sudipta Sarkar; Kumbhakar, Dharmadas; Tiwari, Umesh; Bhatnagar, Randhir

2013-08-01

In this work we present a novel nanomaterial coating technique using evanescent wave (EW). The gradient force in the EW is used as an optical tweezer for tweezing and self-assembling nanoparticles on the source of EW. As a proof of the concept, we have used a laser coupled etched multimode optical fiber, which generates EW for the EW assisted coating. The section-wise etched multimode optical fiber is horizontally and superficially dipped into a silver/gold nanoparticles solution while the laser is switched on. The fiber is left until the solution recedes due to evaporation leaving the fiber in air. The coating time usually takes 40-50 min at room temperature. The scanning electron microscope image shows uniform and thin coating of self-assembled nanoparticles due to EW around the etched section. A coating thickness <200 nm is achieved. The technique could be useful for making surface-plasmon-resonance-based optical fiber probes and other plasmonic circuits.

Development of a near-field/confocal polarization microscope for local measurements of anisotropy in organic films

NASA Astrophysics Data System (ADS)

Kosterin, Andrey Valentinovich

2000-10-01

Polarization microscopy is a powerful technique for imaging structure and stress distributions in many transparent materials, and has been particularly useful in morphology studies of polymer films. Recently the possibility of combining polarization imaging with near-field scanning optical microscopy (NSOM) has been demonstrated, offering new opportunities for studying molecular organization with better than 50 nm resolution. However, there are challenges associated with near-field polarization experiments on organic films: (1) the films are susceptible to damage by the near-field probe; (2) the phase shift or retardation (80) is small, often <0.1 rad; (3) interpretation of near-field images is complicated by topography and probe-sample coupling. To address these challenges, we have developed a new combined near-field/confocal polarization microscope and tested its sensitivity to linear birefringence in thin polymer films. For near-field imaging, the microscope employs a commercially available scanhead with cantilevered (bent) optical fiber probes. To study soft samples (point 1), we have modified the scanhead for tapping mode feedback, which eliminates probe-sample shear forces and prolongs the lifetime of the probe, while minimizing damage to the sample. To achieve sensitivity to small phase shifts (point 2), we have implemented the phase modulation (PM) technique in the optical path. Enhanced sensitivity relative to the standard crossed polarizers scheme is achieved because of the better signal-to-noise discrimination common to lock-in detection and because the detected first harmonic intensity, Io , is linearly proportional to deltatheta instead of (deltatheta) 2. To facilitate interpretation of near-field contrast (point 3), we have incorporated near-field and confocal channels in one instrument. This allows consecutive acquisition of both near-field and far-field images on the same sample area. Since the far-field images do not suffer from the same artifacts, they can be used as a source of independent information on sample optical properties. The combined near-field/confocal polarization microscope is discussed in this thesis as well as some of its applications. Specifically we consider the results of polyethylene oxide (PEO) and crosslinked polybutadiene (PB) thin film imaging.

Endoscopic Optical Coherence Tomography for Clinical Gastroenterology

PubMed Central

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

2014-01-01

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

Stitching Techniques Advance Optics Manufacturing

NASA Technical Reports Server (NTRS)

2010-01-01

Because NASA depends on the fabrication and testing of large, high-quality aspheric (nonspherical) optics for applications like the James Webb Space Telescope, it sought an improved method for measuring large aspheres. Through Small Business Innovation Research (SBIR) awards from Goddard Space Flight Center, QED Technologies, of Rochester, New York, upgraded and enhanced its stitching technology for aspheres. QED developed the SSI-A, which earned the company an R&D 100 award, and also developed a breakthrough machine tool called the aspheric stitching interferometer. The equipment is applied to advanced optics in telescopes, microscopes, cameras, medical scopes, binoculars, and photolithography."

Enhanced weak-signal sensitivity in two-photon microscopy by adaptive illumination.

PubMed

Chu, Kengyeh K; Lim, Daryl; Mertz, Jerome

2007-10-01

We describe a technique to enhance both the weak-signal relative sensitivity and the dynamic range of a laser scanning optical microscope. The technique is based on maintaining a fixed detection power by fast feedback control of the illumination power, thereby transferring high measurement resolution to weak signals while virtually eliminating the possibility of image saturation. We analyze and demonstrate the benefits of adaptive illumination in two-photon fluorescence microscopy.

Characterization of the Polycaprolactone Melt Crystallization: Complementary Optical Microscopy, DSC, and AFM Studies

PubMed Central

Speranza, V.; Sorrentino, A.; De Santis, F.; Pantani, R.

2014-01-01

The first stages of the crystallization of polycaprolactone (PCL) were studied using several techniques. The crystallization exotherms measured by differential scanning calorimetry (DSC) were analyzed and compared with results obtained by polarized optical microscopy (POM), rheology, and atomic force microscope (AFM). The experimental results suggest a strong influence of the observation scale. In particular, the AFM, even if limited on time scale, appears to be the most sensitive technique to detect the first stages of crystallization. On the contrary, at least in the case analysed in this work, rheology appears to be the least sensitive technique. DSC and POM provide closer results. This suggests that the definition of induction time in the polymer crystallization is a vague concept that, in any case, requires the definition of the technique used for its characterization. PMID:24523644

Characterization of the polycaprolactone melt crystallization: complementary optical microscopy, DSC, and AFM studies.

PubMed

Speranza, V; Sorrentino, A; De Santis, F; Pantani, R

2014-01-01

The first stages of the crystallization of polycaprolactone (PCL) were studied using several techniques. The crystallization exotherms measured by differential scanning calorimetry (DSC) were analyzed and compared with results obtained by polarized optical microscopy (POM), rheology, and atomic force microscope (AFM). The experimental results suggest a strong influence of the observation scale. In particular, the AFM, even if limited on time scale, appears to be the most sensitive technique to detect the first stages of crystallization. On the contrary, at least in the case analysed in this work, rheology appears to be the least sensitive technique. DSC and POM provide closer results. This suggests that the definition of induction time in the polymer crystallization is a vague concept that, in any case, requires the definition of the technique used for its characterization.

In vitro monitoring of oxidative processes with self-aggregating gold nanoparticles using all-optical photoacoustic spectroscopy.

PubMed

Yasmin, Zannatul; Khachatryan, Edward; Lee, Yuan-Hao; Maswadi, Saher; Glickman, Randolph; Nash, Kelly L

2015-02-15

In this work, the assembly of gold nanoparticles of (AuNPs) is used to detect the presence of the biomolecule glutathione (GSH) using a novel technique called "all-optical photoacoustic spectroscopy" (AOPAS). The AOPAS technique coupled with AuNPs forms the basis of a biosensing technique capable of probing the dynamic evolution of nano-bio interfaces within a microscopic volume. Dynamic Light Scattering (DLS) and ultraviolet-visible (UV-vis) spectra were measured to describe the kinetics governing the interparticle interactions by monitoring the AuNPs assembly and evolution of the surface plasmon resonance (SPR) band. A comparison of the same dynamic evolution of AuNPs assembly was performed using the AOPAS technique to confirm the validity of this method. The fundamental study is complemented by a demonstration of the performance of this biosensing technique in the presence of cell culture medium containing fetal bovine serum (FBS), which forms a protein corona on the surface of the AuNPs. This work demonstrates that the in vitro monitoring capabilities of the AOPAS provides sensitive measurement at the microscopic level and low nanoparticle concentrations without the artifacts limiting the use of conventional biosensing methods, such as fluorescent indicators. The AOPAS technique not only provides a facile approach for in vitro biosensing, but also shed a light on the real-time detection of thiol containing oxidative stress biomarkers in live systems using AuNPs. Copyright © 2014 Elsevier B.V. All rights reserved.

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

PubMed Central

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

2009-01-01

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

Combined multi-plane phase retrieval and super-resolution optical fluctuation imaging for 4D cell microscopy

NASA Astrophysics Data System (ADS)

Descloux, A.; Grußmayer, K. S.; Bostan, E.; Lukes, T.; Bouwens, A.; Sharipov, A.; Geissbuehler, S.; Mahul-Mellier, A.-L.; Lashuel, H. A.; Leutenegger, M.; Lasser, T.

2018-03-01

Super-resolution fluorescence microscopy provides unprecedented insight into cellular and subcellular structures. However, going `beyond the diffraction barrier' comes at a price, since most far-field super-resolution imaging techniques trade temporal for spatial super-resolution. We propose the combination of a novel label-free white light quantitative phase imaging with fluorescence to provide high-speed imaging and spatial super-resolution. The non-iterative phase retrieval relies on the acquisition of single images at each z-location and thus enables straightforward 3D phase imaging using a classical microscope. We realized multi-plane imaging using a customized prism for the simultaneous acquisition of eight planes. This allowed us to not only image live cells in 3D at up to 200 Hz, but also to integrate fluorescence super-resolution optical fluctuation imaging within the same optical instrument. The 4D microscope platform unifies the sensitivity and high temporal resolution of phase imaging with the specificity and high spatial resolution of fluorescence microscopy.

Magnified hard x-ray microtomography: toward tomography with submicron resolution

NASA Astrophysics Data System (ADS)

Schroer, Christian G.; Benner, Boris; Guenzler, Til F.; Kuhlmann, Marion; Lengeler, Bruno; Rau, Christoph; Weitkamp, Timm; Snigirev, Anatoly A.; Snigireva, Irina

2002-01-01

Parabolic compound refractive lenses (PCRLs) are high quality imaging optics for hard x-rays that can be used as an objective lens in a new type of hard x-ray full field microscope. Using an aluminium PCRL, this new type of microscope has been shown to have a resolution of 350 nm. Further improvement of the resolution down to 50 nm can be expected using beryllium as a lens material. The large depth of field (several mm) of the microscope results in sharp projection images for samples that fit into the field of view of about 300 micrometers. This allows to combine magnified imaging with tomographic techniques. First results of magnified microtomography are shown. Contrast formation in the microscope and the consequences for tomographic reconstruction are discussed. An outlook on further developments is given.

A route for direct retinal input to the preoptic hypothalamus: dendritic projections into the optic chiasm.

PubMed

Silver, J; Brand, S

1979-07-01

With the use of Golgi, horseradish peroxidase, and electron microscopic techniques, neurons within a broad region of the preoptic hypothalamus of the mouse were shown to have dendrites that projected well into the depths of the optic chiasm. Further experimental and ultrastructural investigation demonstrated synapses between these dendrites and retinal axonal boutons within the chiasm. All synapses located in the chiasm were classified as Gray's type I. The possible function of these dendritic projections is discussed.

The measurement of ultrasound scattering from individual micron-sized objects and its application in single cell scattering.

PubMed

Falou, Omar; Rui, Min; El Kaffas, Ahmed; Kumaradas, J Carl; Kolios, Michael C

2010-08-01

The measurement of the ultrasound backscatter from individual micron-sized objects such as cells is required for various applications such as tissue characterization. However, performing such a measurement remains a challenge. For example, the presence of air bubbles in a suspension of cells during the measurements may lead to the incorrect interpretation of the acoustic signals. This work introduces a technique for measuring the ultrasound backscatter from individual micron-sized objects by combining a microinjection system with a co-registered optical microscope and an ultrasound imaging device. This allowed the measurement of the ultrasound backscatter response from a single object under optical microscope guidance. The optical and ultrasonic data were used to determine the size of the object and to deduce its backscatter responses, respectively. In order to calibrate the system, the backscatter frequency responses from polystyrene microspheres were measured and compared to theoretical predictions. A very good agreement was found between the measured backscatter responses of individual microspheres and theoretical predictions of an elastic sphere. The backscatter responses from single OCI-AML-5 cells were also investigated. It was found that the backscatter responses from AML cells are best modeled using the fluid sphere model. The advantages, limitations, and future applications of the developed technique are discussed.

Polarized Light Microscopy

NASA Technical Reports Server (NTRS)

Frandsen, Athela F.

2016-01-01

Polarized light microscopy (PLM) is a technique which employs the use of polarizing filters to obtain substantial optical property information about the material which is being observed. This information can be combined with other microscopy techniques to confirm or elucidate the identity of an unknown material, determine whether a particular contaminant is present (as with asbestos analysis), or to provide important information that can be used to refine a manufacturing or chemical process. PLM was the major microscopy technique in use for identification of materials for nearly a century since its introduction in 1834 by William Fox Talbot, as other techniques such as SEM (Scanning Electron Microscopy), FTIR (Fourier Transform Infrared spectroscopy), XPD (X-ray Powder Diffraction), and TEM (Transmission Electron Microscopy) had not yet been developed. Today, it is still the only technique approved by the Environmental Protection Agency (EPA) for asbestos analysis, and is often the technique first applied for identification of unknown materials. PLM uses different configurations in order to determine different material properties. With each configuration additional clues can be gathered, leading to a conclusion of material identity. With no polarizing filter, the microscope can be used just as a stereo optical microscope, and view qualities such as morphology, size, and number of phases. With a single polarizing filter (single polars), additional properties can be established, such as pleochroism, individual refractive indices, and dispersion staining. With two polarizing filters (crossed polars), even more can be deduced: isotropy vs. anisotropy, extinction angle, birefringence/degree of birefringence, sign of elongation, and anomalous polarization colors, among others. With the use of PLM many of these properties can be determined in a matter of seconds, even for those who are not highly trained. McCrone, a leader in the field of polarized light microscopy, often advised, If you cant determine a specific optical property of a particle after two minutes, move onto another configuration. Since optical properties can be seen so very quickly and easily under polarized light, it is only necessary to spend a maximum of two minutes on a technique to determine a particular property, though often only a few seconds are required.

Imaging Schwarzschild multilayer X-ray microscope

NASA Technical Reports Server (NTRS)

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

1993-01-01

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

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

PubMed

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

2016-09-01

Optoacoustic (OA) microscopy using an all-optical system based on the probe beam deflection technique (PBDT) for detection of laser-induced acoustic signals was investigated as an alternative to conventional piezoelectric transducers. PBDT provides a number of advantages for OA microscopy including (i) efficient coupling of laser excitation energy to the samples being imaged through the probing laser beam, (ii) undistorted coupling of acoustic waves to the detector without the need for separation of the optical and acoustic paths, (iii) high sensitivity and (iv) ultrawide bandwidth. Because of the unimpeded optical path in PBDT, diffraction-limited lateral resolution can be readily achieved. The sensitivity of the current PBDT sensor of 22 μV/Pa and its noise equivalent pressure (NEP) of 11.4 Pa are comparable with these parameters of the optical micro-ring resonator and commercial piezoelectric ultrasonic transducers. Benefits of the present prototype OA microscope were demonstrated by successfully resolving micron-size details in histological sections of cardiac muscle.

Optical Tweezer Assembly and Calibration

NASA Technical Reports Server (NTRS)

Collins, Timothy M.

2004-01-01

An Optical Tweezer, as the name implies, is a useful tool for precision manipulation of micro and nano scale objects. Using the principle of electromagnetic radiation pressure, an optical tweezer employs a tightly focused laser beam to trap and position objects of various shapes and sizes. These devices can trap micrometer and nanometer sized objects. An exciting possibility for optical tweezers is its future potential to manipulate and assemble micro and nano sized sensors. A typical optical tweezer makes use of the following components: laser, mirrors, lenses, a high quality microscope, stage, Charge Coupled Device (CCD) camera, TV monitor and Position Sensitive Detectors (PSDs). The laser wavelength employed is typically in the visible or infrared spectrum. The laser beam is directed via mirrors and lenses into the microscope. It is then tightly focused by a high magnification, high numerical aperture microscope objective into the sample slide, which is mounted on a translating stage. The sample slide contains a sealed, small volume of fluid that the objects are suspended in. The most common objects trapped by optical tweezers are dielectric spheres. When trapped, a sphere will literally snap into and center itself in the laser beam. The PSD s are mounted in such a way to receive the backscatter after the beam has passed through the trap. PSD s used with the Differential Interference Contrast (DIC) technique provide highly precise data. Most optical tweezers employ lasers with power levels ranging from 10 to 100 miliwatts. Typical forces exerted on trapped objects are in the pico-newton range. When PSDs are employed, object movement can be resolved on a nanometer scale in a time range of milliseconds. Such accuracy, however, can only by utilized by calibrating the optical tweezer. Fortunately, an optical tweezer can be modeled accurately as a simple spring. This allows Hook s Law to be used. My goal this summer at NASA Glenn Research Center is the assembly and calibration of an optical tweezer setup in the Instrumentation and Controls Division (5520). I am utilizing a custom LabVIEW Virtual Instrument program for data collection and microscope stage control. Helping me in my assignment are the following people: Mentor Susan Wrbanek (5520), Dr. Baha Jassemnejad (UCO) and Technicians Ken Weiland (7650) and James Williams (7650). Without their help, my task would not be possible.

Optical filters for wavelength selection in fluorescence instrumentation.

PubMed

Erdogan, Turan

2011-04-01

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

Imaging slit-coupled surface plasmon polaritons using conventional optical microscopy.

PubMed

Mehfuz, R; Chowdhury, F A; Chau, K J

2012-05-07

We develop a technique that now enables surface plasmon polaritons (SPPs) coupled by nano-patterned slits in a metal film to be detected using conventional optical microscopy with standard objective lenses. The crux of this method is an ultra-thin polymer layer on the metal surface, whose thickness can be varied over a nanoscale range to enable controllable tuning of the SPP momentum. At an optimal layer thickness for which the SPP momentum matches the momentum of light emerging from the slit, the SPP coupling efficiency is enhanced about six times relative to that without the layer. The enhanced efficiency results in distinctive and bright plasmonic signatures near the slit visible by naked eye under an optical microscope. We demonstrate how this capability can be used for parallel measurement through a simple experiment in which the SPP propagation distance is extracted from a single microscope image of an illuminated array of nano-patterned slits on a metal surface. We also use optical microscopy to image the focal region of a plasmonic lens and obtain results consistent with a previously-reported results using near-field optical microscopy. Measurement of SPPs near a nano-slit using conventional and widely-available optical microscopy is an important step towards making nano-plasmonic device technology highly accessible and easy-to-use.

Effect of 60Co γ-irradiation on structural and optical properties of thin films of Ga10Se80Hg10

NASA Astrophysics Data System (ADS)

Ahmad, Shabir; Asokan, K.; Shahid Khan, Mohd.; Zulfequar, M.

2015-08-01

Thin films of Ga10Se80Hg10 have been deposited onto a chemically cleaned Al2O3 substrates by thermal evaporation technique under vacuum. The investigated thin films are irradiated by 60Co γ-rays in the dose range of 50-150 kGy. X-ray diffraction patterns of the investigated thin films confirm the preferred crystallite growth occurs in the tetragonal phase structure. It also shows, the average crystallite size increases after γ-exposure, which indicates the crystallinity of the material increases after γ-irradiation. These results were further supported by surface morphological analysis carried out by scanning electron microscope and atomic force microscope which also shows the crystallinity of the material increases with increasing the γ-irradiation dose. The optical transmission spectra of the thin films at normal incidence were investigated in the spectral range from 190 to 1100 nm. Using the transmission spectra, the optical constants like refractive index (n) and extinction coefficient (k) were calculated based on Swanepoel's method. The optical band gap (Eg) was also estimated using Tauc's extrapolation procedure. The optical analysis shows: the value of optical band gap of investigated thin films decreases and the corresponding absorption coefficient increases continuously with increasing dose of γ-irradiation.

Science 101: How Does an Electron Microscope Work?

ERIC Educational Resources Information Center

Robertson, Bill

2013-01-01

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

Analysis of the chicken retina with an adaptive optics multiphoton microscope

PubMed Central

Bueno, Juan M.; Giakoumaki, Anastasia; Gualda, Emilio J.; Schaeffel, Frank; Artal, Pablo

2011-01-01

The structure and organization of the chicken retina has been investigated with an adaptive optics multiphoton imaging microscope in a backward configuration. Non-stained flat-mounted retinal tissues were imaged at different depths, from the retinal nerve fiber layer to the outer segment, by detecting the intrinsic nonlinear fluorescent signal. From the stacks of images corresponding to the different retinal layers, volume renderings of the entire retina were reconstructed. The density of photoreceptors and ganglion cells layer were directly estimated from the images as a function of the retinal eccentricity. The maximum anatomical resolving power at different retinal eccentricities was also calculated. This technique could be used for a better characterization of retinal alterations during myopia development, and may be useful for visualization of retinal pathologies and intoxication during pharmacological studies. PMID:21698025

Cryo X-ray microscope with flat sample geometry for correlative fluorescence and nanoscale tomographic imaging.

PubMed

Schneider, Gerd; Guttmann, Peter; Rehbein, Stefan; Werner, Stephan; Follath, Rolf

2012-02-01

X-ray imaging offers a new 3-D view into cells. With its ability to penetrate whole hydrated cells it is ideally suited for pairing fluorescence light microscopy and nanoscale X-ray tomography. In this paper, we describe the X-ray optical set-up and the design of the cryo full-field transmission X-ray microscope (TXM) at the electron storage ring BESSY II. Compared to previous TXM set-ups with zone plate condenser monochromator, the new X-ray optical layout employs an undulator source, a spherical grating monochromator and an elliptically shaped glass capillary mirror as condenser. This set-up improves the spectral resolution by an order of magnitude. Furthermore, the partially coherent object illumination improves the contrast transfer of the microscope compared to incoherent conditions. With the new TXM, cells grown on flat support grids can be tilted perpendicular to the optical axis without any geometrical restrictions by the previously required pinhole for the zone plate monochromator close to the sample plane. We also developed an incorporated fluorescence light microscope which permits to record fluorescence, bright field and DIC images of cryogenic cells inside the TXM. For TXM tomography, imaging with multi-keV X-rays is a straightforward approach to increase the depth of focus. Under these conditions phase contrast imaging is necessary. For soft X-rays with shrinking depth of focus towards 10nm spatial resolution, thin optical sections through a thick specimen might be obtained by deconvolution X-ray microscopy. As alternative 3-D X-ray imaging techniques, the confocal cryo-STXM and the dual beam cryo-FIB/STXM with photoelectron detection are proposed. Copyright © 2012 Elsevier Inc. All rights reserved.

Ultrafast chirped optical waveform recording using referenced heterodyning and a time microscope

DOEpatents

Bennett, Corey Vincent

2010-06-15

A new technique for capturing both the amplitude and phase of an optical waveform is presented. This technique can capture signals with many THz of bandwidths in a single shot (e.g., temporal resolution of about 44 fs), or be operated repetitively at a high rate. That is, each temporal window (or frame) is captured single shot, in real time, but the process may be run repeatedly or single-shot. This invention expands upon previous work in temporal imaging by adding heterodyning, which can be self-referenced for improved precision and stability, to convert frequency chirp (the second derivative of phase with respect to time) into a time varying intensity modulation. By also including a variety of possible demultiplexing techniques, this process is scalable to recoding continuous signals.

Ultrafast chirped optical waveform recorder using referenced heterodyning and a time microscope

DOEpatents

Bennett, Corey Vincent [Livermore, CA

2011-11-22

A new technique for capturing both the amplitude and phase of an optical waveform is presented. This technique can capture signals with many THz of bandwidths in a single shot (e.g., temporal resolution of about 44 fs), or be operated repetitively at a high rate. That is, each temporal window (or frame) is captured single shot, in real time, but the process may be run repeatedly or single-shot. This invention expands upon previous work in temporal imaging by adding heterodyning, which can be self-referenced for improved precision and stability, to convert frequency chirp (the second derivative of phase with respect to time) into a time varying intensity modulation. By also including a variety of possible demultiplexing techniques, this process is scalable to recoding continuous signals.

Quantitative orientation-independent differential interference contrast (DIC) microscopy

NASA Astrophysics Data System (ADS)

Shribak, Michael; LaFountain, James; Biggs, David; Inoué, Shinya

2007-02-01

We describe a new DIC technique, which records phase gradients within microscopic specimens independently of their orientation. The proposed system allows the generation of images representing the distribution of dry mass (optical path difference) in the specimen. Unlike in other forms of interference microscopes, this approach does not require a narrow illuminating cone. The orientation-independent differential interference contrast (OI-DIC) system can also be combined with orientation-independent polarization (OI-Pol) measurements to yield two complementary images: one showing dry mass distribution (which is proportional to refractive index) and the other showing distribution of birefringence (due to structural or internal anisotropy). With a model specimen used for this work -- living spermatocytes from the crane fly, Nephrotoma suturalis --- the OI-DIC image clearly reveals the detailed shape of the chromosomes while the polarization image quantitatively depicts the distribution of the birefringent microtubules in the spindle, both without any need for staining or other modifications of the cell. We present examples of a pseudo-color combined image incorporating both orientation-independent DIC and polarization images of a spermatocyte at diakinesis and metaphase of meiosis I. Those images provide clear evidence that the proposed technique can reveal fine architecture and molecular organization in live cells without perturbation associated with staining or fluorescent labeling. The phase image was obtained using optics having a numerical aperture 1.4, thus achieving a level of resolution never before achieved with any interference microscope.

Calibration of the optical torque wrench.

PubMed

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

2012-02-13

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

Morphology-controllable of Sn doped ZnO nanorods prepared by spray pyrolysis for transparent electrode application

NASA Astrophysics Data System (ADS)

Hameed, M. Shahul; Princice, J. Joseph; Babu, N. Ramesh; Zahirullah, S. Syed; Deshmukh, Sampat G.; Arunachalam, A.

2018-05-01

Transparent conductive Sn doped ZnO nanorods have been deposited at various doping level by spray pyrolysis technique on glass substrate. The structural, surface morphological and optical properties of these films have been investigated with the help of X-ray diffraction (XRD), scanning electron microscope (SEM), atomic force microscope (AFM) and UV-Vis spectrophotometer respectively. XRD patterns revealed a successful high quality growth of single crystal ZnO nanorods with hexagonal wurtzite structure having (002) preferred orientation. The scanning electron microscope (SEM) image of the prepared films exposed the uniform distribution of Sn doped ZnO nanorod shaped grains. All these films were highly transparent in the visible region with average transmittance of 90%.

Multimodality hard-x-ray imaging of a chromosome with nanoscale spatial resolution

DOE PAGES

Yan, Hanfei; Nazaretski, Evgeny; Lauer, Kenneth R.; ...

2016-02-05

Here, we developed a scanning hard x-ray microscope using a new class of x-ray nano-focusing optic called a multilayer Laue lens and imaged a chromosome with nanoscale spatial resolution. The combination of the hard x-ray's superior penetration power, high sensitivity to elemental composition, high spatial-resolution and quantitative analysis creates a unique tool with capabilities that other microscopy techniques cannot provide. Using this microscope, we simultaneously obtained absorption-, phase-, and fluorescence-contrast images of Pt-stained human chromosome samples. The high spatial-resolution of the microscope and its multi-modality imaging capabilities enabled us to observe the internal ultra-structures of a thick chromosome without sectioningmore » it.« less

Multimodality hard-x-ray imaging of a chromosome with nanoscale spatial resolution

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

Yan, Hanfei; Nazaretski, Evgeny; Lauer, Kenneth R.

Here, we developed a scanning hard x-ray microscope using a new class of x-ray nano-focusing optic called a multilayer Laue lens and imaged a chromosome with nanoscale spatial resolution. The combination of the hard x-ray's superior penetration power, high sensitivity to elemental composition, high spatial-resolution and quantitative analysis creates a unique tool with capabilities that other microscopy techniques cannot provide. Using this microscope, we simultaneously obtained absorption-, phase-, and fluorescence-contrast images of Pt-stained human chromosome samples. The high spatial-resolution of the microscope and its multi-modality imaging capabilities enabled us to observe the internal ultra-structures of a thick chromosome without sectioningmore » it.« less

Space-resolved diffusing wave spectroscopy measurements of the macroscopic deformation and the microscopic dynamics in tensile strain tests

NASA Astrophysics Data System (ADS)

Nagazi, Med-Yassine; Brambilla, Giovanni; Meunier, Gérard; Marguerès, Philippe; Périé, Jean-Noël; Cipelletti, Luca

2017-01-01

We couple a laser-based, space-resolved dynamic light scattering apparatus to a universal traction machine for mechanical extensional tests. We perform simultaneous optical and mechanical measurements on polyether ether ketone, a semi-crystalline polymer widely used in the industry. Due to the high turbidity of the sample, light is multiply scattered by the sample and the diffusing wave spectroscopy (DWS) formalism is used to interpret the data. Space-resolved DWS yields spatial maps of the sample strain and of the microscopic dynamics. An excellent agreement is found between the strain maps thus obtained and those measured by a conventional stereo-digital image correlation technique. The microscopic dynamics reveals both affine motion and plastic rearrangements. Thanks to the extreme sensitivity of DWS to displacements as small as 1 nm, plastic activity and its spatial localization can be detected at an early stage of the sample strain, making the technique presented here a valuable complement to existing material characterization methods.

Optical tracking of nanoscale particles in microscale environments

NASA Astrophysics Data System (ADS)

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

2016-03-01

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

Dynamic Measurement of Low Contact Angles by Optical Microscopy.

PubMed

Campbell, James M; Christenson, Hugo K

2018-05-16

Precise measurement of contact angles is an important challenge in surface science, in the design and characterization of materials and in many crystallization experiments. Here we present a novel technique for measuring the contact angles of droplets between about 2° and 30°, with the lowest experimental uncertainty at the lower end of this range, typically ±0.1°. The lensing effect of a droplet interface produces the appearance of bright circles in low-aperture light, whose diameter is related to the contact angle. The technique requires no specialized equipment beyond an ordinary optical microscope, and may be used to study the dynamic evolution of the contact angle in situ during an experiment.

Large-core single-mode rib SU8 waveguide using solvent-assisted microcontact molding.

PubMed

Huang, Cheng-Sheng; Wang, Wei-Chih

2008-09-01

This paper describes a novel fabrication technique for constructing a polymer-based large-core single-mode rib waveguide. A negative tone SU8 photoresist with a high optical transmission over a large wavelength range and stable mechanical properties was used as a waveguide material. A waveguide was constructed by using a polydimethylsiloxane stamp combined with a solvent-assisted microcontact molding technique. The effects on the final pattern's geometry of four different process conditions were investigated. Optical simulations were performed using beam propagation method software. Single-mode beam propagation was observed at the output of the simulated waveguide as well as the actual waveguide through the microscope image.

Time-resolved wide-field optically sectioned fluorescence microscopy

NASA Astrophysics Data System (ADS)

Dupuis, Guillaume; Benabdallah, Nadia; Chopinaud, Aurélien; Mayet, Céline; Lévêque-Fort, Sandrine

2013-02-01

We present the implementation of a fast wide-field optical sectioning technique called HiLo microscopy on a fluorescence lifetime imaging microscope. HiLo microscopy is based on the fusion of two images, one with structured illumination and another with uniform illumination. Optically sectioned images are then digitally generated thanks to a fusion algorithm. HiLo images are comparable in quality with confocal images but they can be acquired faster over larger fields of view. We obtain 4D imaging by combining HiLo optical sectioning, time-gated detection, and z-displacement. We characterize the performances of this set-up in terms of 3D spatial resolution and time-resolved capabilities in both fixed- and live-cell imaging modes.

The Optics of Bruising

NASA Astrophysics Data System (ADS)

Randeberg, Lise Lyngsnes; Skallerud, Bjørn; Langlois, Neil E. I.; Haugen, Olav Anton; Svaasand, Lars Othar

Forensic medicine is a field of medicine where technology plays an increasingly important role in securing and evaluating evidence in, for example, child abuse cases and cases of domestic violence. Methods from chemistry and biological sciences have found a wide application within forensic medicine. Optical technologies like microscopy are also widely used. Despite this, in vivo or post mortem optical diagnostics by spectroscopy have traditionally not had an important role in clinical or forensic examinations. Forensic medical optics as a field might include all kinds of optical analysis for use within forensic science. This includes everything from microscopic techniques to methods for examination of evidence from a crime scene. This chapter will, however, focus on the use of optical diagnostics for examining skin, with a focus on identification, characterization and age determination of minor traumatic injuries like skin bruises.

Current Trends in Intraoperative Optical Imaging for Functional Brain Mapping and Delineation of Lesions of Language Cortex

PubMed Central

Prakash, Neal; Uhleman, Falk; Sheth, Sameer A.; Bookheimer, Susan; Martin, Neil; Toga, Arthur W.

2009-01-01

Resection of a cerebral arteriovenous malformation (AVM), epileptic focus, or glioma, ideally has a prerequisite of microscopic delineation of the lesion borders in relation to the normal gray and white matter that mediate critical functions. Currently, Wada testing and functional magnetic resonance imaging (fMRI) are used for preoperative mapping of critical function, whereas electrical stimulation mapping (ESM) is used for intraoperative mapping. For lesion delineation, MRI and positron emission tomography (PET) are used preoperatively, whereas microscopy and histological sectioning are used intraoperatively. However, for lesions near eloquent cortex, these imaging techniques may lack sufficient resolution to define the relationship between the lesion and language function, and thus not accurately determine which patients will benefit from neurosurgical resection of the lesion without iatrogenic aphasia. Optical techniques such as intraoperative optical imaging of intrinsic signals (iOIS) show great promise for the precise functional mapping of cortices, as well as delineation of the borders of AVMs, epileptic foci, and gliomas. Here we first review the physiology of neuroimaging, and then progress towards the validation and justification of using intraoperative optical techniques, especially in relation to neurosurgical planning of resection AVMs, epileptic foci, and gliomas near or in eloquent cortex. We conclude with a short description of potential novel intraoperative optical techniques. PMID:18786643

An optical clearing technique for plant tissues allowing deep imaging and compatible with fluorescence microscopy.

PubMed

Warner, Cherish A; Biedrzycki, Meredith L; Jacobs, Samuel S; Wisser, Randall J; Caplan, Jeffrey L; Sherrier, D Janine

2014-12-01

We report on a nondestructive clearing technique that enhances transmission of light through specimens from diverse plant species, opening unique opportunities for microscope-enabled plant research. After clearing, plant organs and thick tissue sections are amenable to deep imaging. The clearing method is compatible with immunocytochemistry techniques and can be used in concert with common fluorescent probes, including widely adopted protein tags such as GFP, which has fluorescence that is preserved during the clearing process. © 2014 American Society of Plant Biologists. All Rights Reserved.

Optical Magnification Should Be Mandatory for Microsurgery: Scientific Basis and Clinical Data Contributing to Quality Assurance

PubMed Central

Schoeffl, Harald; Lazzeri, Davide; Schnelzer, Richard; Froschauer, Stefan M.

2013-01-01

Background Microsurgical techniques are considered standard procedures in reconstructive surgery. Although microsurgery by itself is defined as surgery aided by optical magnification, there are no guidelines for determining in which clinical situations a microscope or loupe should be used. Therefore, we conducted standardized experiments to objectively assess the impact of optical magnification in microsurgery. Methods Sixteen participants of microsurgical training courses had to complete 2 sets of experiments. Each set had to be performed with an unaided eye, surgical loupes, and a regular operating microscope. The first set of experiments included coaptation of a chicken femoral nerve, and the second set consisted of anastomosing porcine coronary arteries. Evaluation of the sutured nerves and vessels were performed by 2 experienced microsurgeons using an operating microscope. Results The 16 participants of the study completed all of the experiments. The nerve coaptation and vascular anastomoses exercises showed a direct relationship of error frequency and lower optical magnification, meaning that the highest number of microsurgical errors occurred with the unaided eye. For nerve coaptation, there was a strong relationship (P<0.05) between the number of mistakes and magnification, and this relationship was very strong (P<0.01) for vascular anastomoses. Conclusions We were able to prove that microsurgical success is directly related to optical magnification. The human eye's ability to discriminate potentially important anatomical structures is limited, which might be detrimental for clinical results. Although not legally mandatory, surgeries such as reparative surgery after hand trauma should be conducted with magnifying devices for achieving optimal patient outcomes. PMID:23532716

Optical Microscopy Techniques to Inspect for Metallic Whiskers

NASA Technical Reports Server (NTRS)

Brusse, Jay A.

2006-01-01

Metal surface finishes of tin, zinc and cadmium are often applied to electronic components, mechanical hardware and other structures. These finishes sometimes unpredictably may form metal whiskers over periods that can take from hours to months or even many years. The metal whiskers are crystalline structures commonly having uniform cross sectional area along their entire length. Typical whisker dimensions are nominally on the order of only a few microns (um) across while their lengths can extend from a few microns to several millimeters. Metal whiskers pose a reliability hazard to electronic systems primarily as an electrical shorting hazard. The extremely narrow dimensions of metal whiskers can make observation with optical techniques very challenging. The videos herein were compiled to demonstrate the complexities associated with optical microscope inspection of electronic and mechanical components and assemblies for the presence or absence of metal whiskers. The importance of magnification, light source and angle of illumination play critical roles in being able to detect metal whiskers when present. Furthermore, it is demonstrated how improper techniques can easily obscure detection. It is hoped that these videos will improve the probability of detecting metal whiskers with optical inspection techniques.

Mesoscopic Fluorescence Molecular Tomography for Evaluating Engineered Tissues.

PubMed

Ozturk, Mehmet S; Chen, Chao-Wei; Ji, Robin; Zhao, Lingling; Nguyen, Bao-Ngoc B; Fisher, John P; Chen, Yu; Intes, Xavier

2016-03-01

Optimization of regenerative medicine strategies includes the design of biomaterials, development of cell-seeding methods, and control of cell-biomaterial interactions within the engineered tissues. Among these steps, one paramount challenge is to non-destructively image the engineered tissues in their entirety to assess structure, function, and molecular expression. It is especially important to be able to enable cell phenotyping and monitor the distribution and migration of cells throughout the bulk scaffold. Advanced fluorescence microscopic techniques are commonly employed to perform such tasks; however, they are limited to superficial examination of tissue constructs. Therefore, the field of tissue engineering and regenerative medicine would greatly benefit from the development of molecular imaging techniques which are capable of non-destructive imaging of three-dimensional cellular distribution and maturation within a tissue-engineered scaffold beyond the limited depth of current microscopic techniques. In this review, we focus on an emerging depth-resolved optical mesoscopic imaging technique, termed laminar optical tomography (LOT) or mesoscopic fluorescence molecular tomography (MFMT), which enables longitudinal imaging of cellular distribution in thick tissue engineering constructs at depths of a few millimeters and with relatively high resolution. The physical principle, image formation, and instrumentation of LOT/MFMT systems are introduced. Representative applications in tissue engineering include imaging the distribution of human mesenchymal stem cells embedded in hydrogels, imaging of bio-printed tissues, and in vivo applications.

Microscope sterility during spine surgery.

PubMed

Bible, Jesse E; O'Neill, Kevin R; Crosby, Colin G; Schoenecker, Jonathan G; McGirt, Matthew J; Devin, Clinton J

2012-04-01

Prospective study. Assess the contamination rates of sterile microscope drapes after spine surgery. The use of the operating microscope has become more prevalent in certain spine procedures, providing superior magnification, visualization, and illumination of the operative field. However, it may represent an additional source of bacterial contamination and increase the risk of developing a postoperative infection. This study included 25 surgical spine cases performed by a single spine surgeon that required the use of the operative microscope. Sterile culture swabs were used to obtain samples from 7 defined locations on the microscope drape after its use during the operation. The undraped technician's console was sampled in each case as a positive control, and an additional 25 microscope drapes were swabbed immediately after they were applied to the microscope to obtain negative controls. Swab samples were assessed for bacterial growth on 5% sheep blood Columbia agar plates using a semiquantitative technique. No growth was observed on any of the 25 negative control drapes. In contrast, 100% of preoperative and 96% of postoperative positive controls demonstrated obvious contamination. In the postoperative group, all 7 sites of evaluation were found to be contaminated with rates of 12% to 44%. Four of the 7 evaluated locations were found to have significant contamination rates compared with negative controls, including the shafts of the optic eyepieces on the main surgeon side (24%, P = 0.022), "forehead" portion on both the main surgeon (24%, P = 0.022) and assistant sides (28%, P = 0.010), and "overhead" portion of the drape (44%, P = 0.0002). Bacterial contamination of the operative microscope was found to be significant after spine surgery. Contamination was more common around the optic eyepieces, likely due to inadvertent touching of unsterile portions. Similarly, all regions above the eyepieces also have a propensity for contamination because of unknown contact with unsterile parts of the surgeon. Therefore, we believe that changing gloves after making adjustments to the optic eyepieces and avoid handling any portion of the drape above the eyepieces may decrease the risks of intraoperative contamination and possibly postoperative infection as well.

Simple technique for observing subsurface damage in machining of ceramics

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

Xu, H.H.K.; Jahanmir, S.

1994-05-01

A simple technique is proposed for directly observing subsurface damage in the machining of ceramics. The technique requires two polished specimens and an optical microscope with Nomarski illumination for examination. The subsurface damage created by the grinding of an alumina ceramic is investigated using this technique. The mode of damage is identified as intragrain twinning/slip, and intergranular and transgranular cracking. Chipping along the twinned planes and along the transgranular crack planes, and dislodgement of the intergranularly debonded grains are suggested to be the mechanisms of material removal in the machining of this alumina ceramic.

Laser-assisted marking for toric intraocular lens alignment.

PubMed

Dick, H Burkhard; Schultz, Tim

2016-01-01

We describe a technique of 3-dimensional spectral-domain optical coherence tomography-controlled laser-assisted corneal marking for toric intraocular lens implantation. To facilitate accurate alignment, the technique creates 2 perpendicular intrastromal incisions (width 0.75 mm) using an image-guided femtosecond laser. This was performed in a case series comprising 10 eyes of 10 patients. No posterior corneal perforation or epithelial alterations occurred. The incisions were plainly visible under the operating microscope, and no optical phenomena were reported 6 weeks after surgery. Laser-assisted marking can be performed safely and has the potential to enable precise axis marking. Dr. Dick is a paid consultant to Abbott Medical Optics, Inc. Dr. Schultz has no financial or proprietary interest in any material or method mentioned. Copyright © 2016 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

Effect of etching time on morphological, optical, and electronic properties of silicon nanowires

PubMed Central

2012-01-01

Owing to their interesting electronic, mechanical, optical, and transport properties, silicon nanowires (SiNWs) have attracted much attention, giving opportunities to several potential applications in nanoscale electronic, optoelectronic devices, and silicon solar cells. For photovoltaic application, a superficial film of SiNWs could be used as an efficient antireflection coating. In this work we investigate the morphological, optical, and electronic properties of SiNWs fabricated at different etching times. Characterizations of the formed SiNWs films were performed using a scanning electron microscope, ultraviolet–visible-near-infrared spectroscopy, and light-beam-induced-current technique. The latter technique was used to determine the effective diffusion length in SiNWs films. From these investigations, we deduce that the homogeneity of the SiNWs film plays a key role on the electronic properties. PMID:22799265

Role of coherence in microsphere-assisted nanoscopy

NASA Astrophysics Data System (ADS)

Perrin, Stephane; Lecler, Sylvain; Leong-Hoi, Audrey; Montgomery, Paul C.

2017-06-01

The loss of the information, due to the diffraction and the evanescent waves, limits the resolving power of classical optical microscopy. In air, the lateral resolution of an optical microscope can approximated at half of the wavelength using a low-coherence illumination. Recently, several methods have been developed in order to overcome this limitation and, in 2011, a new far-field and full-field imaging technique was proposed where a sub-diffraction-limit resolution has been achieved using a transparent microsphere. In this article, the phenomenon of super-resolution using microsphere-assisted microscopy is analysed through rigorous electro-magnetic simulations. The performances of the imaging technique are estimated as function of optical and geometrical parameters. Furthermore, the role of coherence is introduced through the temporal coherence of the light source and the phase response of the object.

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

PubMed

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

2018-04-19

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

Site-controlled quantum dots fabricated using an atomic-force microscope assisted technique

PubMed Central

Usuki, T; Ohshima, T; Sakuma, Y; Kawabe, M; Okada, Y; Takemoto, K; Miyazawa, T; Hirose, S; Nakata, Y; Takatsu, M; Yokoyama, N

2006-01-01

An atomic-force microscope assisted technique is developed to control the position and size of self-assembled semiconductor quantum dots (QDs). Presently, the site precision is as good as ± 1.5 nm and the size fluctuation is within ± 5% with the minimum controllable lateral diameter of 20 nm. With the ability of producing tightly packed and differently sized QDs, sophisticated QD arrays can be controllably fabricated for the application in quantum computing. The optical quality of such site-controlled QDs is found comparable to some conventionally self-assembled semiconductor QDs. The single dot photoluminescence of site-controlled InAs/InP QDs is studied in detail, presenting the prospect to utilize them in quantum communication as precisely controlled single photon emitters working at telecommunication bands.

New design of a cryostat-mounted scanning near-field optical microscope for single molecule spectroscopy

NASA Astrophysics Data System (ADS)

Durand, Yannig; Woehl, Jörg C.; Viellerobe, Bertrand; Göhde, Wolfgang; Orrit, Michel

1999-02-01

Due to the weakness of the fluorescence signal from a single fluorophore, a scanning near-field optical microscope for single molecule spectroscopy requires a very efficient setup for the collection and detection of emitted photons. We have developed a home-built microscope for operation in a l-He cryostat which uses a solid parabolic mirror in order to optimize the fluorescence collection efficiency. This microscope works with Al-coated, tapered optical fibers in illumination mode. The tip-sample separation is probed by an optical shear-force detection. First results demonstrate the capability of the microscope to image single molecules and achieve a topographical resolution of a few nanometers vertically and better than 50 nm laterally.

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

PubMed

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

2007-05-15

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

Superhydrophobic, Biomimetic Surfaces with High and Low Adhesion, Optical Transmittance, and Nanoscale Mechanical Wear Resistance

NASA Astrophysics Data System (ADS)

Ebert, Daniel R.

Superhydrophobic surfaces (defined as surfaces having water contact angle greater than 150°) show great promise for use in a rapidly growing number of engineering applications, ranging from biomedical devices to fluid drag reduction in pipelines. In nature, the surfaces of many organisms, such as certain plant leaves, are known to exhibit superhydrophobicity. In some cases, droplet adhesion is very low (droplet rolls away easily), while in other cases adhesion is high (droplet remains adhered when surface is inverted). The recent advent and development of microscopes with resolution down to a few nanometers (such as atomic force microscopes and scanning electron microscopes) has allowed for in-depth understanding of the micro- and nanoscale mechanisms employed by these plant leaves and other natural surfaces to achieve their particular wetting properties. Biomimetics (or "mimicking nature") is therefore a very promising approach for the development of engineering surfaces with desired wetting characteristics. However, research in creating biomimetic surfaces is still in its early stages, and many of the surfaces created thus far are not mechanically robust, which is required for many potential real-world applications. In addition, for applications such as self-cleaning windows and solar panels, optical transparency is required. In this thesis, a set of original studies are presented in which superhydrophobic surfaces were designed based on biomimetics and fabricated using a wide of variety of techniques. The surfaces were characterized with regard to wetting characteristics such as water contact angle and contact angle hysteresis, micro- and nanoscale mechanical durability, and in some cases optical transmittance. Theoretical wetting models served as guides both in the design and in the understanding of experimental results, especially in regard to different wetting regime and regime transition. This work provides important conclusions and valuable insight for identifying materials, techniques, and designs for mechanically durable, optically transparent superhydrophobic surfaces.

Video-Rate Confocal Microscopy for Single-Molecule Imaging in Live Cells and Superresolution Fluorescence Imaging

PubMed Central

Lee, Jinwoo; Miyanaga, Yukihiro; Ueda, Masahiro; Hohng, Sungchul

2012-01-01

There is no confocal microscope optimized for single-molecule imaging in live cells and superresolution fluorescence imaging. By combining the swiftness of the line-scanning method and the high sensitivity of wide-field detection, we have developed a, to our knowledge, novel confocal fluorescence microscope with a good optical-sectioning capability (1.0 μm), fast frame rates (<33 fps), and superior fluorescence detection efficiency. Full compatibility of the microscope with conventional cell-imaging techniques allowed us to do single-molecule imaging with a great ease at arbitrary depths of live cells. With the new microscope, we monitored diffusion motion of fluorescently labeled cAMP receptors of Dictyostelium discoideum at both the basal and apical surfaces and obtained superresolution fluorescence images of microtubules of COS-7 cells at depths in the range 0–85 μm from the surface of a coverglass. PMID:23083712

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

DOE PAGES

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

2017-10-05

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

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

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

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

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

An investigation of nitride precipitates in archaeological iron artefacts from Poland.

PubMed

Kedzierski, Z; Stepiński, J; Zielińska-Lipiec, A

2010-03-01

The paper describes the investigations of nitride precipitates in a spearhead and a sword found in the territory of Poland, in cremation graveyards of the Przeworsk Culture, dated to the Roman Period. Three different techniques of the examination of nitride precipitates were employed: optical microscope, scanning electron microscope (scanning electron microscope with energy dispersive X-ray spectrometer) and transmission electron microscope. Two types of precipitates have been observed, and their plate-like shape was demonstrated. The large precipitate has been confirmed to be gamma'-Fe(4)N, whereas the small one has been identified as alpha''-Fe(16)N(2). The origin of nitride precipitates in archaeological iron artefacts from Poland is probably a result of the manufacturing process or cremation as part of burial rites. An examination of available iron artefacts indicates that nitride precipitates (have only limited effect on mechanical properties) influence the hardness of metal only to a very limited degree.

Diffracting aperture based differential phase contrast for scanning X-ray microscopy.

PubMed

Kaulich, Burkhard; Polack, Francois; Neuhaeusler, Ulrich; Susini, Jean; di Fabrizio, Enzo; Wilhein, Thomas

2002-10-07

It is demonstrated that in a zone plate based scanning X-ray microscope, used to image low absorbing, heterogeneous matter at a mesoscopic scale, differential phase contrast (DPC) can be implemented without adding any additional optical component to the normal scheme of the microscope. The DPC mode is simply generated by an appropriate positioning and alignment of microscope apertures. Diffraction from the apertures produces a wave front with a non-uniform intensity. The signal recorded by a pinhole photo diode located in the intensity gradient is highly sensitive to phase changes introduced by the specimen to be recorded. The feasibility of this novel DPC technique was proven with the scanning X-ray microscope at the ID21 beamline of the European Synchrotron Radiation facility (ESRF) operated at 6 keV photon energy. We observe a differential phase contrast, similar to Nomarski's differential interference contrast for the light microscope, which results in a tremendous increase in image contrast of up to 20 % when imaging low absorbing specimen.

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

PubMed

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

2017-08-01

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

Safely splicing glass optical fibers

NASA Technical Reports Server (NTRS)

Korbelak, K.

1980-01-01

Field-repair technique fuses glass fibers in flammable environment. Apparatus consists of v-groove vacuum chucks on manipulators, high-voltage dc power supply and tungsten electrodes, microscope to observe joint alignment and fusion, means of test transmission through joint. Apparatus is enclosed in gas tight bos filled with inert gas during fusion. About 2 feet of fiber end are necessary for splicing.

[Normal and pathological elastic tissue under the electron microscope on thin and ultrathin sections (author's transl)].

PubMed

Adnet, J J; Pinteaux, A; Pousse, G; Caulet, T

1976-04-01

Three simple methods (adapted from optical techniques) for normal and pathological elastic tissue caracterisation in electron microscopy on thin and ultrathin sections are proposed. Two of these methods (orcein and fuchsin resorcin) seem to have a specificity for arterial and breast cancer elastic tissue. Weigert's method gives the best contrast.

In vivo correlation mapping microscopy

NASA Astrophysics Data System (ADS)

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

2016-04-01

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

Nonlinear vibrational microscopy

DOEpatents

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

2000-01-01

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

In situ TEM near-field optical probing of nanoscale silicon crystallization.

PubMed

Xiang, Bin; Hwang, David J; In, Jung Bin; Ryu, Sang-Gil; Yoo, Jae-Hyuck; Dubon, Oscar; Minor, Andrew M; Grigoropoulos, Costas P

2012-05-09

Laser-based processing enables a wide variety of device configurations comprising thin films and nanostructures on sensitive, flexible substrates that are not possible with more traditional thermal annealing schemes. In near-field optical probing, only small regions of a sample are illuminated by the laser beam at any given time. Here we report a new technique that couples the optical near-field of the laser illumination into a transmission electron microscope (TEM) for real-time observations of the laser-materials interactions. We apply this technique to observe the transformation of an amorphous confined Si volume to a single crystal of Si using laser melting. By confinement of the material volume to nanometric dimensions, the entire amorphous precursor is within the laser spot size and transformed into a single crystal. This observation provides a path for laser processing of single-crystal seeds from amorphous precursors, a potentially transformative technique for the fabrication of solar cells and other nanoelectronic devices.

X-ray laser microscope apparatus

DOEpatents

Suckewer, Szymon; DiCicco, Darrell S.; Hirschberg, Joseph G.; Meixler, Lewis D.; Sathre, Robert; Skinner, Charles H.

1990-01-01

A microscope consisting of an x-ray contact microscope and an optical microscope. The optical, phase contrast, microscope is used to align a target with respect to a source of soft x-rays. The source of soft x-rays preferably comprises an x-ray laser but could comprise a synchrotron or other pulse source of x-rays. Transparent resist material is used to support the target. The optical microscope is located on the opposite side of the transparent resist material from the target and is employed to align the target with respect to the anticipated soft x-ray laser beam. After alignment with the use of the optical microscope, the target is exposed to the soft x-ray laser beam. The x-ray sensitive transparent resist material whose chemical bonds are altered by the x-ray beam passing through the target mater GOVERNMENT LICENSE RIGHTS This invention was made with government support under Contract No. De-FG02-86ER13609 awarded by the Department of Energy. The Government has certain rights in this invention.

Scatter sensitive microscopic techniques to identify contrasting mucosal structures in ultrahigh-resolution optical coherence tomograms of mouse colon

NASA Astrophysics Data System (ADS)

Tumlinson, Alexandre R.; Hariri, Lida P.; Drexler, Wolfgang; Barton, Jennifer K.

2008-02-01

Optical coherence tomography, optical coherence microscopy, reflectance confocal microscopy, and darkfield microscopy all derive contrast from the intensity of endogenous tissue scatter. We have imaged excised mouse colon tissue with these complimentary technologies to make conclusions about structural origins of scatter in the mouse colonic mucosa observed with endoscopic OCT. We find hyperintense scattering both from the cytoplasm of epithelial cells and from the boundary between epithelia and the lamina propria. We find almost no scatter from the portion of epithelial cells containing the nucleus. These observations substantiate explanations for the appearance of colonic crypts and the luminal surface.

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

PubMed Central

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

2007-01-01

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

High-energy proton imaging for biomedical applications

DOE PAGES

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

2016-06-10

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

High-energy proton imaging for biomedical applications

NASA Astrophysics Data System (ADS)

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

2016-06-01

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

In vitro assessments on bacterial adhesion and corrosion performance of TiN coating on Ti6Al4V titanium alloy synthesized by multi-arc ion plating

NASA Astrophysics Data System (ADS)

Lin, Naiming; Huang, Xiaobo; Zhang, Xiangyu; Fan, Ailan; Qin, Lin; Tang, Bin

2012-07-01

TiN coating was synthesized on Ti6Al4V titanium alloy surface by multi-arc ion plating (MIP) technique. Surface morphology, cross sectional microstructure, elemental distributions and phase compositions of the obtained coating were analyzed by means of scanning electron microscope (SEM), optical microscope (OM), glow discharge optical emission spectroscope (GDOES) and X-ray diffraction (XRD). Bacterial adhesion and corrosion performance of Ti6Al4V and the TiN coating were assessed via in vitro bacterial adhesion tests and corrosion experiments, respectively. The results indicated that continuous and compact coating which was built up by pure TiN with a typical columnar crystal structure has reached a thickness of 1.5 μm. This TiN coating could significantly reduce the bacterial adhesion and enhance the corrosion resistance of Ti6Al4V substrate.

High-energy proton imaging for biomedical applications

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

Prall, Matthias; Durante, Marco; Berger, Thomas

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

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

PubMed

Scrimgeour, Jan; Curtis, Jennifer E

2012-06-18

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

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

PubMed Central

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

2017-01-01

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

Tapered fiber coupling of single photons emitted by a deterministically positioned single nitrogen vacancy center

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

Liebermeister, Lars, E-mail: lars.liebermeister@physik.uni-muenchen.de; Petersen, Fabian; Münchow, Asmus v.

2014-01-20

A diamond nano-crystal hosting a single nitrogen vacancy (NV) center is optically selected with a confocal scanning microscope and positioned deterministically onto the subwavelength-diameter waist of a tapered optical fiber (TOF) with the help of an atomic force microscope. Based on this nano-manipulation technique, we experimentally demonstrate the evanescent coupling of single fluorescence photons emitted by a single NV-center to the guided mode of the TOF. By comparing photon count rates of the fiber-guided and the free-space modes and with the help of numerical finite-difference time domain simulations, we determine a lower and upper bound for the coupling efficiency ofmore » (9.5 ± 0.6)% and (10.4 ± 0.7)%, respectively. Our results are a promising starting point for future integration of single photon sources into photonic quantum networks and applications in quantum information science.« less

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

PubMed

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

2005-11-01

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

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

NASA Astrophysics Data System (ADS)

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

2015-02-01

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

Membrane Mirrors With Bimorph Shape Actuators

NASA Technical Reports Server (NTRS)

Yang, Eui-Hyeok

2003-01-01

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

A novel strategy towards designing a CdSe quantum dot-metallohydrogel composite material.

PubMed

Chatterjee, Sayantan; Maitra, Uday

2016-08-11

We have described here an efficient method to disperse hydrophobic CdSe quantum dots (QDs) in an aqueous phase using cetyltrimethylammonium bromide (CTAB) micelles without any surface ligand exchange. The water soluble QDs were then embedded in 3D self assembled fibrillar networks (SAFINs) of a hydrogel showing homogeneous dispersibility as evidenced from optical and electron microscopic techniques. The photophysical studies of the hydrogel-QD composite are reported for the first time. These composite materials may have potential applications in biology, optoelectronics, sensors, non-linear optics and materials science.

Microscopic Imaging and Spectroscopy with Scattered Light

PubMed Central

Boustany, Nada N.; Boppart, Stephen A.; Backman, Vadim

2012-01-01

Optical contrast based on elastic scattering interactions between light and matter can be used to probe cellular structure and dynamics, and image tissue architecture. The quantitative nature and high sensitivity of light scattering signals to subtle alterations in tissue morphology, as well as the ability to visualize unstained tissue in vivo, has recently generated significant interest in optical scatter based biosensing and imaging. Here we review the fundamental methodologies used to acquire and interpret optical scatter data. We report on recent findings in this field and present current advances in optical scatter techniques and computational methods. Cellular and tissue data enabled by current advances in optical scatter spectroscopy and imaging stand to impact a variety of biomedical applications including clinical tissue diagnosis, in vivo imaging, drug discovery and basic cell biology. PMID:20617940

Optical fiber-based full Mueller polarimeter for endoscopic imaging using a two-wavelength simultaneous measurement method

NASA Astrophysics Data System (ADS)

Vizet, Jérémy; Manhas, Sandeep; Tran, Jacqueline; Validire, Pierre; Benali, Abdelali; Garcia-Caurel, Enric; Pierangelo, Angelo; Martino, Antonello De; Pagnoux, Dominique

2016-07-01

This paper reports a technique based on spectrally differential measurement for determining the full Mueller matrix of a biological sample through an optical fiber. In this technique, two close wavelengths were used simultaneously, one for characterizing the fiber and the other for characterizing the assembly of fiber and sample. The characteristics of the fiber measured at one wavelength were used to decouple its contribution from the measurement on the assembly of fiber and sample and then to extract sample Mueller matrix at the second wavelength. The proof of concept was experimentally validated by measuring polarimetric parameters of various calibrated optical components through the optical fiber. Then, polarimetric images of histological cuts of human colon tissues were measured, and retardance, diattenuation, and orientation of the main axes of fibrillar regions were displayed. Finally, these images were successfully compared with images obtained by a free space Mueller microscope. As the reported method does not use any moving component, it offers attractive integration possibilities with an endoscopic probe.

Optical fiber-based full Mueller polarimeter for endoscopic imaging using a two-wavelength simultaneous measurement method.

PubMed

Vizet, Jérémy; Manhas, Sandeep; Tran, Jacqueline; Validire, Pierre; Benali, Abdelali; Garcia-Caurel, Enric; Pierangelo, Angelo; De Martino, Antonello; Pagnoux, Dominique

2016-07-01

This paper reports a technique based on spectrally differential measurement for determining the full Mueller matrix of a biological sample through an optical fiber. In this technique, two close wavelengths were used simultaneously, one for characterizing the fiber and the other for characterizing the assembly of fiber and sample. The characteristics of the fiber measured at one wavelength were used to decouple its contribution from the measurement on the assembly of fiber and sample and then to extract sample Mueller matrix at the second wavelength. The proof of concept was experimentally validated by measuring polarimetric parameters of various calibrated optical components through the optical fiber. Then, polarimetric images of histological cuts of human colon tissues were measured, and retardance, diattenuation, and orientation of the main axes of fibrillar regions were displayed. Finally, these images were successfully compared with images obtained by a free space Mueller microscope. As the reported method does not use any moving component, it offers attractive integration possibilities with an endoscopic probe.

Characterization on RF magnetron sputtered niobium pentoxide thin films

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

Usha, N.; Sivakumar, R., E-mail: krsivakumar1979@yahoo.com; Sanjeeviraja, C.

2014-10-15

Niobium pentoxide (Nb{sub 2}O{sub 5}) thin films with amorphous nature were deposited on microscopic glass substrates at 100°C by rf magnetron sputtering technique. The effect of rf power on the structural, morphological, optical, and vibrational properties of Nb{sub 2}O{sub 5} films have been investigated. Optical study shows the maximum average transmittance of about 87% and the optical energy band gap (indirect allowed) changes between 3.70 eV and 3.47 eV. AFM result indicates the smooth surface nature of the samples. Photoluminescence measurement showed the better optical quality of the deposited films. Raman spectra show the LO-TO splitting of Nb-O stretching ofmore » Nb{sub 2}O{sub 5} films.« less

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

NASA Astrophysics Data System (ADS)

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

2015-05-01

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

High-speed quantitative phase imaging using time-stretch spectral shearing contrast (Conference Presentation)

NASA Astrophysics Data System (ADS)

Bosworth, Bryan; Foster, Mark A.

2017-02-01

Photonic time-stretch microscopy (TSM) provides an ideal platform for high-throughput imaging flow cytometry, affording extremely high shutter speeds and frame rates with high sensitivity. In order to resolve weakly scattering cells in biofluid and solve the issue of signal-to-noise in cell labeling specificity of biomarkers in imaging flow cytometry, several quantitative phase (QP) techniques have recently been adapted to TSM. However, these techniques have relied primarily on sensitive free-space optical configurations to generate full electric field measurements. The present work draws from the field of ultrashort pulse characterization to leverage the coherence of the ultrashort optical pulses integral to all TSM systems in order to do self-referenced single-shot quantitative phase imaging in a TSM system. Self-referencing is achieved via spectral shearing interferometry in an exceptionally stable and straightforward Sagnac loop incorporating an electro-optic phase modulator and polarization-maintaining fiber that produce sheared and unsheared copies of the pulse train with an inter-pulse delay determined by polarization mode dispersion. The spectral interferogram then yields a squared amplitude and a phase derivative image that can be integrated for conventional phase. We apply this spectral shearing contrast microscope to acquire QP images on a high-speed flow microscope at 90-MHz line rates with <400 pixels per line. We also consider the extension of this technique to compressed sensing (CS) acquisition by intensity modulating the interference spectra with pseudorandom binary waveforms to reconstruct the images from a highly sub-Nyquist number of random inner products, providing a path to even higher operating rates and reduced data storage requirements.

Label-free tracking of single extracellular vesicles in a nano-fluidic optical fiber (Conference Presentation)

NASA Astrophysics Data System (ADS)

van der Pol, Edwin; Weidlich, Stefan; Lahini, Yoav; Coumans, Frank A. W.; Sturk, Auguste; Nieuwland, Rienk; Schmidt, Markus A.; Faez, Sanli; van Leeuwen, Ton G.

2016-03-01

Background: Extracellular vesicles, such as exosomes, are abundantly present in human body fluids. Since the size, concentration and composition of these vesicles change during disease, vesicles have promising clinical applications, including cancer diagnosis. However, since ~70% of the vesicles have a diameter <70 nm, detection of single vesicles remains challenging. Thus far, vesicles <70 nm have only be studied by techniques that require the vesicles to be adhered to a surface. Consequently, the majority of vesicles have never been studied in their physiological environment. We present a novel label-free optical technique to track single vesicles <70 nm in suspension. Method: Urinary vesicles were contained within a single-mode light-guiding silica fiber containing a 600 nm nano-fluidic channel. Light from a diode laser (660 nm wavelength) was coupled to the fiber, resulting in a strongly confined optical mode in the nano-fluidic channel, which continuously illuminated the freely diffusing vesicles inside the channel. The elastic light scattering from the vesicles, in the direction orthogonal to the fiber axis, was collected using a microscope objective (NA=0.95) and imaged with a home-built microscope. Results: We have tracked single urinary vesicles as small as 35 nm by elastic light scattering. Please note that vesicles are low-refractive index (n<1.4) particles, which we confirmed by combining data on thermal diffusion and light scattering cross section. Conclusions: For the first time, we have studied vesicles <70 nm freely diffusing in suspension. The ease-of-use and performance of this technique support its potential for vesicle-based clinical applications.

Photoluminescence study of carbon dots from ginger and galangal herbs using microwave technique

NASA Astrophysics Data System (ADS)

Isnaeni; Rahmawati, I.; Intan, R.; Zakaria, M.

2018-03-01

Carbon dots are new type of fluorescent nanoparticle that can be synthesis easily from natural sources. We have synthesized carbon dots from ginger and galangal herbs using microwave technique and studied their optical properties. We synthesized colloidal carbon dots in water solvent by varying microwave processing time. UV-Vis absorbance, photoluminescence, time-resolved photoluminescence, and transmission electron microscope were utilized to study properties of carbon dots. We found that microwave processing time significantly affect optical properties of synthesized carbon dots. UV-Vis absorbance spectra and time-resolved photoluminescence results show that luminescent of carbon dots is dominated by recombination process from n-π* surface energy level. With further development, these carbon dots are potential for several applications.

New Technique for Fabrication of Scanning Single-Electron Transistor Microscopy Tips

NASA Astrophysics Data System (ADS)

Goodwin, Eric; Tessmer, Stuart

Fabrication of glass tips for Scanning Single-Electron Transistor Microscopy (SSETM) can be expensive, time consuming, and inconsistent. Various techniques have been tried, with varying levels of success in regards to cost and reproducibility. The main requirement for SSETM tips is to have a sharp tip ending in a micron-scale flat face to allow for deposition of a quantum dot. Drawing inspiration from methods used to create tips from optical fibers for Near-Field Scanning Optical Microscopes, our group has come up with a quick and cost effective process for creating SSETM tips. By utilizing hydrofluoric acid to etch the tips and oleic acid to guide the etch profile, optical fiber tips with appropriate shaping can be rapidly prepared. Once etched, electric leads are thermally evaporated onto each side of the tip, while an aluminum quantum dot is evaporated onto the face. Preliminary results using various metals, oxide layers, and lead thicknesses have proven promising.

A multimodal imaging platform with integrated simultaneous photoacoustic microscopy, optical coherence tomography, optical Doppler tomography and fluorescence microscopy

NASA Astrophysics Data System (ADS)

Dadkhah, Arash; Zhou, Jun; Yeasmin, Nusrat; Jiao, Shuliang

2018-02-01

Various optical imaging modalities with different optical contrast mechanisms have been developed over the past years. Although most of these imaging techniques are being used in many biomedical applications and researches, integration of these techniques will allow researchers to reach the full potential of these technologies. Nevertheless, combining different imaging techniques is always challenging due to the difference in optical and hardware requirements for different imaging systems. Here, we developed a multimodal optical imaging system with the capability of providing comprehensive structural, functional and molecular information of living tissue in micrometer scale. This imaging system integrates photoacoustic microscopy (PAM), optical coherence tomography (OCT), optical Doppler tomography (ODT) and fluorescence microscopy in one platform. Optical-resolution PAM (OR-PAM) provides absorption-based imaging of biological tissues. Spectral domain OCT is able to provide structural information based on the scattering property of biological sample with no need for exogenous contrast agents. In addition, ODT is a functional extension of OCT with the capability of measurement and visualization of blood flow based on the Doppler effect. Fluorescence microscopy allows to reveal molecular information of biological tissue using autofluoresce or exogenous fluorophores. In-vivo as well as ex-vivo imaging studies demonstrated the capability of our multimodal imaging system to provide comprehensive microscopic information on biological tissues. Integrating all the aforementioned imaging modalities for simultaneous multimodal imaging has promising potential for preclinical research and clinical practice in the near future.

Optical tracking of nanoscale particles in microscale environments

PubMed Central

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

2016-01-01

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

Comparative Mirror Cleaning Study: 'A Study on Removing Particulate Contamination'

NASA Technical Reports Server (NTRS)

Houston, Karrie

2007-01-01

The cleanliness of optical surfaces is recognized as an industry-wide concern for the performance of optical devices such as mirrors and telescopes, microscopes and lenses, lasers and interferometers, and prisms and optical filters. However, no standard has been established for optical cleaning and there is no standard definition of a 'clean' optical element. This study evaluates the effectiveness of commonly used optical cleaning techniques based on wafer configuration, contamination levels, and the number and size of removed particles. It is concluded that cleaning method and exposure time play a significant factor in obtaining a high removal percentage. The detergent bath and solvent rinse method displayed an increase in effective removal percentage as the contamination exposure increased. Likewise, CO2 snow cleaning showed a relatively consistent cleaning effectiveness. The results can help ensure mission success to flight projects developed for the NASA Origins Program. Advantages and disadvantages of each of the optical cleaning methods are described.

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

NASA Astrophysics Data System (ADS)

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

2016-03-01

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

Optical levitation and translation of a microscopic particle by use of multiple beams generated by vertical-cavity surface-emitting laser array sources.

PubMed

Ogura, Yusuke; Shirai, Nobuhiro; Tanida, Jun

2002-09-20

An optical levitation and translation method for a microscopic particle by use of the resultant force induced by multiple light beams is studied. We show dependence of the radiation pressure force on the illuminating distribution by numerical calculation, and we find that the strongest axial force is obtained by a specific spacing period of illuminating beams. Extending the optical manipulation technique by means of vertical-cavity surface-emitting laser (VCSEL) array sources [Appl. Opt. 40, 5430 (2001)], we are the first, to our knowledge, to demonstrate levitation of a particle and its translation while levitated by using a VCSEL array. The vertical position of the target particle can be controlled in a range of a few tens of micrometers with an accuracy of 2 microm or less. The analytical and experimental results suggest that use of multiple beams is an effective method to levitate a particle with low total illumination power. Some issues on the manipulation method that uses multiple beams are discussed.

A review on optical actuators for microfluidic systems

NASA Astrophysics Data System (ADS)

Yang, Tie; Chen, Yue; Minzioni, Paolo

2017-12-01

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

MTF measurements on real time for performance analysis of electro-optical systems

NASA Astrophysics Data System (ADS)

Stuchi, Jose Augusto; Signoreto Barbarini, Elisa; Vieira, Flavio Pascoal; dos Santos, Daniel, Jr.; Stefani, Mário Antonio; Yasuoka, Fatima Maria Mitsue; Castro Neto, Jarbas C.; Linhari Rodrigues, Evandro Luis

2012-06-01

The need of methods and tools that assist in determining the performance of optical systems is actually increasing. One of the most used methods to perform analysis of optical systems is to measure the Modulation Transfer Function (MTF). The MTF represents a direct and quantitative verification of the image quality. This paper presents the implementation of the software, in order to calculate the MTF of electro-optical systems. The software was used for calculating the MTF of Digital Fundus Camera, Thermal Imager and Ophthalmologic Surgery Microscope. The MTF information aids the analysis of alignment and measurement of optical quality, and also defines the limit resolution of optical systems. The results obtained with the Fundus Camera and Thermal Imager was compared with the theoretical values. For the Microscope, the results were compared with MTF measured of Microscope Zeiss model, which is the quality standard of ophthalmological microscope.

Eu3+-doped Gd2O3 nanoparticles as reporters for optical detection and visualization of antibodies patterned by microcontact printing.

PubMed

Nichkova, Mikaela; Dosev, Dosi; Perron, Richard; Gee, Shirley J; Hammock, Bruce D; Kennedy, Ian M

2006-02-01

Lanthanide oxide nanoparticles are promising luminescent probes in bioanalysis, because of their unique spectral properties, photostability, and low-cost synthesis. We report for the first time the application of europium-doped gadolinium oxide (Eu:Gd2O3) nanoparticles to the optical imaging of antibody micropatterns. The nanoparticles were synthesized by spray pyrolysis and coated with antibody (IgG) molecules by physical adsorption. Our experiments showed that the Eu:Gd2O3 is a good biocompatible solid support for antibody immobilization. The antibodies (anti-rabbit IgG) immobilized on the nanoparticles had excellent biological activity in the specific recognition reaction with rabbit IgG patterned in line strips (10 micromx10 microm) on a glass substrate by use of a micro-contact printing technique. The specific immunoreaction was confirmed by two independent microscopic techniques-fluorescence and scanning electron microscopy (SEM). Both microscopic images revealed that the nanoparticles were organized into designated structures as defined by the microcontact printing process with negligible non-specific binding. The nanoparticles can be used as fluorescent markers in a variety of immunosensing applications in a microscale format.

Multimodal label-free ex vivo imaging using a dual-wavelength microscope with axial chromatic aberration compensation.

PubMed

Filippi, Andrea; Dal Sasso, Eleonora; Iop, Laura; Armani, Andrea; Gintoli, Michele; Sandri, Marco; Gerosa, Gino; Romanato, Filippo; Borile, Giulia

2018-03-01

Label-free microscopy is a very powerful technique that can be applied to study samples with no need for exogenous fluorescent probes, keeping the main benefits of multiphoton microscopy, such as longer penetration depths and intrinsic optical sectioning while enabling serial multitechniques examinations on the same specimen. Among the many label-free microscopy methods, harmonic generation (HG) is one of the most intriguing methods due to its generally low photo-toxicity and relative ease of implementation. Today, HG and common two-photon microscopy (TPM) are well-established techniques, and are routinely used in several research fields. However, they require a significant amount of fine-tuning to be fully exploited, making them quite difficult to perform in parallel. Here, we present our designed multimodal microscope, capable of performing simultaneously TPM and HG without any kind of compromise thanks to two, separate, individually optimized laser sources with axial chromatic aberration compensation. We also apply our setup to the examination of a plethora of ex vivo samples to prove its capabilities and the significant advantages of a multimodal approach. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Cathodoluminescence in the scanning transmission electron microscope.

PubMed

Kociak, M; Zagonel, L F

2017-05-01

Cathodoluminescence (CL) is a powerful tool for the investigation of optical properties of materials. In recent years, its combination with scanning transmission electron microscopy (STEM) has demonstrated great success in unveiling new physics in the field of plasmonics and quantum emitters. Most of these results were not imaginable even twenty years ago, due to conceptual and technical limitations. The purpose of this review is to present the recent advances that broke these limitations, and the new possibilities offered by the modern STEM-CL technique. We first introduce the different STEM-CL operating modes and the technical specificities in STEM-CL instrumentation. Two main classes of optical excitations, namely the coherent one (typically plasmons) and the incoherent one (typically light emission from quantum emitters) are investigated with STEM-CL. For these two main classes, we describe both the physics of light production under electron beam irradiation and the physical basis for interpreting STEM-CL experiments. We then compare STEM-CL with its better known sister techniques: scanning electron microscope CL, photoluminescence, and electron energy-loss spectroscopy. We finish by comprehensively reviewing recent STEM-CL applications. Copyright © 2016 Elsevier B.V. All rights reserved.

Cathodoluminescence in the scanning transmission electron microscope.

PubMed

Kociak, M; Zagonel, L F

2016-12-19

Cathodoluminescence (CL) is a powerful tool for the investigation of optical properties of materials. In recent years, its combination with scanning transmission electron microscopy (STEM) has demonstrated great success in unveiling new physics in the field of plasmonics and quantum emitters. Most of these results were not imaginable even twenty years ago, due to conceptual and technical limitations. The purpose of this review is to present the recent advances that broke these limitations, and the new possibilities offered by the modern STEM-CL technique. We first introduce the different STEM-CL operating modes and the technical specificities in STEM-CL instrumentation. Two main classes of optical excitations, namely the coherent one (typically plasmons) and the incoherent one (typically light emission from quantum emitters) are investigated with STEM-CL. For these two main classes, we describe both the physics of light production under electron beam irradiation and the physical basis for interpreting STEM-CL experiments. We then compare STEM-CL with its better known sister techniques: scanning electron microscope CL, photoluminescence, and electron energy-loss spectroscopy. We finish by comprehensively reviewing recent STEM-CL applications. Copyright © 2016 Elsevier B.V. All rights reserved.

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.

Universal method for creating optically active nanostructures on layered materials

NASA Astrophysics Data System (ADS)

Kidd, Tim; He, Rui; Stollenwerk, Andrew; Oshea, Aaron; Beck, Ben; Spurgeon, Kyle; Gu, Genda

2014-03-01

We report a new method for the creating of nanostructures using a scanning electron microscope. Residual organic molecules on the surface of layered materials can be excited by electron beam radiation to burrow into the open spaces between the layers of these materials, and then are broken down further to form photoluminescent carbon nanoclusters. Surface characterization by atomic force microscopy shows the surface is nearly undamaged at the molecular level by this process, and a lack of nanostructure formation in non-layered materials confirms that the structures are created by sub-surface incorporation. The presence of carbon nanoclusters was determined by Raman Spectroscopy and photoluminescence in the visible light range. The nanostructures are react strongly to visible light, making them readily apparent using an optical microscope even for features measuring only a few nanometers tall. This technique can be used on apparently any layered material, with successful results on dichalcogenides, topological insulators, graphite, and high temperature copper oxide superconductors. This technique can create patterned nanostructures with vertical resolution at the nanometer scale and lateral resolution of tens of nanometers depending on beam spot size. This work is funded by University of Northern Iowa, NSF #DMR-1206530, and DOE #DE-AC02-98CH10886.

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

PubMed

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

2013-11-01

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

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.

High-resolution interferometic microscope for traceable dimensional nanometrology in Brazil

NASA Astrophysics Data System (ADS)

Malinovski, I.; França, R. S.; Lima, M. S.; Bessa, M. S.; Silva, C. R.; Couceiro, I. B.

2016-07-01

The double color interferometric microscope is developed for step height standards nanometrology traceable to meter definition via primary wavelength laser standards. The setup is based on two stabilized lasers to provide traceable measurements of highest possible resolution down to the physical limits of the optical instruments in sub-nanometer to micrometer range of the heights. The wavelength reference is He-Ne 633 nm stabilized laser, the secondary source is Blue-Green 488 nm grating laser diode. Accurate fringe portion is measured by modulated phase-shift technique combined with imaging interferometry and Fourier processing. Self calibrating methods are developed to correct systematic interferometric errors.

Fast widefield techniques for fluorescence and phase endomicroscopy

NASA Astrophysics Data System (ADS)

Ford, Tim N.

Endomicroscopy is a recent development in biomedical optics which gives researchers and physicians microscope-resolution views of intact tissue to complement macroscopic visualization during endoscopy screening. This thesis presents HiLo endomicroscopy and oblique back-illumination endomicroscopy, fast wide-field imaging techniques with fluorescence and phase contrast, respectively. Fluorescence imaging in thick tissue is often hampered by strong out-of-focus background signal. Laser scanning confocal endomicroscopy has been developed for optically-sectioned imaging free from background, but reliance on mechanical scanning fundamentally limits the frame rate and represents significant complexity and expense. HiLo is a fast, simple, widefield fluorescence imaging technique which rejects out-of-focus background signal without the need for scanning. It works by acquiring two images of the sample under uniform and structured illumination and synthesizing an optically sectioned result with real-time image processing. Oblique back-illumination microscopy (OBM) is a label-free technique which allows, for the first time, phase gradient imaging of sub-surface morphology in thick scattering tissue with a reflection geometry. OBM works by back-illuminating the sample with the oblique diffuse reflectance from light delivered via off-axis optical fibers. The use of two diametrically opposed illumination fibers allows simultaneous and independent measurement of phase gradients and absorption contrast. Video-rate single-exposure operation using wavelength multiplexing is demonstrated.

Advanced applications of scatterometry based optical metrology

NASA Astrophysics Data System (ADS)

Dixit, Dhairya; Keller, Nick; Kagalwala, Taher; Recchia, Fiona; Lifshitz, Yevgeny; Elia, Alexander; Todi, Vinit; Fronheiser, Jody; Vaid, Alok

2017-03-01

The semiconductor industry continues to drive patterning solutions that enable devices with higher memory storage capacity, faster computing performance, and lower cost per transistor. These developments in the field of semiconductor manufacturing along with the overall minimization of the size of transistors require continuous development of metrology tools used for characterization of these complex 3D device architectures. Optical scatterometry or optical critical dimension (OCD) is one of the most prevalent inline metrology techniques in semiconductor manufacturing because it is a quick, precise and non-destructive metrology technique. However, at present OCD is predominantly used to measure the feature dimensions such as line-width, height, side-wall angle, etc. of the patterned nano structures. Use of optical scatterometry for characterizing defects such as pitch-walking, overlay, line edge roughness, etc. is fairly limited. Inspection of process induced abnormalities is a fundamental part of process yield improvement. It provides process engineers with important information about process errors, and consequently helps optimize materials and process parameters. Scatterometry is an averaging technique and extending it to measure the position of local process induced defectivity and feature-to-feature variation is extremely challenging. This report is an overview of applications and benefits of using optical scatterometry for characterizing defects such as pitch-walking, overlay and fin bending for advanced technology nodes beyond 7nm. Currently, the optical scatterometry is based on conventional spectroscopic ellipsometry and spectroscopic reflectometry measurements, but generalized ellipsometry or Mueller matrix spectroscopic ellipsometry data provides important, additional information about complex structures that exhibit anisotropy and depolarization effects. In addition the symmetry-antisymmetry properties associated with Mueller matrix (MM) elements provide an excellent means of measuring asymmetry present in the structure. The useful additional information as well as symmetry-antisymmetry properties of MM elements is used to characterize fin bending, overlay defects and design improvements in the OCD test structures are used to boost OCDs' sensitivity to pitch-walking. In addition, the validity of the OCD based results is established by comparing the results to the top down critical dimensionscanning electron microscope (CD-SEM) and cross-sectional transmission electron microscope (TEM) images.

Superresolution Imaging with Standard Fluorescent Probes

PubMed Central

Burnette, Dylan T.; Lippincott-Schwartz, Jennifer; Kachar, Bechara

2013-01-01

For more than 100 years, the ultimate resolution of a light microscope (~200 nm) has been constrained by the fundamental physical phenomenon of diffraction, as described by Ernst Abbe in 1873. While this limitation is just as applicable to today’s light microscopes, it is the combination of high-end optics, clever methods of sample illumination, and computational techniques that has enabled researchers to access high-resolution information an order of magnitude greater than once thought possible. This combination, broadly termed superresolution microscopy, has been increasingly practical for many labs to implement from both a hardware and software standpoint, but as with many cutting-edge techniques, it also comes with limitations. One of the current drawbacks to superresolution microscopy is the limited number of probes and conditions that have been suitable for imaging. Here, a technique termed bleaching/blinking assisted localization microscopy (BaLM) makes use of almost all fluorophore’s inherent blinking and bleaching properties as a means to generate superresolution images. PMID:24510788

Optical polymers for laser medical applications

NASA Astrophysics Data System (ADS)

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

2016-01-01

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

Nanocrystalline silicon thin films and grating structures for solar cells

NASA Astrophysics Data System (ADS)

Juneja, Sucheta; Sudhakar, Selvakumar; Khonina, Svetlana N.; Skidanov, Roman V.; Porfirevb, Alexey P.; Moissev, Oleg Y.; Kazanskiy, Nikolay L.; Kumar, Sushil

2016-03-01

Enhancement of optical absorption for achieving high efficiencies in thin film silicon solar cells is a challenge task. Herein, we present the use of grating structure for the enhancement of optical absorption. We have made grating structures and same can be integrated in hydrogenated micro/nanocrystalline silicon (μc/nc-Si: H) thin films based p-i-n solar cells. μc/nc-Si: H thin films were grown using plasma enhanced chemical vapor deposition method. Grating structures integrated with μc/nc-Si: H thin film solar cells may enhance the optical path length and reduce the reflection losses and its characteristics can be probed by spectroscopic and microscopic technique with control design and experiment.

Single-cell optoporation and transfection using femtosecond laser and optical tweezers.

PubMed

Waleed, Muhammad; Hwang, Sun-Uk; Kim, Jung-Dae; Shabbir, Irfan; Shin, Sang-Mo; Lee, Yong-Gu

2013-01-01

In this paper, we demonstrate a new single-cell optoporation and transfection technique using a femtosecond Gaussian laser beam and optical tweezers. Tightly focused near-infrared (NIR) femtosecond laser pulse was employed to transiently perforate the cellular membrane at a single point in MCF-7 cancer cells. A distinct technique was developed by trapping the microparticle using optical tweezers to focus the femtosecond laser precisely on the cell membrane to puncture it. Subsequently, an external gene was introduced in the cell by trapping and inserting the same plasmid-coated microparticle into the optoporated cell using optical tweezers. Various experimental parameters such as femtosecond laser exposure power, exposure time, puncture hole size, exact focusing of the femtosecond laser on the cell membrane, and cell healing time were closely analyzed to create the optimal conditions for cell viability. Following the insertion of plasmid-coated microparticles in the cell, the targeted cells exhibited green fluorescent protein (GFP) under the fluorescent microscope, hence confirming successful transfection into the cell. This new optoporation and transfection technique maximizes the level of selectivity and control over the targeted cell, and this may be a breakthrough method through which to induce controllable genetic changes in the cell.

Optical detection of ultrasound using an apertureless near-field scanning optical microscopy system

NASA Astrophysics Data System (ADS)

Ahn, Phillip; Zhang, Zhen; Sun, Cheng; Balogun, Oluwaseyi

2013-01-01

Laser ultrasonics techniques are power approaches for non-contact generation and detection of high frequency ultrasound on a local scale. In these techniques, optical diffraction limits the spatial information that can be accessed from a measurement. In order to improve the lateral spatial resolution, we incorporate an apertureless near-field scanning optical microscope (aNSOM) into laser ultrasonics setup for local detection of laser generated ultrasound. The aNSOM technique relies on the measurement of a weak backscattered near-field light intensity resulting from the oblique illumination of a nanoscale probe-tip positioned close to a sample surface. We enhance the optical near-field intensity by coupling light to surface plasmon polaritons (SPPs) on the shaft of an atomic force microscopy (AFM) cantilever. The SPPs propagate down the AFM shaft, localize at the tip apex, and are backscattered to the far-field when the separation distance between the probe tip and the sample surface is comparable to the probe-tip radius. The backscattered near-field intensity is dynamically modulated when an ultrasonic wave arrives at the sample surface leading to a transient change in the tip-sample separation distance. We present experimental results detailing measurement of broadband and narrowband laser generated ultrasound in solids with frequencies reaching up to 180 MHz range.

Pulse-Shaping-Based Nonlinear Microscopy: Development and Applications

NASA Astrophysics Data System (ADS)

Flynn, Daniel Christopher

The combination of optical microscopy and ultrafast spectroscopy make the spatial characterization of chemical kinetics on the femtosecond time scale possible. Commercially available octave-spanning Ti:Sapphire oscillators with sub-8 fs pulse durations can drive a multitude of nonlinear transitions across a significant portion of the visible spectrum with minimal average power. Unfortunately, dispersion from microscope objectives broadens pulse durations, decreases temporal resolution and lowers the peak intensities required for driving nonlinear transitions. In this dissertation, pulse shaping is used to compress laser pulses after the microscope objective. By using a binary genetic algorithm, pulse-shapes are designed to enable selective two-photon excitation. The pulse-shapes are demonstrated in two-photon fluorescence of live COS-7 cells expressing GFP-variants mAmetrine and tdTomato. The pulse-shaping approach is applied to a new multiphoton fluorescence resonance energy transfer (FRET) stoichiometry method that quantifies donor and acceptor molecules in complex, as well as the ratio of total donor to acceptor molecules. Compared to conventional multi-photon imaging techniques that require laser tuning or multiple laser systems to selectively excite individual fluorophores, the pulse-shaping approach offers rapid selective multifluorphore imaging at biologically relevant time scales. By splitting the laser beam into two beams and building a second pulse shaper, a pulse-shaping-based pump-probe microscope is developed. The technique offers multiple imaging modalities, such as excited state absorption (ESA), ground state bleach (GSB), and stimulated emission (SE), enhancing contrast of structures via their unique quantum pathways without the addition of contrast agents. Pulse-shaping based pump-probe microscopy is demonstrated for endogenous chemical-contrast imaging of red blood cells. In the second section of this dissertation, ultrafast spectroscopic techniques are used to characterize structure-function relationships of two-photon absorbing GFP-type probes and optical limiting materials. Fluorescence lifetimes of GFP-type probes are shown to depend on functional group substitution position, therefore, enabling the synthesis of designer probes for the possible study of conformation changes and aggregation in biological systems. Similarly, it is determined that small differences in the structure and dimensionality of organometallic macrocycles result in a diverse set of optical properties, which serves as a basis for the molecular level design of nonlinear optical materials.

Hybrid setup for micro- and nano-computed tomography in the hard X-ray range

NASA Astrophysics Data System (ADS)

Fella, Christian; Balles, Andreas; Hanke, Randolf; Last, Arndt; Zabler, Simon

2017-12-01

With increasing miniaturization in industry and medical technology, non-destructive testing techniques are an area of ever-increasing importance. In this framework, X-ray microscopy offers an efficient tool for the analysis, understanding, and quality assurance of microscopic samples, in particular as it allows reconstructing three-dimensional data sets of the whole sample's volume via computed tomography (CT). The following article describes a compact X-ray microscope in the hard X-ray regime around 9 keV, based on a highly brilliant liquid-metal-jet source. In comparison to commercially available instruments, it is a hybrid that works in two different modes. The first one is a micro-CT mode without optics, which uses a high-resolution detector to allow scans of samples in the millimeter range with a resolution of 1 μm. The second mode is a microscope, which contains an X-ray optical element to magnify the sample and allows resolving 150 nm features. Changing between the modes is possible without moving the sample. Thus, the instrument represents an important step towards establishing high-resolution laboratory-based multi-mode X-ray microscopy as a standard investigation method.

Hyperlens-array-implemented optical microscopy

NASA Astrophysics Data System (ADS)

Iwanaga, Masanobu

2014-08-01

Limit of resolution of conventional optical microscopes has never reached below 100 nm under visible light illumination. We show that numerically designed high-transmittance hyperlens array (HLA) is implemented in an optical microscope and works in practice for achieving one-shot-recording optical images of in-situ placed objects with sub 50 nm resolution in lateral direction. Direct resolution test employing well-defined nanopatterns proves that the HLA-implemented imaging is super-resolution optical microscopy, which works even under nW/mm2 visible illumination for objects. The HLA implementation makes the resolution of conventional microscopes one-scale higher, leading to the 1/10 illumination wavelength range, that is, mesoscopic range.

Recent Progress in Optical Biosensors Based on Smartphone Platforms

PubMed Central

Geng, Zhaoxin; Zhang, Xiong; Fan, Zhiyuan; Lv, Xiaoqing; Su, Yue; Chen, Hongda

2017-01-01

With a rapid improvement of smartphone hardware and software, especially complementary metal oxide semiconductor (CMOS) cameras, many optical biosensors based on smartphone platforms have been presented, which have pushed the development of the point-of-care testing (POCT). Imaging-based and spectrometry-based detection techniques have been widely explored via different approaches. Combined with the smartphone, imaging-based and spectrometry-based methods are currently used to investigate a wide range of molecular properties in chemical and biological science for biosensing and diagnostics. Imaging techniques based on smartphone-based microscopes are utilized to capture microscale analysts, while spectrometry-based techniques are used to probe reactions or changes of molecules. Here, we critically review the most recent progress in imaging-based and spectrometry-based smartphone-integrated platforms that have been developed for chemical experiments and biological diagnosis. We focus on the analytical performance and the complexity for implementation of the platforms. PMID:29068375

Rapid, High-Throughput Tracking of Bacterial Motility in 3D via Phase-Contrast Holographic Video Microscopy

PubMed Central

Cheong, Fook Chiong; Wong, Chui Ching; Gao, YunFeng; Nai, Mui Hoon; Cui, Yidan; Park, Sungsu; Kenney, Linda J.; Lim, Chwee Teck

2015-01-01

Tracking fast-swimming bacteria in three dimensions can be extremely challenging with current optical techniques and a microscopic approach that can rapidly acquire volumetric information is required. Here, we introduce phase-contrast holographic video microscopy as a solution for the simultaneous tracking of multiple fast moving cells in three dimensions. This technique uses interference patterns formed between the scattered and the incident field to infer the three-dimensional (3D) position and size of bacteria. Using this optical approach, motility dynamics of multiple bacteria in three dimensions, such as speed and turn angles, can be obtained within minutes. We demonstrated the feasibility of this method by effectively tracking multiple bacteria species, including Escherichia coli, Agrobacterium tumefaciens, and Pseudomonas aeruginosa. In addition, we combined our fast 3D imaging technique with a microfluidic device to present an example of a drug/chemical assay to study effects on bacterial motility. PMID:25762336

Recent Progress in Optical Biosensors Based on Smartphone Platforms.

PubMed

Geng, Zhaoxin; Zhang, Xiong; Fan, Zhiyuan; Lv, Xiaoqing; Su, Yue; Chen, Hongda

2017-10-25

With a rapid improvement of smartphone hardware and software, especially complementary metal oxide semiconductor (CMOS) cameras, many optical biosensors based on smartphone platforms have been presented, which have pushed the development of the point-of-care testing (POCT). Imaging-based and spectrometry-based detection techniques have been widely explored via different approaches. Combined with the smartphone, imaging-based and spectrometry-based methods are currently used to investigate a wide range of molecular properties in chemical and biological science for biosensing and diagnostics. Imaging techniques based on smartphone-based microscopes are utilized to capture microscale analysts, while spectrometry-based techniques are used to probe reactions or changes of molecules. Here, we critically review the most recent progress in imaging-based and spectrometry-based smartphone-integrated platforms that have been developed for chemical experiments and biological diagnosis. We focus on the analytical performance and the complexity for implementation of the platforms.

Rotary-scanning optical resolution photoacoustic microscopy

NASA Astrophysics Data System (ADS)

Qi, Weizhi; Xi, Lei

2016-10-01

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

Pressure-assisted melt-filling and optical characterization of Au nano-wires in microstructured fibers.

PubMed

Lee, H W; Schmidt, M A; Russell, R F; Joly, N Y; Tyagi, H K; Uebel, P; Russell, P St J

2011-06-20

We report a novel splicing-based pressure-assisted melt-filling technique for creating metallic nanowires in hollow channels in microstructured silica fibers. Wires with diameters as small as 120 nm (typical aspect ration 50:1) could be realized at a filling pressure of 300 bar. As an example we investigate a conventional single-mode step-index fiber with a parallel gold nanowire (wire diameter 510 nm) running next to the core. Optical transmission spectra show dips at wavelengths where guided surface plasmon modes on the nanowire phase match to the glass core mode. By monitoring the side-scattered light at narrow breaks in the nanowire, the loss could be estimated. Values as low as 0.7 dB/mm were measured at resonance, corresponding to those of an ultra-long-range eigenmode of the glass-core/nanowire system. By thermal treatment the hollow channel could be collapsed controllably, permitting creation of a conical gold nanowire, the optical properties of which could be monitored by side-scattering. The reproducibility of the technique and the high optical quality of the wires suggest applications in fields such as nonlinear plasmonics, near-field scanning optical microscope tips, cylindrical polarizers, optical sensing and telecommunications.

Augmented microscopy: real-time overlay of bright-field and near-infrared fluorescence images.

PubMed

Watson, Jeffrey R; Gainer, Christian F; Martirosyan, Nikolay; Skoch, Jesse; Lemole, G Michael; Anton, Rein; Romanowski, Marek

2015-10-01

Intraoperative applications of near-infrared (NIR) fluorescent contrast agents can be aided by instrumentation capable of merging the view of surgical field with that of NIR fluorescence. We demonstrate augmented microscopy, an intraoperative imaging technique in which bright-field (real) and electronically processed NIR fluorescence (synthetic) images are merged within the optical path of a stereomicroscope. Under luminance of 100,000 lx, representing typical illumination of the surgical field, the augmented microscope detects 189 nM concentration of indocyanine green and produces a composite of the real and synthetic images within the eyepiece of the microscope at 20 fps. Augmentation described here can be implemented as an add-on module to visualize NIR contrast agents, laser beams, or various types of electronic data within the surgical microscopes commonly used in neurosurgical, cerebrovascular, otolaryngological, and ophthalmic procedures.

Augmented microscopy: real-time overlay of bright-field and near-infrared fluorescence images

NASA Astrophysics Data System (ADS)

Watson, Jeffrey R.; Gainer, Christian F.; Martirosyan, Nikolay; Skoch, Jesse; Lemole, G. Michael, Jr.; Anton, Rein; Romanowski, Marek

2015-10-01

Intraoperative applications of near-infrared (NIR) fluorescent contrast agents can be aided by instrumentation capable of merging the view of surgical field with that of NIR fluorescence. We demonstrate augmented microscopy, an intraoperative imaging technique in which bright-field (real) and electronically processed NIR fluorescence (synthetic) images are merged within the optical path of a stereomicroscope. Under luminance of 100,000 lx, representing typical illumination of the surgical field, the augmented microscope detects 189 nM concentration of indocyanine green and produces a composite of the real and synthetic images within the eyepiece of the microscope at 20 fps. Augmentation described here can be implemented as an add-on module to visualize NIR contrast agents, laser beams, or various types of electronic data within the surgical microscopes commonly used in neurosurgical, cerebrovascular, otolaryngological, and ophthalmic procedures.

Sensing cantilever beam bending by the optical lever technique and its application to surface stress.

PubMed

Evans, Drew R; Craig, Vincent S J

2006-03-23

Cantilever beams, both microscopic and macroscopic, are used as sensors in a great variety of applications. An optical lever system is commonly employed to determine the deflection and thereby the profile of the cantilever under load. The sensitivity of the optical lever must be calibrated, and this is usually achieved by application of a known load or deflection to the free end of the cantilever. When the sensing operation involves a different type of load or a combination of types of loadings, the calibration and the deflection values derived from it become invalid. Here we develop a master equation that permits the true deflection of the cantilever to be obtained simply from the measurement of the apparent deflection for uniformly distributed loadings and end-moment loadings. These loadings are relevant to the uniform adsorption or application of material to the cantilever or the application of a surface stress to the cantilever and should assist experimentalists using the optical lever, such as in the atomic force microscope, to measure cantilever deflections in a great variety of sensing applications. We then apply this treatment to the experimental evaluation of surface stress. Three forms of Stoney's equation that relate the apparent deflection to the surface stress, which is valid for both macroscopic and microscopic experiments, are derived. Analysis of the errors arising from incorrect modeling of the loading conditions of the cantilever currently applied in experiments is also presented. It is shown that the reported literature values for surface stress in microscopic experiments are typically 9% smaller than their true value. For macroscopic experiments, we demonstrate that the added mass of the film or coating generally dominates the measured deflection and must be accounted for accurately if surface stress measurements are to be made. Further, the reported measurements generally use a form of Stoney's equation that is in error, resulting in an overestimation of surface stress by a factor >5.

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

NASA Astrophysics Data System (ADS)

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

2018-03-01

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

Nanoroughened plasmonic films for enhanced biosensing detection

NASA Astrophysics Data System (ADS)

LeMoal, Eric; Lévêque-Fort, Sandrine; Potier, Marie-Claude; Fort, Emmanuel

2009-06-01

Although fluorescence is the prevailing labeling technique in biosensing applications, sensitivity improvement is still a striving challenge. We show that coating standard microscope slides with nanoroughened silver films provides a high fluorescence signal enhancement due to plasmonic interactions. As a proof of concept, we applied these films with tailored plasmonic properties to DNA microarrays. Using common optical scanning devices, we achieved signal amplifications of more than 40-fold.

Time-resolved molecular imaging

NASA Astrophysics Data System (ADS)

Xu, Junliang; Blaga, Cosmin I.; Agostini, Pierre; DiMauro, Louis F.

2016-06-01

Time-resolved molecular imaging is a frontier of ultrafast optical science and physical chemistry. In this article, we review present and future key spectroscopic and microscopic techniques for ultrafast imaging of molecular dynamics and show their differences and connections. The advent of femtosecond lasers and free electron x-ray lasers bring us closer to this goal, which eventually will extend our knowledge about molecular dynamics to the attosecond time domain.

Highly sensitive mode mapping of whispering-gallery modes by scanning thermocouple-probe microscopy.

PubMed

Klein, Angela E; Schmidt, Carsten; Liebsch, Mattes; Janunts, Norik; Dobynde, Mikhail; Tünnermann, Andreas; Pertsch, Thomas

2014-03-01

We propose a method for mapping optical near-fields with the help of a thermocouple scanning-probe microscope tip. As the tip scans the sample surface, its apex is heated by light absorption, generating a thermovoltage. The thermovoltage map represents the intensity distribution of light at the sample surface. The measurement technique has been employed to map optical whispering-gallery modes in fused silica microdisk resonators operating at near-infrared wavelengths. The method could potentially be employed for near-field imaging of a variety of systems in the near-infrared and visible spectral range.

Theoretical study of ferroelectric nanoparticles using phase reconstructed electron microscopy

NASA Astrophysics Data System (ADS)

Phatak, C.; Petford-Long, A. K.; Beleggia, M.; De Graef, M.

2014-06-01

Ferroelectric nanostructures are important for a variety of applications in electronic and electro-optical devices, including nonvolatile memories and thin-film capacitors. These applications involve stability and switching of polarization using external stimuli, such as electric fields. We present a theoretical model describing how the shape of a nanoparticle affects its polarization in the absence of screening charges, and quantify the electron-optical phase shift for detecting ferroelectric signals with phase-sensitive techniques in a transmission electron microscope. We provide an example phase shift computation for a uniformly polarized prolate ellipsoid with varying aspect ratio in the absence of screening charges.

Three-dimensional imaging of micro-specimen by optical scanning holography

NASA Astrophysics Data System (ADS)

Liu, Jung-Ping; Tsou, Cheng-Hao

2017-04-01

Optical scanning holography (OSH) is a scanning-type digital holographic technique. In OSH, a heterodyne interference pattern is generated to raster scan the object. OSH can be operated in the incoherent mode and thus is able to record a fluorescence hologram. In addition, resolution of the OSH is proportional to the density of the interference pattern. Here we use a high-NA microscope objective to generate a dynamic Fresnel zone plate to record a hologram of micro-specimen. The achieved transverse resolution and longitudinal resolution are 0.78μm and 3.1μm, respectively.

Ultrafast IR detector response in high Tc superconducting thin films

NASA Technical Reports Server (NTRS)

Lindgren, Mikael; Ahlberg, Henrik; Danerud, Martin; Larsson, Anders; Eng, Sverre T.

1991-01-01

The response from a high Tc superconducting multielement optical detector made of a laser deposited Y-Ba-Cu-O thin film has been evaluated. Several microscopic and spectroscopic techniques were used to establish the presence of the correct phase of the thin film. Optical pulses from a laser diode at 830 nm and from a Q-switched CO2-laser at 10.6 microns were used. The detector responded to 50 ps (FWHM) pulses. A comparison between dR/dT of the film and the response amplitude as a function of temperature indicated a bolometric response.

Surface topography characterization using 3D stereoscopic reconstruction of SEM images

NASA Astrophysics Data System (ADS)

Vedantha Krishna, Amogh; Flys, Olena; Reddy, Vijeth V.; Rosén, B. G.

2018-06-01

A major drawback of the optical microscope is its limitation to resolve finer details. Many microscopes have been developed to overcome the limitations set by the diffraction of visible light. The scanning electron microscope (SEM) is one such alternative: it uses electrons for imaging, which have much smaller wavelength than photons. As a result high magnification with superior image resolution can be achieved. However, SEM generates 2D images which provide limited data for surface measurements and analysis. Often many research areas require the knowledge of 3D structures as they contribute to a comprehensive understanding of microstructure by allowing effective measurements and qualitative visualization of the samples under study. For this reason, stereo photogrammetry technique is employed to convert SEM images into 3D measurable data. This paper aims to utilize a stereoscopic reconstruction technique as a reliable method for characterization of surface topography. Reconstructed results from SEM images are compared with coherence scanning interferometer (CSI) results obtained by measuring a roughness reference standard sample. This paper presents a method to select the most robust/consistent surface texture parameters that are insensitive to the uncertainties involved in the reconstruction technique itself. Results from the two-stereoscopic reconstruction algorithms are also documented in this paper.

Dictionary of Microscopy

NASA Astrophysics Data System (ADS)

Heath, Julian

2005-10-01

The past decade has seen huge advances in the application of microscopy in all areas of science. This welcome development in microscopy has been paralleled by an expansion of the vocabulary of technical terms used in microscopy: terms have been coined for new instruments and techniques and, as microscopes reach even higher resolution, the use of terms that relate to the optical and physical principles underpinning microscopy is now commonplace. The Dictionary of Microscopy was compiled to meet this challenge and provides concise definitions of over 2,500 terms used in the fields of light microscopy, electron microscopy, scanning probe microscopy, x-ray microscopy and related techniques. Written by Dr Julian P. Heath, Editor of Microscopy and Analysis, the dictionary is intended to provide easy navigation through the microscopy terminology and to be a first point of reference for definitions of new and established terms. The Dictionary of Microscopy is an essential, accessible resource for: students who are new to the field and are learning about microscopes equipment purchasers who want an explanation of the terms used in manufacturers' literature scientists who are considering using a new microscopical technique experienced microscopists as an aide mémoire or quick source of reference librarians, the press and marketing personnel who require definitions for technical reports.

Video-rate confocal microscopy for single-molecule imaging in live cells and superresolution fluorescence imaging.

PubMed

Lee, Jinwoo; Miyanaga, Yukihiro; Ueda, Masahiro; Hohng, Sungchul

2012-10-17

There is no confocal microscope optimized for single-molecule imaging in live cells and superresolution fluorescence imaging. By combining the swiftness of the line-scanning method and the high sensitivity of wide-field detection, we have developed a, to our knowledge, novel confocal fluorescence microscope with a good optical-sectioning capability (1.0 μm), fast frame rates (<33 fps), and superior fluorescence detection efficiency. Full compatibility of the microscope with conventional cell-imaging techniques allowed us to do single-molecule imaging with a great ease at arbitrary depths of live cells. With the new microscope, we monitored diffusion motion of fluorescently labeled cAMP receptors of Dictyostelium discoideum at both the basal and apical surfaces and obtained superresolution fluorescence images of microtubules of COS-7 cells at depths in the range 0-85 μm from the surface of a coverglass. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

MIDAS: Lessons learned from the first spaceborne atomic force microscope

NASA Astrophysics Data System (ADS)

Bentley, Mark Stephen; Arends, Herman; Butler, Bart; Gavira, Jose; Jeszenszky, Harald; Mannel, Thurid; Romstedt, Jens; Schmied, Roland; Torkar, Klaus

2016-08-01

The Micro-Imaging Dust Analysis System (MIDAS) atomic force microscope (AFM) onboard the Rosetta orbiter was the first such instrument launched into space in 2004. Designed only a few years after the technique was invented, MIDAS is currently orbiting comet 67P Churyumov-Gerasimenko and producing the highest resolution 3D images of cometary dust ever made in situ. After more than a year of continuous operation much experience has been gained with this novel instrument. Coupled with operations of the Flight Spare and advances in terrestrial AFM a set of "lessons learned" has been produced, cumulating in recommendations for future spaceborne atomic force microscopes. The majority of the design could be reused as-is, or with incremental upgrades to include more modern components (e.g. the processor). Key additional recommendations are to incorporate an optical microscope to aid the search for particles and image registration, to include a variety of cantilevers (with different spring constants) and a variety of tip geometries.

Semi-automated sorting using holographic optical tweezers remotely controlled by eye/hand tracking camera

NASA Astrophysics Data System (ADS)

Tomori, Zoltan; Keša, Peter; Nikorovič, Matej; Kaůka, Jan; Zemánek, Pavel

2016-12-01

We proposed the improved control software for the holographic optical tweezers (HOT) proper for simple semi-automated sorting. The controller receives data from both the human interface sensors and the HOT microscope camera and processes them. As a result, the new positions of active laser traps are calculated, packed into the network format and sent to the remote HOT. Using the photo-polymerization technique, we created a sorting container consisting of two parallel horizontal walls where one wall contains "gates" representing a place where the trapped particle enters into the container. The positions of particles and gates are obtained by image analysis technique which can be exploited to achieve the higher level of automation. Sorting is documented on computer game simulation and the real experiment.

Optimized optical clearing method for imaging central nervous system

NASA Astrophysics Data System (ADS)

Yu, Tingting; Qi, Yisong; Gong, Hui; Luo, Qingming; Zhu, Dan

2015-03-01

The development of various optical clearing methods provides a great potential for imaging entire central nervous system by combining with multiple-labelling and microscopic imaging techniques. These methods had made certain clearing contributions with respective weaknesses, including tissue deformation, fluorescence quenching, execution complexity and antibody penetration limitation that makes immunostaining of tissue blocks difficult. The passive clarity technique (PACT) bypasses those problems and clears the samples with simple implementation, excellent transparency with fine fluorescence retention, but the passive tissue clearing method needs too long time. In this study, we not only accelerate the clearing speed of brain blocks but also preserve GFP fluorescence well by screening an optimal clearing temperature. The selection of proper temperature will make PACT more applicable, which evidently broaden the application range of this method.

Optical spectroscopies diagnose cancer

NASA Astrophysics Data System (ADS)

Alfano, Robert R.; Das, Bidyut B.; Glassman, Wenling S.; Pradhan, Asima; Tang, Gui C.

1992-02-01

Today's medical professional is looking beyond the conventional procedures of X-rays, nuclear radiation, magnetic resonance, chemical analysis, and ultrasound to diagnose diseases ranging from cancer to heart ailments. In view of the possible dangerous side effects of X-rays and nuclear radiation, a need exists for novel techniques in disease detection that can either eliminate or reduce their use in examinations. For more than half a century, fluorescence, absorption, and light scattering spectroscopies have been widely used as probes to acquire fundamental knowledge about various physical, chemical, and biological processes. Light may offer alternatives to X-rays and nuclear approaches, and in some cases is non-invasive. Optical spectroscopy and laser technology may offer techniques for the detection and characterization of physical and chemical changes that occur in diseased tissue on a microscopic level.

Nondestructive analysis of automotive paints with spectral domain optical coherence tomography.

PubMed

Dong, Yue; Lawman, Samuel; Zheng, Yalin; Williams, Dominic; Zhang, Jinke; Shen, Yao-Chun

2016-05-01

We have demonstrated for the first time, to our knowledge, the use of optical coherence tomography (OCT) as an analytical tool for nondestructively characterizing the individual paint layer thickness of multiple layered automotive paints. A graph-based segmentation method was used for automatic analysis of the thickness distribution for the top layers of solid color paints. The thicknesses measured with OCT were in good agreement with the optical microscope and ultrasonic techniques that are the current standard in the automobile industry. Because of its high axial resolution (5.5 μm), the OCT technique was shown to be able to resolve the thickness of individual paint layers down to 11 μm. With its high lateral resolution (12.4 μm), the OCT system was also able to measure the cross-sectional area of the aluminum flakes in a metallic automotive paint. The range of values measured was 300-1850  μm². In summary, the proposed OCT is a noncontact, high-resolution technique that has the potential for inclusion as part of the quality assurance process in automobile coating.

Superresolution Microscopy of the Nuclear Envelope and Associated Proteins.

PubMed

Xie, Wei; Horn, Henning F; Wright, Graham D

2016-01-01

Superresolution microscopy is undoubtedly one of the most exciting technologies since the invention of the optical microscope. Capable of nanometer-scale resolution to surpass the diffraction limit and coupled with the versatile labeling techniques available, it is revolutionizing the study of cell biology. Our understanding of the nucleus, the genetic and architectural center of the cell, has gained great advancements through the application of various superresolution microscopy techniques. This chapter describes detailed procedures of multichannel superresolution imaging of the mammalian nucleus, using structured illumination microscopy and single-molecule localization microscopy.

Crystallography of ordered colloids using optical microscopy. 2. Divergent-beam technique.

PubMed

Rogers, Richard B; Lagerlöf, K Peter D

2008-04-10

A technique has been developed to extract quantitative crystallographic data from randomly oriented colloidal crystals using a divergent-beam approach. This technique was tested on a series of diverse experimental images of colloidal crystals formed from monodisperse suspensions of sterically stabilized poly-(methyl methacrylate) spheres suspended in organic index-matching solvents. Complete sets of reciprocal lattice basis vectors were extracted in all but one case. When data extraction was successful, results appeared to be accurate to about 1% for lattice parameters and to within approximately 2 degrees for orientation. This approach is easier to implement than a previously developed parallel-beam approach with the drawback that the divergent-beam approach is not as robust in certain situations with random hexagonal close-packed crystals. The two techniques are therefore complimentary to each other, and between them it should be possible to extract quantitative crystallographic data with a conventional optical microscope from any closely index-matched colloidal crystal whose lattice parameters are compatible with visible wavelengths.

High-throughput automatic defect review for 300mm blank wafers with atomic force microscope

NASA Astrophysics Data System (ADS)

Zandiatashbar, Ardavan; Kim, Byong; Yoo, Young-kook; Lee, Keibock; Jo, Ahjin; Lee, Ju Suk; Cho, Sang-Joon; Park, Sang-il

2015-03-01

While feature size in lithography process continuously becomes smaller, defect sizes on blank wafers become more comparable to device sizes. Defects with nm-scale characteristic size could be misclassified by automated optical inspection (AOI) and require post-processing for proper classification. Atomic force microscope (AFM) is known to provide high lateral and the highest vertical resolution by mechanical probing among all techniques. However, its low throughput and tip life in addition to the laborious efforts for finding the defects have been the major limitations of this technique. In this paper we introduce automatic defect review (ADR) AFM as a post-inspection metrology tool for defect study and classification for 300 mm blank wafers and to overcome the limitations stated above. The ADR AFM provides high throughput, high resolution, and non-destructive means for obtaining 3D information for nm-scale defect review and classification.

Spirally-patterned pinhole arrays for long-term fluorescence cell imaging.

PubMed

Koo, Bon Ung; Kang, YooNa; Moon, SangJun; Lee, Won Gu

2015-11-07

Fluorescence cell imaging using a fluorescence microscope is an extensively used technique to examine the cell nucleus, internal structures, and other cellular molecules with fluorescence response time and intensity. However, it is difficult to perform high resolution cell imaging for a long period of time with this technique due to necrosis and apoptosis depending on the type and subcellular location of the damage caused by phototoxicity. A large number of studies have been performed to resolve this problem, but researchers have struggled to meet the challenge between cellular viability and image resolution. In this study, we employ a specially designed disc to reduce cell damage by controlling total fluorescence exposure time without deterioration of the image resolution. This approach has many advantages such as, the apparatus is simple, cost-effective, and easily integrated into the optical pathway through a conventional fluorescence microscope.

Probing plasmons in three dimensions by combining complementary spectroscopies in a scanning transmission electron microscope

DOE PAGES

Hachtel, Jordan A.; Marvinney, Claire; Mouti, Anas; ...

2016-03-02

The nanoscale optical response of surface plasmons in three-dimensional metallic nanostructures plays an important role in many nanotechnology applications, where precise spatial and spectral characteristics of plasmonic elements control device performance. Electron energy loss spectroscopy (EELS) and cathodoluminescence (CL) within a scanning transmission electron microscope have proven to be valuable tools for studying plasmonics at the nanoscale. Each technique has been used separately, producing three-dimensional reconstructions through tomography, often aided by simulations for complete characterization. Here we demonstrate that the complementary nature of the two techniques, namely that EELS probes beam-induced electronic excitations while CL probes radiative decay, allows usmore » to directly obtain a spatially- and spectrally-resolved picture of the plasmonic characteristics of nanostructures in three dimensions. Furthermore, the approach enables nanoparticle-by-nanoparticle plasmonic analysis in three dimensions to aid in the design of diverse nanoplasmonic applications.« less

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

Investigation of skin structures based on infrared wave parameter indirect microscopic imaging

NASA Astrophysics Data System (ADS)

Zhao, Jun; Liu, Xuefeng; Xiong, Jichuan; Zhou, Lijuan

2017-02-01

Detailed imaging and analysis of skin structures are becoming increasingly important in modern healthcare and clinic diagnosis. Nanometer resolution imaging techniques such as SEM and AFM can cause harmful damage to the sample and cannot measure the whole skin structure from the very surface through epidermis, dermis to subcutaneous. Conventional optical microscopy has the highest imaging efficiency, flexibility in onsite applications and lowest cost in manufacturing and usage, but its image resolution is too low to be accepted for biomedical analysis. Infrared parameter indirect microscopic imaging (PIMI) uses an infrared laser as the light source due to its high transmission in skins. The polarization of optical wave through the skin sample was modulated while the variation of the optical field was observed at the imaging plane. The intensity variation curve of each pixel was fitted to extract the near field polarization parameters to form indirect images. During the through-skin light modulation and image retrieving process, the curve fitting removes the blurring scattering from neighboring pixels and keeps only the field variations related to local skin structures. By using the infrared PIMI, we can break the diffraction limit, bring the wide field optical image resolution to sub-200nm, in the meantime of taking advantage of high transmission of infrared waves in skin structures.

Optical path difference microscopy with a Shack-Hartmann wavefront sensor.

PubMed

Gong, Hai; Agbana, Temitope E; Pozzi, Paolo; Soloviev, Oleg; Verhaegen, Michel; Vdovin, Gleb

2017-06-01

In this Letter, we show that a Shack-Hartmann wavefront sensor can be used for the quantitative measurement of the specimen optical path difference (OPD) in an ordinary incoherent optical microscope, if the spatial coherence of the illumination light in the plane of the specimen is larger than the microscope resolution. To satisfy this condition, the illumination numerical aperture should be smaller than the numerical aperture of the imaging lens. This principle has been successfully applied to build a high-resolution reference-free instrument for the characterization of the OPD of micro-optical components and microscopic biological samples.

Design of small confocal endo-microscopic probe working under multiwavelength environment

NASA Astrophysics Data System (ADS)

Kim, Young-Duk; Ahn, MyoungKi; Gweon, Dae-Gab

2010-02-01

Recently, optical imaging system is widely used in medical purpose. By using optical imaging system specific diseases can be easily diagnosed at early stage because optical imaging system has high resolution performance and various imaging method. These methods are used to get high resolution image of human body and can be used to verify whether the cell is infected by virus. Confocal microscope is one of the famous imaging systems which is used for in-vivo imaging. Because most of diseases are accompanied with cellular level changes, doctors can diagnosis at early stage by observing the cellular image of human organ. Current research is focused in the development of endo-microscope that has great advantage in accessibility to human body. In this research, I designed small probe that is connected to confocal microscope through optical fiber bundle and work as endo-microscope. And this small probe is mainly designed to correct chromatic aberration to use various laser sources for both fluorescence type and reflection type confocal images. By using two kinds of laser sources at the same time we demonstrated multi-modality confocal endo-microscope.

Spectro-microscopy of living plant cells.

PubMed

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

2012-01-01

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

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.

Simultaneous imaging of cellular morphology and multiple biomarkers using an acousto-optic tunable filter-based bright field microscope.

PubMed

Wachman, Elliot S; Geyer, Stanley J; Recht, Joel M; Ward, Jon; Zhang, Bill; Reed, Murray; Pannell, Chris

2014-05-01

An acousto-optic tunable filter (AOTF)-based multispectral imaging microscope system allows the combination of cellular morphology and multiple biomarker stainings on a single microscope slide. We describe advances in AOTF technology that have greatly improved spectral purity, field uniformity, and image quality. A multispectral imaging bright field microscope using these advances demonstrates pathology results that have great potential for clinical use.

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

PubMed

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

2009-07-01

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

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

PubMed

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

2010-12-10

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

Images from Phoenix's MECA Instruments

NASA Technical Reports Server (NTRS)

2008-01-01

The image on the upper left is from NASA's Phoenix Mars Lander's Optical Microscope after a sample informally called 'Sorceress' was delivered to its silicon substrate on the 38th Martian day, or sol, of the mission (July 2, 2008).

A 3D representation of the same sample is on the right, as seen by Phoenix's Atomic Force Microscope. This is 100 times greater magnification than the view from the Optical Microscope, and the most highly magnified image ever seen from another world.

The Optical Microscope and the Atomic Force Microscope are part of Phoenix's Microscopy, Electrochemistry and Conductivity Analyzer instrument.

The Atomic Force Microscope was developed by a Swiss-led consortium in collaboration with Imperial College London.

The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

Investigation on the growth, spectral, lifetime, mechanical analysis and third-order nonlinear optical studies of L-methionine admixtured D-mandelic acid single crystal: A promising material for nonlinear optical applications

NASA Astrophysics Data System (ADS)

Jayaprakash, P.; Sangeetha, P.; Kumari, C. Rathika Thaya; Caroline, M. Lydia

2017-08-01

A nonlinear optical bulk single crystal of L-methionine admixtured D-mandelic acid (LMDMA) has been grown by slow solvent evaporation technique using water as solvent at ambient temperature. The crystallized LMDMA single crystal subjected to single crystal X-ray diffraction study confirmed monoclinic system with the acentric space group P21. The FTIR analysis gives information about the modes of vibration in the various functional groups present in LMDMA. The UV-visible spectral analysis assessed the optical quality and linear optical properties such as extinction coefficient, reflectance, refractive index and from which optical conductivity and electric susceptibility were also evaluated. The frequency doubling efficiency was observed using Kurtz Perry powder technique. A multiple shot laser was utilized to evaluate the laser damage threshold energy of the crystal. Discrete thermodynamic properties were carried out by TG-DTA studies. The hardness, Meyer's index, yield strength, elastic stiffness constant, Knoop hardness, fracture toughness and brittleness index were analyzed using Vickers microhardness tester. Layer growth pattern and the surface defect were examined by chemical etching studies using optical microscope. Fluorescence emission spectrum was recorded and lifetime was also studied. The electric field response of crystal was investigated from the dielectric studies at various temperatures at different frequencies. The third-order nonlinear optical response in LMDMA has been investigated using Z-scan technique with He-Ne laser at 632.8 nm and nonlinear parameters such as refractive index (n2), absorption coefficient (β) and susceptibility (χ3) investigated extensively for they are in optical phase conjucation, high-speed optical switches and optical dielectric devices.

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.

Non-destructive optical clearing technique enhances optical coherence tomography (OCT) for real-time, 3D histomorphometry of brain tissue (Conference Presentation)

NASA Astrophysics Data System (ADS)

Paul, Akshay; Chang, Theodore H.; Chou, Li-Dek; Ramalingam, Tirunelveli S.

2016-03-01

Evaluation of neurodegenerative disease often requires examination of brain morphology. Volumetric analysis of brain regions and structures can be used to track developmental changes, progression of disease, and the presence of transgenic phenotypes. Current standards for microscopic investigation of brain morphology are limited to detection of superficial structures at a maximum depth of 300μm. While histological techniques can provide detailed cross-sections of brain structures, they require complicated tissue preparation and the ultimate destruction of the sample. A non-invasive, label-free imaging modality known as Optical Coherence Tomography (OCT) can produce 3-dimensional reconstructions through high-speed, cross-sectional scans of biological tissue. Although OCT allows for the preservation of intact samples, the highly scattering and absorbing properties of biological tissue limit imaging depth to 1-2mm. Optical clearing agents have been utilized to increase imaging depth by index matching and lipid digestion, however, these contemporary techniques are expensive and harsh on tissues, often irreversibly denaturing proteins. Here we present an ideal optical clearing agent that offers ease-of-use and reversibility. Similar to how SeeDB has been effective for microscopy, our fructose-based, reversible optical clearing technique provides improved OCT imaging and functional immunohistochemical mapping of disease. Fructose is a natural, non-toxic sugar with excellent water solubility, capable of increasing tissue transparency and reducing light scattering. We will demonstrate the improved depth-resolving performance of OCT for enhanced whole-brain imaging of normal and diseased murine brains following a fructose clearing treatment. This technique potentially enables rapid, 3-dimensional evaluation of biological tissues at axial and lateral resolutions comparable to histopathology.

Multimodal biophotonic workstation for live cell analysis.

PubMed

Esseling, Michael; Kemper, Björn; Antkowiak, Maciej; Stevenson, David J; Chaudet, Lionel; Neil, Mark A A; French, Paul W; von Bally, Gert; Dholakia, Kishan; Denz, Cornelia

2012-01-01

A reliable description and quantification of the complex physiology and reactions of living cells requires a multimodal analysis with various measurement techniques. We have investigated the integration of different techniques into a biophotonic workstation that can provide biological researchers with these capabilities. The combination of a micromanipulation tool with three different imaging principles is accomplished in a single inverted microscope which makes the results from all the techniques directly comparable. Chinese Hamster Ovary (CHO) cells were manipulated by optical tweezers while the feedback was directly analyzed by fluorescence lifetime imaging, digital holographic microscopy and dynamic phase-contrast microscopy. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Multidimensional deconvolution of optical microscope and ultrasound imaging using adaptive least-mean-square (LMS) inverse filtering

NASA Astrophysics Data System (ADS)

Sapia, Mark Angelo

2000-11-01

Three-dimensional microscope images typically suffer from reduced resolution due to the effects of convolution, optical aberrations and out-of-focus blurring. Two- dimensional ultrasound images are also degraded by convolutional bluffing and various sources of noise. Speckle noise is a major problem in ultrasound images. In microscopy and ultrasound, various methods of digital filtering have been used to improve image quality. Several methods of deconvolution filtering have been used to improve resolution by reversing the convolutional effects, many of which are based on regularization techniques and non-linear constraints. The technique discussed here is a unique linear filter for deconvolving 3D fluorescence microscopy or 2D ultrasound images. The process is to solve for the filter completely in the spatial-domain using an adaptive algorithm to converge to an optimum solution for de-blurring and resolution improvement. There are two key advantages of using an adaptive solution: (1)it efficiently solves for the filter coefficients by taking into account all sources of noise and degraded resolution at the same time, and (2)achieves near-perfect convergence to the ideal linear deconvolution filter. This linear adaptive technique has other advantages such as avoiding artifacts of frequency-domain transformations and concurrent adaptation to suppress noise. Ultimately, this approach results in better signal-to-noise characteristics with virtually no edge-ringing. Many researchers have not adopted linear techniques because of poor convergence, noise instability and negative valued data in the results. The methods presented here overcome many of these well-documented disadvantages and provide results that clearly out-perform other linear methods and may also out-perform regularization and constrained algorithms. In particular, the adaptive solution is most responsible for overcoming the poor performance associated with linear techniques. This linear adaptive approach to deconvolution is demonstrated with results of restoring blurred phantoms for both microscopy and ultrasound and restoring 3D microscope images of biological cells and 2D ultrasound images of human subjects (courtesy of General Electric and Diasonics, Inc.).

Plasmonic Imaging of Electrochemical Reactions of Single Nanoparticles.

PubMed

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

2016-11-15

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

Generation of multiple Bessel beams for a biophotonics workstation.

PubMed

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

2008-09-01

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

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

NASA Astrophysics Data System (ADS)

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

2014-09-01

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

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

PubMed

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

2014-09-01

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

Measurement of anchoring coefficient of homeotropically aligned nematic liquid crystal using a polarizing optical microscope in reflective mode

NASA Astrophysics Data System (ADS)

Baek, Sang-In; Kim, Sung-Jo; Kim, Jong-Hyun

2015-09-01

Although the homeotropic alignment of liquid crystals is widely used in LCD TVs, no easy method exists to measure its anchoring coefficient. In this study, we propose an easy and convenient measurement technique in which a polarizing optical microscope is used in the reflective mode with an objective lens having a low depth of focus. All measurements focus on the reflection of light near the interface between the liquid crystal and alignment layer. The change in the reflected light is measured by applying an electric field. We model the response of the director of the liquid crystal to the electric field and, thus, the change in reflectance. By adjusting the extrapolation length in the calculation, we match the experimental and calculated results and obtain the anchoring coefficient. In our experiment, the extrapolation lengths were 0.31 ± 0.04 μm, 0.32 ± 0.08 μm, and 0.23 ± 0.05 μm for lecithin, AL-64168, and SE-5662, respectively.

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

NASA Astrophysics Data System (ADS)

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

2012-10-01

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

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.

Noninvasive determination of optical lever sensitivity in atomic force microscopy

NASA Astrophysics Data System (ADS)

Higgins, M. J.; Proksch, R.; Sader, J. E.; Polcik, M.; Mc Endoo, S.; Cleveland, J. P.; Jarvis, S. P.

2006-01-01

Atomic force microscopes typically require knowledge of the cantilever spring constant and optical lever sensitivity in order to accurately determine the force from the cantilever deflection. In this study, we investigate a technique to calibrate the optical lever sensitivity of rectangular cantilevers that does not require contact to be made with a surface. This noncontact approach utilizes the method of Sader et al. [Rev. Sci. Instrum. 70, 3967 (1999)] to calibrate the spring constant of the cantilever in combination with the equipartition theorem [J. L. Hutter and J. Bechhoefer, Rev. Sci. Instrum. 64, 1868 (1993)] to determine the optical lever sensitivity. A comparison is presented between sensitivity values obtained from conventional static mode force curves and those derived using this noncontact approach for a range of different cantilevers in air and liquid. These measurements indicate that the method offers a quick, alternative approach for the calibration of the optical lever sensitivity.

Simulation and experimental results of optical and thermal modeling of gold nanoshells.

PubMed

Ghazanfari, Lida; Khosroshahi, Mohammad E

2014-09-01

This paper proposes a generalized method for optical and thermal modeling of synthesized magneto-optical nanoshells (MNSs) for biomedical applications. Superparamagnetic magnetite nanoparticles with diameter of 9.5 ± 1.4 nm are fabricated using co-precipitation method and subsequently covered by a thin layer of gold to obtain 15.8 ± 3.5 nm MNSs. In this paper, simulations and detailed analysis are carried out for different nanoshell geometry to achieve a maximum heat power. Structural, magnetic and optical properties of MNSs are assessed using vibrating sample magnetometer (VSM), X-ray diffraction (XRD), UV-VIS spectrophotometer, dynamic light scattering (DLS), and transmission electron microscope (TEM). Magnetic saturation of synthesized magnetite nanoparticles are reduced from 46.94 to 11.98 emu/g after coating with gold. The performance of the proposed optical-thermal modeling technique is verified by simulation and experimental results. Copyright © 2014 Elsevier B.V. All rights reserved.

Zr-doped SnO2 thin films synthesized by spray pyrolysis technique for barrier layers in solar cells

NASA Astrophysics Data System (ADS)

Reddy, N. Nanda Kumar; Akkera, Harish Sharma; Sekhar, M. Chandra; Park, Si-Hyun

2017-12-01

In the present work, we investigated the effect of Zr doping (0-6 at%) on the structural, electrical, and optical properties of tin oxide (SnO2) thin films deposited onto glass substrates using a spray pyrolysis technique. The room-temperature X-ray diffraction pattern shows that all deposited films exhibit polycrystalline tetragonal structure. The pure SnO2 film is grown along a preferred (200) direction, whereas Zr-doped SnO2 (Zr:SnO2) films started growing along the (220) orientation along with a high intensity peak of (200). Scanning electron microscope (SEM) and atomic force microscope (AFM) images showed that the grains of the films are spherical in structure, and the grain size decreased with increasing of Zr concentration. The optical transmission spectra of deposited films as a function of wavelength confirm that the average optical transmittance is > 85% for Zr:SnO2 films. The value of the optical bandgap is significantly decreased from 3.94 to 3.68 eV with increasing Zr concentration. Furthermore, the electrical measurements found that the sheet resistance ( R sh) and resistivity ( ρ) values are decreased with increasing of Zr doping. The lowest values of R sh = 6.82 Ω and ρ = 0.4 × 10- 3 Ω cm are found in 6-at% Zr-doped SnO2 film. In addition, a good efficiency value of the figure of merit ( ɸ = 3.35 × 10- 3 Ω-1) is observed in 6-at% Zr-doped SnO2 film. These outstanding properties of Zr-doped SnO2 films make them useful for several optoelectronic device applications.

Dynamic light scattering microscopy

NASA Astrophysics Data System (ADS)

Dzakpasu, Rhonda

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

Lensfree On-Chip Microscopy and Tomography for Bio-Medical Applications

PubMed Central

Isikman, Serhan O.; Bishara, Waheb; Mudanyali, Onur; Sencan, Ikbal; Su, Ting-Wei; Tseng, Derek; Yaglidere, Oguzhan; Sikora, Uzair; Ozcan, Aydogan

2012-01-01

Lensfree on-chip holographic microscopy is an emerging technique that offers imaging of biological specimens over a large field-of-view without using any lenses or bulky optical components. Lending itself to a compact, cost-effective and mechanically robust architecture, lensfree on-chip holographic microscopy can offer an alternative toolset addressing some of the emerging needs of microscopic analysis and diagnostics in low-resource settings, especially for telemedicine applications. In this review, we summarize the latest achievements in lensfree optical microscopy based on partially coherent on-chip holography, including portable telemedicine microscopy, cell-phone based microscopy and field-portable optical tomographic microscopy. We also discuss some of the future directions for telemedicine microscopy and its prospects to help combat various global health challenges. PMID:24478572

Structure and optical properties of noble metal and oxide nanoparticles dispersed in various polysaccharide biopolymers

NASA Astrophysics Data System (ADS)

Djoković, V.; Božanic, D. K.; Vodnik, V. V.; Krsmanović, R. M.; Trandafilovic, L. V.; Dimitrijević-Branković, S.

2011-10-01

We present the results on the structure and the optical properties of noble metal (Ag, Au) and oxide (ZnO) nanoparticles synthesized by various methods in different polysaccharide matrices such as chitosan, glycogen, alginate and starch. The structure of the obtained nanoparticles was studied in detail with microscopic techniques (TEM, SEM), while the XPS spectroscopy was used to investigate the effects at the nanoparticle-biomolecule interfaces. The antimicrobial activity of the nanocomposite films with Ag nanoparticles was tested against the Staphylococcus aureus, Escherichia coli and Candida albicans pathogens. In addition, we will present the results on the structure and optical properties of the tryptophan amino acid functionalized silver nanoparticles dispersed in water soluble polymer matrices.

The microscopic protein structure of the lens with a theory for cataract formation as determined by Raman spectroscopy of intact bovine lenses.

PubMed

Schachar, R A; Solin, S A

1975-05-01

Intact bovine lenses have been studied using the polarized Raman spectroscopic technique. A brief theoretical and experimental review of Raman spectroscopy is presented. From the dependence of the Raman depolarization ratio on the propagation direction of the incident radiation we have determined that the uniaxial qualities of the lens result from microscopic anisotropy and have established the quantitative positional correlation of specific chemical bonds with respect to the lens optic axis. In particular, the hydrogen bonded linear CONH groups of the antiparallel beta-pleated sheet are preferentially oriented in directions orthogonal to the lens optic axis. The Raman spectra of intact lenses do not exhibit bands at positions characteristic of either the alpha-helix or the random coil protein structure. The antiparallel beta-pleated sheet protein microstructure and the lens fiber cross-sectional macrostructure exhibit a remarkable similarity. This similarity may be causal and is consistent with the protein concentration of the lens, the birefringent properties observed by both Lenhard and Brewster, the CONH bond angle distribution with respect to the optic axis, and the lens anatomy. It is suggested that cortical cataracts are caused by fluctuations in protein orientational order.

Sub-diffraction nano manipulation using STED AFM.

PubMed

Chacko, Jenu Varghese; Canale, Claudio; Harke, Benjamin; Diaspro, Alberto

2013-01-01

In the last two decades, nano manipulation has been recognized as a potential tool of scientific interest especially in nanotechnology and nano-robotics. Contemporary optical microscopy (super resolution) techniques have also reached the nanometer scale resolution to visualize this and hence a combination of super resolution aided nano manipulation ineluctably gives a new perspective to the scenario. Here we demonstrate how specificity and rapid determination of structures provided by stimulated emission depletion (STED) microscope can aid another microscopic tool with capability of mechanical manoeuvring, like an atomic force microscope (AFM) to get topological information or to target nano scaled materials. We also give proof of principle on how high-resolution real time visualization can improve nano manipulation capability within a dense sample, and how STED-AFM is an optimal combination for this job. With these evidences, this article points to future precise nano dissections and maybe even to a nano-snooker game with an AFM tip and fluorospheres.

Augmented microscopy: real-time overlay of bright-field and near-infrared fluorescence images

PubMed Central

Watson, Jeffrey R.; Gainer, Christian F.; Martirosyan, Nikolay; Skoch, Jesse; Lemole, G. Michael; Anton, Rein; Romanowski, Marek

2015-01-01

Abstract. Intraoperative applications of near-infrared (NIR) fluorescent contrast agents can be aided by instrumentation capable of merging the view of surgical field with that of NIR fluorescence. We demonstrate augmented microscopy, an intraoperative imaging technique in which bright-field (real) and electronically processed NIR fluorescence (synthetic) images are merged within the optical path of a stereomicroscope. Under luminance of 100,000 lx, representing typical illumination of the surgical field, the augmented microscope detects 189 nM concentration of indocyanine green and produces a composite of the real and synthetic images within the eyepiece of the microscope at 20 fps. Augmentation described here can be implemented as an add-on module to visualize NIR contrast agents, laser beams, or various types of electronic data within the surgical microscopes commonly used in neurosurgical, cerebrovascular, otolaryngological, and ophthalmic procedures. PMID:26440760

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.

Optical analysis of electro-optical systems by MTF calculus

NASA Astrophysics Data System (ADS)

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

2011-08-01

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

Biological applications of near-field scanning optical microscopy

NASA Astrophysics Data System (ADS)

Moers, Marco H. P.; Ruiter, A. G. T.; Jalocha, Alain; van Hulst, Niko F.; Kalle, W. H. J.; Wiegant, J. C. A. G.; Raap, A. K.

1995-09-01

Near-field Scanning Optical Microscopy (NSOM) is a true optical microscopic technique allowing fluorescence, absorption, reflection and polarization contrast with the additional advantage of nanometer lateral resolution, unlimited by diffraction and operation at ambient conditions. NSOM based on metal coated adiabatically tapered fibers, combined with shear force feedback and operated in illumination mode, has proven to be the most powerful NSOM arrangement, because of its true localization of the optical interaction, its various optical contrast possibilities and its sensitivity down to the single molecular level. In this paper applications of `aperture' NSOM to Fluorescence In Situ Hybridization of human metaphase chromosomes are presented, where the localized fluorescence allows to identify specific DNA sequences. All images are accompanied by the simultaneously acquired force image, enabling direct comparison of the optical contrast with the sample topography on nanometer scale, far beyond the diffraction limit. Thus the unique combination of high resolution, specific optical contrast and ambient operation offers many new direction possibilities in biological studies.

The Optical Properties of Ion Implanted Silica

NASA Technical Reports Server (NTRS)

Smith, Cydale C.; Ila, D.; Sarkisov, S.; Williams, E. K.; Poker, D. B.; Hensley, D. K.

1997-01-01

We will present our investigation on the change in the optical properties of silica, 'suprasil', after keV through MeV implantation of copper, tin, silver and gold and after annealing. Suprasil-1, name brand of silica glass produced by Hereaus Amerisil, which is chemically pure with well known optical properties. Both linear nonlinear optical properties of the implanted silica were investigated before and after thermal annealing. All implants, except for Sn, showed strong optical absorption bands in agreement with Mie's theory. We have also used Z-scan to measure the strength of the third order nonlinear optical properties of the produced thin films, which is composed of the host material and the metallic nanoclusters. For implants with a measurable optical absorption band we used Doyle's theory and the full width half maximum of the absorption band to calculate the predicted size of the formed nanoclusters at various heat treatment temperatures. These results are compared with those obtained from direct observation using transmission electron microscopic techniques.

In Vivo Near Infrared Virtual Intraoperative Surgical Photoacoustic Optical Coherence Tomography

PubMed Central

Lee, Donghyun; Lee, Changho; Kim, Sehui; Zhou, Qifa; Kim, Jeehyun; Kim, Chulhong

2016-01-01

Since its first implementation in otolaryngological surgery nearly a century ago, the surgical microscope has improved the accuracy and the safety of microsurgeries. However, the microscope shows only a magnified surface view of the surgical region. To overcome this limitation, either optical coherence tomography (OCT) or photoacoustic microscopy (PAM) has been independently combined with conventional surgical microscope. Herein, we present a near-infrared virtual intraoperative photoacoustic optical coherence tomography (NIR-VISPAOCT) system that combines both PAM and OCT with a conventional surgical microscope. Using optical scattering and absorption, the NIR-VISPAOCT system simultaneously provides surgeons with real-time comprehensive biological information such as tumor margins, tissue structure, and a magnified view of the region of interest. Moreover, by utilizing a miniaturized beam projector, it can back-project 2D cross-sectional PAM and OCT images onto the microscopic view plane. In this way, both microscopic and cross-sectional PAM and OCT images are concurrently displayed on the ocular lens of the microscope. To verify the usability of the NIR-VISPAOCT system, we demonstrate simulated surgeries, including in vivo image-guided melanoma resection surgery and in vivo needle injection of carbon particles into a mouse thigh. The proposed NIR-VISPAOCT system has potential applications in neurosurgery, ophthalmological surgery, and other microsurgeries. PMID:27731390

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.

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

PubMed Central

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

2008-01-01

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

Investigation of the structural, surface, optical and electrical properties of the Indium doped CuxO thin films deposited by a thermionic vacuum arc

NASA Astrophysics Data System (ADS)

Musaoğlu, Caner; Pat, Suat; Özen, Soner; Korkmaz, Şadan; Mohammadigharehbagh, Reza

2018-03-01

In this study, investigation of some physical properties of In-doped CuxO thin films onto amorphous glass substrates were done. The thin films were depsoied by thermionic vacuum arc technique (TVA). TVA technique gives a thin film with lower precursor impurity according to the other chemical and physical depsoition methods. The microstructural properties of the produced thin films was determined by x-ray diffraction device (XRD). The thickness values were measured as to be 30 nm and 60 nm, respectively. The miller indices of the thin films’ crystalline planes were determined as to be Cu (111), CuO (\\bar{1} 12), CuInO2 (107) and Cu2O (200), Cu (111), CuO (\\bar{1} 12), CuO (\\bar{2} 02), CuInO2 (015) for sample C1 and C2, respectively. The produced In-doped CuO thin films are in polycrystalline structure. The surface properties of produced In doped CuO thin films were determined by using an atomic force microscope (AFM) and field emission scanning electron microscope (FESEM) tools. The optical properties of the In doped CuO thin films were determined by UV–vis spectrophotometer, interferometer, and photoluminescence devices. p-type semiconductor thin film was obtained by TVA depsoition.

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

PubMed

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

2017-07-01

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

Optical planar waveguides in photo-thermal-refractive glasses fabricated by single- or double-energy carbon ion implantation

NASA Astrophysics Data System (ADS)

Wang, Yue; Shen, Xiao-Liang; Zheng, Rui-Lin; Guo, Hai-Tao; Lv, Peng; Liu, Chun-Xiao

2018-01-01

Ion implantation has demonstrated to be an efficient and reliable technique for the fabrication of optical waveguides in a diversity of transparent materials. Photo-thermal-refractive glass (PTR) is considered to be durable and stable holographic recording medium. Optical planar waveguide structures in the PTR glasses were formed, for the first time to our knowledge, by the C3+-ion implantation with single-energy (6.0 MeV) and double-energy (5.5+6.0 MeV), respectively. The process of the carbon ion implantation was simulated by the stopping and range of ions in matter code. The morphologies of the waveguides were recorded by a microscope operating in transmission mode. The guided beam distributions of the waveguides were measured by the end-face coupling technique. Comparing with the single-energy implantation, the double-energy implantation improves the light confinement for the dark-mode spectrum. The guiding properties suggest that the carbon-implanted PTR glass waveguides have potential for the manufacture of photonic devices.

The Conformation of Thermoresponsive Polymer Brushes Probed by Optical Reflectivity.

PubMed

Varma, Siddhartha; Bureau, Lionel; Débarre, Delphine

2016-04-05

We describe a microscope-based optical setup that allows us to perform space- and time-resolved measurements of the spectral reflectance of transparent substrates coated with ultrathin films. This technique is applied to investigate the behavior in water of thermosensitive polymer brushes made of poly(N-isopropylacrylamide) grafted on glass. We show that spectral reflectance measurements yield quantitative information about the conformation and axial structure of the brushes as a function of temperature. We study how parameters such as grafting density and chain length affect the hydration state of a brush, and provide one of the few experimental evidences for the occurrence of vertical phase separation in the vicinity of the lower critical solution temperature of the polymer. The origin of the hysteretic behavior of poly(N-isopropylacrylamide) brushes upon cycling the temperature is also clarified. We thus demonstrate that our optical technique allows for in-depth characterization of stimuli-responsive polymer layers, which is crucial for the rational design of smart polymer coatings in actuation, gating, or sensing applications.

Analytical model of the optical vortex microscope.

PubMed

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

2016-04-20

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

Fibre-optic nonlinear optical microscopy and endoscopy.

PubMed

Fu, L; Gu, M

2007-06-01

Nonlinear optical microscopy has been an indispensable laboratory tool of high-resolution imaging in thick tissue and live animals. Rapid developments of fibre-optic components in terms of growing functionality and decreasing size provide enormous opportunities for innovations in nonlinear optical microscopy. Fibre-based nonlinear optical endoscopy is the sole instrumentation to permit the cellular imaging within hollow tissue tracts or solid organs that are inaccessible to a conventional optical microscope. This article reviews the current development of fibre-optic nonlinear optical microscopy and endoscopy, which includes crucial technologies for miniaturized nonlinear optical microscopy and their embodiments of endoscopic systems. A particular attention is given to several classes of photonic crystal fibres that have been applied to nonlinear optical microscopy due to their unique properties for ultrashort pulse delivery and signal collection. Furthermore, fibre-optic nonlinear optical imaging systems can be classified into portable microscopes suitable for imaging behaving animals, rigid endoscopes that allow for deep tissue imaging with minimally invasive manners, and flexible endoscopes enabling imaging of internal organs. Fibre-optic nonlinear optical endoscopy is coming of age and a paradigm shift leading to optical microscope tools for early cancer detection and minimally invasive surgery.

Optical, structural and nonlinear optical properties of laser ablation synthesized Ag nanoparticles and photopolymer nanocomposites based on them

NASA Astrophysics Data System (ADS)

Zulina, Natalia A.; Pavlovetc, Ilia M.; Baranov, Mikhail A.; Denisyuk, Igor Yu.

2017-03-01

In this work Ag nanoparticles (NPs) stable colloid solution were prepared by laser ablation of chemically pure silver rod in liquid monomer isodecyl acrylate (IDA). Sizes of obtained nanoparticles were determined by scanning electron microscope and vary from 27 to 120 nm. Nanocomposites films were prepared from obtained stable colloid solution of AgNPs by photocuring. For aliphatic polymer IDA long molecules cross-linking Diurethane dimethacrylate, 1,6-Hexandiol diacrylate and Tetra (ethylene glycol) diacrylate were used. Prepared nanomaterials exhibit strong third-order nonlinear optical responses, which was estimated by using z-scan technique. The third-order nonlinear optical coefficients of the studied nanocomposites were found to be up to Reχ(3)=1.31×10-5 (esu) and Imχ(3)=7.64×10-5 (esu).

Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.

PubMed

Ma, Qian; Khademhosseinieh, Bahar; Huang, Eric; Qian, Haoliang; Bakowski, Malina A; Troemel, Emily R; Liu, Zhaowei

2016-08-16

The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume.

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

NASA Technical Reports Server (NTRS)

Lauer, James L.; Abel, Phillip B.

1988-01-01

The characteristics of the scanning tunneling microscope and atomic force microscope (AFM) are briefly reviewed, and optical methods, mainly interferometry, of sufficient resolution to measure AFM deflections are discussed. The methods include optical resonators, laser interferometry, multiple-beam interferometry, and evanescent wave detection. Experimental results using AFM are reviewed.

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

PubMed

Majima, K

1998-01-01

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

Multiparallel Three-Dimensional Optical Microscopy

NASA Technical Reports Server (NTRS)

Nguyen, Lam K.; Price, Jeffrey H.; Kellner, Albert L.; Bravo-Zanoquera, Miguel

2010-01-01

Multiparallel three-dimensional optical microscopy is a method of forming an approximate three-dimensional image of a microscope sample as a collection of images from different depths through the sample. The imaging apparatus includes a single microscope plus an assembly of beam splitters and mirrors that divide the output of the microscope into multiple channels. An imaging array of photodetectors in each channel is located at a different distance along the optical path from the microscope, corresponding to a focal plane at a different depth within the sample. The optical path leading to each photodetector array also includes lenses to compensate for the variation of magnification with distance so that the images ultimately formed on all the photodetector arrays are of the same magnification. The use of optical components common to multiple channels in a simple geometry makes it possible to obtain high light-transmission efficiency with an optically and mechanically simple assembly. In addition, because images can be read out simultaneously from all the photodetector arrays, the apparatus can support three-dimensional imaging at a high scanning rate.

Acoustical nanometre-scale vibrations of live cells detected by a near-field optical setup

NASA Astrophysics Data System (ADS)

Piga, Rosaria; Micheletto, Ruggero; Kawakami, Yoichi

2007-04-01

The Scanning Near-field Optical Microscope (SNOM) is able to detect tiny vertical movement on the cell membrane in the range of only 1 nanometer or less, about 3 orders of magnitude better than conventional optical microscopes. Here we show intriguing data of cell membrane nanometer-scale dynamics associated to different phenomena of the cell’s The Scanning Near-field Optical Microscope (SNOM) is able to detect tiny vertical movement on the cell membrane in the range of only 1 nanometer or less, about 3 orders of magnitude better than conventional optical microscopes. Here we show intriguing data of cell membrane nanometer-scale dynamics associated to different phenomena of the cell’s life, such as cell cycle and cell death, on rat pheochromocytoma line PC12. Working in culture medium with alive and unperturbed samples, we could detect nanometer-sized movements; Fourier components revealed a clear distinct behavior associated to regulation of neurite outgrowth and changes on morphology after necrotic stimulus.

Improving high resolution retinal image quality using speckle illumination HiLo imaging

PubMed Central

Zhou, Xiaolin; Bedggood, Phillip; Metha, Andrew

2014-01-01

Retinal image quality from flood illumination adaptive optics (AO) ophthalmoscopes is adversely affected by out-of-focus light scatter due to the lack of confocality. This effect is more pronounced in small eyes, such as that of rodents, because the requisite high optical power confers a large dioptric thickness to the retina. A recently-developed structured illumination microscopy (SIM) technique called HiLo imaging has been shown to reduce the effect of out-of-focus light scatter in flood illumination microscopes and produce pseudo-confocal images with significantly improved image quality. In this work, we adopted the HiLo technique to a flood AO ophthalmoscope and performed AO imaging in both (physical) model and live rat eyes. The improvement in image quality from HiLo imaging is shown both qualitatively and quantitatively by using spatial spectral analysis. PMID:25136486

Improving high resolution retinal image quality using speckle illumination HiLo imaging.

PubMed

Zhou, Xiaolin; Bedggood, Phillip; Metha, Andrew

2014-08-01

Retinal image quality from flood illumination adaptive optics (AO) ophthalmoscopes is adversely affected by out-of-focus light scatter due to the lack of confocality. This effect is more pronounced in small eyes, such as that of rodents, because the requisite high optical power confers a large dioptric thickness to the retina. A recently-developed structured illumination microscopy (SIM) technique called HiLo imaging has been shown to reduce the effect of out-of-focus light scatter in flood illumination microscopes and produce pseudo-confocal images with significantly improved image quality. In this work, we adopted the HiLo technique to a flood AO ophthalmoscope and performed AO imaging in both (physical) model and live rat eyes. The improvement in image quality from HiLo imaging is shown both qualitatively and quantitatively by using spatial spectral analysis.

PDMS microfludic device for optical detection of protein immunoassay using gold nanoparticles.

PubMed

Luo, Chunxiong; Fu, Qiang; Li, Hao; Xu, Luping; Sun, Manhui; Ouyang, Qi; Chen, Yong; Ji, Hang

2005-07-01

A simple but highly specific immunoassay system for goat anti-human IgG has been developed using gold nanoparticles and microfluidic techniques. The assay is based on the deposition of gold nanoparticles that are coated with protein antigens in the presence of their corresponding antibodies to microfluidic channel surface. The effects of time accumulation, the flow velocity, and the concentration of antibodies to the red light absorption percentage (RAP) of deposition were investigated with an ordinary optical microscope. By controlling the reaction time and flow velocity, a dynamic range of 3 orders of magnitude and a detection sensitivity of 10 ng ml(-1) of goat anti-human IgG were achieved. Because of its simplicity and flexibility, this new technique should be useful for fast, highthroughput screening of antibodies in clinical diagnostic applications.

Simultaneous optical and electrical recording of a single ion-channel.

PubMed

Ide, Toru; Takeuchi, Yuko; Aoki, Takaaki; Yanagida, Toshio

2002-10-01

In recent years, the single-molecule imaging technique has proven to be a valuable tool in solving many basic problems in biophysics. The technique used to measure single-molecule functions was initially developed to study electrophysiological properties of channel proteins. However, the technology to visualize single channels at work has not received as much attention. In this study, we have for the first time, simultaneously measured the optical and electrical properties of single-channel proteins. The large conductance calcium-activated potassium channel (BK-channel) labeled with fluorescent dye molecules was incorporated into a planar bilayer membrane and the fluorescent image captured with a total internal reflection fluorescence microscope simultaneously with single-channel current recording. This innovative technology will greatly advance the study of channel proteins as well as signal transduction processes that involve ion permeation processes.

Structural and optical properties of WTe2 single crystals synthesized by DVT technique

NASA Astrophysics Data System (ADS)

Dixit, Vijay; Vyas, Chirag; Pathak, V. M.; Soalanki, G. K.; Patel, K. D.

2018-05-01

Layered transition metal di-chalcogenide (LTMDCs) crystals have attracted much attention due to their potential in optoelectronic device applications recently due to realization of their monolayer based structures. In the present investigation we report growth of WTe2 single crystals by direct vapor transport (DVT) technique. These crystals are then characterized by energy dispersive analysis of x-rays (EDAX) to study stoichiometric composition after growth. The structural properties are studied by x-ray diffraction (XRD) and selected area electron diffraction (SAED) is used to confirm orthorhombic structure of grown WTe2 crystal. Surface morphological properties of the crystals are also studied by scanning electron microscope (SEM). The optical properties of the grown crystals are studied by UV-Visible spectroscopy which gives direct band gap of 1.44 eV for grown WTe2 single crystals.

Micro Structural, Optical and Magnetic Properties of Co-SiO2 Nanocomposite Synthesized by Sol-Gel Technique

NASA Astrophysics Data System (ADS)

Saha, Mrinal; Mukherjee, Soumya; Gayen, Arup; Mukherjee, Siddhartha

2017-04-01

Co-SiO2 nano composite has been synthesized via sol-gel technique using dextrose [C6H12O6] as a reducer and tetraethyl orthosilicate [Si(OCH2CH3)4] as oxide forming agent, respectively. The dried gel has been subsequently calcined at different temperature (850 and 900 °C) for 30 min in an inert atmosphere by N2 purging. The synthesized materials have been characterized by X-ray diffraction, high resolution transmission microscope, Fourier transform infrared spectroscopy, UV-Vis spectroscopy, scanning electron microscope, field emission microscope, atomic force microscope and physical property measurement system. The crystallite sizes of the face centered cubic cobalt nano composite materials have been found to be in the range of 14-28 nm. The crystallite size of the material found to be increased at higher calcination temperature due to the grain growth. The surface morphology of the obtained material has been found to be agglomerated but spherical in nature. This agglomeration tendency could be attributed to magnetic interaction between particles, large surface area as well as high surface energy. The band gap value of the obtained material has been determined to be 1.92 eV. The saturation magnetization (Ms) and the coercivity (Hc) of the composite material were found to be 29.45 emu g-1and 23.2 Oe, respectively. This technique has thus been found to be a convenient and effective method to prepare pure metallic cobalt nanoparticles with uniform size and homogeneous distribution throughout the matrix.

Ion Engine Grid Gap Measurements

NASA Technical Reports Server (NTRS)

Soulas, Gerge C.; Frandina, Michael M.

2004-01-01

A simple technique for measuring the grid gap of an ion engine s ion optics during startup and steady-state operation was demonstrated with beam extraction. The grid gap at the center of the ion optics assembly was measured with a long distance microscope that was focused onto an alumina pin that protruded through the center accelerator grid aperture and was mechanically attached to the screen grid. This measurement technique was successfully applied to a 30 cm titanium ion optics assembly mounted onto an NSTAR engineering model ion engine. The grid gap and each grid s movement during startup from room temperature to both full and low power were measured. The grid gaps with and without beam extraction were found to be significantly different. The grid gaps at the ion optics center were both significantly smaller than the cold grid gap and different at the two power levels examined. To avoid issues associated with a small grid gap during thruster startup with titanium ion optics, a simple method was to operate the thruster initially without beam extraction to heat the ion optics. Another possible method is to apply high voltage to the grids prior to igniting the discharge because power deposition to the grids from the plasma is lower with beam extraction than without. Further testing would be required to confirm this approach.

Coherent anti-Stokes Raman scattering microscope with a high-signal-to-noise ratio, high stability, and high-speed imaging for live cell observation

NASA Astrophysics Data System (ADS)

Hayashi, Shinichi; Takimoto, Shinichi; Hashimoto, Takeshi

2007-02-01

Coherent anti-Stokes Raman scattering (CARS) microscopy, which can produce images of specific molecules without staining, has attracted the attention of researchers, as it matches the need for molecular imaging and pathway analysis of live cells. In particular, there have been an increasing number of CARS experimental results regarding lipids in live cells, which cannot be fluorescently tagged while keeping the cells alive. One of the important applications of lipid research is for the metabolic syndrome. Since the metabolic syndrome is said to be related to the lipids in lipocytes, blood, arterial vessels, and so on, the CARS technique is expected to find application in this field. However, CARS microscopy requires a pair of picosecond laser pulses, which overlap both temporally and spatially. This makes the optical adjustments of a CARS microscope challenging. The authors developed a CARS unit that includes optics for easy and stable adjustment of the overlap of these laser pulses. Adding the CARS unit to a laser scanning microscope provides CARS images of a high signal-to-noise ratio, with an acquisition rate as high as 2 microseconds per pixel. Thus, images of fast-moving lipid droplets in Hela cells were obtained.

Analysis of Phase Separation in Czochralski Grown Single Crystal Ilmenite

NASA Technical Reports Server (NTRS)

Wilkins, R.; Powell, Kirk St. A.; Loregnard, Kieron R.; Lin, Sy-Chyi; Muthusami, Jayakumar; Zhou, Feng; Pandey, R. K.; Brown, Geoff; Hawley, M. E.

1998-01-01

Ilmenite (FeTiOs) is a wide bandgap semiconductor with an energy gap of 2.58 eV. Ilmenite has properties suited for radiation tolerant applications, as well as a variety of other electronic applications. Single crystal ilmenite has been grown from the melt using the Czochralski method. Growth conditions have a profound effect on the microstructure of the samples. Here we present data from a variety of analytical techniques which indicate that some grown crystals exhibit distinct phase separation during growth. This phase separation is apparent for both post-growth annealed and unannealed samples. Under optical microscopy, there appear two distinct areas forming a matrix with an array of dots on order of 5 pm diameter. While appearing bright in the optical micrograph, atomic force microscope (AFM) shows the dots to be shallow pits on the surface. Magnetic force microscope (MFM) shows the dots to be magnetic. Phase identification via electron microprobe analysis (EMPA) indicates two major phases in the unannealed samples and four in the annealed samples, where the dots appear to be almost pure iron. This is consistent with micrographs taken with a scanning probe microscope used in the magnetic force mode. Samples that do not exhibit the phase separation have little or no discernible magnetic structure detectable by the MFM.

Effect of process parameters on microstructure and mechanical properties of friction stir welded joints: A review

NASA Astrophysics Data System (ADS)

Wanare, S. P.; Kalyankar, V. D.

2018-04-01

Friction stir welding is emerging as a promising technique for joining of lighter metal alloys due to its several advantages over conventional fusion welding processes such as low thermal distortion, good mechanical properties, fine weld joint microstructure, etc. This review article mainly focuses on analysis of microstructure and mechanical properties of friction stir welded joints. Various microstructure characterization techniques used by previous researchers such as optical microscopes, x-ray diffraction, electron probe microscope, transmission electron microscope, scanning electron microscopes with electron back scattered diffraction, electron dispersive microscopy, etc. are thoroughly overviewed and their results are discussed. The effects of friction stir welding process parameters such as tool rotational speed, welding speed, tool plunge depth, axial force, tool shoulder diameter to tool pin diameter ratio, tool geometry etc. on microstructure and mechanical properties of welded joints are studied and critical observations are noted down. The microstructure examination carried out by previous researchers on various zones of welded joints such as weld zone, heat affected zone and base metal are studied and critical remarks have been presented. Mechanical performances of friction stir welded joints based on tensile test, micro-hardness test, etc. are discussed. This article includes exhaustive literature review of standard research articles which may become ready information for subsequent researchers to establish their line of action.

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.

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.

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.

EDITORIAL: Nanotechnology in motion Nanotechnology in motion

NASA Astrophysics Data System (ADS)

Demming, Anna

2012-02-01

Microscopes provide tools of inimitable value for probing the building blocks of the world around us. The identity of the inventor of the first microscope remains under debate, but a name unequivocally linked with early developments in microscopy is Robert Hooke. His Micrographia published in 1665, was the first ever bestseller in science and brought topics in microscopy to the broader public eye with pages of detailed micrographs, most famously the fly's eye and plant cells. Since the first microscopes in the late 16th century, ingenious alternatives to the original optical microscopes have been developed to create images of the world at ever smaller dimensions. Innovations include scanning probe techniques such as the atomic force microscope [1]. As Toshio Ando describes in a review in this issue [2], these devices have also entered a new era in the past decade with the development of high-speed atomic force microscopy. Now, we can not only see the nanoscale components that make up the world around us, but we can watch them at work. One of the first innovations in optical microscopy was the use of dyes. This principle first came into practice with the use of ultraviolet light to reveal previously indistinguishable features. As explained by a researcher in the early 1930s, 'It is obvious that if the dyes used for selective staining in ordinary microscopical work are supplemented by substances which cause a particular detail of the structure to fluoresce with a specific colour in ultraviolet light, then many strings will be added to the bow of the practical microscopist' [3]. More recently, emphasis on the role of plasmons—collective oscillations of electrons in nanoscale metal structures—has received considerable research attention. Plasmons enhance the local electromagnetic field and can lead to increased fluorescence rates from nearby fluorophores depending on the efficiency of the counteracting process, non-radiative transfer [4]. The 1930s also saw the development of the electron microscope, which aimed to exceed the resolving power of diffraction-limited optical microscopes. Since the diffraction limit is proportional to the incident wavelength, the shorter wavelength electron beam allows smaller features to be resolved than optical light. Ernst Ruska shared the Nobel Prize for Physics in 1986 for his work in developing the transmission electron microscope [5]. The technique continues to provide an invaluable tool in nanotechnology studies, as demonstrated recently by a collaboration of researchers in the US, Singapore and Korea used electron and atomic force microscopy in their investigation of the deposition of gold nanoparticles on graphene and the enhanced conductivity of the doped film [6]. The other half of the 1986 Nobel Prize was awarded jointly to Gerd Binnig and Heinrich Rohrer 'for their design of the scanning tunnelling microscope'. The scanning tunnelling microscope offered the first glimpses of atomic scale features, galvanizing research in nanoscale science and technology into a burst of fruitful activity that persists to this day. Instead of using the diffraction and scattering of beams to 'see' nanoscale structures, the atomic force microscope developed by Binnig, Quate and Gerber in the 1980s [1] determines the surface topology 'by touch'. The device uses nanoscale changes in the forces exerted on a tip as it scans the sample surface to generate an image. As might be expected, innovations on the original atomic force microscope have now been developed achieving ever greater sensitivities for imaging soft matter without destroying it. Recent work by collaborators at the University of Bristol and the University of Glasgow used a cigar-shaped nanoparticle held in optical tweezers as the scanning tip. The technique is not diffraction limited, imparts less force on samples than contact scanning probe microscopy techniques, and allows highly curved and strongly scattering samples to be imaged [7]. In this issue, Toshio Ando from the University of Kanazawa provides an overview of developments that have allowed atomic force microscopy to move from rates of the order of one frame a minute to over a thousand frames per second in constant height mode, as reported by Mervyn Miles and colleagues at Bristol University and University College London [8]. Among the pioneers in the field, Ando's group demonstrated the ability to record the Brownian motion of myosin V molecules on mica with image capture rates of 100 x 100 pixels in 80 ms over a decade ago [9]. The developments unleash the potential of atomic force microscopy to observe the dynamics of biological and materials systems. If seeing is believing, the ability to present real motion pictures of the nanoworld cannot fail to capture the public imagination and stimulate burgeoning new avenues of scientific endeavour. Nearly 350 years on from the publication Micrographia, images in microscopy have moved from the page to the movies. References [1] Binnig G, Quate C F, and Gerber Ch 1986 Phys. Rev. Lett. 56 930-3 [2] Ando T 2012 Nanotechnology 23 062001 [3] J G 1934 Nature 134 635-6 [4] Bharadwaj P, Anger P and Novotny L 2007 Nanotechnology 18 044017 [5] The Nobel Prize in Physics 1986 Nobelprize.org [6] Kim K K, Reina A, Shi Y, Park H, Li L-J, Lee Y H and Kong J 2010 Nanotechnology 21 285205 [7] Phillips D B, Grieve J A, Olof S N, Kocher S J, Bowman R, Padgett M J, Miles M J and Carberry D M 2011 Nanotechnology 22 285503 [8] Picco L M, Bozec L, Ulcinas A, Engledew D J, Antognozzi M, Horton M A and Miles M J 2007 Nanotechnology 18 044030 [9] Ando T, Kodera N, Takai E, Maruyama D, Saito K and Toda A 2001 Proc. Natl. Acad. Sci. 98 12468

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

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

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

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

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

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

Array microscopy technology and its application to digital detection of Mycobacterium tuberculosis

NASA Astrophysics Data System (ADS)

McCall, Brian P.

Tuberculosis causes more deaths worldwide than any other curable infectious disease. This is the case despite tuberculosis appearing to be on the verge of eradication midway through the last century. Efforts at reversing the spread of tuberculosis have intensified since the early 1990s. Since then, microscopy has been the primary frontline diagnostic. In this dissertation, advances in clinical microscopy towards array microscopy for digital detection of Mycobacterium tuberculosis are presented. Digital array microscopy separates the tasks of microscope operation and pathogen detection and will reduce the specialization needed in order to operate the microscope. Distributing the work and reducing specialization will allow this technology to be deployed at the point of care, taking the front-line diagnostic for tuberculosis from the microscopy center to the community health center. By improving access to microscopy centers, hundreds of thousands of lives can be saved. For this dissertation, a lens was designed that can be manufactured as 4x6 array of microscopes. This lens design is diffraction limited, having less than 0.071 waves of aberration (root mean square) over the entire field of view. A total area imaged onto a full-frame digital image sensor is expected to be 3.94 mm2, which according to tuberculosis microscopy guidelines is more than sufficient for a sensitive diagnosis. The design is tolerant to single point diamond turning manufacturing errors, as found by tolerance analysis and by fabricating a prototype. Diamond micro-milling, a fabrication technique for lens array molds, was applied to plastic plano-concave and plano-convex lens arrays, and found to produce high quality optical surfaces. The micro-milling technique did not prove robust enough to produce bi-convex and meniscus lens arrays in a variety of lens shapes, however, and it required lengthy fabrication times. In order to rapidly prototype new lenses, a new diamond machining technique was developed called 4-axis single point diamond machining. This technique is 2-10x faster than micro-milling, depending on how advanced the micro-milling equipment is. With array microscope fabrication still in development, a single prototype of the lens designed for an array microscope was fabricated using single point diamond turning. The prototype microscope objective was validated in a pre-clinical trial. The prototype was compared with a standard clinical microscope objective in diagnostic tests. High concordance, a Fleiss's kappa of 0.88, was found between diagnoses made using the prototype and standard microscope objectives and a reference test. With the lens designed and validated and an advanced fabrication process developed, array microscopy technology is advanced to the point where it is feasible to rapidly prototype an array microscope for detection of tuberculosis and translate array microscope from an innovative concept to a device that can save lives.

Quantum coherent optical phase modulation in an ultrafast transmission electron microscope.

PubMed

Feist, Armin; Echternkamp, Katharina E; Schauss, Jakob; Yalunin, Sergey V; Schäfer, Sascha; Ropers, Claus

2015-05-14

Coherent manipulation of quantum systems with light is expected to be a cornerstone of future information and communication technology, including quantum computation and cryptography. The transfer of an optical phase onto a quantum wavefunction is a defining aspect of coherent interactions and forms the basis of quantum state preparation, synchronization and metrology. Light-phase-modulated electron states near atoms and molecules are essential for the techniques of attosecond science, including the generation of extreme-ultraviolet pulses and orbital tomography. In contrast, the quantum-coherent phase-modulation of energetic free-electron beams has not been demonstrated, although it promises direct access to ultrafast imaging and spectroscopy with tailored electron pulses on the attosecond scale. Here we demonstrate the coherent quantum state manipulation of free-electron populations in an electron microscope beam. We employ the interaction of ultrashort electron pulses with optical near-fields to induce Rabi oscillations in the populations of electron momentum states, observed as a function of the optical driving field. Excellent agreement with the scaling of an equal-Rabi multilevel quantum ladder is obtained, representing the observation of a light-driven 'quantum walk' coherently reshaping electron density in momentum space. We note that, after the interaction, the optically generated superposition of momentum states evolves into a train of attosecond electron pulses. Our results reveal the potential of quantum control for the precision structuring of electron densities, with possible applications ranging from ultrafast electron spectroscopy and microscopy to accelerator science and free-electron lasers.

Quantum coherent optical phase modulation in an ultrafast transmission electron microscope

NASA Astrophysics Data System (ADS)

Feist, Armin; Echternkamp, Katharina E.; Schauss, Jakob; Yalunin, Sergey V.; Schäfer, Sascha; Ropers, Claus

2015-05-01

Coherent manipulation of quantum systems with light is expected to be a cornerstone of future information and communication technology, including quantum computation and cryptography. The transfer of an optical phase onto a quantum wavefunction is a defining aspect of coherent interactions and forms the basis of quantum state preparation, synchronization and metrology. Light-phase-modulated electron states near atoms and molecules are essential for the techniques of attosecond science, including the generation of extreme-ultraviolet pulses and orbital tomography. In contrast, the quantum-coherent phase-modulation of energetic free-electron beams has not been demonstrated, although it promises direct access to ultrafast imaging and spectroscopy with tailored electron pulses on the attosecond scale. Here we demonstrate the coherent quantum state manipulation of free-electron populations in an electron microscope beam. We employ the interaction of ultrashort electron pulses with optical near-fields to induce Rabi oscillations in the populations of electron momentum states, observed as a function of the optical driving field. Excellent agreement with the scaling of an equal-Rabi multilevel quantum ladder is obtained, representing the observation of a light-driven `quantum walk' coherently reshaping electron density in momentum space. We note that, after the interaction, the optically generated superposition of momentum states evolves into a train of attosecond electron pulses. Our results reveal the potential of quantum control for the precision structuring of electron densities, with possible applications ranging from ultrafast electron spectroscopy and microscopy to accelerator science and free-electron lasers.

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

NASA Astrophysics Data System (ADS)

Kumler, James J.; Buss, Christian

2017-08-01

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

Thin film optical coatings for the ultraviolet spectral region

NASA Astrophysics Data System (ADS)

Torchio, P.; Albrand, G.; Alvisi, M.; Amra, C.; Rauf, H.; Cousin, B.; Otrio, G.

2017-11-01

The applications and innovations related to the ultraviolet field are today in strong growth. To satisfy these developments which go from biomedical to the large equipment like the Storage Ring Free Electron Laser, it is crucial to control with an extreme precision the optical performances, in using the substrates and the thin film materials impossible to circumvent in this spectral range. In particular, the reduction of the losses by electromagnetic diffusion, Joule effect absorption, or the behavior under UV luminous flows of power, resistance to surrounding particulate flows... become top priority which concerns a broad European and international community. Our laboratory has the theoretical, experimental and technological tools to design and fabricate numerous multilayer coatings with desirable optical properties in the visible and infrared spectral ranges. We have extended our expertise to the ultraviolet. We present here some results on high reflectivity multidielectric mirrors towards 250 nm in wavelength, produced by Ion Plating Deposition. The latter technique allows us to obtain surface treatments with low absorption and high resistance. We give in this study the UV transparent materials and the manufacturing technology which have been the best suited to meet requirements. Single UV layers were deposited and characterized. HfO2/SiO2 mirrors with a reflectance higher than 99% at 300 nm were obtained. Optical and non-optical characterizations such as UV spectrophotometric measurements, X-Ray Diffraction spectra, Scanning Electron Microscope and Atomic Force Microscope images were performed

HIGH TEMPERATURE MICROSCOPE AND FURNACE

DOEpatents

Olson, D.M.

1961-01-31

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

Emerging fiber optic endomicroscopy technologies towards noninvasive real-time visualization of histology in situ

NASA Astrophysics Data System (ADS)

Xi, Jiefeng; Zhang, Yuying; Huo, Li; Chen, Yongping; Jabbour, Toufic; Li, Ming-Jun; Li, Xingde

2010-09-01

This paper reviews our recent developments of ultrathin fiber-optic endomicroscopy technologies for transforming high-resolution noninvasive optical imaging techniques to in vivo and clinical applications such as early disease detection and guidance of interventions. Specifically we describe an all-fiber-optic scanning endomicroscopy technology, which miniaturizes a conventional bench-top scanning laser microscope down to a flexible fiber-optic probe of a small footprint (i.e. ~2-2.5 mm in diameter), capable of performing two-photon fluorescence and second harmonic generation microscopy in real time. This technology aims to enable realtime visualization of histology in situ without the need for tissue removal. We will also present a balloon OCT endoscopy technology which permits high-resolution 3D imaging of the entire esophagus for detection of neoplasia, guidance of biopsy and assessment of therapeutic outcome. In addition we will discuss the development of functional polymeric fluorescent nanocapsules, which use only FAD approved materials and potentially enable fast track clinical translation of optical molecular imaging and targeted therapy.

Effects of neutron irradiation on optical and chemical properties of CR-39: Potential application in neutron dosimetry.

PubMed

Sahoo, G S; Paul, S; Tripathy, S P; Sharma, S C; Jena, S; Rout, S; Joshi, D S; Bandyopadhyay, T

2014-12-01

Effects of high-dose neutron irradiation on chemical and optical properties of CR-39 were studied using FTIR (Fourier Transform Infrared) and UV-vis (Ultraviolet-Visible) spectroscopy. The primary goal was to find a correlation between the neutron dose and the corresponding changes in the optical and chemical properties of CR-39 resulted from the neutron irradiation. The neutrons were produced by bombarding a thick Be target with 22-MeV protons. In the FTIR spectra, prominent absorbance peaks were observed at 1735cm(-1) (C=O stretching), 1230cm(-1)(C-O-C stretching), and 783cm(-1)(=C-H bending), the intensities of which decreased with increasing neutron dose. The optical absorbance in the visible range increased linearly with the neutron dose. Empirical relations were established to estimate neutron doses from these optical properties. This technique is particularly useful in measuring high doses, where track analysis with an optical microscope is difficult because of track overlapping. Copyright © 2014 Elsevier Ltd. All rights reserved.

Sedimentological Investigations of the Martian Surface using the Mars 2001 Robotic Arm Camera and MECA Optical Microscope

NASA Technical Reports Server (NTRS)

Rice, J. W., Jr.; Smith, P. H.; Marshall, J. R.

1999-01-01

The first microscopic sedimentological studies of the Martian surface will commence with the landing of the Mars Polar Lander (MPL) December 3, 1999. The Robotic Arm Camera (RAC) has a resolution of 25 um/p which will permit detailed micromorphological analysis of surface and subsurface materials. The Robotic Ann will be able to dig up to 50 cm below the surface. The walls of the trench will also be inspected by RAC to look for evidence of stratigraphic and / or sedimentological relationships. The 2001 Mars Lander will build upon and expand the sedimentological research begun by the RAC on MPL. This will be accomplished by: (1) Macroscopic (dm to cm): Descent Imager, Pancam, RAC; (2) Microscopic (mm to um RAC, MECA Optical Microscope (Figure 2), AFM This paper will focus on investigations that can be conducted by the RAC and MECA Optical Microscope.

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

ScienceCinema

Nazaretski, Evgeny

2018-06-13

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

Multilayer mounting enables long-term imaging of zebrafish development in a light sheet microscope.

PubMed

Kaufmann, Anna; Mickoleit, Michaela; Weber, Michael; Huisken, Jan

2012-09-01

Light sheet microscopy techniques, such as selective plane illumination microscopy (SPIM), are ideally suited for time-lapse imaging of developmental processes lasting several hours to a few days. The success of this promising technology has mainly been limited by the lack of suitable techniques for mounting fragile samples. Embedding zebrafish embryos in agarose, which is common in conventional confocal microscopy, has resulted in severe growth defects and unreliable results. In this study, we systematically quantified the viability and mobility of zebrafish embryos mounted under more suitable conditions. We found that tubes made of fluorinated ethylene propylene (FEP) filled with low concentrations of agarose or methylcellulose provided an optimal balance between sufficient confinement of the living embryo in a physiological environment over 3 days and optical clarity suitable for fluorescence imaging. We also compared the effect of different concentrations of Tricaine on the development of zebrafish and provide guidelines for its optimal use depending on the application. Our results will make light sheet microscopy techniques applicable to more fields of developmental biology, in particular the multiview long-term imaging of zebrafish embryos and other small organisms. Furthermore, the refinement of sample preparation for in toto and in vivo imaging will promote other emerging optical imaging techniques, such as optical projection tomography (OPT).

Acetone sensor based on zinc oxide hexagonal tubes

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

Hastir, Anita, E-mail: anitahastir@gmail.com; Singh, Onkar, E-mail: anitahastir@gmail.com; Anand, Kanika, E-mail: anitahastir@gmail.com

2014-04-24

In this work hexagonal tubes of zinc oxide have been synthesized by co-precipitation method. For structural, morphological, elemental and optical analysis synthesized powders were characterized by using x-ray diffraction, field emission scanning microscope, EDX, UV-visible and FTIR techniques. For acetone sensing thick films of zinc oxide have been deposited on alumina substrate. The fabricated sensors exhibited maximum sensing response towards acetone vapour at an optimum operating temperature of 400°C.

Biological imaging with coherent Raman scattering microscopy: a tutorial

PubMed Central

Alfonso-García, Alba; Mittal, Richa; Lee, Eun Seong; Potma, Eric O.

2014-01-01

Abstract. Coherent Raman scattering (CRS) microscopy is gaining acceptance as a valuable addition to the imaging toolset of biological researchers. Optimal use of this label-free imaging technique benefits from a basic understanding of the physical principles and technical merits of the CRS microscope. This tutorial offers qualitative explanations of the principles behind CRS microscopy and provides information about the applicability of this nonlinear optical imaging approach for biological research. PMID:24615671

High-speed transport-of-intensity phase microscopy with an electrically tunable lens.

PubMed

Zuo, Chao; Chen, Qian; Qu, Weijuan; Asundi, Anand

2013-10-07

We present a high-speed transport-of-intensity equation (TIE) quantitative phase microscopy technique, named TL-TIE, by combining an electrically tunable lens with a conventional transmission microscope. This permits the specimen at different focus position to be imaged in rapid succession, with constant magnification and no physically moving parts. The simplified image stack collection significantly reduces the acquisition time, allows for the diffraction-limited through-focus intensity stack collection at 15 frames per second, making dynamic TIE phase imaging possible. The technique is demonstrated by profiling of microlens array using optimal frequency selection scheme, and time-lapse imaging of live breast cancer cells by inversion the defocused phase optical transfer function to correct the phase blurring in traditional TIE. Experimental results illustrate its outstanding capability of the technique for quantitative phase imaging, through a simple, non-interferometric, high-speed, high-resolution, and unwrapping-free approach with prosperous applications in micro-optics, life sciences and bio-photonics.

Bulk heterojunction thin film formation by single and dual feed ultrasonic spray method for application in organic solar cells

NASA Astrophysics Data System (ADS)

Marathe, D. M.; Tarkas, H. S.; Mahajan, M. S.; Lonkar, G. S.; Tak, S. R.; Sali, J. V.

2016-09-01

We here present a way of preparing the polymer: fullerene BHJ using dual feed method which can lead to formation of pure phases. In this report, we present results of our initial experiments in this direction. The effect of process parameters on the thickness and surface roughness of the active layer has been discussed. The structural and optical properties have been studied using the optical microscope, UV—visible spectroscopy and photoluminescence spectroscopy. Significant PL quenching indicates efficient charge separation in the BHJ formed using this technique. We have also compared the BHJ thin films prepared with this dual feed ultrasonic technique with the single feed spray method. The BHJ formed using this technique has been used as an active layer in OSC. supported by the University Grants Commission, New Delhi, under Faculty Improvement Programme (No. 33-02/12(WRO) Dt.19.03.2013) and the Special Assistance Programme (530/2/DRS/2010(SAP-I)) Phase-II.

White matter segmentation by estimating tissue optical attenuation from volumetric OCT massive histology of whole rodent brains

NASA Astrophysics Data System (ADS)

Lefebvre, Joël.; Castonguay, Alexandre; Lesage, Frédéric

2017-02-01

A whole rodent brain was imaged using an automated massive histology setup and an Optical Coherence Tomography (OCT) microscope. Thousands of OCT volumetric tiles were acquired, each covering a size of about 2.5x2.5x0.8 mm3 with a sampling resolution of 4.9x4.9x6.5 microns. This paper shows the techniques for reconstruction, attenuation compensation and segmentation of the sliced brains. The tile positions within the mosaic were evaluated using a displacement model of the motorized stage and pairwise coregistration. Volume blending was then performed by solving the 3D Laplace equation, and consecutive slices were assembled using the cross-correlation of their 2D image gradient. This reconstruction algorithm resulted in a 3D map of optical reflectivity for the whole brain at micrometric resolution. OCT tissue slices were then used to estimate the local attenuation coefficient based on a single scattering photon model. The attenuation map obtained exhibits a high contrast for all white matter fibres, regardless of their orientation. The tissue optical attenuation from the intrinsic OCT reflectivity contributes to better white matter tissue segmentation. The combined 3D maps of reflectivity and attenuation is a step toward the study of white matter at a microscopic scale for the whole brain in small animals.

Mirror-based broadband scanner with minimized aberration

NASA Astrophysics Data System (ADS)

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

2009-02-01

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

A compact CCD-monitored atomic force microscope with optical vision and improved performances.

PubMed

Mingyue, Liu; Haijun, Zhang; Dongxian, Zhang

2013-09-01

A novel CCD-monitored atomic force microscope (AFM) with optical vision and improved performances has been developed. Compact optical paths are specifically devised for both tip-sample microscopic monitoring and cantilever's deflection detecting with minimized volume and optimal light-amplifying ratio. The ingeniously designed AFM probe with such optical paths enables quick and safe tip-sample approaching, convenient and effective tip-sample positioning, and high quality image scanning. An image stitching method is also developed to build a wider-range AFM image under monitoring. Experiments show that this AFM system can offer real-time optical vision for tip-sample monitoring with wide visual field and/or high lateral optical resolution by simply switching the objective; meanwhile, it has the elegant performances of nanometer resolution, high stability, and high scan speed. Furthermore, it is capable of conducting wider-range image measurement while keeping nanometer resolution. Copyright © 2013 Wiley Periodicals, Inc.

Optical anisotropy of the human cornea determined with a polarizing microscope.

PubMed

Bone, Richard A; Draper, Grenville

2007-12-01

We have investigated the optical anisotropy of the human cornea using a polarizing microscope normally used for optical mineralogy studies. The central part of the cornea was removed from 14 eyes (seven donors). With the sample placed on the microscope stage, we consistently observed hyperbolic isogyres characteristic of a negative biaxial material. The angle between the optic axes, generally similar in both eyes, ranged from 12 degrees to 40 degrees (mean+/-SD=31 degrees +/-8 degrees ). The optic axial plane always inclined downward in the nasal direction at 1 degrees -45 degrees below the horizontal (mean+/-SD=22+/-13 degrees ). The retardance produced by the corneas was estimated to be less than 200 nm. In conclusion, the human cornea possesses the anisotropy of a negative biaxial material. Both the angle between the optic axes and the retardance were fairly constant among the majority of samples, suggestive of uniformity in corneal structure.

Enhanced coupling of light into a turbid medium through microscopic interface engineering

PubMed Central

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

2017-01-01

There are many optical detection and sensing methods used today that provide powerful ways to diagnose, characterize, and study materials. For example, the measurement of spontaneous Raman scattering allows for remote detection and identification of chemicals. Many other optical techniques provide unique solutions to learn about biological, chemical, and even structural systems. However, when these systems exist in a highly scattering or turbid medium, the optical scattering effects reduce the effectiveness of these methods. In this article, we demonstrate a method to engineer the geometry of the optical interface of a turbid medium, thereby drastically enhancing the coupling efficiency of light into the material. This enhanced optical coupling means that light incident on the material will penetrate deeper into (and through) the medium. It also means that light thus injected into the material will have an enhanced interaction time with particles contained within the material. These results show that, by using the multiple scattering of light in a turbid medium, enhanced light–matter interaction can be achieved; this has a direct impact on spectroscopic methods such as Raman scattering and fluorescence detection in highly scattering regimes. Furthermore, the enhanced penetration depth achieved by this method will directly impact optical techniques that have previously been limited by the inability to deposit sufficient amounts of optical energy below or through highly scattering layers. PMID:28701381

Raman spectroscopy for medical diagnostics--From in-vitro biofluid assays to in-vivo cancer detection.

PubMed

Kong, Kenny; Kendall, Catherine; Stone, Nicholas; Notingher, Ioan

2015-07-15

Raman spectroscopy is an optical technique based on inelastic scattering of light by vibrating molecules and can provide chemical fingerprints of cells, tissues or biofluids. The high chemical specificity, minimal or lack of sample preparation and the ability to use advanced optical technologies in the visible or near-infrared spectral range (lasers, microscopes, fibre-optics) have recently led to an increase in medical diagnostic applications of Raman spectroscopy. The key hypothesis underpinning this field is that molecular changes in cells, tissues or biofluids, that are either the cause or the effect of diseases, can be detected and quantified by Raman spectroscopy. Furthermore, multivariate calibration and classification models based on Raman spectra can be developed on large "training" datasets and used subsequently on samples from new patients to obtain quantitative and objective diagnosis. Historically, spontaneous Raman spectroscopy has been known as a low signal technique requiring relatively long acquisition times. Nevertheless, new strategies have been developed recently to overcome these issues: non-linear optical effects and metallic nanoparticles can be used to enhance the Raman signals, optimised fibre-optic Raman probes can be used for real-time in-vivo single-point measurements, while multimodal integration with other optical techniques can guide the Raman measurements to increase the acquisition speed and spatial accuracy of diagnosis. These recent efforts have advanced Raman spectroscopy to the point where the diagnostic accuracy and speed are compatible with clinical use. This paper reviews the main Raman spectroscopy techniques used in medical diagnostics and provides an overview of various applications. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Design of an imaging microscope for soft X-ray applications

NASA Astrophysics Data System (ADS)

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

1988-01-01

An imaging soft X-ray microscope with a spatial resolution of 0.1 micron and normal incidence multilayer optics is discussed. The microscope has a Schwarzschild configuration, which consists of two concentric spherical mirrors with radii of curvature which minimize third-order spherical aberration, coma, and astigmatism. The performance of the Stanford/MSFC Cassegrain X-ray telescope and its relevance to the present microscope are addressed. A ray tracing analysis of the optical system indicates that diffraction-limited performance can be expected for an object height of 0.2 mm.

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.

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

Cholesterol-Based Grafted Polymer Brushes as Alignment Coating with Temperature-Tuned Anchoring for Nematic Liquid Crystals.

PubMed

Stetsyshyn, Yurij; Raczkowska, Joanna; Budkowski, Andrzej; Awsiuk, Kamil; Kostruba, Andriy; Nastyshyn, Svyatoslav; Harhay, Khrystyna; Lychkovskyy, Edward; Ohar, Halyna; Nastishin, Yuriy

2016-10-11

Novel alignment coating with temperature-tuned anchoring for nematic liquid crystals (NLCs) was successfully fabricated in three step process, involving polymerization of poly(cholesteryl methacrylate) (PChMa) from oligoproxide grafted to the glass surface premodified with 3-aminopropyltriethoxysilane. Molecular composition, thickness, wettability of the PChMa coating and its alignment action for a NLC were examined with time of flight-secondary ion mass spectrometry, ellipsometry, contact angle measurements, polarization optical microscopy and commercially produced PolScope technique allowing for mapping of the optic axis and optical retardance within the microscope field view. We find that the PChMa coating provides a specific monotonous increase (decrease) in the tilt angle of the NLC director with respect to the substrates normal upon heating (cooling) referred to as anchoring tuning.

Enhanced and selective optical trapping in a slot-graphite photonic crystal.

PubMed

Krishnan, Aravind; Huang, Ningfeng; Wu, Shao-Hua; Martínez, Luis Javier; Povinelli, Michelle L

2016-10-03

Applicability of optical trapping tools for nanomanipulation is limited by the available laser power and trap efficiency. We utilized the strong confinement of light in a slot-graphite photonic crystal to develop high-efficiency parallel trapping over a large area. The stiffness is 35 times higher than our previously demonstrated on-chip, near field traps. We demonstrate the ability to trap both dielectric and metallic particles of sub-micron size. We find that the growth kinetics of nanoparticle arrays on the slot-graphite template depends on particle size. This difference is exploited to selectively trap one type of particle out of a binary colloidal mixture, creating an efficient optical sieve. This technique has rich potential for analysis, diagnostics, and enrichment and sorting of microscopic entities.

Classification of Salmonella serotypes with hyperspectral microscope imagery

USDA-ARS?s Scientific Manuscript database

Previous research has demonstrated an optical method with acousto-optic tunable filter (AOTF) based hyperspectral microscope imaging (HMI) had potential for classifying gram-negative from gram-positive foodborne pathogenic bacteria rapidly and nondestructively with a minimum sample preparation. In t...

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

NASA Astrophysics Data System (ADS)

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

2018-06-01

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

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.

U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

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

Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.

2016-10-01

The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

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

Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.

2016-03-30

The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

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

Micropaleontological studies of lunar and terrestrial precambrian materials

NASA Technical Reports Server (NTRS)

Schope, J. W.

1974-01-01

Optical microscopic and scanning electron microscopic studies of rock chips and dust returned by Apollo 14, 15, 16, and 17 are analyzed along with optical microscopic studies of petrographic thin sections of breccias and basalts returned by Apollo 14, 15, and 16. Results show no evidence of modern or fossil lunar organisms. The lunar surface is now, and apparently has been throughout the geologic past, inimical to known biologic systems.

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

PubMed Central

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

2012-01-01

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

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

PubMed

Stemmer, A

1995-04-01

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

Optical levitation and manipulation of stuck particles with pulsed optical tweezers.

PubMed

Ambardekar, Amol Ashok; Li, Yong-Qing

2005-07-15

We report on optical levitation and manipulation of microscopic particles that are stuck on a glass surface with pulsed optical tweezers. An infrared pulse laser at 1.06 microm was used to generate a large gradient force (up to 10(-9) N) within a short duration (approximately 45 micros) that overcomes the adhesive interaction between the particles and the glass surface. Then a low-power continuous-wave diode laser at 785 nm was used to capture and manipulate the levitated particle. We have demonstrated that both stuck dielectric and biological micrometer-sized particles, including polystyrene beads, yeast cells, and Bacillus cereus bacteria, can be levitated and manipulated with this technique. We measured the single-pulse levitation efficiency for 2.0 microm polystyrene beads as a function of the pulse energy and of the axial displacement from the stuck particle to the pulsed laser focus, which was as high as 88%.

Plastic fiber optics for micro-imaging of fluorescence signals in living cells

NASA Astrophysics Data System (ADS)

Sakurai, Takashi; Natsume, Mitsuo; Koida, Kowa

2015-03-01

The fiber-coupled microscope (FCM) enables in vivo imaging at deep sites in the tissues or organs that other optical techniques are unable to reach. To develop FCM-based intravital imaging, we employed a plastic optical fiber (POF) bundle that included more than 10,000-units of polystyrene core and polymethyl methacrylate cladding. Each POF had a diameter of less than 5 μm the tip of the bundle was less than 0.5 mm wide, and the flexible wire had a length of 1,000 mm. The optical performance of the plastic FCM was sufficient for detection of significant signal changes in an acinus of rat pancreas labeled with a calcium ion-sensitive fluorescent dye. In the future, the potential power of plastic FCM is expected to increase, enabling analysis of structure and organization of specific functions in live cells within vulnerable organs.

Giant Kerr response of ultrathin gold films from quantum size effect.

PubMed

Qian, Haoliang; Xiao, Yuzhe; Liu, Zhaowei

2016-10-10

With the size of plasmonic devices entering into the nanoscale region, the impact of quantum physics needs to be considered. In the past, the quantum size effect on linear material properties has been studied extensively. However, the nonlinear aspects have not been explored much so far. On the other hand, much effort has been put into the field of integrated nonlinear optics and a medium with large nonlinearity is desirable. Here we study the optical nonlinear properties of a nanometre scale gold quantum well by using the z-scan method and nonlinear spectrum broadening technique. The quantum size effect results in a giant optical Kerr susceptibility, which is four orders of magnitude higher than the intrinsic value of bulk gold and several orders larger than traditional nonlinear media. Such high nonlinearity enables efficient nonlinear interaction within a microscopic footprint, making quantum metallic films a promising candidate for integrated nonlinear optical applications.

Photonic engineering for biological study

NASA Astrophysics Data System (ADS)

Wu, Fei

My dissertation focuses on designing and developing prototypes of optical tools in the laboratory that can facilitate practical medical therapies. More specifically, this dissertation examines two novel biophotonic techniques: (1) a frequency multiplexed confocal microscope with the potential to provide rational therapy of congestive heart failure (CHF), and (2) the "optical comb" with the potential to improve results of retina reattachment surgery and accelerate post surgical recovery. Next, I will discuss the background, design and initial experimental results of each study individually. Part I: The Frequency Multiplexed Confocal Microscope. To overcome the limitations of existing confocal microscope technology, this dissertation proposes a non-scanning, real-time, high resolution technique (a multi-point frequency multiplexed confocal microscope) to measure 3-D intracellular calcium ion concentration in a living cardiac myocyte. This method can be also applied to measure the intracellular sodium ion concentration, or other ions in which high quantum-yield fluorescent probes are available. The novelty of the proposed research lies in the introduction of carrier frequency multiplexing techniques which can differentiate fluorescence emitted at different spatial locations in cardiac myocyte by their modulated frequency. It therefore opens the possibility to visualize the transient dynamics of intracellular dynamics at multiple locations in cells simultaneously, which will shine a new light on our understanding of CHF. The procedure for frequency multiplexing proposed is described below. Multiple incident laser beams are focused onto different locations in an isolated rat cardiac myocyte with each beam modulated at a different carrier frequency. The fluorescence emission at each location therefore bears the same modulated frequency as the stimulation laser beam. Each fluorescence signal is sent to the photo multiplier tube (PMT) after being spatially filtered by a single mode fiber (functioning as a pinhole). Since each signal has a different carrier frequency, only one signal detector is required to collect multiple signal streams which eliminates the errors introduced by difference of multiple detectors. After taking the Fourier Transform of the collected data, multiple peaks can be found in the frequency domain. Each peak refers to a corresponding location in the sample. The temporal information of the fluorescence signal variation at each location can be obtained by demodulating the low frequency information from the carrier frequency, followed by an inverse Fourier transform. Part II: The "Optical Comb". Retinal detachment refers to separation of the inner layers of the retina from the underlying retinal pigment epithelium. It can cause degeneration of the retina and may lead to permanent vision loss if not promptly treated and hence is considered an ocular emergency. Currently, the only treatment available for retinal detachment is surgical reattachment. The idea of an "optical comb" is developed from the general working principle of the well known "optical tweezers" in the optical literature, which can pull micro-objects through the trapping force produced by a focused laser beam. If we can manage to incident the focused laser beam onto the misaligned photoreceptors and further scan it back and forth, trapping forces that produced may be able to "comb" the photoreceptors to be aligned, and thereby help with post surgery recovery. A series of experiments have been carried out to demonstrate the plausibility of this idea. First, several micro glass rods with size similar to human's photoreceptors (6 microns in diameter and 30 microns in length) were used. We observed that when the laser beam is focused close to one end of the micro rod originally laid on a glass coverslip, the rod is pulled to stand upright successfully, and we can manipulate the direction it faces by controlling its relative position to the laser beam. We are now experimenting with this combing technique with detached bovine retina samples to further verify its feasibility over live animal cells. (Abstract shortened by UMI.)

Influence of graphene quantum dots on electrical properties of polymer composites

NASA Astrophysics Data System (ADS)

Arthisree, D.; Joshi, Girish M.

2017-07-01

We successfully prepared synthetic nanocomposite (SNC) by dispersing graphene quantum dots (GQD) in cellulose acetate (CA) polymer system. The dispersion and occupied network of GQD were foreseen by microscopic techniques. The variation of plane to crossed linked array network was observed by the polarizing optical microscopic (POM) technique. The scanning electron microscopy (SEM) revealed the leaves like impressions of GQD in host polymer system. The series network of GQD occupied in CA at higher resolution was confirmed by transmission electron microscopy (TEM). The two dimensional (2D) topographic images demonstrated an entangled polymer network to plane morphology. The variation in surface roughness was evaluated from the dimensional (3D) topography. The influence of temperature on AC conductivity with highest value (4  ×  10-5 S cm-1), contributes to the decrease in activation energy. The DC conductivity obeys the percolation criteria co-related to the GQD loading by weight fraction. Furthermore, this synthetic nanocomposite is feasible for the development of sensing and electrical applications.

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.

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.

Wafer defect detection by a polarization-insensitive external differential interference contrast module.

PubMed

Nativ, Amit; Feldman, Haim; Shaked, Natan T

2018-05-01

We present a system that is based on a new external, polarization-insensitive differential interference contrast (DIC) module specifically adapted for detecting defects in semiconductor wafers. We obtained defect signal enhancement relative to the surrounding wafer pattern when compared with bright-field imaging. The new DIC module proposed is based on a shearing interferometer that connects externally at the output port of an optical microscope and enables imaging thin samples, such as wafer defects. This module does not require polarization optics (such as Wollaston or Nomarski prisms) and is insensitive to polarization, unlike traditional DIC techniques. In addition, it provides full control of the DIC shear and orientation, which allows obtaining a differential phase image directly on the camera (with no further digital processing) while enhancing defect detection capabilities, even if the size of the defect is smaller than the resolution limit. Our technique has the potential of future integration into semiconductor production lines.

Investigation of tin oxide nanofibers synthesized via bio-template technique

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

Kundu, Virender Singh, E-mail: vskundu-kuk@rediffmail.com; Dhiman, Jonny; Kumar, Suresh

In the present paper tin oxide nanofibers have been by synthesized using cotton as bio-template via sol-gel route. This is comparatively a new synthesis technique. The structure and morphology of the obtained SnO{sub 2} nanofibers were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy (EDX). The optical properties of the same have been studied by using UV-Vis spectroscopy. The observed XRD pattern showed that peaks are very narrow and sharp which indicates crystalline nature of samples. SEM images gave an idea about the sample morphology and confirm that the obtained sample were nanofibers. The optical absorbancemore » spectrum of the sample under study was recorded in UV-visible region (200nm- 800nm). The band gap of the sample was found to be 3.95 eV which is higher than their bulk counterpart.« less

Photoacoustic imaging of teeth for dentine imaging and enamel characterization

NASA Astrophysics Data System (ADS)

Periyasamy, Vijitha; Rangaraj, Mani; Pramanik, Manojit

2018-02-01

Early detection of dental caries, cracks and lesions is needed to prevent complicated root canal treatment and tooth extraction procedures. Resolution of clinically used x-ray imaging is low, hence optical imaging techniques such as optical coherence tomography, fluorescence imaging, and Raman imaging are widely experimented for imaging dental structures. Photoacoustic effect is used in photon induced photoacoustic streaming technique to debride the root canal. In this study, the extracted teeth were imaged using photoacoustic tomography system at 1064 nm. The degradation of enamel and dentine is an indicator of onset of dental caries. Photoacoustic microscopy (PAM) was used to study the tooth enamel. Images were acquired using acoustic resolution PAM system. This was done to identify microscopic cracks and dental lesion at different anatomical sites (crown and cementum). The PAM tooth profile is an indicator of calcium distribution which is essential for demineralization studies.

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

NASA Astrophysics Data System (ADS)

Grover, Ginni

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

Study of UV surface plasmons on metallic nanostructures and its applications to nanophotonics

NASA Astrophysics Data System (ADS)

Zhou, Liangcheng

Modern nanotechnology requires the characterization ability in the order of 100 nm or smaller. This resolution requirement cannot be met by using conventional optical microscopy. Nowadays, the mainstream technique that is universally adopted to characterize optical properties on this length scale is Near-field Scanning Optical Microscopy (NSOM). In the effort to improve the resolution and efficiency of NSOM techniques, both nanoscopic fabrication and imaging techniques are critical because the light field strongly intereacts with the metallic NSOM probe or other surfaces to form surface plasmons (SPs). However, much is still unknown about the behavior of light interacting with metallic nanostructures. This calls for research that develops the tool set, methodology and that includes both experimental characterization, and numerical simulations, for the investigation of SPs. The short wavelength of UV light makes it particularly desirable for many industrial processes. So far, little research has been carried out to understand surface plasmon in the UV spectral region. Like conventional optics, UV SPs have unique properties and optical behavior. For this purpose, we modified our existing NSOM into a Photon Scanning Tunneling Microscope (PTSM) and demonstrate its power for the imaging of UV SPs. We present what we believe to be the first direct mapping of the UV SPs on an Al2O3/Al surface. UV SP modes launched by one-dimensional slits or two-dimensional groove arrays and corresponding interference phenomenon were both observed. We then use the same methodology in the engineering of optimized nano aperture such as UV bowtie nanoantenna. For the latter, we find a strong UV intensity profile which is localized to less than 50nm caused by a localized surface plasmon resonance. The relationship of optical field enhancement and antenna geometric shape is studied using numerical simulations and NSOM experiments. In another project, we examine the propagation of light from near-field to far-field. For that purpose, a micro-lens with bull's-eye ring structure, similar to a Fresnel zone plate, is fabricated. We mapped the far-field light distribution from the micro-lens' output by using confocal microscope, which shows that this ring structure exhibit focusing ability as well. Furthermore, we study the ultraviolet (UV) extraordinary optical transmission through nanoslit structures into the far field as well as the localized field enhancement in the near field. The experimental results are compared to numerical modeling results showing good agreement.

Parallel mapping of optical near-field interactions by molecular motor-driven quantum dots.

PubMed

Groß, Heiko; Heil, Hannah S; Ehrig, Jens; Schwarz, Friedrich W; Hecht, Bert; Diez, Stefan

2018-04-30

In the vicinity of metallic nanostructures, absorption and emission rates of optical emitters can be modulated by several orders of magnitude 1,2 . Control of such near-field light-matter interaction is essential for applications in biosensing 3 , light harvesting 4 and quantum communication 5,6 and requires precise mapping of optical near-field interactions, for which single-emitter probes are promising candidates 7-11 . However, currently available techniques are limited in terms of throughput, resolution and/or non-invasiveness. Here, we present an approach for the parallel mapping of optical near-field interactions with a resolution of <5 nm using surface-bound motor proteins to transport microtubules carrying single emitters (quantum dots). The deterministic motion of the quantum dots allows for the interpolation of their tracked positions, resulting in an increased spatial resolution and a suppression of localization artefacts. We apply this method to map the near-field distribution of nanoslits engraved into gold layers and find an excellent agreement with finite-difference time-domain simulations. Our technique can be readily applied to a variety of surfaces for scalable, nanometre-resolved and artefact-free near-field mapping using conventional wide-field microscopes.

Effect of reduction time on third order optical nonlinearity of reduced graphene oxide

NASA Astrophysics Data System (ADS)

Sreeja, V. G.; Vinitha, G.; Reshmi, R.; Anila, E. I.; Jayaraj, M. K.

2017-04-01

We report the influence of reduction time on structural, linear and nonlinear optical properties of reduced graphene oxide (rGO) thin films synthesized by spin coating method. We observed that the structural, linear and nonlinear optical properties can be tuned with reduction time in GO is due to the increased structural ordering because of the restoration of sp2 carbon atoms with the time of reduction. The nonlinear absorption studies by open aperture Z-scan technique exhibited a saturable absorption. The nonlinear refraction studies showed the self de focusing nature of rGO by closed aperture Z scan technique. The nonlinear absorption coefficient and saturation intensity varies with the time for reduction of GO which is attributed to the depletion of valence band and the conduction band filling effect. Our results emphasize duration for reduction of GO dependent optical nonlinearity of rGO thin films to a great extent and explore its applications Q switched mode locking laser systems for generating ultra short laser pulses and in optical sensors. The rGO coated films were characterized by X-Ray diffraction method (XRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, UV-Vis absorption spectroscopy (UV-Vis), Photoluminescence (PL) and Scanning electron microscope (SEM) measurements.

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

PubMed Central

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

2014-01-01

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

(Gene sequencing by scanning molecular exciton microscopy)

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

Not Available

1991-01-01

This report details progress made in setting up a laboratory for optical microscopy of genes. The apparatus including a fluorescence microscope, a scanning optical microscope, various spectrometers, and supporting computers is described. Results in developing photon and exciton tips, and in preparing samples are presented. (GHH)

Design, assembly, and optical bench testing of a high-numerical-aperture miniature injection-molded objective for fiber-optic confocal reflectance microscopy.

PubMed

Chidley, Matthew D; Carlson, Kristen D; Richards-Kortum, Rebecca R; Descour, Michael R

2006-04-10

The design, analysis, assembly methods, and optical-bench test results for a miniature injection-molded plastic objective lens used in a fiber-optic confocal reflectance microscope are presented. The five-lens plastic objective was tested as a stand-alone optical system before its integration into a confocal microscope for in vivo imaging of cells and tissue. Changing the spacing and rotation of the individual optical elements can compensate for fabrication inaccuracies and improve performance. The system performance of the miniature objective lens is measured by use of an industry-accepted slanted-edge modulation transfer function (MTF) metric. An estimated Strehl ratio of 0.61 and a MTF value of 0.66 at the fiber-optic bundle Nyquist frequency have been obtained. The optical bench testing system is configured to permit interactive optical alignment during testing to optimize performance. These results are part of an effort to demonstrate the manufacturability of low-cost, high-performance biomedical optics for high-resolution in vivo imaging. Disposable endoscopic microscope objectives could help in vivo confocal microscopy technology mature to permit wide-scale clinical screening and detection of early cancers and precancerous lesions.

Site-Resolved Imaging with the Fermi Gas Microscope

NASA Astrophysics Data System (ADS)

Huber, Florian Gerhard

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

Optical and Probe Diagnostics Applied to Reacting Flows

NASA Technical Reports Server (NTRS)

Ticich, Thomas M.

2003-01-01

The general theme of the research my NASA colleague and I have planned is "Optical and probe diagnostics applied to reacting flows". We plan to explore three major threads during the fellowship period. The first interrogates the flame synthesis of carbon nanotubes using aerosol catalysts. Having demonstrated the viability of the technique for nanotube synthesis, we seek to understand the details of this reacting system which are important to its practical application. Laser light scattering will reveal changes in particle size at various heights above the burner. Analysis of the flame gas by mass spectroscopy will reveal the chemical composition of the mixture. Finally, absorption measurements will map the nanotube concentration within the flow. The second thread explores soot oxidation kinetics. Despite the impact of soot on engine performance, fire safety and pollution, models for its oxidation are inhibited by uncertainty in the values of the oxidation rate. We plan to employ both optical and microscopic measurements to refine this rate. Cavity ring-down absorption measurements of the carbonaceous aerosol can provide a measure of the mass concentration with time and, hence, an oxidation rate. Spectroscopic and direct probe measurements will provide the temperature of the system needed for subsequent modeling. These data will be benchmarked against changes in soot nanostructures as revealed by transmission electron microscopic images from directly sampled material.

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.

The Mechanosensitive Ca2+ Channel as a Central Regulator of Prostate Tumor Cell Migration and Invasiveness

DTIC Science & Technology

2010-01-01

Use time-lapse videomicroscopy and patch-clamp techniques to characterize the motility of eGFP-transfected PC-3 cells in which MScCa/TRPC1 has been...except for GsmTx-4 (Peptides International, Louisville, KY) and fluorescent agents (Invitrogen/Molecular Probes, Carlsbad, CA). Videomicroscopy ...and Ca2+-imaging. Cell migration was monitored at 37oC by time-lapse videomicroscopy using Nomarski optics with an Epifluorescent microscope (Nikon

Fabrication of miniature elastomer lenses with programmable liquid mold for smartphone microscopy: curing polydimethylsiloxane with in situ curvature control.

PubMed

Karunakaran, Bhuvaneshwari; Tharion, Joseph; Dhawangale, Arvind Ramrao; Paul, Debjani; Mukherji, Soumyo

2018-02-01

Miniature lenses can transform commercial imaging systems, e.g., smartphones and webcams, into powerful, low-cost, handheld microscopes. To date, the reproducible fabrication of polymer lenses is still a challenge as they require controlled dispensing of viscous liquid. This paper reports a reproducible lens fabrication technique using liquid mold with programmable curvature and off-the-shelf materials. The lens curvature is controlled during fabrication by tuning the curvature of an interface of two immiscible liquids [polydimethylsiloxane (PDMS) and glycerol]. The curvature control is implemented using a visual feedback system, which includes a software-based guiding system to produce lenses of desired curvature. The technique allows PDMS lens fabrication of a wide range of sizes and focal lengths, within 20 min. The fabrication of two lens diameters: 1 and 5 mm with focal lengths ranging between 1.2 and 11 mm are demonstrated. The lens surface and bulk quality check performed using X-ray microtomography and atomic force microscopy reveal that the lenses are suitable for optical imaging. Furthermore, a smartphone microscope with ∼1.4-μm resolution is developed using a self-assembly of a single high power fabricated lens and microaperture. The lenses have various potential applications, e.g., optofluidics, diagnostics, forensics, and surveillance. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Sensorless adaptive optics for isoSTED nanoscopy

NASA Astrophysics Data System (ADS)

Antonello, Jacopo; Hao, Xiang; Allgeyer, Edward S.; Bewersdorf, Joerg; Rittscher, Jens; Booth, Martin J.

2018-02-01

The presence of aberrations is a major concern when using fluorescence microscopy to image deep inside tissue. Aberrations due to refractive index mismatch and heterogeneity of the specimen under investigation cause severe reduction in the amount of fluorescence emission that is collected by the microscope. Furthermore, aberrations adversely affect the resolution, leading to loss of fine detail in the acquired images. These phenomena are particularly troublesome for super-resolution microscopy techniques such as isotropic stimulated-emission-depletion microscopy (isoSTED), which relies on accurate control of the shape and co-alignment of multiple excitation and depletion foci to operate as expected and to achieve the super-resolution effect. Aberrations can be suppressed by implementing sensorless adaptive optics techniques, whereby aberration correction is achieved by maximising a certain image quality metric. In confocal microscopy for example, one can employ the total image brightness as an image quality metric. Aberration correction is subsequently achieved by iteratively changing the settings of a wavefront corrector device until the metric is maximised. This simplistic approach has limited applicability to isoSTED microscopy where, due to the complex interplay between the excitation and depletion foci, maximising the total image brightness can lead to introducing aberrations in the depletion foci. In this work we first consider the effects that different aberration modes have on isoSTED microscopes. We then propose an iterative, wavelet-based aberration correction algorithm and evaluate its benefits.

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.

Investigation of cladding and coating stripping methods for specialty optical fibers

NASA Astrophysics Data System (ADS)

Lee, Jung-Ryul; Dhital, Dipesh; Yoon, Dong-Jin

2011-03-01

Fiber optic sensing technology is used extensively in several engineering fields, including smart structures, health and usage monitoring, non-destructive testing, minimum invasive sensing, safety monitoring, and other advanced measurement fields. A general optical fiber consists of a core, cladding, and coating layers. Many sensing principles require that the cladding or coating layer should be removed or modified. In addition, since different sensing systems are needed for different types of optical fibers, it is very important to find and sort out the suitable cladding or coating removal method for a particular fiber. This study focuses on finding the cladding and coating stripping methods for four recent specialty optical fibers, namely: hard polymer-clad fiber, graded-index plastic optical fiber, copper/carbon-coated optical fiber, and aluminum-coated optical fiber. Several methods, including novel laser stripping and conventional chemical and mechanical stripping, were tried to determine the most suitable and efficient technique. Microscopic investigation of the fiber surfaces was used to visually evaluate the mechanical reliability. Optical time domain reflectometric signals of the successful removal cases were investigated to further examine the optical reliability. Based on our results, we describe and summarize the successful and unsuccessful methods.

Optical Scatter Imaging with a digital micromirror device.

PubMed

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

2009-10-26

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

Liquid crystal optics for communications, signal processing and 3-D microscopic imaging

NASA Astrophysics Data System (ADS)

Khan, Sajjad Ali

This dissertation proposes, studies and experimentally demonstrates novel liquid crystal (LC) optics to solve challenging problems in RF and photonic signal processing, freespace and fiber optic communications and microscopic imaging. These include free-space optical scanners for military and optical wireless applications, variable fiber-optic attenuators for optical communications, photonic control techniques for phased array antennas and radar, and 3-D microscopic imaging. At the heart of the applications demonstrated in this thesis are LC devices that are non-pixelated and can be controlled either electrically or optically. Instead of the typical pixel-by-pixel control as is custom in LC devices, the phase profile across the aperture of these novel LC devices is varied through the use of high impedance layers. Due to the presence of the high impedance layer, there forms a voltage gradient across the aperture of such a device which results in a phase gradient across the LC layer which in turn is accumulated by the optical beam traversing through this LC device. The geometry of the electrical contacts that are used to apply the external voltage will define the nature of the phase gradient present across the optical beam. In order to steer a laser beam in one angular dimension, straight line electrical contacts are used to form a one dimensional phase gradient while an annular electrical contact results in a circularly symmetric phase profile across the optical beam making it suitable for focusing the optical beam. The geometry of the electrical contacts alone is not sufficient to form the linear and the quadratic phase profiles that are required to either deflect or focus an optical beam. Clever use of the phase response of a typical nematic liquid crystal (NLC) is made such that the linear response region is used for the angular beam deflection while the high voltage quadratic response region is used for focusing the beam. Employing an NLC deflector, a device that uses the linear angular deflection, laser beam steering is demonstrated in two orthogonal dimensions whereas an NLC lens is used to address the third dimension to complete a three dimensional (3-D) scanner. Such an NLC deflector was then used in a variable optical attenuator (VOA), whereby a laser beam coupled between two identical single mode fibers (SMF) was mis-aligned away from the output fiber causing the intensity of the output coupled light to decrease as a function of the angular deflection. Since the angular deflection is electrically controlled, hence the VOA operation is fairly simple and repeatable. An extension of this VOA for wavelength tunable operation is also shown in this dissertation. (Abstract shortened by UMI.)

Annealing Temperature Dependent Structural and Optical Properties of RF Sputtered ZnO Thin Films.

PubMed

Sharma, Shashikant; Varma, Tarun; Asokan, K; Periasamy, C; Boolchandani, Dharmendar

2017-01-01

This work investigates the effect of annealing temperature on structural and optical properties of ZnO thin films grown over Si 100 and glass substrates using RF sputtering technique. Annealing temperature has been varied from 300 °C to 600 °C in steps of 100, and different microstructural parameters such as grain size, dislocation density, lattice constant, stress and strain have been evaluated. The structural and surface morphological characterization has been done using X-ray Diffraction (XRD) and Scanning Electron Microscope (SEM). XRD analysis reveals that the peak intensity of 002 crystallographic orientation increases with increased annealing temperature. Optical characterization of deposited films have been done using UV-Vis-NIR spectroscopy and photoluminescence spectrometer. An increase in optical bandgap of deposited ZnO thin films with increasing annealing temperature has been observed. The average optical transmittance was found to be more than 85% for all deposited films. Photoluminiscense spectra (PL) suggest that the crystalline quality of deposited film has increased at higher annealing temperature.

DMD-based LED-illumination super-resolution and optical sectioning microscopy.

PubMed

Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

2013-01-01

Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×10(7) pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

DMD-based LED-illumination Super-resolution and optical sectioning microscopy

PubMed Central

Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

2013-01-01

Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×107 pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens. PMID:23346373

Dual-beam optical trapping of cells in an optofluidic device fabricated by femtosecond lasers

NASA Astrophysics Data System (ADS)

Bellini, N.; Bragheri, F.; Vishnubhatla, K. C.; Ferrara, L.; Minzioni, P.; Cerullo, G.; Ramponi, R.; Cristiani, I.; Osellame, R.

2010-02-01

We present design and optimization of an optofluidic monolithic chip, able to provide optical trapping and controlled stretching of single cells. The chip is fabricated in a fused silica glass substrate by femtosecond laser micromachining, which can produce both optical waveguides and microfluidic channels with great accuracy. Versatility and three-dimensional capabilities of this fabrication technology provide the possibility to fabricate circular cross-section channels with enlarged access holes for an easy connection with an external fluidic circuit. Moreover, a new fabrication procedure adopted allows the demonstration of microchannels with a square cross-section, thus guaranteeing an improved quality of the trapped cell images. Optical trapping and stretching of single red blood cells are demonstrated, thus proving the effectiveness of the proposed device as a monolithic optical stretcher. We believe that femtosecond laser micromachining represents a promising technique for the development of multifunctional integrated biophotonic devices that can be easily coupled to a microscope platform, thus enabling a complete characterization of the cells under test.

Wide spectral range confocal microscope based on endlessly single-mode fiber.

PubMed

Hubbard, R; Ovchinnikov, Yu B; Hayes, J; Richardson, D J; Fu, Y J; Lin, S D; See, P; Sinclair, A G

2010-08-30

We report an endlessly single mode, fiber-optic confocal microscope, based on a large mode area photonic crystal fiber. The microscope confines a very broad spectral range of excitation and emission wavelengths to a single spatial mode in the fiber. Single-mode operation over an optical octave is feasible. At a magnification of 10 and λ = 900 nm, its resolution was measured to be 1.0 μm (lateral) and 2.5 μm (axial). The microscope's use is demonstrated by imaging single photons emitted by individual InAs quantum dots in a pillar microcavity.

Investigations on structural and optical properties of starch capped ZnS nanoparticles synthesized by microwave irradiation method

NASA Astrophysics Data System (ADS)

Lalithadevi, B.; Mohan Rao, K.; Ramananda, D.

2018-05-01

Following a green synthesis method, zinc sulfide (ZnS) nanoparticles were prepared by chemical co-precipitation technique using starch as capping agent. Microwave irradiation was used as heating source. X-ray diffraction studies indicated that nanopowders obtained were polycrystalline possessing ZnS simple cubic structure. Transmission electron microscopic studies indicated that starch limits the agglomeration by steric stabilization. Interaction between ZnS and starch was confirmed by Fourier transform infrared spectroscopy as well as Raman scattering studies. Quantum size effects were observed in optical absorption studies while quenching of defect states on nanoparticles was improved with increase in starch addition as indicated by photoluminescence spectra.

Crystalline phase transformation of colloidal cadmium sulfide nanocrystals

NASA Astrophysics Data System (ADS)

Ghali, M.; Eissa, A. M.; Mosaad, M. M.

2017-03-01

In this paper, we give a microscopic view concerning influence of the growth conditions on the physical properties of nanocrystals (NCs) thin films made of CdS, prepared using chemical bath deposition CBD technique. We show a crystalline phase transformation of CdS NCs from hexagonal wurtzite (W) structure to cubic zincblende (ZB) when the growth conditions change, particularly the solution pH values. This effect was confirmed using X-ray diffraction (XRD), transmission electron microscopy (TEM), optical absorption and photoluminescence (PL) measurements. The optical absorption spectra allow calculation of the bandgap value, Eg, where significant increase ˜200 meV in the CdS bandgap when transforming from Hexagonal to Cubic phase was found.

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.

On soft clipping of Zernike moments for deblurring and enhancement of optical point spread functions

NASA Astrophysics Data System (ADS)

Becherer, Nico; Jödicke, Hanna; Schlosser, Gregor; Hesser, Jürgen; Zeilfelder, Frank; Männer, Reinhard

2006-02-01

Blur and noise originating from the physical imaging processes degrade the microscope data. Accurate deblurring techniques require, however, an accurate estimation of the underlying point-spread function (PSF). A good representation of PSFs can be achieved by Zernike Polynomials since they offer a compact representation where low-order coefficients represent typical aberrations of optical wavefronts while noise is represented in higher order coefficients. A quantitative description of the noise distribution (Gaussian) over the Zernike moments of various orders is given which is the basis for the new soft clipping approach for denoising of PSFs. Instead of discarding moments beyond a certain order, those Zernike moments that are more sensitive to noise are dampened according to the measured distribution and the present noise model. Further, a new scheme to combine experimental and theoretical PSFs in Zernike space is presented. According to our experimental reconstructions, using the new improved PSF the correlation between reconstructed and original volume is raised by 15% on average cases and up to 85% in the case of thin fibre structures, compared to reconstructions where a non improved PSF was used. Finally, we demonstrate the advantages of our approach on 3D images of confocal microscopes by generating visually improved volumes. Additionally, we are presenting a method to render the reconstructed results using a new volume rendering method that is almost artifact-free. The new approach is based on a Shear-Warp technique, wavelet data encoding techniques and a recent approach to approximate the gray value distribution by a Super spline model.

Generation-3 programmable array microscope (PAM) with digital micro-mirror device (DMD)

NASA Astrophysics Data System (ADS)

De Beule, Pieter A. A.; de Vries, Anthony H. B.; Arndt-Jovin, Donna J.; Jovin, Thomas M.

2011-03-01

We report progress on the construction of an optical sectioning programmable array microscope (PAM) implemented with a digital micro-mirror device (DMD) spatial light modulator (SLM) utilized for both fluorescence illumination and detection. The introduction of binary intensity modulation at the focal plane of a microscope objective in a computer controlled pixilated mode allows the recovery of an optically sectioned image. Illumination patterns can be changed very quickly, in contrast to static Nipkow disk or aperture correlation implementations, thereby creating an optical system that can be optimized to the optical specimen in a convenient manner, e.g. for patterned photobleaching, photobleaching reduction, or spatial superresolution. We present a third generation (Gen-3) dual path PAM module incorporating the 25 kHz binary frame rate TI 1080p DMD and a newly developed optical system that offers diffraction limited imaging with compensation of tilt angle distortion.

Thickness dependent structural, optical and electrical properties of Se85In12Bi3 nanochalcogenide thin films

NASA Astrophysics Data System (ADS)

Tripathi, Ravi P.; Zulfequar, M.; Khan, Shamshad A.

2018-04-01

Our aim is to study the thickness dependent effects on structure, electrical and optical properties of Se85In12Bi3 nanochalcogenide thin films. Bulk alloy of Se85In12Bi3 was synthesized by melt-quenching technique. The amorphous as well as glassy nature of Se85In12Bi3 chalcogenide was confirmed by non-isothermal Differential Scanning Calorimetry (DSC) measurements. The nanochalcogenide thin films of thickness 30, 60 and 90 nm were prepared on glass/Si wafer substrate using Physical Vapour Condensation Technique (PVCT). From XRD studies it was found that thin films have amorphous texture. The surface morphology and particle size of films were studied by Field Emission Scanning Electron Microscope (FESEM). From optical studies, different optical parameters were estimated for Se85In12Bi3 thin films at different thickness. It was found that the absorption coefficient (α) and extinction coefficient (k) increases with photon energy and decreases with film thickness. The optical absorption process followed the rule of indirect transitions and optical band gap were found to be increase with film thickness. The value of Urbach energy (Et) and steepness parameter (σ) were also calculated for different film thickness. For electrical studies, dc-conductivity measurement was done at different temperature and activation energy (ΔEc) were determined and found to be increase with film thickness.