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Sample records for techniques optical microscopy

  1. Optical Imaging and Microscopy Techniques and Advanced Systems

    CERN Document Server

    Török, Peter

    2007-01-01

    This text on contemporary optical systems is intended for optical researchers and engineers, graduate students and optical microscopists in the biological and biomedical sciences. This second edition contains two completely new chapters. In addition most of the chapters from the first edition have been revised and updated. The book consists of three parts: The first discusses high-aperture optical systems, which form the backbone of optical microscopes. An example is a chapter new in the second edition on the emerging field of high numerical aperture diffractive lenses which seems to have particular promise in improving the correction of lenses. In this part particular attention is paid to optical data storage. The second part is on the use of non-linear optical techniques, including nonlinear optical excitation (total internal reflection fluorescence, second and third harmonic generation and two photon microscopy) and non-linear spectroscopy (CARS). The final part of the book presents miscellaneous technique...

  2. Gabor-based fusion technique for Optical Coherence Microscopy.

    Science.gov (United States)

    Rolland, Jannick P; Meemon, Panomsak; Murali, Supraja; Thompson, Kevin P; Lee, Kye-sung

    2010-02-15

    We recently reported on an Optical Coherence Microscopy technique, whose innovation intrinsically builds on a recently reported - 2 microm invariant lateral resolution by design throughout a 2 mm cubic full-field of view - liquid-lens-based dynamic focusing optical probe [Murali et al., Optics Letters 34, 145-147, 2009]. We shall report in this paper on the image acquisition enabled by this optical probe when combined with an automatic data fusion method developed and described here to produce an in-focus high resolution image throughout the imaging depth of the sample. An African frog tadpole (Xenopus laevis) was imaged with the novel probe and the Gabor-based fusion technique, demonstrating subcellular resolution in a 0.5 mm (lateral) x 0.5 mm (axial) without the need, for the first time, for x-y translation stages, depth scanning, high-cost adaptive optics, or manual intervention. In vivo images of human skin are also presented.

  3. Comparison between optical techniques and confocal microscopy for defect detection on thin wires

    Energy Technology Data Exchange (ETDEWEB)

    Siegmann, Philip; Sanchez-Brea, Luis Miguel; Martinez-Anton, Juan Carlos; Bernabeu, Eusebio

    2004-11-15

    Conventional microscopy techniques, such as atomic force microscopy (AFM), scanning electron microscopy (SEM), and confocal microscopy (CM) are not suitable for on-line surface inspection of fine metallic wires. In the recent years, some optical techniques have been developed to be used for those tasks. However, they need a rigorous validation. In this work, we have used confocal microscopy to obtain the topography z(x,y) of wires with longitudinal defects, such as dielines. The topography has been used to predict the light scattered by the wire. These simulations have been compared with experimental results, showing a good agreement.

  4. The development of optical microscopy techniques for the advancement of single-particle studies

    Science.gov (United States)

    Marchuk, Kyle

    Single particle orientation and rotational tracking (SPORT) has recently become a powerful optical microscopy tool that can expose many molecular motions. Unfortunately, there is not yet a single microscopy technique that can decipher all particle motions in all environmental conditions, thus there are limitations to current technologies. Within, the two powerful microscopy tools of total internal reflection and interferometry are advanced to determine the position, orientation, and optical properties of metallic nanoparticles in a variety of environments. Total internal reflection is an optical phenomenon that has been applied to microscopy to produce either fluorescent or scattered light. The non-invasive far-field imaging technique is coupled with a near-field illumination scheme that allows for better axial resolution than confocal microscopy and epi-fluorescence microscopy. By controlling the incident illumination angle using total internal reflection fluorescence (TIRF) microscopy, a new type of imaging probe called "non-blinking" quantum dots (NBQDs) were super-localized in the axial direction to sub-10-nm precision. These particles were also used to study the rotational motion of microtubules being propelled by the motor protein kinesin across the substrate surface. The same instrument was modified to function under total internal reflection scattering (TIRS) microscopy to study metallic anisotropic nanoparticles and their dynamic interactions with synthetic lipid bilayers. Utilizing two illumination lasers with opposite polarization directions at wavelengths corresponding to the short and long axis surface plasmon resonance (SPR) of the nanoparticles, both the in-plane and out-of-plane movements of many particles could be tracked simultaneously. When combined with Gaussian point spread function (PSF) fitting for particle super-localization, the binding status and rotational movement could be resolved without degeneracy. TIRS microscopy was also used to

  5. The development of optical microscopy techniques for the advancement of single-particle studies

    Energy Technology Data Exchange (ETDEWEB)

    Marchuk, Kyle [Iowa State Univ., Ames, IA (United States)

    2013-05-15

    Single particle orientation and rotational tracking (SPORT) has recently become a powerful optical microscopy tool that can expose many molecular motions. Unfortunately, there is not yet a single microscopy technique that can decipher all particle motions in all environmental conditions, thus there are limitations to current technologies. Within, the two powerful microscopy tools of total internal reflection and interferometry are advanced to determine the position, orientation, and optical properties of metallic nanoparticles in a variety of environments. Total internal reflection is an optical phenomenon that has been applied to microscopy to produce either fluorescent or scattered light. The non-invasive far-field imaging technique is coupled with a near-field illumination scheme that allows for better axial resolution than confocal microscopy and epi-fluorescence microscopy. By controlling the incident illumination angle using total internal reflection fluorescence (TIRF) microscopy, a new type of imaging probe called “non-blinking” quantum dots (NBQDs) were super-localized in the axial direction to sub-10-nm precision. These particles were also used to study the rotational motion of microtubules being propelled by the motor protein kinesin across the substrate surface. The same instrument was modified to function under total internal reflection scattering (TIRS) microscopy to study metallic anisotropic nanoparticles and their dynamic interactions with synthetic lipid bilayers. Utilizing two illumination lasers with opposite polarization directions at wavelengths corresponding to the short and long axis surface plasmon resonance (SPR) of the nanoparticles, both the in-plane and out-of-plane movements of many particles could be tracked simultaneously. When combined with Gaussian point spread function (PSF) fitting for particle super-localization, the binding status and rotational movement could be resolved without degeneracy. TIRS microscopy was also used to

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

    Directory of Open Access Journals (Sweden)

    Saher M. Maswadi

    2016-09-01

    Full Text Available 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.

  7. Dual beam light profile microscopy: a new technique for optical absorption depth profilometry.

    Science.gov (United States)

    Power, J F; Fu, S W

    2004-02-01

    Light profile microscopy (LPM) is a recently developed technique of optical inspection that is used to record micrometer-scale images of thin-film cross-sections on a direct basis. In single beam mode, LPM provides image contrast based on luminescence, elastic, and/or inelastic scatter. However, LPM may also be used to depth profile the optical absorption coefficient of a thin film based on a method of dual beam irradiation presented in this work. The method uses a pair of collimated laser beams to consecutively irradiate a film from two opposing directions along the depth axis. An average profile of the beam's light intensity variation through the material is recovered for each direction and used to compute a depth-dependent differential absorbance profile. This latter quantity is shown from theory to be related to the film's depth-dependent optical absorption coefficient through a simple linear model that may be inverted by standard methods of numerical linear algebra. The inverse problem is relatively well posed, showing good immunity to data errors. This profilometry method is experimentally applied to a set of well-characterized materials with known absorption properties over a scale of tens of micrometers, and the reconstructed absorption profiles were found to be highly consistent with the reference data.

  8. Finite-difference time-domain-based optical microscopy simulation of dispersive media facilitates the development of optical imaging techniques

    Science.gov (United States)

    Zhang, Di; Capoglu, Ilker; Li, Yue; Cherkezyan, Lusik; Chandler, John; Spicer, Graham; Subramanian, Hariharan; Taflove, Allen; Backman, Vadim

    2016-06-01

    Combining finite-difference time-domain (FDTD) methods and modeling of optical microscopy modalities, we previously developed an open-source software package called Angora, which is essentially a "microscope in a computer." However, the samples being simulated were limited to nondispersive media. Since media dispersions are common in biological samples (such as cells with staining and metallic biomarkers), we have further developed a module in Angora to simulate samples having complicated dispersion properties, thereby allowing the synthesis of microscope images of most biological samples. We first describe a method to integrate media dispersion into FDTD, and we validate the corresponding Angora dispersion module by applying Mie theory, as well as by experimentally imaging gold microspheres. Then, we demonstrate how Angora can facilitate the development of optical imaging techniques with a case study.

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

    Science.gov (United States)

    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.

  10. Near-field Optical Microscopy

    NARCIS (Netherlands)

    Ruiter, A.G.T.

    1997-01-01

    Near-field scanning optical microscopy (NSOM) is one of the most recent scanning probe techniques. In this technique, an optical probe is brought in the vicinity of the sample surface, in the near-field zone. The microscope can either work in illumination mode, in which the probe consists of a

  11. Near-field Optical Microscopy

    NARCIS (Netherlands)

    Ruiter, Anthonius Gerardus Theodorus

    1997-01-01

    Near-field scanning optical microscopy (NSOM) is one of the most recent scanning probe techniques. In this technique, an optical probe is brought in the vicinity of the sample surface, in the near-field zone. The microscope can either work in illumination mode, in which the probe consists of a sub-w

  12. Axial Phase-Darkfield-Contrast (APDC), a new technique for variable optical contrasting in light microscopy.

    Science.gov (United States)

    Piper, T; Piper, J

    2012-09-01

    Axial phase-darkfield-contrast (APDC) has been developed as an illumination technique in light microscopy which promises significant improvements and a higher variability in imaging of several transparent 'problem specimens'. With this method, a phase contrast image is optically superimposed on an axial darkfield image so that a partial image based on the principal zeroth order maximum (phase contrast) interferes with an image, which is based on the secondary maxima (axial darkfield). The background brightness and character of the resulting image can be continuously modulated from a phase contrast-dominated to a darkfield-dominated character. In order to achieve this illumination mode, normal objectives for phase contrast have to be fitted with an additional central light stopper needed for axial (central) darkfield illumination. In corresponding condenser light masks, a small perforation has to be added in the centre of the phase contrast providing light annulus. These light modulating elements are properly aligned when the central perforation is congruent with the objective's light stop and the light annulus is conjugate with the phase ring. The breadth of the condenser light annulus and thus the intensity of the phase contrast partial image can be regulated with the aperture diaphragm. Additional contrast effects can be achieved when both illuminating light components are filtered at different colours. In this technique, the axial resolution (depth of field) is significantly enhanced and the specimen's three-dimensional appearance is accentuated with improved clarity as well as fine details at the given resolution limit. Typical artefacts associated with phase contrast and darkfield illumination are reduced in our methods.

  13. Revealing t-tubules in striated muscle with new optical super-resolution microscopy techniques

    Directory of Open Access Journals (Sweden)

    Isuru D. Jayasinghe

    2014-12-01

    Full Text Available The t-tubular system plays a central role in the synchronisation of calcium signalling and excitation-contraction coupling in most striated muscle cells. Light microscopy has been used for imaging t-tubules for well over 100 years and together with electron microscopy (EM, has revealed the three-dimensional complexities of the t-system topology within cardiomyocytes and skeletal muscle fibres from a range of species. The emerging super-resolution single molecule localisation microscopy (SMLM techniques are offering a near 10-fold improvement over the resolution of conventional fluorescence light microscopy methods, with the ability to spectrally resolve nanometre scale distributions of multiple molecular targets. In conjunction with the next generation of electron microscopy, SMLM has allowed the visualisation and quantification of intricate t-tubule morphologies within large areas of muscle cells at an unprecedented level of detail. In this paper, we review recent advancements in the t-tubule structural biology with the utility of various microscopy techniques. We outline the technical considerations in adapting SMLM to study t-tubules and its potential to further our understanding of the molecular processes that underlie the sub-micron scale structural alterations observed in a range of muscle pathologies.

  14. Microscopy techniques in flavivirus research.

    Science.gov (United States)

    Chong, Mun Keat; Chua, Anthony Jin Shun; Tan, Terence Tze Tong; Tan, Suat Hoon; Ng, Mah Lee

    2014-04-01

    The Flavivirus genus is composed of many medically important viruses that cause high morbidity and mortality, which include Dengue and West Nile viruses. Various molecular and biochemical techniques have been developed in the endeavour to study flaviviruses. However, microscopy techniques still have irreplaceable roles in the identification of novel virus pathogens and characterization of morphological changes in virus-infected cells. Fluorescence microscopy contributes greatly in understanding the fundamental viral protein localizations and virus-host protein interactions during infection. Electron microscopy remains the gold standard for visualizing ultra-structural features of virus particles and infected cells. New imaging techniques and combinatory applications are continuously being developed to push the limit of resolution and extract more quantitative data. Currently, correlative live cell imaging and high resolution three-dimensional imaging have already been achieved through the tandem use of optical and electron microscopy in analyzing biological specimens. Microscopy techniques are also used to measure protein binding affinities and determine the mobility pattern of proteins in cells. This chapter will consolidate on the applications of various well-established microscopy techniques in flavivirus research, and discuss how recently developed microscopy techniques can potentially help advance our understanding in these membrane viruses.

  15. All-optical photoacoustic microscopy

    Directory of Open Access Journals (Sweden)

    Sung-Liang Chen

    2015-12-01

    Full Text Available Three-dimensional photoacoustic microscopy (PAM has gained considerable attention within the biomedical imaging community during the past decade. Detecting laser-induced photoacoustic waves by optical sensing techniques facilitates the idea of all-optical PAM (AOPAM, which is of particular interest as it provides unique advantages for achieving high spatial resolution using miniaturized embodiments of the imaging system. The review presents the technology aspects of optical-sensing techniques for ultrasound detection, such as those based on optical resonators, as well as system developments of all-optical photoacoustic systems including PAM, photoacoustic endoscopy, and multi-modality microscopy. The progress of different AOPAM systems and their representative applications are summarized.

  16. Optical imaging. Expansion microscopy.

    Science.gov (United States)

    Chen, Fei; Tillberg, Paul W; Boyden, Edward S

    2015-01-30

    In optical microscopy, fine structural details are resolved by using refraction to magnify images of a specimen. We discovered that by synthesizing a swellable polymer network within a specimen, it can be physically expanded, resulting in physical magnification. By covalently anchoring specific labels located within the specimen directly to the polymer network, labels spaced closer than the optical diffraction limit can be isotropically separated and optically resolved, a process we call expansion microscopy (ExM). Thus, this process can be used to perform scalable superresolution microscopy with diffraction-limited microscopes. We demonstrate ExM with apparent ~70-nanometer lateral resolution in both cultured cells and brain tissue, performing three-color superresolution imaging of ~10(7) cubic micrometers of the mouse hippocampus with a conventional confocal microscope.

  17. Resolution enhancement techniques in microscopy

    Science.gov (United States)

    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.

  18. Emerging optical nanoscopy techniques

    Directory of Open Access Journals (Sweden)

    Montgomery PC

    2015-09-01

    Full Text Available Paul C Montgomery, Audrey Leong-Hoi Laboratoire des Sciences de l'Ingénieur, de l'Informatique et de l'Imagerie (ICube, Unistra-CNRS, Strasbourg, France Abstract: To face the challenges of modern health care, new imaging techniques with subcellular resolution or detection over wide fields are required. Far field optical nanoscopy presents many new solutions, providing high resolution or detection at high speed. We present a new classification scheme to help appreciate the growing number of optical nanoscopy techniques. We underline an important distinction between superresolution techniques that provide improved resolving power and nanodetection techniques for characterizing unresolved nanostructures. Some of the emerging techniques within these two categories are highlighted with applications in biophysics and medicine. Recent techniques employing wider angle imaging by digital holography and scattering lens microscopy allow superresolution to be achieved for subcellular and even in vivo, imaging without labeling. Nanodetection techniques are divided into four subcategories using contrast, phase, deconvolution, and nanomarkers. Contrast enhancement is illustrated by means of a polarized light-based technique and with strobed phase-contrast microscopy to reveal nanostructures. Very high sensitivity phase measurement using interference microscopy is shown to provide nanometric surface roughness measurement or to reveal internal nanometric structures. Finally, the use of nanomarkers is illustrated with stochastic fluorescence microscopy for mapping intracellular structures. We also present some of the future perspectives of optical nanoscopy. Keywords: microscopy, imaging, superresolution, nanodetection, biophysics, medical imaging

  19. Emerging optical nanoscopy techniques

    Science.gov (United States)

    Montgomery, Paul C; Leong-Hoi, Audrey

    2015-01-01

    To face the challenges of modern health care, new imaging techniques with subcellular resolution or detection over wide fields are required. Far field optical nanoscopy presents many new solutions, providing high resolution or detection at high speed. We present a new classification scheme to help appreciate the growing number of optical nanoscopy techniques. We underline an important distinction between superresolution techniques that provide improved resolving power and nanodetection techniques for characterizing unresolved nanostructures. Some of the emerging techniques within these two categories are highlighted with applications in biophysics and medicine. Recent techniques employing wider angle imaging by digital holography and scattering lens microscopy allow superresolution to be achieved for subcellular and even in vivo, imaging without labeling. Nanodetection techniques are divided into four subcategories using contrast, phase, deconvolution, and nanomarkers. Contrast enhancement is illustrated by means of a polarized light-based technique and with strobed phase-contrast microscopy to reveal nanostructures. Very high sensitivity phase measurement using interference microscopy is shown to provide nanometric surface roughness measurement or to reveal internal nanometric structures. Finally, the use of nanomarkers is illustrated with stochastic fluorescence microscopy for mapping intracellular structures. We also present some of the future perspectives of optical nanoscopy. PMID:26491270

  20. High-Contrast Fluorescence Microscopy for a Biomolecular Analysis Based on Polarization Techniques Using an Optical Interference Mirror Slide

    Directory of Open Access Journals (Sweden)

    Mitsuru Yasuda

    2014-12-01

    Full Text Available Fluorescence microscopy with an improved contrast for fluorescence images is developed using an optical interference mirror (OIM slide, which can enhance the fluorescence from a fluorophore as a result of the double interference of the excitation light and emission light. To improve the contrast of a fluorescence image using an OIM slide, a linearly-polarized excitation light was employed, and the fluorescence emission polarized perpendicular to the polarization of the excitation light was detected. The image contrast with this optical system was improved 110-fold for rhodamine B spotted on the OIM, in comparison with a glass slide using a general fluorescence microscopy optical system. Moreover, a 24-fold improvement of the image contrast was achieved for the detection of Cy3-labeled streptavidin bound to immobilize biotin.

  1. Applicability of quantitative optical imaging techniques for intraoperative perfusion diagnostics: a comparison of laser speckle contrast imaging, sidestream dark-field microscopy, and optical coherence tomography.

    Science.gov (United States)

    Jansen, Sanne M; de Bruin, Daniel M; Faber, Dirk J; Dobbe, Iwan J G G; Heeg, Erik; Milstein, Dan M J; Strackee, Simon D; van Leeuwen, Ton G

    2017-08-01

    Patient morbidity and mortality due to hemodynamic complications are a major problem in surgery. Optical techniques can image blood flow in real-time and high-resolution, thereby enabling perfusion monitoring intraoperatively. We tested the feasibility and validity of laser speckle contrast imaging (LSCI), optical coherence tomography (OCT), and sidestream dark-field microscopy (SDF) for perfusion diagnostics in a phantom model using whole blood. Microvessels with diameters of 50, 100, and 400  μm were constructed in a scattering phantom. Perfusion was simulated by pumping heparinized human whole blood at five velocities (0 to 20  mm/s). Vessel diameter and blood flow velocity were assessed with LSCI, OCT, and SDF. Quantification of vessel diameter was feasible with OCT and SDF. LSCI could only visualize the 400-μm vessel, perfusion units scaled nonlinearly with blood velocity. OCT could assess blood flow velocity in terms of inverse OCT speckle decorrelation time. SDF was not feasible to measure blood flow; however, for diluted blood the measurements were linear with the input velocity up to 1  mm/s. LSCI, OCT, and SDF were feasible to visualize blood flow. Validated blood flow velocity measurements intraoperatively in the desired parameter (mL·min-1·g-1) remain challenging. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

  2. Rotary-scanning optical resolution photoacoustic microscopy

    Science.gov (United States)

    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.

  3. Applicability of quantitative optical imaging techniques for intraoperative perfusion diagnostics: a comparison of laser speckle contrast imaging, sidestream dark-field microscopy, and optical coherence tomography

    Science.gov (United States)

    Jansen, Sanne M.; de Bruin, Daniel M.; Faber, Dirk J.; Dobbe, Iwan J. G. G.; Heeg, Erik; Milstein, Dan M. J.; Strackee, Simon D.; van Leeuwen, Ton G.

    2017-08-01

    Patient morbidity and mortality due to hemodynamic complications are a major problem in surgery. Optical techniques can image blood flow in real-time and high-resolution, thereby enabling perfusion monitoring intraoperatively. We tested the feasibility and validity of laser speckle contrast imaging (LSCI), optical coherence tomography (OCT), and sidestream dark-field microscopy (SDF) for perfusion diagnostics in a phantom model using whole blood. Microvessels with diameters of 50, 100, and 400 μm were constructed in a scattering phantom. Perfusion was simulated by pumping heparinized human whole blood at five velocities (0 to 20 mm/s). Vessel diameter and blood flow velocity were assessed with LSCI, OCT, and SDF. Quantification of vessel diameter was feasible with OCT and SDF. LSCI could only visualize the 400-μm vessel, perfusion units scaled nonlinearly with blood velocity. OCT could assess blood flow velocity in terms of inverse OCT speckle decorrelation time. SDF was not feasible to measure blood flow; however, for diluted blood the measurements were linear with the input velocity up to 1 mm/s. LSCI, OCT, and SDF were feasible to visualize blood flow. Validated blood flow velocity measurements intraoperatively in the desired parameter (mL·g-1) remain challenging.

  4. Super-resolution optical microscopy: multiple choices.

    Science.gov (United States)

    Huang, Bo

    2010-02-01

    The recent invention of super-resolution optical microscopy enables the visualization of fine features in biological samples with unprecedented clarity. It creates numerous opportunities in biology because vast amount of previously obscured subcellular processes now can be directly observed. Rapid development in this field in the past two years offers many imaging modalities that address different needs but they also complicates the choice of the 'perfect' method for answering a specific question. Here I will briefly describe the principles of super-resolution optical microscopy techniques and then focus on comparing their characteristics in various aspects of practical applications.

  5. Single spin stochastic optical reconstruction microscopy

    CERN Document Server

    Pfender, Matthias; Waldherr, Gerald; Wrachtrup, Jörg

    2014-01-01

    We experimentally demonstrate precision addressing of single quantum emitters by combined optical microscopy and spin resonance techniques. To this end we utilize nitrogen-vacancy (NV) color centers in diamond confined within a few ten nanometers as individually resolvable quantum systems. By developing a stochastic optical reconstruction microscopy (STORM) technique for NV centers we are able to simultaneously perform sub diffraction-limit imaging and optically detected spin resonance (ODMR) measurements on NV spins. This allows the assignment of spin resonance spectra to individual NV center locations with nanometer scale resolution and thus further improves spatial discrimination. For example, we resolved formerly indistinguishable emitters by their spectra. Furthermore, ODMR spectra contain metrology information allowing for sub diffraction-limit sensing of, for instance, magnetic or electric fields with inherently parallel data acquisition. As an example, we have detected nuclear spins with nanometer sca...

  6. Digital Holographic Microscopy Principles, Techniques, and Applications

    CERN Document Server

    Kim, Myung K

    2011-01-01

    Digital holography is an emerging field of new paradigm in general imaging applications. By replacing the photochemical procedures with electronic imaging and having a direct numerical access to the complex optical field, a wide range of new imaging capabilities become available, many of them difficult or infeasible in conventional holography. An increasing number of researchers—not only in optical physics and optical engineering, but also in diverse applications areas such as microbiology, medicine, marine science, particle analysis, microelectromechanics, and metrology—are realizing and exploiting the new capabilities of digital holography. Digital Holographic Microscopy: Principles, Techniques, and Applications, by Dr. Myung K. Kim, is intended to provide a brief but consistent introduction to the principles of digital holography as well as to give an organized overview of the large number of techniques and applications being developed. This will also shed some light on the range of possibilities for f...

  7. Optical Photon Reassignment Microscopy (OPRA)

    CERN Document Server

    Roth, Stephan; Wicker, Kai; Heintzmann, Rainer

    2013-01-01

    To enhance the resolution of a confocal laser scanning microscope the additional information of a pinhole plane image taken at every excitation scan position can be used [C. J. R. Sheppard, Super-resolution in confocal imaging, Optik 80, 5354 (1988)]. This photon reassignment principle is based on the fact that the most probable position of an emitter is at half way between the nominal focus of the excitation laser and the position corresponding to the (off centre) detection position. Therefore, by reassigning the detected photons to this place, an image with enhanced detection efficiency and resolution is obtained. Here we present optical photon reassignment microscopy (OPRA) which realises this concept in an all-optical way obviating the need for image-processing. With the help of an additional intermediate optical beam expansion between descanning and a further rescanning of the detected light, an image with the advantages of photon reassignment can be acquired. Due to its simplicity and flexibility this m...

  8. Visual-servoing optical microscopy

    Science.gov (United States)

    Callahan, Daniel E.; Parvin, Bahram

    2009-06-09

    The present invention provides methods and devices for the knowledge-based discovery and optimization of differences between cell types. In particular, the present invention provides visual servoing optical microscopy, as well as analysis methods. The present invention provides means for the close monitoring of hundreds of individual, living cells over time: quantification of dynamic physiological responses in multiple channels; real-time digital image segmentation and analysis; intelligent, repetitive computer-applied cell stress and cell stimulation; and the ability to return to the same field of cells for long-term studies and observation. The present invention further provides means to optimize culture conditions for specific subpopulations of cells.

  9. Swept source optical coherence tomography Gabor fusion splicing technique for microscopy of thick samples using a deformable mirror

    Science.gov (United States)

    Costa, Christopher; Bradu, Adrian; Rogers, John; Phelan, Pauline; Podoleanu, Adrian

    2015-01-01

    We present a swept source optical coherence tomography (OCT) system at 1060 nm equipped with a wavefront sensor at 830 nm and a deformable mirror in a closed-loop adaptive optics (AO) system. Due to the AO correction, the confocal profile of the interface optics becomes narrower than the OCT axial range, restricting the part of the B-scan (cross section) with good contrast. By actuating on the deformable mirror, the depth of the focus is changed and the system is used to demonstrate Gabor filtering in order to produce B-scan OCT images with enhanced sensitivity throughout the axial range from a Drosophila larvae. The focus adjustment is achieved by manipulating the curvature of the deformable mirror between two user-defined limits. Particularities of controlling the focus for Gabor filtering using the deformable mirror are presented.

  10. Correlative stochastic optical reconstruction microscopy and electron microscopy.

    Directory of Open Access Journals (Sweden)

    Doory Kim

    Full Text Available Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets.

  11. Disposable optics for microscopy diagnostics.

    Science.gov (United States)

    Vilmi, Pauliina; Varjo, Sami; Sliz, Rafal; Hannuksela, Jari; Fabritius, Tapio

    2015-11-20

    The point-of-care testing (POCT) is having increasing role on modern health care systems due to a possibility to perform tests for patients conveniently and immediately. POCT includes lot of disposable devices because of the environment they are often used. For a disposable system to be reasonably utilized, it needs to be high in quality but low in price. Optics based POCT systems are interesting approach to be developed, and here we describe a low-cost fabrication process for microlens arrays for microscopy. Lens arrays having average lens diameter of 222 μm with 300 μm lens pitch were fabricated. The lenses were characterized to have standard deviation of 0.06 μm in height and 4.61 μm in diameter. The resolution limit of 3.9μm is demonstrated with real images, and the images were compared with ones made with glass and polycarbonate lens arrays. The image quality is at the same level than with the glass lenses and the manufacturing costs are very low, thus making them suitable for POCT applications.

  12. Disposable optics for microscopy diagnostics

    Science.gov (United States)

    Vilmi, Pauliina; Varjo, Sami; Sliz, Rafal; Hannuksela, Jari; Fabritius, Tapio

    2015-11-01

    The point-of-care testing (POCT) is having increasing role on modern health care systems due to a possibility to perform tests for patients conveniently and immediately. POCT includes lot of disposable devices because of the environment they are often used. For a disposable system to be reasonably utilized, it needs to be high in quality but low in price. Optics based POCT systems are interesting approach to be developed, and here we describe a low-cost fabrication process for microlens arrays for microscopy. Lens arrays having average lens diameter of 222 μm with 300 μm lens pitch were fabricated. The lenses were characterized to have standard deviation of 0.06 μm in height and 4.61 μm in diameter. The resolution limit of 3.9μm is demonstrated with real images, and the images were compared with ones made with glass and polycarbonate lens arrays. The image quality is at the same level than with the glass lenses and the manufacturing costs are very low, thus making them suitable for POCT applications.

  13. Development of emulsion track expansion techniques for optical-microscopy-observation of low-velocity ion tracks with ranges beyond optical resolution limit

    Energy Technology Data Exchange (ETDEWEB)

    Naka, T. [F-lab., Department of Physics, Nagoya University, Nagoya 464-8602 (Japan); Natsume, M. [F-lab., Department of Physics, Nagoya University, Nagoya 464-8602 (Japan)], E-mail: natsume@flab.phys.nagoya-u.ac.jp; Niwa, K.; Hoshino, K.; Nakamura, M.; Nakano, T.; Sato, O. [F-lab., Department of Physics, Nagoya University, Nagoya 464-8602 (Japan)

    2007-11-01

    We succeeded to observe tracks of low-velocity Kr ions, having originally ranges below optical resolution, in a fine grain nuclear emulsion with an optical microscope after expanding the emulsion along the incident direction. This opens up the possibility of tracking low-velocity nuclear recoils from massive dark matter particles using optical microscope scanning systems.

  14. Using electron microscopy to calculate optical properties of biological samples

    OpenAIRE

    Wu, Wenli; Radosevich, Andrew J.; Eshein, Adam; Nguyen, The-Quyen; Yi, Ji; Cherkezyan, Lusik; Roy, Hemant K.; Szleifer, Igal; Backman, Vadim

    2016-01-01

    The microscopic structural origins of optical properties in biological media are still not fully understood. Better understanding these origins can serve to improve the utility of existing techniques and facilitate the discovery of other novel techniques. We propose a novel analysis technique using electron microscopy (EM) to calculate optical properties of specific biological structures. This method is demonstrated with images of human epithelial colon cell nuclei. The spectrum of anisotropy...

  15. Photoinduced force microscopy: A technique for hyperspectral nanochemical mapping

    Science.gov (United States)

    Murdick, Ryan A.; Morrison, William; Nowak, Derek; Albrecht, Thomas R.; Jahng, Junghoon; Park, Sung

    2017-08-01

    Advances in nanotechnology have intensified the need for tools that can characterize newly synthesized nanomaterials. A variety of techniques has recently been shown which combines atomic force microscopy (AFM) with optical illumination including tip-enhanced Raman spectroscopy (TERS), scattering-type scanning near-field optical microscopy (sSNOM), and photothermal induced resonance microscopy (PTIR). To varying degrees, these existing techniques enable optical spectroscopy with the nanoscale spatial resolution inherent to AFM, thereby providing nanochemical interrogation of a specimen. Here we discuss photoinduced force microscopy (PiFM), a recently developed technique for nanoscale optical spectroscopy that exploits image forces acting between an AFM tip and sample to detect wavelength-dependent polarization within the sample to generate absorption spectra. This approach enables ∼10 nm spatial resolution with spectra that show correlation with macroscopic optical absorption spectra. Unlike other techniques, PiFM achieves this high resolution with virtually no constraints on sample or substrate properties. The applicability of PiFM to a variety of archetypal systems is reported here, highlighting the potential of PiFM as a useful tool for a wide variety of industrial and academic investigations, including semiconducting nanoparticles, nanocellulose, block copolymers, and low dimensional systems, as well as chemical and morphological mixing at interfaces.

  16. Fibre-optic nonlinear optical microscopy and endoscopy.

    Science.gov (United States)

    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.

  17. Characterization of Polymer Blends: Optical Microscopy (*Polarized, Interference and Phase Contrast Microscopy*) and Confocal Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ramanathan, Nathan Muruganathan [ORNL; Darling, Seth B. [Argonne National Laboratory (ANL)

    2015-01-01

    Chapter 15 surveys the characterization of macro, micro and meso morphologies of polymer blends by optical microscopy. Confocal Microscopy offers the ability to view the three dimensional morphology of polymer blends, popular in characterization of biological systems. Confocal microscopy uses point illumination and a spatial pinhole to eliminate out-of focus light in samples that are thicker than the focal plane.

  18. Optical tweezers for confocal microscopy

    Science.gov (United States)

    Hoffmann, A.; Meyer zu Hörste, G.; Pilarczyk, G.; Monajembashi, S.; Uhl, V.; Greulich, K. O.

    2000-11-01

    In confocal laser scanning microscopes (CLSMs), lasers can be used for image formation as well as tools for the manipulation of microscopic objects. In the latter case, in addition to the imaging lasers, the light of an extra laser has to be focused into the object plane of the CLSM, for example as optical tweezers. Imaging as well as trapping by optical tweezers can be done using the same objective lens. In this case, z-sectioning for 3D imaging shifts the optical tweezers with the focal plane of the objective along the optical axis, so that a trapped object remains positioned in the focal plane. Consequently, 3D imaging of trapped objects is impossible without further measures. We present an experimental set-up keeping the axial trapping position of the optical tweezers at its intended position whilst the focal plane can be axially shifted over a distance of about 15 μm. It is based on fast-moving correctional optics synchronized with the objective movement. First examples of application are the 3D imaging of chloroplasts of Elodea densa (Canadian waterweed) in a vigorous cytoplasmic streaming and the displacement of zymogen granules in pancreatic cancer cells (AR42 J).

  19. Scanning tunneling microscopy II further applications and related scanning techniques

    CERN Document Server

    Güntherodt, Hans-Joachim

    1992-01-01

    Scanning Tunneling Microscopy II, like its predecessor, presents detailed and comprehensive accounts of the basic principles and broad range of applications of STM and related scanning probe techniques. The applications discussed in this volume come predominantly from the fields of electrochemistry and biology. In contrast to those described in Vol. I, these sudies may be performed in air and in liquids. The extensions of the basic technique to map other interactions are described inchapters on scanning force microscopy, magnetic force microscopy, scanning near-field optical microscopy, together with a survey of other related techniques. Also described here is the use of a scanning proximal probe for surface modification. Togehter, the two volumes give a comprehensive account of experimental aspcets of STM. They provide essentialreading and reference material for all students and researchers involvedin this field.

  20. Scanning tunneling microscopy II further applications and related scanning techniques

    CERN Document Server

    Güntherodt, Hans-Joachim

    1995-01-01

    Scanning Tunneling Microscopy II, like its predecessor, presents detailed and comprehensive accounts of the basic principles and broad range of applications of STM and related scanning probe techniques. The applications discussed in this volume come predominantly from the fields of electrochemistry and biology. In contrast to those described in STM I, these studies may be performed in air and in liquids. The extensions of the basic technique to map other interactions are described in chapters on scanning force microscopy, magnetic force microscopy, and scanning near-field optical microscopy, together with a survey of other related techniques. Also described here is the use of a scanning proximal probe for surface modification. Together, the two volumes give a comprehensive account of experimental aspects of STM. They provide essential reading and reference material for all students and researchers involved in this field. In this second edition the text has been updated and new methods are discussed.

  1. Volumetric optical coherence microscopy enabled by aberrated optics (Conference Presentation)

    Science.gov (United States)

    Mulligan, Jeffrey A.; Liu, Siyang; Adie, Steven G.

    2017-02-01

    Optical coherence microscopy (OCM) is an interferometric imaging technique that enables high resolution, non-invasive imaging of 3D cell cultures and biological tissues. Volumetric imaging with OCM suffers a trade-off between high transverse resolution and poor depth-of-field resulting from defocus, optical aberrations, and reduced signal collection away from the focal plane. While defocus and aberrations can be compensated with computational methods such as interferometric synthetic aperture microscopy (ISAM) or computational adaptive optics (CAO), reduced signal collection must be physically addressed through optical hardware. Axial scanning of the focus is one approach, but comes at the cost of longer acquisition times, larger datasets, and greater image reconstruction times. Given the capabilities of CAO to compensate for general phase aberrations, we present an alternative method to address the signal collection problem without axial scanning by using intentionally aberrated optical hardware. We demonstrate the use of an astigmatic spectral domain (SD-)OCM imaging system to enable single-acquisition volumetric OCM in 3D cell culture over an extended depth range, compared to a non-aberrated SD-OCM system. The transverse resolution of the non-aberrated and astigmatic imaging systems after application of CAO were 2 um and 2.2 um, respectively. The depth-range of effective signal collection about the nominal focal plane was increased from 100 um in the non-aberrated system to over 300 um in the astigmatic system, extending the range over which useful data may be acquired in a single OCM dataset. We anticipate that this method will enable high-throughput cellular-resolution imaging of dynamic biological systems over extended volumes.

  2. Aberrations and adaptive optics in super-resolution microscopy.

    Science.gov (United States)

    Booth, Martin; Andrade, Débora; Burke, Daniel; Patton, Brian; Zurauskas, Mantas

    2015-08-01

    As one of the most powerful tools in the biological investigation of cellular structures and dynamic processes, fluorescence microscopy has undergone extraordinary developments in the past decades. The advent of super-resolution techniques has enabled fluorescence microscopy - or rather nanoscopy - to achieve nanoscale resolution in living specimens and unravelled the interior of cells with unprecedented detail. The methods employed in this expanding field of microscopy, however, are especially prone to the detrimental effects of optical aberrations. In this review, we discuss how super-resolution microscopy techniques based upon single-molecule switching, stimulated emission depletion and structured illumination each suffer from aberrations in different ways that are dependent upon intrinsic technical aspects. We discuss the use of adaptive optics as an effective means to overcome this problem.

  3. Optical Property Analyses of Plant Cells for Adaptive Optics Microscopy

    Science.gov (United States)

    Tamada, Yosuke; Murata, Takashi; Hattori, Masayuki; Oya, Shin; Hayano, Yutaka; Kamei, Yasuhiro; Hasebe, Mitsuyasu

    2014-04-01

    In astronomy, adaptive optics (AO) can be used to cancel aberrations caused by atmospheric turbulence and to perform diffraction-limited observation of astronomical objects from the ground. AO can also be applied to microscopy, to cancel aberrations caused by cellular structures and to perform high-resolution live imaging. As a step toward the application of AO to microscopy, here we analyzed the optical properties of plant cells. We used leaves of the moss Physcomitrella patens, which have a single layer of cells and are thus suitable for optical analysis. Observation of the cells with bright field and phase contrast microscopy, and image degradation analysis using fluorescent beads demonstrated that chloroplasts provide the main source of optical degradations. Unexpectedly, the cell wall, which was thought to be a major obstacle, has only a minor effect. Such information provides the basis for the application of AO to microscopy for the observation of plant cells.

  4. Near-field reflection backscattering apertureless optical microscopy: Application to spectroscopy experiments on opaque samples, comparison between lock-in and digital photon counting detection techniques

    Energy Technology Data Exchange (ETDEWEB)

    Diziain, S. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Bijeon, J.-L. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France)]. E-mail: bijeon@utt.fr; Adam, P.-M. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Lamy de la Chapelle, M. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Thomas, B. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Deturche, R. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France); Royer, P. [Institut Charles Delaunay, CNRS FRE 2848, Laboratoire de Nanotechnologie et d' Instrumentation Optique, Universite de technologie de Troyes, 12 rue Marie Curie, BP 2060, 10010 Troyes cedex (France)

    2007-01-15

    An apertureless scanning near-field optical microscope (ASNOM) in reflection backscattering configuration is designed to conduct spectroscopic experiments on opaque samples constituted of latex beads. The ASNOM proposed takes advantage of the depth-discrimination properties of confocal microscopes to efficiently extract the near-field optical signal. Given their importance in a spectroscopic experiment, we systematically compare the lock-in and synchronous photon counting detection methods. Some results of Rayleigh's scattering in the near field of the test samples are used to illustrate the possibilities of this technique for reflection backscattering spectroscopy.

  5. Adaptive optics in digital micromirror based confocal microscopy

    Science.gov (United States)

    Pozzi, P.; Wilding, D.; Soloviev, O.; Vdovin, G.; Verhaegen, M.

    2016-03-01

    This proceeding reports early results in the development of a new technique for adaptive optics in confocal microscopy. The term adaptive optics refers to the branch of optics in which an active element in the optical system is used to correct inhomogeneities in the media through which light propagates. In its most classical form, mostly used in astronomical imaging, adaptive optics is achieved through a closed loop in which the actuators of a deformable mirror are driven by a wavefront sensor. This approach is severely limited in fluorescence microscopy, as the use of a wavefront sensor requires the presence of a bright, point like source in the field of view, a condition rarely satisfied in microscopy samples. Previously reported approaches to adaptive optics in fluorescence microscopy are therefore limited to the inclusion of fluorescent microspheres in the sample, to use as bright stars for wavefront sensors, or time consuming sensorless optimization procedures, requiring several seconds of optimization before the acquisition of a single image. We propose an alternative approach to the problem, implementing sensorless adaptive optics in a Programmable array microscope. A programmable array microscope is a microscope based on a digital micromirror device, in which the single elements of the micromirror act both as point sources and pinholes.

  6. Where Do We Stand with Super-Resolution Optical Microscopy?

    Science.gov (United States)

    Nienhaus, Karin; Nienhaus, G Ulrich

    2016-01-29

    Super-resolution fluorescence microscopy has become an invaluable, powerful approach to study biomolecular dynamics and interactions via selective labeling and observation of specific molecules in living cells, tissues and even entire organisms. In this perspective, we present a brief overview of the main techniques and their application to cellular biophysics. We place special emphasis on super-resolution imaging via single-molecule localization microscopy and stimulated emission depletion/reversible saturable optical fluorescence transitions microscopy, and we also briefly address fluorescence fluctuation approaches, notably raster image correlation spectroscopy, as tools to record fast diffusion and transport.

  7. Optical super-resolution microscopy in neurobiology.

    Science.gov (United States)

    Sigrist, Stephan J; Sabatini, Bernardo L

    2012-02-01

    Understanding the highly plastic nature of neurons requires the dynamic visualization of their molecular and cellular organization in a native context. However, due to the limited resolution of standard light microscopy, many of the structural specializations of neurons cannot be resolved. A recent revolution in light microscopy has given rise to several super-resolution light microscopy methods yielding 2-10-fold higher resolution than conventional microscopy. We here describe the principles behind these techniques as well as their application to the analysis of the molecular architecture of the synapse. Furthermore, we discuss the potential for continued development of super-resolution microscopy as necessary for live imaging of neuronal structure and function in the brain.

  8. Optical techniques in optogenetics

    Science.gov (United States)

    Mohanty, Samarendra K.; Lakshminarayananan, Vasudevan

    2015-07-01

    Optogenetics is an innovative technique for optical control of cells. This field has exploded over the past decade or so and has given rise to great advances in neuroscience. A variety of applications both from the basic and applied research have emerged, turning the early ideas into a powerful paradigm for cell biology, neuroscience, and medical research. This review aims at highlighting the basic concepts that are essential for a comprehensive understanding of optogenetics and some important biological/biomedical applications. Further, emphasis is placed on advancement in optogenetics-associated light-based methods for controlling gene expression, spatially controlled optogenetic stimulation and detection of cellular activities.

  9. Capturing the surface texture and shape of pollen: a comparison of microscopy techniques.

    Directory of Open Access Journals (Sweden)

    Mayandi Sivaguru

    Full Text Available Research on the comparative morphology of pollen grains depends crucially on the application of appropriate microscopy techniques. Information on the performance of microscopy techniques can be used to inform that choice. We compared the ability of several microscopy techniques to provide information on the shape and surface texture of three pollen types with differing morphologies. These techniques are: widefield, apotome, confocal and two-photon microscopy (reflected light techniques, and brightfield and differential interference contrast microscopy (DIC (transmitted light techniques. We also provide a first view of pollen using super-resolution microscopy. The three pollen types used to contrast the performance of each technique are: Croton hirtus (Euphorbiaceae, Mabea occidentalis (Euphorbiaceae and Agropyron repens (Poaceae. No single microscopy technique provided an adequate picture of both the shape and surface texture of any of the three pollen types investigated here. The wavelength of incident light, photon-collection ability of the optical technique, signal-to-noise ratio, and the thickness and light absorption characteristics of the exine profoundly affect the recovery of morphological information by a given optical microscopy technique. Reflected light techniques, particularly confocal and two-photon microscopy, best capture pollen shape but provide limited information on very fine surface texture. In contrast, transmitted light techniques, particularly differential interference contrast microscopy, can resolve very fine surface texture but provide limited information on shape. Texture comprising sculptural elements that are spaced near the diffraction limit of light (~250 nm; NDL presents an acute challenge to optical microscopy. Super-resolution structured illumination microscopy provides data on the NDL texture of A. repens that is more comparable to textural data from scanning electron microscopy than any other optical

  10. Quantitative Topographical Characterization of Thermally Sprayed Coatings by Optical Microscopy

    Science.gov (United States)

    Schwaller, P.; Züst, R.; Michler, J.

    2009-03-01

    Topography measurements and roughness calculations for different rough surfaces (Rugotest surface comparator and thermally sprayed coatings) are presented. The surfaces are measured with a novel quantitative topography measurement technique based on optical stereomicroscopy and a comparison is made with established scanning stylus and optical profilometers. The results show that for most cases the different methods yield similar results. Stereomicroscopy is therefore a valuable method for topographical investigations in both quality control and research. On the other hand, the method based on optical microscopy demands a careful optimization of the experimental settings like the magnification and the illumination to achieve satisfactory results.

  11. Using electron microscopy to calculate optical properties of biological samples.

    Science.gov (United States)

    Wu, Wenli; Radosevich, Andrew J; Eshein, Adam; Nguyen, The-Quyen; Yi, Ji; Cherkezyan, Lusik; Roy, Hemant K; Szleifer, Igal; Backman, Vadim

    2016-11-01

    The microscopic structural origins of optical properties in biological media are still not fully understood. Better understanding these origins can serve to improve the utility of existing techniques and facilitate the discovery of other novel techniques. We propose a novel analysis technique using electron microscopy (EM) to calculate optical properties of specific biological structures. This method is demonstrated with images of human epithelial colon cell nuclei. The spectrum of anisotropy factor g, the phase function and the shape factor D of the nuclei are calculated. The results show strong agreement with an independent study. This method provides a new way to extract the true phase function of biological samples and provides an independent validation for optical property measurement techniques.

  12. Imaging photothermal microscopy for absorption measurements of optical coatings

    Institute of Scientific and Technical Information of China (English)

    Chunxian Tao; Yuanan Zhao; Hongbo He; Dawei Li; Jianda Shao; Zhengxiu Fan

    2009-01-01

    @@ For absorption measurement of large-aperture optical coatings, a novel method of imaging photothermal microscopy based on image lock-in technique is presented.Detailed theoretical analysis and numerical calculation are made based on the image photothermal technique.The feasibility of this imaging method is proved through the coincidence between the theoretical results of single spot method and multi-channel method.The measuring speed of this imaging method can be increased hundreds of times compared with that of the raster scanning.This technique can expand the applications of photothermal technique.

  13. Virtual k -Space Modulation Optical Microscopy

    Science.gov (United States)

    Kuang, Cuifang; Ma, Ye; Zhou, Renjie; Zheng, Guoan; Fang, Yue; Xu, Yingke; Liu, Xu; So, Peter T. C.

    2016-07-01

    We report a novel superresolution microscopy approach for imaging fluorescence samples. The reported approach, termed virtual k -space modulation optical microscopy (VIKMOM), is able to improve the lateral resolution by a factor of 2, reduce the background level, improve the optical sectioning effect and correct for unknown optical aberrations. In the acquisition process of VIKMOM, we used a scanning confocal microscope setup with a 2D detector array to capture sample information at each scanned x -y position. In the recovery process of VIKMOM, we first modulated the captured data by virtual k -space coding and then employed a ptychography-inspired procedure to recover the sample information and correct for unknown optical aberrations. We demonstrated the performance of the reported approach by imaging fluorescent beads, fixed bovine pulmonary artery endothelial (BPAE) cells, and living human astrocytes (HA). As the VIKMOM approach is fully compatible with conventional confocal microscope setups, it may provide a turn-key solution for imaging biological samples with ˜100 nm lateral resolution, in two or three dimensions, with improved optical sectioning capabilities and aberration correcting.

  14. A near-field optical microscopy nanoarray

    Energy Technology Data Exchange (ETDEWEB)

    Semin, D.J.; Ambrose, W.P.; Goodwin, P.M.; Kwller, A. [Los Alamos National Lab., NM (United States); Wendt, J.R. [Sandia National Labs., Albuquerque, NM (United States)

    1996-12-31

    Multiplexing near-field scanning optical microscopy (NSOM) by the use of a nanoarray with parallel imaging is studied. The fabrication, characterization, and utilization of nanoarrays with {approximately} 100 nm diameter apertures spaced 500 nm center-to- center is presented. Extremely uniform nanoarrays with {approximately} 10{sup 8} apertures were fabricated by electron beam lithography and reactive ion etching. The nanoarrays were characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM). In this paper we utilize these nanoarrays in a laser-illuminated microscope with parallel detection on a charge- coupled device (CCD). Detection of B-phycoerythrin (B-PE) molecules using near-field illumination is presented. In principle, our system can be used to obtain high lateral resolution NSOM images over a wide-field of view (e.g. 50-100 {mu}m) within seconds.

  15. Integrated structural and functional optical imaging combining spectral-domain optical coherence and multiphoton microscopy

    CERN Document Server

    Vinegoni, C; Luo, W; Marks, D L; Ralston, T; Tan, W

    2005-01-01

    An integrated microscope that combines different optical techniques for simultaneous imaging is demonstrated. The microscope enables spectral-domain optical coherence microscopy based on optical backscatter, and multi-photon microscopy for the detection of two-photon fluorescence and second harmonic generation signals. The unique configuration of this integrated microscope allows for the simultaneous acquisition of both anatomical (structural) and functional imaging information with particular emphasis for applications in the fields of tissue engineering and cell biology. In addition, the contemporary analysis of the spectroscopic features can enhance contrast by differentiating among different tissue components.

  16. A correlative optical microscopy and scanning electron microscopy approach to locating nanoparticles in brain tumors.

    Science.gov (United States)

    Kempen, Paul J; Kircher, Moritz F; de la Zerda, Adam; Zavaleta, Cristina L; Jokerst, Jesse V; Mellinghoff, Ingo K; Gambhir, Sanjiv S; Sinclair, Robert

    2015-01-01

    The growing use of nanoparticles in biomedical applications, including cancer diagnosis and treatment, demands the capability to exactly locate them within complex biological systems. In this work a correlative optical and scanning electron microscopy technique was developed to locate and observe multi-modal gold core nanoparticle accumulation in brain tumor models. Entire brain sections from mice containing orthotopic brain tumors injected intravenously with nanoparticles were imaged using both optical microscopy to identify the brain tumor, and scanning electron microscopy to identify the individual nanoparticles. Gold-based nanoparticles were readily identified in the scanning electron microscope using backscattered electron imaging as bright spots against a darker background. This information was then correlated to determine the exact location of the nanoparticles within the brain tissue. The nanoparticles were located only in areas that contained tumor cells, and not in the surrounding healthy brain tissue. This correlative technique provides a powerful method to relate the macro- and micro-scale features visible in light microscopy with the nanoscale features resolvable in scanning electron microscopy.

  17. Super-resolution microscopy of single atoms in optical lattices

    CERN Document Server

    Alberti, Andrea; Alt, Wolfgang; Brakhane, Stefan; Karski, Michał; Reimann, René; Widera, Artur; Meschede, Dieter

    2015-01-01

    We report on image processing techniques and experimental procedures to determine the lattice-site positions of single atoms in an optical lattice with high reliability, even for limited acquisition time or optical resolution. Determining the positions of atoms beyond the diffraction limit relies on parametric deconvolution in close analogy to methods employed in super-resolution microscopy. We develop a deconvolution method that makes effective use of the prior knowledge of the optical transfer function, noise properties, and discreteness of the optical lattice. We show that accurate knowledge of the image formation process enables a dramatic improvement on the localization reliability. This is especially relevant for closely packed ensembles of atoms where the separation between particles cannot be directly optically resolved. Furthermore, we demonstrate experimental methods to precisely reconstruct the point spread function with sub-pixel resolution from fluorescence images of single atoms, and we give a m...

  18. Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

    OpenAIRE

    Doory Kim; Deerinck, Thomas J.; Sigal, Yaron M.; Babcock, Hazen P.; Ellisman, Mark H.; Xiaowei Zhuang

    2015-01-01

    Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and ima...

  19. Multiparallel Three-Dimensional Optical Microscopy

    Science.gov (United States)

    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.

  20. Microscopy Techniques for Analysis of Nickel Metal Hydride Batteries Constituents.

    Science.gov (United States)

    Carpenter, Graham J C; Wronski, Zbigniew

    2015-12-01

    With the need for improvements in the performance of rechargeable batteries has come the necessity to better characterize cell electrodes and their component materials. Electron microscopy has been shown to reveal many important features of microstructure that are becoming increasingly important for understanding the behavior of the components during the many charge/discharge cycles required in modern applications. The aim of this paper is to present an overview of how the full suite of techniques available using transmission electron microscopy (TEM) and scanning transmission electron microscopy was applied to the case of materials for the positive electrode in nickel metal hydride rechargeable battery electrodes. Embedding and sectioning of battery-grade powders with an ultramicrotome was used to produce specimens that could be readily characterized by TEM. Complete electrodes were embedded after drying, and also after dehydration from the original wet state, for examination by optical microscopy and using focused ion beam techniques. Results of these studies are summarized to illustrate the significance of the microstructural information obtained.

  1. Optical techniques in regenerative medicine

    CERN Document Server

    Morgan, Stephen P

    2013-01-01

    In regenerative medicine, tissue engineers largely rely on destructive and time-consuming techniques that do not allow in situ and spatial monitoring of tissue growth. Furthermore, once the therapy is implanted in the patient, clinicians are often unable to monitor what is happening in the body. To tackle these barriers, optical techniques have been developed to image and characterize many tissue properties, fabricate tissue engineering scaffolds, and characterize the properties of the scaffolds. Optical Techniques in Regenerative Medicine illustrates how to use optical imaging techniques and

  2. Portable fiber-optic taper coupled optical microscopy platform

    Science.gov (United States)

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

    2017-04-01

    The optical fiber taper coupled with CMOS has advantages of high sensitivity, compact structure and low distortion in the imaging platform. So it is widely used in low light, high speed and X-ray imaging systems. In the meanwhile, the peculiarity of the coupled structure can meet the needs of the demand in microscopy imaging. Toward this end, we developed a microscopic imaging platform based on the coupling of cellphone camera module and fiber optic taper for the measurement of the human blood samples and ascaris lumbricoides. The platform, weighing 70 grams, is based on the existing camera module of the smartphone and a fiber-optic array which providing a magnification factor of 6x.The top facet of the taper, on which samples are placed, serves as an irregular sampling grid for contact imaging. The magnified images of the sample, located on the bottom facet of the fiber, are then projected onto the CMOS sensor. This paper introduces the portable medical imaging system based on the optical fiber coupling with CMOS, and theoretically analyzes the feasibility of the system. The image data and process results either can be stored on the memory or transmitted to the remote medical institutions for the telemedicine. We validate the performance of this cell-phone based microscopy platform using human blood samples and test target, achieving comparable results to a standard bench-top microscope.

  3. Holographic fluorescence microscopy with incoherent digital holographic adaptive optics.

    Science.gov (United States)

    Jang, Changwon; Kim, Jonghyun; Clark, David C; Lee, Seungjae; Lee, Byoungho; Kim, Myung K

    2015-01-01

    Introduction of adaptive optics technology into astronomy and ophthalmology has made great contributions in these fields, allowing one to recover images blurred by atmospheric turbulence or aberrations of the eye. Similar adaptive optics improvement in microscopic imaging is also of interest to researchers using various techniques. Current technology of adaptive optics typically contains three key elements: a wavefront sensor, wavefront corrector, and controller. These hardware elements tend to be bulky, expensive, and limited in resolution, involving, for example, lenslet arrays for sensing or multiactuator deformable mirrors for correcting. We have previously introduced an alternate approach based on unique capabilities of digital holography, namely direct access to the phase profile of an optical field and the ability to numerically manipulate the phase profile. We have also demonstrated that direct access and compensation of the phase profile are possible not only with conventional coherent digital holography, but also with a new type of digital holography using incoherent light: selfinterference incoherent digital holography (SIDH). The SIDH generates a complex—i.e., amplitude plus phase—hologram from one or several interferograms acquired with incoherent light, such as LEDs, lamps, sunlight, or fluorescence. The complex point spread function can be measured using guide star illumination and it allows deterministic deconvolution of the full-field image. We present experimental demonstration of aberration compensation in holographic fluorescence microscopy using SIDH. Adaptive optics by SIDH provides new tools for improved cellular fluorescence microscopy through intact tissue layers or other types of aberrant media.

  4. Holographic fluorescence microscopy with incoherent digital holographic adaptive optics

    Science.gov (United States)

    Jang, Changwon; Kim, Jonghyun; Clark, David C.; Lee, Seungjae; Lee, Byoungho; Kim, Myung K.

    2015-11-01

    Introduction of adaptive optics technology into astronomy and ophthalmology has made great contributions in these fields, allowing one to recover images blurred by atmospheric turbulence or aberrations of the eye. Similar adaptive optics improvement in microscopic imaging is also of interest to researchers using various techniques. Current technology of adaptive optics typically contains three key elements: a wavefront sensor, wavefront corrector, and controller. These hardware elements tend to be bulky, expensive, and limited in resolution, involving, for example, lenslet arrays for sensing or multiactuator deformable mirrors for correcting. We have previously introduced an alternate approach based on unique capabilities of digital holography, namely direct access to the phase profile of an optical field and the ability to numerically manipulate the phase profile. We have also demonstrated that direct access and compensation of the phase profile are possible not only with conventional coherent digital holography, but also with a new type of digital holography using incoherent light: self­interference incoherent digital holography (SIDH). The SIDH generates a complex-i.e., amplitude plus phase-hologram from one or several interferograms acquired with incoherent light, such as LEDs, lamps, sunlight, or fluorescence. The complex point spread function can be measured using guide star illumination and it allows deterministic deconvolution of the full-field image. We present experimental demonstration of aberration compensation in holographic fluorescence microscopy using SIDH. Adaptive optics by SIDH provides new tools for improved cellular fluorescence microscopy through intact tissue layers or other types of aberrant media.

  5. Nonlinear optical microscopy improvement by focal-point axial modulation

    Science.gov (United States)

    Dashtabi, Mahdi Mozdoor; Massudi, Reza

    2016-05-01

    Among the most important challenges of microscopy-even more important than the resolution enhancement, especially in biological and neuroscience applications-is noninvasive and label-free imaging deeper into live scattering samples. However, the fundamental limitation on imaging depth is the signal-to-background ratio in scattering biological tissues. Here, using a vibrating microscope objective in conjunction with a lock-in amplifier, we demonstrate the background cancellation in imaging the samples surrounded by turbid and scattering media, which leads to more clear images deeper into the samples. Furthermore, this technique offers the localization and resolution enhancement as well as resolves ambiguities in signal interpretation, using a single-color laser. This technique is applicable to most nonlinear as well as some linear point-scanning optical microscopies.

  6. Super-resolution microscopy of single atoms in optical lattices

    Science.gov (United States)

    Alberti, Andrea; Robens, Carsten; Alt, Wolfgang; Brakhane, Stefan; Karski, Michał; Reimann, René; Widera, Artur; Meschede, Dieter

    2016-05-01

    We report on image processing techniques and experimental procedures to determine the lattice-site positions of single atoms in an optical lattice with high reliability, even for limited acquisition time or optical resolution. Determining the positions of atoms beyond the diffraction limit relies on parametric deconvolution in close analogy to methods employed in super-resolution microscopy. We develop a deconvolution method that makes effective use of the prior knowledge of the optical transfer function, noise properties, and discreteness of the optical lattice. We show that accurate knowledge of the image formation process enables a dramatic improvement on the localization reliability. This allows us to demonstrate super-resolution of the atoms’ position in closely packed ensembles where the separation between particles cannot be directly optically resolved. Furthermore, we demonstrate experimental methods to precisely reconstruct the point spread function with sub-pixel resolution from fluorescence images of single atoms, and we give a mathematical foundation thereof. We also discuss discretized image sampling in pixel detectors and provide a quantitative model of noise sources in electron multiplying CCD cameras. The techniques developed here are not only beneficial to neutral atom experiments, but could also be employed to improve the localization precision of trapped ions for ultra precise force sensing.

  7. Immobilization Techniques of Bacteria for Live Super-resolution Imaging Using Structured Illumination Microscopy.

    Science.gov (United States)

    Bottomley, Amy L; Turnbull, Lynne; Whitchurch, Cynthia B; Harry, Elizabeth J

    2017-01-01

    Advancements in optical microscopy technology have allowed huge progression in the ability to understand protein structure and dynamics in live bacterial cells using fluorescence microscopy. Paramount to high-quality microscopy is good sample preparation to avoid bacterial cell movement that can result in motion blur during image acquisition. Here, we describe two techniques of sample preparation that reduce unwanted cell movement and are suitable for application to a number of bacterial species and imaging methods.

  8. Optical pulses, lasers, measuring techniques

    CERN Document Server

    Früngel, Frank B A

    1965-01-01

    High Speed Pulse Technology: Volume II: Optical Pulses - Lasers - Measuring Techniques focuses on the theoretical and engineering problems that result from the capacitor discharge technique.This book is organized into three main topics: light flash production from a capacitive energy storage; signal transmission and ranging systems by capacitor discharges and lasers; and impulse measuring technique. This text specifically discusses the air spark under atmospheric conditions, industrial equipment for laser flashing, and claims for light transmitting system. The application of light impulse sign

  9. Correlative super-resolution fluorescence microscopy combined with optical coherence microscopy

    Science.gov (United States)

    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.

  10. Automated seeding-based nuclei segmentation in nonlinear optical microscopy.

    Science.gov (United States)

    Medyukhina, Anna; Meyer, Tobias; Heuke, Sandro; Vogler, Nadine; Dietzek, Benjamin; Popp, Jürgen

    2013-10-01

    Nonlinear optical (NLO) microscopy based, e.g., on coherent anti-Stokes Raman scattering (CARS) or two-photon-excited fluorescence (TPEF) is a fast label-free imaging technique, with a great potential for biomedical applications. However, NLO microscopy as a diagnostic tool is still in its infancy; there is a lack of robust and durable nuclei segmentation methods capable of accurate image processing in cases of variable image contrast, nuclear density, and type of investigated tissue. Nonetheless, such algorithms specifically adapted to NLO microscopy present one prerequisite for the technology to be routinely used, e.g., in pathology or intraoperatively for surgical guidance. In this paper, we compare the applicability of different seeding and boundary detection methods to NLO microscopic images in order to develop an optimal seeding-based approach capable of accurate segmentation of both TPEF and CARS images. Among different methods, the Laplacian of Gaussian filter showed the best accuracy for the seeding of the image, while a modified seeded watershed segmentation was the most accurate in the task of boundary detection. The resulting combination of these methods followed by the verification of the detected nuclei performs high average sensitivity and specificity when applied to various types of NLO microscopy images.

  11. Environmental monitoring using optical techniques

    Science.gov (United States)

    Svanberg, Sune

    2003-11-01

    An overview of optical techniques for environmental monitoring is presented. Range-resolved measurements of atmospheric pollutants can be performed using the differential absorption lidar technique. Fluorescence lidar allows assessment of vegetation status and also the conditions of the facades of historical buildings. Diode lasers provide particularly realistic schemes for atmospheric gas analysis, where certain wavelength ranges, which are not easily directly assessed, can be reached by sum- and difference frequency generation. Finally, the gas correlation principle can be used for real-time imaging of hydrocarbons. Several types of such optical environmental monitoring are illustrated with examples from research at the Lund Institute of Technology, Sweden.

  12. Metal alloys, matrix inclusions and manufacturing techniques of Moinhos de Golas collection (North Portugal): a study by micro-EDXRF, SEM-EDS, optical microscopy and X-ray radiography

    Science.gov (United States)

    Loureiro, Joana; Figueiredo, Elin; Silva, Rui J. C.; Araújo, M. Fátima; Fonte, João; Bettencourt, Ana M. S.

    2016-09-01

    A collection of 35 metallic artefacts comprising various typologies, some of which can be attributed to the Bronze Age and others to later periods, were studied to provide detailed information on elemental composition, manufacturing techniques and preservation state. Elemental analysis by micro-EDXRF and SEM-EDS was performed to investigate the use of different alloys and to study the presence of microstructural heterogeneities, as inclusions. X-ray radiography, optical microscopy and SEM-EDS were used to investigate manufacturing techniques and degradation features. Results showed that most of the artefacts were produced in a binary bronze alloy (Cu-Sn) with 10-15 wt% Sn and a low concentration of impurities. Other artefacts were produced in copper or in brass, the latest with varying contents of Zn, Sn and Pb. A variety of inclusions in the metal matrices were also found, some related to specific types of alloys, as (Cu-Ni)S2 in coppers, or ZnS in brasses. Microstructural observations revealed that the majority of the artefacts were subjected to cycles of thermomechanical processing after casting, being evident that among some artefacts different parts were subjected to distinct treatments. The radiographic images revealed structural heterogeneities related to local corrosion processes and fissures that seem to have developed in wear-tension zones, as in the handle of some daggers. Radiographic images were also useful to detect the use of different materials in one particular brass artefact, revealing the presence of a possible Cu-Sn solder.

  13. Absolute Position Total Internal Reflection Microscopy with an Optical Tweezer

    CERN Document Server

    Liu, Lulu; Rodriguez, Alejandro W; Capasso, Federico

    2014-01-01

    A non-invasive, in-situ calibration method for Total Internal Reflection Microscopy (TIRM) based on optical tweezing is presented which greatly expands the capabilities of this technique. We show that by making only simple modifications to the basic TIRM sensing setup and procedure, a probe particle's absolute position relative to a dielectric interface may be known with better than 10 nm precision out to a distance greater than 1 $\\mu$m from the surface. This represents an approximate 10x improvement in error and 3x improvement in measurement range over conventional TIRM methods. The technique's advantage is in the direct measurement of the probe particle's scattering intensity vs. height profile in-situ, rather than relying on calculations or inexact system analogs for calibration. To demonstrate the improved versatility of the TIRM method in terms of tunability, precision, and range, we show our results for the hindered near-wall diffusion coefficient for a spherical dielectric particle.

  14. Multimodal nonlinear optical microscopy used to discriminate epithelial ovarian cancer

    Science.gov (United States)

    Adur, J.; Pelegati, V. B.; de Thomaz, A. A.; Almeida, D. B.; Bottcher-Luiz, F.; Andrade, L. A. L. A.; Cesar, C. L.

    2011-07-01

    We used human specimens of epithelial ovarian cancer (serous type) to test the feasibility of nonlinear imaging as complementary tools for ovarian cancer diagnosis. Classical hematoxylin-and-eosin stained sections were applied to combining two-photon excitation fluorescence (TPEF), second (SHG), and third (THG) harmonic microscopy within the same imaging platform. We show that strong TPEF + SHG + THG signals can be obtained in fixed samples stained with Hematoxylin & Eosin (H&E) stored for a very long time and that H&E staining enhanced the THG signal. We demonstrate using anisotropy and morphological measurements, that SHG and THG of stained optical sections allow reproducible identification of neoplastic features such as architectural alterations of collagen fibrils at different stages of the neoplastic transformation and cellular atypia. Taken together, these results suggest that, with our viable imaging system, we can qualitatively and quantitatively assess endogenous optical biomarkers of the ovarian tissue with SHG and THG microscopy. This imaging capability may prove to be highly valuable in aiding to determine structural changes at the cellular and tissue levels, which may contribute to the development of new diagnostic techniques.

  15. Low-temperature electron microscopy: techniques and protocols.

    Science.gov (United States)

    Fleck, Roland A

    2015-01-01

    Low-temperature electron microscopy endeavors to provide "solidification of a biological specimen by cooling with the aim of minimal displacement of its components through the use of low temperature as a physical fixation strategy" (Steinbrecht and Zierold, Cryotechniques in biological electron microscopy. Springer-Verlag, Berlin, p 293, 1987). The intention is to maintain confidence that the tissue observed retains the morphology and dimensions of the living material while also ensuring soluble cellular components are not displaced. As applied to both scanning and transmission electron microscopy, cryo-electron microscopy is a strategy whereby the application of low-temperature techniques are used to reduce or remove processing artifacts which are commonly encountered in more conventional room temperature electron microscopy techniques which rely heavily on chemical fixation and heavy metal staining. Often, cryo-electron microscopy allows direct observation of specimens, which have not been stained or chemically fixed.

  16. Axial range of conjugate adaptive optics in two-photon microscopy

    CERN Document Server

    Paudel, Hari P; Mertz, Jerome; Bifano, Thomas

    2015-01-01

    We describe an adaptive optics technique for two-photon microscopy in which the deformable mirror used for aberration compensation is positioned in a plane conjugate to the plane of the aberration. We demonstrate in a proof-of-principle experiment that this technique yields a large field of view advantage in comparison to standard pupil-conjugate adaptive optics. Further, we show that the extended field of view in conjugate AO is maintained over a relatively large axial translation of the deformable mirror with respect to the conjugate plane. We conclude with a discussion of limitations and prospects for the conjugate AO technique in two-photon biological microscopy.

  17. Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy

    Science.gov (United States)

    Yu, Jiun-Yann; Kuo, Chun-Hung; Holland, Daniel B.; Chen, Yenyu; Ouyang, Mingxing; Blake, Geoffrey A.; Zadoyan, Ruben; Guo, Chin-Lin

    2011-11-01

    Optical sectioning provides three-dimensional (3D) information in biological tissues. However, most imaging techniques implemented with optical sectioning are either slow or deleterious to live tissues. Here, we present a simple design for wide-field multiphoton microscopy, which provides optical sectioning at a reasonable frame rate and with a biocompatible laser dosage. The underlying mechanism of optical sectioning is diffuser-based temporal focusing. Axial resolution comparable to confocal microscopy is theoretically derived and experimentally demonstrated. To achieve a reasonable frame rate without increasing the laser power, a low-repetition-rate ultrafast laser amplifier was used in our setup. A frame rate comparable to that of epifluorescence microscopy was demonstrated in the 3D imaging of fluorescent protein expressed in live epithelial cell clusters. In this report, our design displays the potential to be widely used for video-rate live-tissue and embryo imaging with axial resolution comparable to laser scanning microscopy.

  18. Backscattered Electron Microscopy as an Advanced Technique in Petrography.

    Science.gov (United States)

    Krinsley, David Henry; Manley, Curtis Robert

    1989-01-01

    Three uses of this method with sandstone, desert varnish, and granite weathering are described. Background information on this technique is provided. Advantages of this type of microscopy are stressed. (CW)

  19. Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy

    Science.gov (United States)

    Neuman, Keir C.; Nagy, Attila

    2012-01-01

    Single-molecule force spectroscopy has emerged as a powerful tool to investigate the forces and motions associated with biological molecules and enzymatic activity. The most common force spectroscopy techniques are optical tweezers, magnetic tweezers and atomic force microscopy. These techniques are described and illustrated with examples highlighting current capabilities and limitations. PMID:18511917

  20. Recent Advances in Biological Single-Molecule Applications of Optical Tweezers and Fluorescence Microscopy

    NARCIS (Netherlands)

    Hashemi Shabestari, M; Meijering, A E C; Roos, W H; Wuite, G J L; Peterman, E J G

    2017-01-01

    Over the past two decades, single-molecule techniques have evolved into robust tools to study many fundamental biological processes. The combination of optical tweezers with fluorescence microscopy and microfluidics provides a powerful single-molecule manipulation and visualization technique that

  1. Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy

    OpenAIRE

    Neuman, Keir C.; Nagy, Attila

    2008-01-01

    Single-molecule force spectroscopy has emerged as a powerful tool to investigate the forces and motions associated with biological molecules and enzymatic activity. The most common force spectroscopy techniques are optical tweezers, magnetic tweezers and atomic force microscopy. These techniques are described and illustrated with examples highlighting current capabilities and limitations.

  2. Hyperspectral Dark Field Optical Microscopy of Single Silver Nanospheres

    Energy Technology Data Exchange (ETDEWEB)

    El-Khoury, Patrick Z.; Joly, Alan G.; Hess, Wayne P.

    2016-04-07

    We record spectrally (400 nm ≤ λ ≤ 675 nm, Δλ < 4.69 nm) and spatially (diffraction-limited, sampled at 85 nm2/pixel) resolved dark field (DF) scattering from single silver nanospheres of 100 nm in diameter. Hyperspectral DF optical microscopy is achieved by coupling a hyperspectral detector to an optical microscope, whereby spectrally resolved diffraction-limited images of hundreds of silver nanoparticles can be recorded in ~30 seconds. We demonstrate how the centers and edges of individual particles can be localized in 2D to within a single pixel (85 nm2), using a statistical method for examining texture based on a co-occurrence matrix. Subsequently, spatial averaging of the spectral response in a 3x3 pixel area around the particle centers affords ample signal-to-noise to resolve the plasmon resonance of a single silver nanosphere. A close inspection of the scattering spectra of 31 different nanospheres reveals that each particle has its unique (i) relative scattering efficiency, and (ii) plasmon resonance maximum and dephasing time. These observations are suggestive of nanometric structural variations over length scales much finer than the spatial resolution attainable using the all-optical technique described herein.

  3. Automated control of optical polarization for nonlinear microscopy

    Science.gov (United States)

    Brideau, Craig; Stys, Peter K.

    2012-03-01

    Laser-scanning non-linear optical techniques such as multi-photon fluorescence excitation microscopy (MPM), Second/ Third Harmonic Generation (SHG/THG), and Coherent Anti-Stokes Raman Scattering (CARS) are being utilized in research laboratories worldwide. The efficiencies of these non-linear effects are dependent on the polarization state of the excitation light relative to the orientation of the sample being imaged. In highly ordered anisotropic biological samples this effect can become pronounced and the excitation polarization can have a dramatic impact on imaging experiments. Therefore, controlling the polarization state of the exciting light is important; however this is challenging when the excitation light passes through a complex optical system. In a typical laser-scanning microscope, components such as the dichroic filters, lenses, and even mirrors can alter the polarization state of a laser beam before it reaches the sample. We present an opto-mechanical solution to compensate for the polarization effects of an optical path, and to precisely program the polarization state of the exciting laser light. The device and accompanying procedures allow the delivery of precise laser polarization states at constant average power levels to a sample during an imaging experiment.

  4. Synergizing superresolution optical fluctuation imaging with single molecule localization microscopy

    CERN Document Server

    Schidorsky, Shachar; Razvag, Yair; Golan, Yonatan; Weiss, Shimon; Sherman, Eilon

    2016-01-01

    Single molecule localization microscopy (SMLM) techniques enable imaging biological samples well beyond the diffraction limit of light, but they vary significantly in their spatial and temporal resolutions. High-order statistical analysis of temporal fluctuations as in superresolution optical fluctuation imaging (SOFI) also enable imaging beyond diffraction limit, but usually at a lower resolution as compared to SMLM. Since the same data format is acquired for both methods, their algorithms can be applied to the same data set, and thus may be combined synergistically to improve overall imaging performance. Here, we find that SOFI converges much faster than SMLM, provides additive information to SMLM, and can efficiently reject background. We then show how SOFI-assisted SMLM imaging can improve SMLM image reconstruction by rejecting common sources of background, especially under low signal-to-background conditions. The performance of our approach was evaluated using a realistic simulation of fluorescence imagi...

  5. Exploring lipids with nonlinear optical microscopy in multiple biological systems

    Science.gov (United States)

    Alfonso-Garcia, Alba

    Lipids are crucial biomolecules for the well being of humans. Altered lipid metabolism may give rise to a variety of diseases that affect organs from the cardiovascular to the central nervous system. A deeper understanding of lipid metabolic processes would spur medical research towards developing precise diagnostic tools, treatment methods, and preventive strategies for reducing the impact of lipid diseases. Lipid visualization remains a complex task because of the perturbative effect exerted by traditional biochemical assays and most fluorescence markers. Coherent Raman scattering (CRS) microscopy enables interrogation of biological samples with minimum disturbance, and is particularly well suited for label-free visualization of lipids, providing chemical specificity without compromising on spatial resolution. Hyperspectral imaging yields large datasets that benefit from tailored multivariate analysis. In this thesis, CRS microscopy was combined with Raman spectroscopy and other label-free nonlinear optical techniques to analyze lipid metabolism in multiple biological systems. We used nonlinear Raman techniques to characterize Meibum secretions in the progression of dry eye disease, where the lipid and protein contributions change in ratio and phase segregation. We employed similar tools to examine lipid droplets in mice livers aboard a spaceflight mission, which lose their retinol content contributing to the onset of nonalcoholic fatty-liver disease. We also focused on atherosclerosis, a disease that revolves around lipid-rich plaques in arterial walls. We examined the lipid content of macrophages, whose variable phenotype gives rise to contrasting healing and inflammatory activities. We also proposed new label-free markers, based on lifetime imaging, for macrophage phenotype, and to detect products of lipid oxidation. Cholesterol was also detected in hepatitis C virus infected cells, and in specific strains of age-related macular degeneration diseased cells by

  6. Quantitative photoacoustic microscopy of optical absorption coefficients from acoustic spectra in the optical diffusive regime

    OpenAIRE

    Guo, Zijian; Favazza, Christopher; Garcia-Uribe, Alejandro; Lihong V. Wang

    2012-01-01

    Photoacoustic (PA) microscopy (PAM) can image optical absorption contrast with ultrasonic spatial resolution in the optical diffusive regime. Conventionally, accurate quantification in PAM requires knowledge of the optical fluence attenuation, acoustic pressure attenuation, and detection bandwidth. We circumvent this requirement by quantifying the optical absorption coefficients from the acoustic spectra of PA signals acquired at multiple optical wavelengths. With the acoustic spectral method...

  7. Ex vivo imaging of human thyroid pathology using integrated optical coherence tomography and optical coherence microscopy

    Science.gov (United States)

    Zhou, Chao; Wang, Yihong; Aguirre, Aaron D.; Tsai, Tsung-Han; Cohen, David W.; Connolly, James L.; Fujimoto, James G.

    2010-01-01

    We evaluate the feasibility of optical coherence tomography (OCT) and optical coherence microscopy (OCM) for imaging of benign and malignant thyroid lesions ex vivo using intrinsic optical contrast. 34 thyroid gland specimens are imaged from 17 patients, covering a spectrum of pathology ranging from normal thyroid to benign disease/neoplasms (multinodular colloid goiter, Hashimoto's thyroiditis, and follicular adenoma) and malignant thyroid tumors (papillary carcinoma and medullary carcinoma). Imaging is performed using an integrated OCT and OCM system, with sections. Characteristic features that suggest malignant lesions, such as complex papillary architecture, microfollicules, psammomatous calcifications, or replacement of normal follicular architecture with sheets/nests of tumor cells, can be identified from OCT and OCM images and are clearly differentiable from normal or benign thyroid tissues. With further development of needle-based imaging probes, OCT and OCM could be promising techniques to use for the screening of thyroid nodules and to improve the diagnostic specificity of fine needle aspiration evaluation.

  8. Internal Defect Measurement of Scattering Media by Optical Coherence Microscopy

    Institute of Scientific and Technical Information of China (English)

    ZHU Yong-kai; ZHAO Hong; WANG Zhao; WANG Jun-li

    2005-01-01

    Optical coherence microscopy is applied to measure scattering media's internal defect, which based on low coherence interferometry and confocal microscopy. Optical coherence microscopy is more effective in the rejection of out of focus and multiple scattered photons originating further away of the focal plane. With the three-dimension scanning, the internal defect is detected by measuring the thickness of different points on the sample. The axial resolution is 6 μm and lateral resolution is 1.2 μm. This method is possessed of the advantages over the other measurement method of scattering media, such as non-destruction and highresolution.

  9. Optimization-based wavefront sensorless adaptive optics for multiphoton microscopy

    NARCIS (Netherlands)

    Antonello, J.; Werkhoven, T. van; Verhaegen, M.; Truong, H.H.; Keller, C.U.; Gerritsen, H.C.

    2014-01-01

    Optical aberrations have detrimental effects in multiphoton microscopy. These effects can be curtailed by implementing model-based wavefront sensorless adaptive optics, which only requires the addition of a wavefront shaping device, such as a deformable mirror (DM) to an existing microscope. The abe

  10. Focus defect and dispersion mismatch in full-field optical coherence microscopy.

    Science.gov (United States)

    Dubois, Arnaud

    2017-03-20

    Full-field optical coherence microscopy (FFOCM) is an optical technique, based on low-coherence interference microscopy, for tomographic imaging of semi-transparent samples with micrometer-scale spatial resolution. The differences in refractive index between the sample and the immersion medium of the microscope objectives may degrade the FFOCM image quality because of focus defect and optical dispersion mismatch. These phenomena and their consequences are discussed in this theoretical paper. Experimental methods that have been implemented in FFOCM to minimize the adverse effects of these phenomena are summarized and compared.

  11. Nonlinear optical microscopy for imaging thin films and surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Smilowitz, L.B.; McBranch, D.W.; Robinson, J.M.

    1995-03-01

    We have used the inherent surface sensitivity of second harmonic generation to develop an instrument for nonlinear optical microscopy of surfaces and interfaces. We have demonstrated the use of several nonlinear optical responses for imaging thin films. The second harmonic response of a thin film of C{sub 60} has been used to image patterned films. Two photon absorption light induced fluorescence has been used to image patterned thin films of Rhodamine 6G. Applications of nonlinear optical microscopy include the imaging of charge injection and photoinduced charge transfer between layers in semiconductor heterojunction devices as well as across membranes in biological systems.

  12. Review of near-field optical microscopy

    Institute of Scientific and Technical Information of China (English)

    WU Shi-fa

    2006-01-01

    This review has introduced a new near-field optical microscope (NOM)-atomic force microscope combined with photon scanning tunneling microscope (AF/PSTM).During scanning,AF/PSTM could get two optical images of refractive index image and transmissivity image,and two AFM images of topography image and phase image.A reflected near-field optical microscope (AF/RSNOM) has also been developed on AF/PSTM platform.The NOM has been reviewed in this paper and the comparison between AF/PSTM & RSNOM and the commercial A-SNOM & RNOM has also been discussed.The functions of AF/PSTM & RSNOM are much better than A-SNOM & RNOM.

  13. Particles and waves in electron optics and microscopy

    CERN Document Server

    Pozzi, Giulio

    2016-01-01

    Advances in Imaging and Electron Physics merges two long-running serials, Advances in Electronics and Electron Physics and Advances in Optical and Electron Microscopy. The series features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science, digital image processing, electromagnetic wave propagation, electron microscopy, and the computing methods used in all these domains. * Contains contributions from leading authorities on the subject matter* Informs and updates all the latest developments in the field of imaging and electron physics* Provides practitioners interested in microscopy, optics, image processing, mathematical morphology, electromagnetic fields, electron, and ion emission with a valuable resource* Features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science, and digital image pro...

  14. Non-linear optical microscopy sheds light on cardiovascular disease.

    Directory of Open Access Journals (Sweden)

    Valentina Caorsi

    Full Text Available Many cardiac diseases have been associated with increased fibrosis and changes in the organization of fibrillar collagen. The degree of fibrosis is routinely analyzed with invasive histological and immunohistochemical methods, giving a limited and qualitative understanding of the tissue's morphological adaptation to disease. Our aim is to quantitatively evaluate the increase in fibrosis by three-dimensional imaging of the collagen network in the myocardium using the non-linear optical microscopy techniques Two-Photon Excitation microscopy (TPE and Second Harmonic signal Generation (SHG. No sample staining is needed because numerous endogenous fluorophores are excited by a two-photon mechanism and highly non-centrosymmetric structures such as collagen generate strong second harmonic signals. We propose for the first time a 3D quantitative analysis to carefully evaluate the increased fibrosis in tissue from a rat model of heart failure post myocardial infarction. We show how to measure changes in fibrosis from the backward SHG (B(SHG alone, as only backward-propagating SHG is accessible for true in vivo applications. A 5-fold increase in collagen I fibrosis is detected in the remote surviving myocardium measured 20 weeks after infarction. The spatial distribution is also shown to change markedly, providing insight into the morphology of disease progression.

  15. Non-Linear Optical Microscopy Sheds Light on Cardiovascular Disease

    Science.gov (United States)

    Caorsi, Valentina; Toepfer, Christopher; Sikkel, Markus B.; Lyon, Alexander R.; MacLeod, Ken; Ferenczi, Mike A.

    2013-01-01

    Many cardiac diseases have been associated with increased fibrosis and changes in the organization of fibrillar collagen. The degree of fibrosis is routinely analyzed with invasive histological and immunohistochemical methods, giving a limited and qualitative understanding of the tissue's morphological adaptation to disease. Our aim is to quantitatively evaluate the increase in fibrosis by three-dimensional imaging of the collagen network in the myocardium using the non-linear optical microscopy techniques Two-Photon Excitation microscopy (TPE) and Second Harmonic signal Generation (SHG). No sample staining is needed because numerous endogenous fluorophores are excited by a two-photon mechanism and highly non-centrosymmetric structures such as collagen generate strong second harmonic signals. We propose for the first time a 3D quantitative analysis to carefully evaluate the increased fibrosis in tissue from a rat model of heart failure post myocardial infarction. We show how to measure changes in fibrosis from the backward SHG (BSHG) alone, as only backward-propagating SHG is accessible for true in vivo applications. A 5-fold increase in collagen I fibrosis is detected in the remote surviving myocardium measured 20 weeks after infarction. The spatial distribution is also shown to change markedly, providing insight into the morphology of disease progression. PMID:23409139

  16. SCANNING TUNNELING MICROSCOPY STUDIES ON OPTICAL DISC

    Institute of Scientific and Technical Information of China (English)

    徐磊; 顾冬红; 等

    1994-01-01

    Scanning tunneling microscope(STM) is used to investigate the optical dise.The areas with and without data stampers are all observedcarefully.Three-dimensional images of the disc surface clearly demonstrate the period.depth of the grooves and the shape of data stampers.Some phenomena of STM imaging are also discussed.

  17. Optical pump-probe microscopy for biomedicine and art conservation

    Science.gov (United States)

    Fischer, Martin

    2013-03-01

    Nonlinear optical microscopy can provide contrast in highly heterogeneous media and a wide range of applications has emerged, primarily in biology, medicine, and materials science. Compared to linear microscopy methods, the localized nature of nonlinear interactions leads to high spatial resolution, optical sectioning, and larger possible imaging depth in scattering media. However, nonlinear contrast (other than fluorescence, harmonic generation or CARS) is generally difficult to measure because it is overwhelmed by the large background of detected illumination light. This background can be suppressed by using femtosecond pulse or pulse train shaping to encode nonlinear interactions in background-free regions of the frequency spectrum. We have developed this shaping technology to study novel intrinsic structural and molecular contrast in biological tissue, generally using less power than a laser pointer. For example we have recently been able to sensitively measure detailed transient absorption dynamics of melanin sub-types in a variety of skin lesions, showing clinically relevant differences of melanin type and distribution between cancerous and benign tissue.[1] Recently we have also applied this technology to paint samples and to historic artwork in order to provide detailed, depth-resolved pigment identification. Initial studies in different inorganic and organic pigments have shown a rich and pigment-specific nonlinear absorption signature.[2] Some pigments, for example lapis lazuli (natural ultramarine), even show marked differences in signal depending on its geographic origin and on age, demonstrating the potential of this technique to determine authenticity, provenance, technology of manufacture, or state of preservation of historic works of art.

  18. Optical-sectioning microscopy by patterned illumination

    Science.gov (United States)

    Saavedra, G.; Martinez-Corral, M.; Sanchez-Ortiga, E.; Doblas, A.

    2010-02-01

    We propose a very simple method for the flexible production of 1D structured illumination for high resolution 3D microscopy. Specifically, we propose the insertion of a Fresnel biprism after a monochromatic point source for producing a pair of twin, fully coherent, virtual point sources. The resulting interference fringes are projected into the 3D sample and, by simply varying the distance between the biprism and the point source, one can tune the period of the fringes, while keeping their contrast, in a very versatile and efficient way.

  19. Image correction in magneto-optical microscopy

    DEFF Research Database (Denmark)

    Paturi, P.; Larsen, B.H.; Jacobsen, B.A.

    2003-01-01

    An image-processing procedure that assures correct determination of the magnetic field distribution of magneto-optical images is presented. The method remedies image faults resulting from sources that are proportional to the incident light intensity, such as different types of defects in the indi......An image-processing procedure that assures correct determination of the magnetic field distribution of magneto-optical images is presented. The method remedies image faults resulting from sources that are proportional to the incident light intensity, such as different types of defects...... in the indicator film and unevenness of light, as well as additive signals from detector bias, external light sources, etc. When properly corrected a better measurement of the local magnetic field can be made, even in the case of heavily damaged films. For superconductors the magnetic field distributions may...

  20. Full-color structured illumination optical sectioning microscopy

    Science.gov (United States)

    Qian, Jia; Lei, Ming; Dan, Dan; Yao, Baoli; Zhou, Xing; Yang, Yanlong; Yan, Shaohui; Min, Junwei; Yu, Xianghua

    2015-09-01

    In merits of super-resolved resolution and fast speed of three-dimensional (3D) optical sectioning capability, structured illumination microscopy (SIM) has found variety of applications in biomedical imaging. So far, most SIM systems use monochrome CCD or CMOS cameras to acquire images and discard the natural color information of the specimens. Although multicolor integration scheme are employed, multiple excitation sources and detectors are required and the spectral information is limited to a few of wavelengths. Here, we report a new method for full-color SIM with a color digital camera. A data processing algorithm based on HSV (Hue, Saturation, and Value) color space is proposed, in which the recorded color raw images are processed in the Hue, Saturation, Value color channels, and then reconstructed to a 3D image with full color. We demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and the surface of coins. The presented technique is applicable to some circumstance where color information plays crucial roles, such as in materials science and surface morphology.

  1. Non-iterative adaptive optical microscopy using wavefront sensing

    Science.gov (United States)

    Tao, X.; Azucena, O.; Kubby, J.

    2016-03-01

    This paper will review the development of wide-field and confocal microscopes with wavefront sensing and adaptive optics for correcting refractive aberrations and compensating scattering when imaging through thick tissues (Drosophila embryos and mouse brain tissue). To make wavefront measurements in biological specimens we have modified the laser guide-star techniques used in astronomy for measuring wavefront aberrations that occur as star light passes through Earth's turbulent atmosphere. Here sodium atoms in Earth's mesosphere, at an altitude of 95 km, are excited to fluoresce at resonance by a high-power sodium laser. The fluorescent light creates a guide-star reference beacon at the top of the atmosphere that can be used for measuring wavefront aberrations that occur as the light passes through the atmosphere. We have developed a related approach for making wavefront measurements in biological specimens using cellular structures labeled with fluorescent proteins as laser guide-stars. An example is a fluorescently labeled centrosome in a fruit fly embryo or neurons and dendrites in mouse brains. Using adaptive optical microscopy we show that the Strehl ratio, the ratio of the peak intensity of an aberrated point source relative to the diffraction limited image, can be improved by an order of magnitude when imaging deeply into live dynamic specimens, enabling near diffraction limited deep tissue imaging.

  2. Optical biomarkers of serous and mucinous human ovarian tumor assessed with nonlinear optics microscopies.

    Directory of Open Access Journals (Sweden)

    Javier Adur

    Full Text Available BACKGROUND: Nonlinear optical (NLO microscopy techniques have potential to improve the early detection of epithelial ovarian cancer. In this study we showed that multimodal NLO microscopies, including two-photon excitation fluorescence (TPEF, second-harmonic generation (SHG, third-harmonic generation (THG and fluorescence lifetime imaging microscopy (FLIM can detect morphological and metabolic changes associated with ovarian cancer progression. METHODOLOGY/PRINCIPAL FINDINGS: We obtained strong TPEF + SHG + THG signals from fixed samples stained with Hematoxylin & Eosin (H&E and robust FLIM signal from fixed unstained samples. Particularly, we imaged 34 ovarian biopsies from different patients (median age, 49 years including 5 normal ovarian tissue, 18 serous tumors and 11 mucinous tumors with the multimodal NLO platform developed in our laboratory. We have been able to distinguish adenomas, borderline, and adenocarcinomas specimens. Using a complete set of scoring methods we found significant differences in the content, distribution and organization of collagen fibrils in the stroma as well as in the morphology and fluorescence lifetime from epithelial ovarian cells. CONCLUSIONS/SIGNIFICANCE: NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for serous and mucinous ovarian tumors. The results provide a basis to interpret future NLO images of ovarian tissue and lay the foundation for future in vivo optical evaluation of premature ovarian lesions.

  3. Integrated optical coherence tomography and optical coherence microscopy imaging of human pathology

    Science.gov (United States)

    Lee, Hsiang-Chieh; Zhou, Chao; Wang, Yihong; Aquirre, Aaron D.; Tsai, Tsung-Han; Cohen, David W.; Connolly, James L.; Fujimoto, James G.

    2010-02-01

    Excisional biopsy is the current gold standard for disease diagnosis; however, it requires a relatively long processing time and it may also suffer from unacceptable false negative rates due to sampling errors. Optical coherence tomography (OCT) is a promising imaging technique that provide real-time, high resolution and three-dimensional (3D) images of tissue morphology. Optical coherence microscopy (OCM) is an extension of OCT, combining both the coherence gating and the confocal gating techniques. OCM imaging achieves cellular resolution with deeper imaging depth compared to confocal microscopy. An integrated OCT/OCM imaging system can provide co-registered multiscale imaging of tissue morphology. 3D-OCT provides architectural information with a large field of view and can be used to find regions of interest; while OCM provides high magnification to enable cellular imaging. The integrated OCT/OCM system has an axial resolution of kidney (19), were imaged with OCT and OCM within 2 to 6 hours after excision. The images were compared with H & E histology to identify characteristic features useful for disease diagnosis. The feasibility of visualizing human pathology using integrated OCT/OCM was demonstrated in the pathology laboratory settings.

  4. Recent Advances in Biological Single-Molecule Applications of Optical Tweezers and Fluorescence Microscopy.

    Science.gov (United States)

    Hashemi Shabestari, M; Meijering, A E C; Roos, W H; Wuite, G J L; Peterman, E J G

    2017-01-01

    Over the past two decades, single-molecule techniques have evolved into robust tools to study many fundamental biological processes. The combination of optical tweezers with fluorescence microscopy and microfluidics provides a powerful single-molecule manipulation and visualization technique that has found widespread application in biology. In this combined approach, the spatial (~nm) and temporal (~ms) resolution, as well as the force scale (~pN) accessible to optical tweezers is complemented with the power of fluorescence microscopy. Thereby, it provides information on the local presence, identity, spatial dynamics, and conformational dynamics of single biomolecules. Together, these techniques allow comprehensive studies of, among others, molecular motors, protein-protein and protein-DNA interactions, biomolecular conformational changes, and mechanotransduction pathways. In this chapter, recent applications of fluorescence microscopy in combination with optical trapping are discussed. After an introductory section, we provide a description of instrumentation together with the current capabilities and limitations of the approaches. Next we summarize recent studies that applied this combination of techniques in biological systems and highlight some representative biological assays to mark the exquisite opportunities that optical tweezers combined with fluorescence microscopy provide. © 2017 Elsevier Inc. All rights reserved.

  5. Optical far-field super-resolution microscopy using nitrogen vacancy center ensemble in bulk diamond

    Science.gov (United States)

    Li, Shen; Chen, Xiang-dong; Zhao, Bo-Wen; Dong, Yang; Zou, Chong-Wen; Guo, Guang-Can; Sun, Fang-Wen

    2016-09-01

    We demonstrate optical far-field super-resolution microscopy using an array of nitrogen vacancy centers in bulk diamond as near-field optical probes. The local optical field, which transmits through the nanostructures on the diamond surface, is measured by detecting the charge state conversion of the nitrogen vacancy center. Locating the nitrogen vacancy center with a spatial resolution of 6.1 nm is realized with charge state depletion nanoscopy. The nanostructures on the surface of a diamond are then imaged with a resolution below the optical diffraction limit. The results offer an approach to build a general-purpose optical super-resolution microscopy technique and a convenient platform for high spatial resolution quantum sensing with nitrogen vacancy centers.

  6. Optical and digital microscopic imaging techniques and applications in pathology.

    Science.gov (United States)

    Chen, Xiaodong; Zheng, Bin; Liu, Hong

    2011-01-01

    The conventional optical microscope has been the primary tool in assisting pathological examinations. The modern digital pathology combines the power of microscopy, electronic detection, and computerized analysis. It enables cellular-, molecular-, and genetic-imaging at high efficiency and accuracy to facilitate clinical screening and diagnosis. This paper first reviews the fundamental concepts of microscopic imaging and introduces the technical features and associated clinical applications of optical microscopes, electron microscopes, scanning tunnel microscopes, and fluorescence microscopes. The interface of microscopy with digital image acquisition methods is discussed. The recent developments and future perspectives of contemporary microscopic imaging techniques such as three-dimensional and in vivo imaging are analyzed for their clinical potentials.

  7. Super-resolution optical microscopy based on scannable cantilever-combined microsphere.

    Science.gov (United States)

    Wang, Shuying; Zhang, Dongxian; Zhang, Haijun; Han, Xu; Xu, Rui

    2015-12-01

    We report an ingenious method of super-resolution optical microscopy utilizing scannable cantilever-combined microsphere. By scanning the microsphere over the sample surface in a cantilever-combined microsphere-sample contact state, super-resolution images can be acquired at arbitrary sample regions through near-field information collection by the microsphere. In addition, such a state can effectively reduce the possibility of breaking the cantilever and damaging the microsphere or sample surface. This work has developed a new method and technique of sub-diffraction-limit optical microscopy, and can be practically applied in various fields of micro/nanoscopy.

  8. Perfect optical vortex enhanced surface plasmon excitation for plasmonic structured illumination microscopy imaging

    Science.gov (United States)

    Zhang, Chonglei; Min, Changjun; Du, Luping; Yuan, X.-C.

    2016-05-01

    We propose an all-optical technique for plasmonic structured illumination microscopy (PSIM) with perfect optical vortex (POV). POV can improve the efficiency of the excitation of surface plasma and reduce the background noise of the excited fluorescence. The plasmonic standing wave patterns are excited by POV with fractional topological charges for accurate phase shift of {-2π/3, 0, and 2π/3}. The imaging resolution of less than 200 nm was produced. This PSIM technique is expected to be used as a wide field, super resolution imaging technique in dynamic biological imaging.

  9. Cognitive optical networks: architectures and techniques

    Science.gov (United States)

    Grebeshkov, Alexander Y.

    2017-04-01

    This article analyzes architectures and techniques of the optical networks with taking into account the cognitive methodology based on continuous cycle "Observe-Orient-Plan-Decide-Act-Learn" and the ability of the cognitive systems adjust itself through an adaptive process by responding to new changes in the environment. Cognitive optical network architecture includes cognitive control layer with knowledge base for control of software-configurable devices as reconfigurable optical add-drop multiplexers, flexible optical transceivers, software-defined receivers. Some techniques for cognitive optical networks as flexible-grid technology, broker-oriented technique, machine learning are examined. Software defined optical network and integration of wireless and optical networks with radio over fiber technique and fiber-wireless technique in the context of cognitive technologies are discussed.

  10. Quantitative optical microscopy and micromanipulation studies on the lipid bilayer membranes of giant unilamellar vesicles

    DEFF Research Database (Denmark)

    Bagatolli, Luis; Needham, David

    2014-01-01

    some of their most important contributions to our understanding of lipid bilayer membranes; and (iii) outline studies that would utilize both techniques simultaneously on the same vesicle thus bringing the ability to characterize structure and strain responses together with the direct application......This manuscript discusses basic methodological aspects of optical microscopy and micromanipulation methods to study membranes and reviews methods to generate giant unilamellar vesicles (GUVs). In particular, we focus on the use of fluorescence microscopy and micropipette manipulation techniques...... to study composition-structure-property materials relationships of free-standing lipid bilayer membranes. Because their size (~5 to 100 m diameter) that is well above the resolution limit of regular light microscopes, GUVs are suitable membrane models for optical microscopy and micromanipulation...

  11. Hybrid microscopy of human carotid atheroma by means of optical-resolution optoacoustic and non-linear optical microscopy

    Science.gov (United States)

    Seeger, Markus; Karlas, Angelos; Soliman, Dominik; Pelisek, Jaroslav; Ntziachristos, Vasilis

    2017-03-01

    Carotid atheromatosis is causally related to stroke, a leading cause of disability and death. We present the analysis of a human carotid atheroma using a novel hybrid microscopy system that combines optical-resolution optoacoustic (photoacoustic) microscopy and several non-linear optical microscopy modalities (second and third harmonic generation, as well as, two-photon excitation fluorescence) to achieve a multimodal examination of the extracted tissue within the same imaging framework. Our system enables the label-free investigation of atheromatous human carotid tissue with a resolution of about 1 μm and allows for the congruent interrogation of plaque morphology and clinically relevant constituents such as red blood cells, collagen, and elastin. Our data reveal mutual interactions between blood embeddings and connective tissue within the atheroma, offering comprehensive insights into its stage of evolution and severity, and potentially facilitating the further development of diagnostic tools, as well as treatment strategies.

  12. In vivo switchable optical- and acoustic-resolution photoacoustic microscopy

    Science.gov (United States)

    Jeon, Seungwan; Kim, Jaewoo; Kim, Chulhong

    2016-03-01

    Photoacoustic microscopy (PAM) provides high resolution and large penetration depth by utilizing the high optical sensitivity and low scattering of ultrasound. Hybrid PAM systems can be classified into two categories: opticalresolution photoacoustic microscopy (OR-PAM) and acoustic-resolution photoacoustic microscopy (AR-PAM). ORPAM provides a very high lateral resolution with a strong optical focus, but the penetration depth is limited to one optical transport mean free path. AR-PAM provides a relatively greater penetration depth using diffused light in biological tissues. The resolution of AR-PAM is determined by its ultrasonic parameters. In this study, we performed an in vivo testing of a switchable OR-/AR-PAM system. In this system, two modes can be switched by changing its collimator lens and optical fiber. The lateral resolution of OR-PAM was measured using a resolution test target, and the full width at half maximum (FWHM) of the edge spread function was 2.5 μm. To calculate the lateral resolution of ARPAM, a 6-μm-diameter carbon fiber was used, and the FWHM of the line spread function was 80.2 μm. We successfully demonstrated the multiscale imaging capability of the switchable OR-/AR-PAM system by visualizing microvascular networks in mouse ears, brain, legs, skin, and eyes.

  13. Vibrational and optical spectroscopies integrated with environmental transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Picher, Matthieu; Mazzucco, Stefano [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899-6203 (United States); Institute for Research in Electronics and Applied Physics, University of Maryland, College Park, MD 20740 (United States); Blankenship, Steve [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899-6203 (United States); Sharma, Renu, E-mail: renu.sharma@nist.gov [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899-6203 (United States)

    2015-03-15

    Here, we present a measurement platform for collecting multiple types of spectroscopy data during high-resolution environmental transmission electron microscopy observations of dynamic processes. Such coupled measurements are made possible by a broadband, high-efficiency, free-space optical system. The critical element of the system is a parabolic mirror, inserted using an independent hollow rod and placed below the sample holder which can focus a light on the sample and/or collect the optical response. We demonstrate the versatility of this optical setup by using it to combine in situ atomic-scale electron microscopy observations with Raman spectroscopy. The Raman data is also used to measure the local temperature of the observed sample area. Other applications include, but are not limited to: cathodo- and photoluminescence spectroscopy, and use of the laser as a local, high-rate heating source. - Highlights: • Broadband, high-efficiency design adaptable to other electron microscopes. • Raman spectroscopy integrated with environmental transmission electron microscopy. • Raman spectra peak frequency shifts enable measurement of local sample temperature. • Multiple types of optical spectroscopy enabled, e.g. cathodoluminescence.

  14. Optical near-field microscopy of light focusing through a photonic crystal flat lens.

    Science.gov (United States)

    Fabre, Nathalie; Lalouat, Loïc; Cluzel, Benoit; Mélique, Xavier; Lippens, Didier; de Fornel, Frédérique; Vanbésien, Olivier

    2008-08-15

    We report here the direct observation by using a scanning near-field microscopy technique of the light focusing through a photonic crystal flat lens designed and fabricated to operate at optical frequencies. The lens is fabricated using a III-V semiconductor slab, and we directly visualize the propagation of the electromagnetic waves by using a scanning near-field optical microscope. We directly evidence spatially, as well as spectrally, the focusing operating regime of the lens. At last, in light of the experimental scanning near-field optical microscope pictures, we discuss the lens ability to focus light at a subwavelength scale.

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

    Science.gov (United States)

    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.

  16. Colour metallography in commercial Al-Si alloys. Optimization of the microstructural characterization techniques in light optical microscopy; Metalografia a color en aleaciones Al-Si comerciales. Optimizacion de las tecnicas de caracterizacion microestructural mediante microscopia optica de reflexion

    Energy Technology Data Exchange (ETDEWEB)

    Suarez-Pena, B.; Asensio-Lozano, J.; Vander-Voort, G.F.

    2010-07-01

    The present demand on alloy production with improved quality requires the optimization of the metallographic procedures used on its characterization. Traditional etching techniques ommonly employed for phase identification by optical metallography in aluminium alloys are not always suitable for a detailed analysis of existing phases, nor to accurately predict the mechanisms that govern the solidification process in certain detail. This work explores the potential of colour metallography to reveal at its best as-cast microstructures in Al-Si 12 alloys. For this purpose a colour etching technique, specifically developed for aluminium alloys and based on theWeck reagent[1]. The application of such etchant has allowed the qualitative characterization of the microstructure. And it has also shown the advantages of colour metallography over black and white (B&W) etching techniques. (Author).

  17. Quantitative photoacoustic microscopy of optical absorption coefficients from acoustic spectra in the optical diffusive regime.

    Science.gov (United States)

    Guo, Zijian; Favazza, Christopher; Garcia-Uribe, Alejandro; Wang, Lihong V

    2012-06-01

    Photoacoustic (PA) microscopy (PAM) can image optical absorption contrast with ultrasonic spatial resolution in the optical diffusive regime. Conventionally, accurate quantification in PAM requires knowledge of the optical fluence attenuation, acoustic pressure attenuation, and detection bandwidth. We circumvent this requirement by quantifying the optical absorption coefficients from the acoustic spectra of PA signals acquired at multiple optical wavelengths. With the acoustic spectral method, the absorption coefficients of an oxygenated bovine blood phantom at 560, 565, 570, and 575 nm were quantified with errors of acoustic spectral method provides greater quantification accuracy in the optical diffusive regime. The limitations of the acoustic spectral method was also discussed.

  18. All-optically integrated multimodality imaging system: combined photoacoustic microscopy, optical coherence tomography, and fluorescence imaging

    Science.gov (United States)

    Chen, Zhongjiang; Yang, Sihua; Xing, Da

    2016-10-01

    We have developed a multimodality imaging system by optically integrating all-optical photoacoustic microscopy (AOPAM), optical coherence tomography (OCT) and fluorescence microscopy (FLM) to provide complementary information including optical absorption, optical back-scattering and fluorescence contrast of biological tissue. By sharing the same low-coherence Michelson interferometer, AOPAM and OCT could be organically optically combined to obtain the absorption and scattering information of the biological tissues. Also, owing to using the same laser source and objective lens, intrinsically registered photoacoustic and fluorescence signals are obtained to present the radiative and nonradiative transition process of absorption. Simultaneously photoacoustic angiography, tissue structure and fluorescence molecular in vivo images of mouse ear were acquired to demonstrate the capabilities of the optically integrated trimodality imaging system, which can present more information to study tumor angiogenesis, vasculature, anatomical structure and microenvironments in vivo.

  19. Advanced 3D Optical Microscopy in ENS Research.

    Science.gov (United States)

    Vanden Berghe, Pieter

    2016-01-01

    Microscopic techniques are among the few approaches that have survived the test of time. Being invented half way the seventeenth century by Antonie van Leeuwenhoek and Robert Hooke, this technology is still essential in modern biomedical labs. Many microscopy techniques have been used in ENS research to guide researchers in their dissections and later to enable electrode recordings. Apart from this, microscopy has been instrumental in the identification of subpopulations of cells in the ENS, using a variety of staining methods. A significant step forward in the use of microscopy was the introduction of fluorescence approaches. Due to the fact that intense excitation light is now filtered away from the longer wavelength emission light, the contrast can be improved drastically, which helped to identify subpopulations of enteric neurons in a variety of species. Later functionalized fluorescent probes were used to measure and film activity in muscle and neuronal cells. Another important impetus to the use of microscopy was the discovery and isolation of the green fluorescent protein (GFP), as it gave rise to the development of many different color variants and functionalized constructs. Recent advances in microscopy are the result of a continuous search to enhance contrast between the item of interest and its background but also to improve resolving power to tell two small objects apart. In this chapter three different microscopy approaches will be discussed that can aid to improve our understanding of ENS function within the gut wall.

  20. Resolution enhancement techniques in optical lithography

    CERN Document Server

    Wong, Alfred K

    2001-01-01

    Ever-smaller IC devices are pushing the optical lithography envelope, increasing the importance of resolution enhancement techniques. This tutorial encompasses two decades of research. It discusses theoretical and practical aspects of commonly used techniques, including optical imaging and resolution, modified illumination, optical proximity correction, alternating and attenuating phase-shifting masks, selecting RETs, and second-generation RETs. Useful for students and practicing lithographers.

  1. Transfer functions in collection scanning near-field optical microscopy

    DEFF Research Database (Denmark)

    Bozhevolnyi, Sergey I.; Vohnsen, Brian; Bozhevolnaya, Elena A.

    1999-01-01

    It is generally accepted that, if in collection near-field optical microscopy the probe-sample coupling can be disregarded, a fiber probe can be considered as a detector of the near-field intensity whose size can be accounted for via an intensity transfer function. We show that, in general...... are considered with respect to the relation between near-field optical images and the corresponding intensity distributions. Our conclusions are supported with numerical simulations and experimental results obtained by using a photon scanning tunneling microscope with an uncoated fiber tip....

  2. Fluorescent Nanodiamond-Gold Hybrid Particles for Multimodal Optical and Electron Microscopy Cellular Imaging.

    Science.gov (United States)

    Liu, Weina; Naydenov, Boris; Chakrabortty, Sabyasachi; Wuensch, Bettina; Hübner, Kristina; Ritz, Sandra; Cölfen, Helmut; Barth, Holger; Koynov, Kaloian; Qi, Haoyuan; Leiter, Robert; Reuter, Rolf; Wrachtrup, Jörg; Boldt, Felix; Scheuer, Jonas; Kaiser, Ute; Sison, Miguel; Lasser, Theo; Tinnefeld, Philip; Jelezko, Fedor; Walther, Paul; Wu, Yuzhou; Weil, Tanja

    2016-10-12

    There is a continuous demand for imaging probes offering excellent performance in various microscopy techniques for comprehensive investigations of cellular processes by more than one technique. Fluorescent nanodiamond-gold nanoparticles (FND-Au) constitute a new class of "all-in-one" hybrid particles providing unique features for multimodal cellular imaging including optical imaging, electron microscopy, and, and potentially even quantum sensing. Confocal and optical coherence microscopy of the FND-Au allow fast investigations inside living cells via emission, scattering, and photothermal imaging techniques because the FND emission is not quenched by AuNPs. In electron microscopy, transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) analysis of FND-Au reveals greatly enhanced contrast due to the gold particles as well as an extraordinary flickering behavior in three-dimensional cellular environments originating from the nanodiamonds. The unique multimodal imaging characteristics of FND-Au enable detailed studies inside cells ranging from statistical distributions at the entire cellular level (micrometers) down to the tracking of individual particles in subcellular organelles (nanometers). Herein, the processes of endosomal membrane uptake and release of FNDs were elucidated for the first time by the imaging of individual FND-Au hybrid nanoparticles with single-particle resolution. Their convenient preparation, the availability of various surface groups, their flexible detection modalities, and their single-particle contrast in combination with the capability for endosomal penetration and low cytotoxicity make FND-Au unique candidates for multimodal optical-electronic imaging applications with great potential for emerging techniques, such as quantum sensing inside living cells.

  3. Cytology 3D structure formation based on optical microscopy images

    Science.gov (United States)

    Pronichev, A. N.; Polyakov, E. V.; Shabalova, I. P.; Djangirova, T. V.; Zaitsev, S. M.

    2017-01-01

    The article the article is devoted to optimization of the parameters of imaging of biological preparations in optical microscopy using a multispectral camera in visible range of electromagnetic radiation. A model for the image forming of virtual preparations was proposed. The optimum number of layers was determined for the object scan in depth and holistic perception of its switching according to the results of the experiment.

  4. Optical coherence tomography: Technique and applications

    DEFF Research Database (Denmark)

    Thomsen, Jakob Borup; Sander, Birgit; Mogensen, Mette;

    2009-01-01

    Optical coherence tomography (OCT) is a noninvasive optical imaging modality providing real-time video rate images in two and three dimensions of biological tissues with micrometer resolution. OCT fills the gap between ultrasound and confocal microscopy, since it has a higher resolution than ultr...... of retinal diseases. The potential of OCT in many other applications is currently being explored, such as in developmental biology, skin cancer diagnostics, vulnerable plaque detection in cardiology, esophageal diagnostics and a number of other applications within oncology....

  5. Super-resolution optical microscopy of lipid plasma membrane dynamics.

    Science.gov (United States)

    Eggeling, Christian

    2015-01-01

    Plasma membrane dynamics are an important ruler of cellular activity, particularly through the interaction and diffusion dynamics of membrane-embedded proteins and lipids. FCS (fluorescence correlation spectroscopy) on an optical (confocal) microscope is a popular tool for investigating such dynamics. Unfortunately, its full applicability is constrained by the limited spatial resolution of a conventional optical microscope. The present chapter depicts the combination of optical super-resolution STED (stimulated emission depletion) microscopy with FCS, and why it is an important tool for investigating molecular membrane dynamics in living cells. Compared with conventional FCS, the STED-FCS approach demonstrates an improved possibility to distinguish free from anomalous molecular diffusion, and thus to give new insights into lipid-protein interactions and the traditional lipid 'raft' theory.

  6. The OPFOS microscopy family: High-resolution optical-sectioning of biomedical specimens

    CERN Document Server

    Buytaert, Jan A N; Adriaens, Dominique; Dirckx, Joris J J

    2011-01-01

    We report on the recently emerging (Laser) Light Sheet based Fluorescence Microscopy field (LSFM). The techniques used in this field allow to study and visualize biomedical objects non-destructively in high-resolution through virtual optical sectioning with sheets of laser light. Fluorescence originating in the cross section of the sheet and sample is recorded orthogonally with a camera. In this paper, the first implementation of LSFM to image biomedical tissue in three dimensions - Orthogonal-Plane Fluorescence Optical Sectioning microscopy (OPFOS) - is discussed. Since then many similar and derived methods have surfaced (SPIM, Ultramicroscopy, HR-OPFOS, mSPIM, DSLM, TSLIM...) which we all briefly discuss. All these optical sectioning methods create images showing histological detail. We illustrate the applicability of LSFM on several specimen types with application in biomedical and life sciences.

  7. Fast benchtop fabrication of laminar flow chambers for advanced microscopy techniques.

    Directory of Open Access Journals (Sweden)

    David S Courson

    Full Text Available BACKGROUND: Fluid handling technology is acquiring an ever more prominent place in laboratory science whether it is in simple buffer exchange systems, perfusion chambers, or advanced microfluidic devices. Many of these applications remain the providence of laboratories at large institutions with a great deal of expertise and specialized equipment. Even with the expansion of these techniques, limitations remain that frequently prevent the coupling of controlled fluid flow with other technologies, such as coupling microfluidics and high-resolution position and force measurements by optical trapping microscopy. METHOD: Here we present a method for fabrication of multiple-input laminar flow devices that are optically clear [glass] on each face, chemically inert, reusable, inexpensive, and can be fabricated on the benchtop in approximately one hour. Further these devices are designed to allow flow regulation by a simple gravity method thus requiring no specialized equipment to drive flow. Here we use these devices to perform total internal reflection fluorescence microscopy measurements as well as position sensitive optical trapping experiments. SIGNIFICANCE: Flow chamber technology needs to be more accessible to the general scientific community. The method presented here is versatile and robust. These devices use standard slides and coverslips making them compatible with nearly all types and models of light microscopes. These devices meet the needs of groups doing advanced optical trapping experiments, but could also be adapted by nearly any lab that has a function for solution flow coupled with microscopy.

  8. GPU-based computational adaptive optics for volumetric optical coherence microscopy

    Science.gov (United States)

    Tang, Han; Mulligan, Jeffrey A.; Untracht, Gavrielle R.; Zhang, Xihao; Adie, Steven G.

    2016-03-01

    Optical coherence tomography (OCT) is a non-invasive imaging technique that measures reflectance from within biological tissues. Current higher-NA optical coherence microscopy (OCM) technologies with near cellular resolution have limitations on volumetric imaging capabilities due to the trade-offs between resolution vs. depth-of-field and sensitivity to aberrations. Such trade-offs can be addressed using computational adaptive optics (CAO), which corrects aberration computationally for all depths based on the complex optical field measured by OCT. However, due to the large size of datasets plus the computational complexity of CAO and OCT algorithms, it is a challenge to achieve high-resolution 3D-OCM reconstructions at speeds suitable for clinical and research OCM imaging. In recent years, real-time OCT reconstruction incorporating both dispersion and defocus correction has been achieved through parallel computing on graphics processing units (GPUs). We add to these methods by implementing depth-dependent aberration correction for volumetric OCM using plane-by-plane phase deconvolution. Following both defocus and aberration correction, our reconstruction algorithm achieved depth-independent transverse resolution of 2.8 um, equal to the diffraction-limited focal plane resolution. We have translated the CAO algorithm to a CUDA code implementation and tested the speed of the software in real-time using two GPUs - NVIDIA Quadro K600 and Geforce TITAN Z. For a data volume containing 4096×256×256 voxels, our system's processing speed can keep up with the 60 kHz acquisition rate of the line-scan camera, and takes 1.09 seconds to simultaneously update the CAO correction for 3 en face planes at user-selectable depths.

  9. DMD-based LED-illumination super-resolution and optical sectioning microscopy.

    Science.gov (United States)

    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.

  10. DMD-based LED-illumination Super-resolution and optical sectioning microscopy

    Science.gov (United States)

    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.

  11. Scanning second-harmonic optical microscopy of self-assembled InAlGaAs quantum dots

    DEFF Research Database (Denmark)

    Vohnsen, B.; Bozhevolnyi, S. I.; Pedersen, K.

    2001-01-01

    or in the illumination itself. Thus, a combination of scanning microscopy with SH detection may be a highly suitable candidate to reveal the presence of QD's embedded in an otherwise isotropic material. We have used scanning far-field (SFOM) and scanning near field optical microscopy (SNOM) techniques to locally probe......Microscopy provides a suitable technique for local probing of small ensembles of (or even individual) QD's, and when combined with the detection of second-harmonic (SH) generation the technique becomes suitable to reveal tiny changes of symmetry originating either in the material structures...

  12. All-optical signal processing technique for secure optical communication

    Science.gov (United States)

    Qian, Feng-chen; Su, Bing; Ye, Ya-lin; Zhang, Qian; Lin, Shao-feng; Duan, Tao; Duan, Jie

    2015-10-01

    Secure optical communication technologies are important means to solve the physical layer security for optical network. We present a scheme of secure optical communication system by all-optical signal processing technique. The scheme consists of three parts, as all-optical signal processing unit, optical key sequence generator, and synchronous control unit. In the paper, all-optical signal processing method is key technology using all-optical exclusive disjunction (XOR) gate based on optical cross-gain modulation effect, has advantages of wide dynamic range of input optical signal, simple structure and so on. All-optical XOR gate composed of two semiconductor optical amplifiers (SOA) is a symmetrical structure. By controlling injection current, input signal power, delay and filter bandwidth, the extinction ratio of XOR can be greater than 8dB. Finally, some performance parameters are calculated and the results are analyzed. The simulation and experimental results show that the proposed method can be achieved over 10Gbps optical signal encryption and decryption, which is simple, easy to implement, and error-free diffusion.

  13. An integrated instrumental setup for the combination of atomic force microscopy with optical spectroscopy.

    Science.gov (United States)

    Owen, R J; Heyes, C D; Knebel, D; Röcker, C; Nienhaus, G U

    2006-07-01

    In recent years, the study of single biomolecules using fluorescence microscopy and atomic force microscopy (AFM) techniques has resulted in a plethora of new information regarding the physics underlying these complex biological systems. It is especially advantageous to be able to measure the optical, topographical, and mechanical properties of single molecules simultaneously. Here an AFM is used that is especially designed for integration with an inverted optical microscope and that has a near-infrared light source (850 nm) to eliminate interference between the optical experiment and the AFM operation. The Tip Assisted Optics (TAO) system consists of an additional 100 x 100-microm(2) X-Y scanner for the sample, which can be independently and simultaneously used with the AFM scanner. This allows the offset to be removed between the confocal optical image obtained with the sample scanner and the simultaneously acquired AFM topography image. The tip can be positioned exactly into the optical focus while the user can still navigate within the AFM image for imaging or manipulation of the sample. Thus the tip-enhancement effect can be maximized and it becomes possible to perform single molecule manipulation experiments within the focus of a confocal optical image. Here this is applied to simultaneous measurement of single quantum dot fluorescence and topography with high spatial resolution.

  14. Surface plasmon resonance microscopy: Achieving a quantitative optical response

    Science.gov (United States)

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

    2016-09-01

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

  15. Electron microscopy of primary cell cultures in solution and correlative optical microscopy using ASEM

    Energy Technology Data Exchange (ETDEWEB)

    Hirano, Kazumi; Kinoshita, Takaaki [Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577 (Japan); Uemura, Takeshi [Department of Molecular Neurobiology and Pharmacology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Department of Molecular and Cellular Physiology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621 (Japan); Motohashi, Hozumi [Department of Gene Expression Regulation, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryo-cho, Aoba-ku, Sendai 980-8575 (Japan); Watanabe, Yohei; Ebihara, Tatsuhiko [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Nishiyama, Hidetoshi [JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558 (Japan); Sato, Mari [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Suga, Mitsuo [JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558 (Japan); Maruyama, Yuusuke; Tsuji, Noriko M. [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Yamamoto, Masayuki [Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai 980-8575 (Japan); Nishihara, Shoko, E-mail: shoko@soka.ac.jp [Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577 (Japan); Sato, Chikara, E-mail: ti-sato@aist.go.jp [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan)

    2014-08-01

    Correlative light-electron microscopy of cells in a natural environment of aqueous liquid facilitates high-throughput observation of protein complex formation. ASEM allows the inverted SEM to observe the wet sample from below, while an optical microscope observes it from above quasi-simultaneously. The disposable ASEM dish with a silicon nitride (SiN) film window can be coated variously to realize the primary-culture of substrate-sensitive cells in a few milliliters of culture medium in a stable incubator environment. Neuron differentiation, neural networking, proplatelet-formation and phagocytosis were captured by optical or fluorescence microscopy, and imaged at high resolution by gold-labeled immuno-ASEM with/without metal staining. Fas expression on the cell surface was visualized, correlated to the spatial distribution of F-actin. Axonal partitioning was studied using primary-culture neurons, and presynaptic induction by GluRδ2-N-terminus-linked fluorescent magnetic beads was correlated to the presynaptic-marker Bassoon. Further, megakaryocytes secreting proplatelets were captured, and P-selectins with adherence activity were localized to some of the granules present by immuno-ASEM. The phagocytosis of lactic acid bacteria by dendritic cells was also imaged. Based on these studies, ASEM correlative microscopy promises to allow the study of various mesoscopic-scale dynamics in the near future. - Highlights: • In situ correlative light electron microscopy of samples in open solution by ASEM. • Primary cultures for in-solution CLEM by developing SiN-film coating methods • First visualization of fluorescent magnetic beads in aqueous solution by CLEM. • Presynaptic induction of neurons by GluRδ2-N-terminus-coated beads studied by CLEM. • Axonal partitioning, bacterial phagocytosis, platelet formation imaged by CLEM.

  16. Adaptive optics in spinning disk microscopy: improved contrast and brightness by a simple and fast method.

    Science.gov (United States)

    Fraisier, V; Clouvel, G; Jasaitis, A; Dimitrov, A; Piolot, T; Salamero, J

    2015-09-01

    Multiconfocal microscopy gives a good compromise between fast imaging and reasonable resolution. However, the low intensity of live fluorescent emitters is a major limitation to this technique. Aberrations induced by the optical setup, especially the mismatch of the refractive index and the biological sample itself, distort the point spread function and further reduce the amount of detected photons. Altogether, this leads to impaired image quality, preventing accurate analysis of molecular processes in biological samples and imaging deep in the sample. The amount of detected fluorescence can be improved with adaptive optics. Here, we used a compact adaptive optics module (adaptive optics box for sectioning optical microscopy), which was specifically designed for spinning disk confocal microscopy. The module overcomes undesired anomalies by correcting for most of the aberrations in confocal imaging. Existing aberration detection methods require prior illumination, which bleaches the sample. To avoid multiple exposures of the sample, we established an experimental model describing the depth dependence of major aberrations. This model allows us to correct for those aberrations when performing a z-stack, gradually increasing the amplitude of the correction with depth. It does not require illumination of the sample for aberration detection, thus minimizing photobleaching and phototoxicity. With this model, we improved both signal-to-background ratio and image contrast. Here, we present comparative studies on a variety of biological samples.

  17. Optical metrology techniques for dimensional stability measurements

    NARCIS (Netherlands)

    Ellis, Jonathan David

    2010-01-01

    This thesis work is optical metrology techniques to determine material stability. In addition to displacement interferometry, topics such as periodic nonlinearity, Fabry-Perot interferometry, refractometry, and laser stabilization are covered.

  18. Optical metrology techniques for dimensional stability measurements

    NARCIS (Netherlands)

    Ellis, Jonathan David

    2010-01-01

    This thesis work is optical metrology techniques to determine material stability. In addition to displacement interferometry, topics such as periodic nonlinearity, Fabry-Perot interferometry, refractometry, and laser stabilization are covered.

  19. Quantitative interferometric microscopy cytometer based on regularized optical flow algorithm

    Science.gov (United States)

    Xue, Liang; Vargas, Javier; Wang, Shouyu; Li, Zhenhua; Liu, Fei

    2015-09-01

    Cell detections and analysis are important in various fields, such as medical observations and disease diagnoses. In order to analyze the cell parameters as well as observe the samples directly, in this paper, we present an improved quantitative interferometric microscopy cytometer, which can monitor the quantitative phase distributions of bio-samples and realize cellular parameter statistics. The proposed system is able to recover the phase imaging of biological samples in the expanded field of view via a regularized optical flow demodulation algorithm. This algorithm reconstructs the phase distribution with high accuracy with only two interferograms acquired at different time points simplifying the scanning system. Additionally, the method is totally automatic, and therefore it is convenient for establishing a quantitative phase cytometer. Moreover, the phase retrieval approach is robust against noise and background. Excitingly, red blood cells are readily investigated with the quantitative interferometric microscopy cytometer system.

  20. Nonlinear Optical Microscopy Signal Processing Strategies in Cancer

    Science.gov (United States)

    Adur, Javier; Carvalho, Hernandes F; Cesar, Carlos L; Casco, Víctor H

    2014-01-01

    This work reviews the most relevant present-day processing methods used to improve the accuracy of multimodal nonlinear images in the detection of epithelial cancer and the supporting stroma. Special emphasis has been placed on methods of non linear optical (NLO) microscopy image processing such as: second harmonic to autofluorescence ageing index of dermis (SAAID), tumor-associated collagen signatures (TACS), fast Fourier transform (FFT) analysis, and gray level co-occurrence matrix (GLCM)-based methods. These strategies are presented as a set of potential valuable diagnostic tools for early cancer detection. It may be proposed that the combination of NLO microscopy and informatics based image analysis approaches described in this review (all carried out on free software) may represent a powerful tool to investigate collagen organization and remodeling of extracellular matrix in carcinogenesis processes. PMID:24737930

  1. Assessment of fibrotic liver disease with multimodal nonlinear optical microscopy

    Science.gov (United States)

    Lu, Fake; Zheng, Wei; Tai, Dean C. S.; Lin, Jian; Yu, Hanry; Huang, Zhiwei

    2010-02-01

    Liver fibrosis is the excessive accumulation of extracellular matrix proteins such as collagens, which may result in cirrhosis, liver failure, and portal hypertension. In this study, we apply a multimodal nonlinear optical microscopy platform developed to investigate the fibrotic liver diseases in rat models established by performing bile duct ligation (BDL) surgery. The three nonlinear microscopy imaging modalities are implemented on the same sectioned tissues of diseased model sequentially: i.e., second harmonic generation (SHG) imaging quantifies the contents of the collagens, the two-photon excitation fluorescence (TPEF) imaging reveals the morphology of hepatic cells, while coherent anti-Stokes Raman scattering (CARS) imaging maps the distributions of fats or lipids quantitatively across the tissue. Our imaging results show that during the development of liver fibrosis (collagens) in BDL model, fatty liver disease also occurs. The aggregated concentrations of collagen and fat constituents in liver fibrosis model show a certain correlationship between each other.

  2. Quantitative optical coherence microscopy for the in situ investigation of the biofilm

    Science.gov (United States)

    Meleppat, Ratheesh Kumar; Shearwood, Christopher; Keey, Seah Leong; Matham, Murukeshan Vadakke

    2016-12-01

    This paper explores the potential of optical coherence microscopy (OCM) for the in situ monitoring of biofilm growth. The quantitative imaging of the early developmental biology of a representative biofilm, Klebsiella pneumonia (KP-1), was performed using a swept source-based Fourier domain OCM system. The growth dynamics of the KP-1 biofilms and their transient response under perturbation was investigated using the enface visualization of microcolonies and their spatial localization. Furthermore, the optical density (OD) and planar density of the biofilms are calculated using an OCM technique and compared with OD and colony forming units measured using standard procedures via the sampling of the flow-cell effluent.

  3. Optical mapping of a rice B AC clone using restriction endonuclease and imaging with fluorescent microscopy at single molecule level

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A method of constructing restriction map by optical mapping and single molecule fluorescent microscopy is described. DNA molecules were aligned and adsorbed on a glass coverslip surface by a mbdified "molecular combing"technique, and then the surface-immobilized DNAs were cleaved in situ with a restriction endonuclease. Individual DNA molecules digested by the endonuclease EcoR I were observable with fluorescent microscopy. Using optical mapping, a physical map of a rice bacterial artificial chromosome clone was constructed. This method will facilitate genomic mapping and tracing the dynamic process in real time at a single molecule level with fluorescence microscopy.

  4. High speed sub-micrometric microscopy using optical polymer microlens

    Institute of Scientific and Technical Information of China (English)

    X.H.Zeng; J.Plain; S.Jradi; P.Renaud Goud; R.Deturche; P.Royer; R.Bachelot

    2009-01-01

    We report the high speed scanning submicronic microscopy (SSM) using a low cost polymer microlens integrated at the extremity of an optical fiber.These microlenses are fabricated by a free-radical photopolymerization method.Using a polymer microlens with a radius of curvature of 250 nm,a sub-micrometric gold pattern is imaged experimentally by SSM.Different distances between the tip and the sample are used with a high scanning speed of 200 cm/s.In particular,metallic absorption contrasts are described with an optical spatial resolution of 250 nm at the wavelength of 532 nm.Moreover,finite-difference time-domain (FDTD) simulations concerning the focal lengths of microlenses with different geometries and heights support the experimental data.

  5. Extending single-molecule microscopy using optical Fourier processing.

    Science.gov (United States)

    Backer, Adam S; Moerner, W E

    2014-07-17

    This article surveys the recent application of optical Fourier processing to the long-established but still expanding field of single-molecule imaging and microscopy. A variety of single-molecule studies can benefit from the additional image information that can be obtained by modulating the Fourier, or pupil, plane of a widefield microscope. After briefly reviewing several current applications, we present a comprehensive and computationally efficient theoretical model for simulating single-molecule fluorescence as it propagates through an imaging system. Furthermore, we describe how phase/amplitude-modulating optics inserted in the imaging pathway may be modeled, especially at the Fourier plane. Finally, we discuss selected recent applications of Fourier processing methods to measure the orientation, depth, and rotational mobility of single fluorescent molecules.

  6. Imaging of apoptotic HeLa cells by using scanning near-field optical microscopy

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    By using scanning near-field optical microscopy (SNOM), HeLa cells in apoptosis process are imaged with a higher optical resolution beyond the diffraction limit. Since SNOM provides both topographic and transmitted light intensity information of a cell, it can correlate the structural characteristics and optical properties with the spatial position of the apoptotic cells. Wavelength imaging by using near-field spectroscopy shows that there is a great difference in light propagation and absorption in the cell. This unique technique can be applied to the super high resolution imaging of different components in the cell. The observations by near-field optical imaging and near-field spectroscopy indicate an inhomogeneous aggregation of the inner structure in the apoptotic HeLa cells and the change of transmission intensity of light with the apoptosis status.

  7. Simultaneous imaging of magnetic field and temperature distributions by magneto optical indicator microscopy

    Science.gov (United States)

    Lee, Hanju; Jeon, Sunghoon; Friedman, Barry; Lee, Kiejin

    2017-01-01

    We report a simultaneous imaging method of the temperature and the magnetic field distributions based on the magneto optical indicator microscopy. The present method utilizes an optical indicator composed of a bismuth-substituted yttrium iron garnet thin film, and visualizes the magnetic field and temperature distributions through the magneto-optical effect and the temperature dependent optical absorption of the garnet thin film. By using a printed circuit board that carries an electric current as a device under test, we showed that the present method can visualize the magnetic field and temperature distribution simultaneously with a comparable temperature sensitivity (0.2 K) to that of existing conventional thermal imagers. The present technique provides a practical way to get a high resolution magnetic and thermal image at the same time, which is valuable in investigating how thermal variation results in a change of the operation state of a micrometer sized electronic device or material. PMID:28252018

  8. Combined Use of Electron and Light Microscopy Techniques Reveals False Secondary Shell Units in Megaloolithidae Eggshells.

    Science.gov (United States)

    Moreno-Azanza, Miguel; Bauluz, Blanca; Canudo, José Ignacio; Gasca, José Manuel; Torcida Fernández-Baldor, Fidel

    2016-01-01

    Abnormalities in the histo- and ultrastructure of the amniote eggshell are often related to diverse factors, such as ambient stress during egg formation, pathologies altering the physiology of the egg-laying females, or evolutionarily selected modifications of the eggshell structure that vary the physical properties of the egg, for example increasing its strength so as to avoid fracture during incubation. When dealing with fossil materials, all the above hypotheses are plausible, but a detailed taphonomical study has to be performed to rule out the possibility that secondary processes of recrystallization have occurred during fossilization. Traditional analyses, such as optical microscopy inspection and cathodoluminescence, have proven not to be enough to understand the taphonomic story of some eggshells. Recently, electron backscatter diffraction has been used, in combination with other techniques, to better understand the alteration of fossil eggshells. Here we present a combined study using scanning electron microscopy, optical microscopy, cathodoluminescence and electron backscatter diffraction of eggshell fragments assigned to Megaloolithus cf. siruguei from the Upper Cretaceous outcrops of the Cameros Basin. We focus our study on the presence of secondary shell units that mimic most aspects of the ultrastructure of the eggshell mammillae, but grow far from the inner surface of the eggshell. We call these structures extra-spherulites, describe their crystal structure and demonstrate their secondary origin. Our study has important implications for the interpretation of secondary shell units as biological or pathological structures. Thus, electron backscatter diffraction complements other microscope techniques as a useful tool for understanding taphonomical alterations in fossil eggshells.

  9. Recognition of serous ovarian tumors in human samples by multimodal nonlinear optical microscopy

    Science.gov (United States)

    Adur, Javier; Pelegati, Vitor B.; Costa, Leverson F. L.; Pietro, Luciana; de Thomaz, Andre A.; Almeida, Diogo B.; Bottcher-Luiz, Fatima; Andrade, Liliana A. L. A.; Cesar, Carlos L.

    2011-09-01

    We used a multimodal nonlinear optics microscopy, specifically two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG/THG) microscopies, to observe pathological conditions of ovarian tissues obtained from human samples. We show that strong TPEF + SHG + THG signals can be obtained in fixed samples stained with hematoxylin and eosin (H&E) stored for a very long time, and that H&E staining enhanced the THG signal. We then used the multimodal TPEF-SHG-THG microscopies in a stored file of H&E stained samples of human ovarian cancer to obtain complementary information about the epithelium/stromal interface, such as the transformation of epithelium surface (THG) and the overall fibrillary tissue architecture (SHG). This multicontrast nonlinear optics microscopy is able to not only differentiate between cancerous and healthy tissue, but can also distinguish between normal, benign, borderline, and malignant specimens according to their collagen disposition and compression levels within the extracellular matrix. The dimensions of the layers of epithelia can also be measured precisely and automatically. Our data demonstrate that optical techniques can detect pathological changes associated with ovarian cancer.

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

    Science.gov (United States)

    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.

  11. Optical and Digital Microscopic Imaging Techniques and Applications in Pathology

    Directory of Open Access Journals (Sweden)

    Xiaodong Chen

    2011-01-01

    Full Text Available The conventional optical microscope has been the primary tool in assisting pathological examinations. The modern digital pathology combines the power of microscopy, electronic detection, and computerized analysis. It enables cellular-, molecular-, and genetic-imaging at high efficiency and accuracy to facilitate clinical screening and diagnosis. This paper first reviews the fundamental concepts of microscopic imaging and introduces the technical features and associated clinical applications of optical microscopes, electron microscopes, scanning tunnel microscopes, and fluorescence microscopes. The interface of microscopy with digital image acquisition methods is discussed. The recent developments and future perspectives of contemporary microscopic imaging techniques such as three-dimensional and in vivo imaging are analyzed for their clinical potentials.

  12. Confocal microscopy through a multimode fiber using optical correlation

    CERN Document Server

    Loterie, Damien; Psaltis, Demetri; Moser, Christophe

    2015-01-01

    We report on a method to obtain confocal imaging through multimode fibers using optical correlation. First, we measure the fiber's transmission matrix in a calibration step. This allows us to create focused spots at one end of the fiber by shaping the wavefront sent into it from the opposite end. These spots are scanned over a sample, and the light coming back from the sample via the fiber is optically correlated with the input pattern. We show that this achieves spatial selectivity in the detection. The technique is demonstrated on microbeads, a dried epithelial cell, and a cover glass.

  13. Confocal microscopy through a multimode fiber using optical correlation

    Science.gov (United States)

    Loterie, Damien; Goorden, Sebastianus A.; Psaltis, Demetri; Moser, Christophe

    2015-12-01

    We report on a method to obtain confocal imaging through multimode fibers using optical correlation. First, we measure the fiber's transmission matrix in a calibration step. This allows us to create focused spots at one end of the fiber by shaping the wavefront sent into it from the opposite end. These spots are scanned over a sample, and the light coming back from the sample via the fiber is optically correlated with the input pattern. We show that this achieves spatial selectivity in the detection. The technique is demonstrated on microbeads, a dried epithelial cell, and a cover glass.

  14. Electrical characterization of grain boundaries of CZTS thin films using conductive atomic force microscopy techniques

    Energy Technology Data Exchange (ETDEWEB)

    Muhunthan, N.; Singh, Om Pal [Compound Semiconductor Solar Cell, Physics of Energy Harvesting Division, New Delhi 110012 (India); Toutam, Vijaykumar, E-mail: toutamvk@nplindia.org [Quantum Phenomena and Applications Division, CSIR-National Physical Laboratory, Dr. K.S. Krishnan Marg, New Delhi 110012 (India); Singh, V.N., E-mail: singhvn@nplindia.org [Compound Semiconductor Solar Cell, Physics of Energy Harvesting Division, New Delhi 110012 (India)

    2015-10-15

    Graphical abstract: Experimental setup for conducting AFM (C-AFM). - Highlights: • Cu{sub 2}ZnSnS{sub 4} (CZTS) thin film was grown by reactive co-sputtering. • The electronic properties were probed using conducting atomic force microscope, scanning Kelvin probe microscopy and scanning capacitance microscopy. • C-AFM current flow mainly through grain boundaries rather than grain interiors. • SKPM indicated higher potential along the GBs compared to grain interiors. • The SCM explains that charge separation takes place at the interface of grain and grain boundary. - Abstract: Electrical characterization of grain boundaries (GB) of Cu-deficient CZTS (Copper Zinc Tin Sulfide) thin films was done using atomic force microscopic (AFM) techniques like Conductive atomic force microscopy (CAFM), Kelvin probe force microscopy (KPFM) and scanning capacitance microscopy (SCM). Absorbance spectroscopy was done for optical band gap calculations and Raman, XRD and EDS for structural and compositional characterization. Hall measurements were done for estimation of carrier mobility. CAFM and KPFM measurements showed that the currents flow mainly through grain boundaries (GB) rather than grain interiors. SCM results showed that charge separation mainly occurs at the interface of grain and grain boundaries and not all along the grain boundaries.

  15. Conjugate adaptive optics in widefield microscopy with an extended-source wavefront sensor

    CERN Document Server

    Li, Jiang; Paudel, Hari; Barankov, Roman; Bifano, Thomas; Mertz, Jerome

    2015-01-01

    Adaptive optics is a strategy to compensate for sample-induced aberrations in microscopy applications. Generally, it requires the presence of "guide stars" in the sample to serve as localized reference targets. We describe an implementation of conjugate adaptive optics that is amenable to widefield (i.e. non-scanning) microscopy, and can provide aberration corrections over potentially large fields of view without the use of guide stars. A unique feature of our implementation is that it is based on wavefront sensing with a single-shot partitioned-aperture sensor that provides large dynamic range compatible with extended samples. Combined information provided by this sensor and the imaging camera enable robust image de-blurring based on a rapid estimation of sample and aberrations obtained by closed-loop feedback. We present the theoretical principle of our technique and proof of concept experimental demonstrations.

  16. Nanoscale optical properties of metal nanoparticles probed by Second Harmonic Generation microscopy.

    Science.gov (United States)

    Shen, Hong; Nguyen, Ngoc; Gachet, David; Maillard, Vincent; Toury, Timothée; Brasselet, Sophie

    2013-05-20

    We report spatial and vectorial imaging of local fields' confinement properties in metal nanoparticles with branched shapes, using Second Harmonic Generation (SHG) microscopy. Taking advantage of the coherent nature of this nonlinear process, the technique provides a direct evidence of the coupling between the excitation polarization and both localization and polarization specificities of local fields at the sub-diffraction scale. These combined features, which are governed by the nanoparticles' symmetry, are not accessible using other contrasts such as linear optical techniques or two-photon luminescence.

  17. Fluorescence microscopy techniques for quantitative evaluation of organic biocide distribution in antifouling paint coatings: Application to model antifouling coatings

    NARCIS (Netherlands)

    Goodes, L.R.; Dennington, S.P.; Schuppe, H.; Wharton, J.A.; Bakker, M.; Klijnstra, J.W.; Stokes, K.R.

    2012-01-01

    A test matrix of antifouling (AF) coatings including pMMA, an erodible binder and a novel trityl copolymer incorporating Cu 2O and a furan derivative (FD) natural product, were subjected to pontoon immersion and accelerated rotor tests. Fluorescence and optical microscopy techniques were applied to

  18. Stochastic optical reconstruction microscopy (STORM) in comparison with stimulated emission depletion (STED) and other imaging methods.

    Science.gov (United States)

    Tam, Johnny; Merino, David

    2015-11-01

    Stochastic optical reconstruction microscopy (STORM) and stimulated emission depletion (STED) microscopy are two super-resolution optical microscopy approaches that have rapidly gained popularity in recent years. Both modalities offer super-resolution imaging capabilities with the potential for imaging in multiple colors, three-dimensions, and the possibility to image in live cells. In this review, we focus on the specific advantages and disadvantages of each technique in the context of each other. STORM has been reported to achieve higher spatial resolution when compared to STED, but a lengthy acquisition may be required. STED utilizes relatively higher laser intensities, but is able to generate a super-resolution image immediately after acquisition without the need for any additional data processing. Ultimately, the choice between STORM and STED will depend not only on the specific application, but also on the users' ability to understand and optimize the various parameters ranging from sample preparation to image acquisition, which determine the quality of the final image. Stochastic optical reconstruction microscopy (STORM) and stimulated emission depletion (STED) are two super-resolution microscopy approaches that have rapidly gained popularity in recent years. STORM is based on the precise localization of a large number of individual molecules that together form a super-resolved image (bottom), whereas STED is based on the scanning of two super-imposed light sources which together allow for a super-resolved spot on the sample to be imaged (top). We discuss the specific advantages and disadvantages of each technique and explain the various parameters that affect image quality, which should be taken into consideration when planning experiments.

  19. Performance Monitoring Techniques Supporting Cognitive Optical Networking

    DEFF Research Database (Denmark)

    Caballero Jambrina, Antonio; Borkowski, Robert; Zibar, Darko

    2013-01-01

    to solve this issue by realizing a network that can observe, act, learn and optimize its performance, taking into account end-to-end goals. In this letter we present the approach of cognition applied to heterogeneous optical networks developed in the framework of the EU project CHRON: Cognitive...... Heterogeneous Reconfigurable Optical Network. We focus on the approaches developed in the project for optical performance monitoring, which enable the feedback from the physical layer to the cognitive decision system by providing accurate description of the performance of the established lightpaths.......High degree of heterogeneity of future optical networks, such as services with different quality-of-transmission requirements, modulation formats and switching techniques, will pose a challenge for the control and optimization of different parameters. Incorporation of cognitive techniques can help...

  20. Toward optical-tweezers-based force microscopy for airborne microparticles.

    Science.gov (United States)

    Power, Rory M; Burnham, Daniel R; Reid, Jonathan P

    2014-12-20

    Optical tweezers have found widespread application in biological and colloidal physics for the measurement of pN forces over nanometer to micrometer length scales. Similar aerosol-phase measurements of interparticle force have not been reported in spite of the potential to better resolve particle coagulation kinetics. Various refractive index mismatches in the beam path as well as the need to explicitly account for gravity and inertial particle motion provide a number of challenges that must be overcome to make such measurements tractable. In this regard, we demonstrate schemes by which the particle position and trap stiffness may be unambiguously measured using bright-field microscopy with resolution comparable with analogous condensed-phase measurements. Moreover, some of the challenges of working with highly dynamic aqueous particles are introduced and exploited to observe size-dependent phenomena in aerosol optical tweezers. Notably, when combined with cavity-enhanced Raman spectroscopy, this provides a unique opportunity to explore trapping forces over a continuum of particle size and refractive index. It is expected that the methods developed will provide a basis for the measurement of pairwise interaction forces in aerosol optical tweezers while providing a probe of fundamental airborne particle trapping dynamics.

  1. Bow-tie optical antenna probes for single-emitter scanning near-field optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Farahani, Javad N [Nano-Optics Group, National Center of Competence for Research in Nanoscale Science, Institute of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel (Switzerland); Eisler, Hans-Juergen [Nano-Optics Group, National Center of Competence for Research in Nanoscale Science, Institute of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel (Switzerland); Pohl, Dieter W [Nano-Optics Group, National Center of Competence for Research in Nanoscale Science, Institute of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel (Switzerland); Pavius, Michael [Center of MicroNanoTechnology (CMI), Ecole Polytechnique Federale de Lausanne (EPFL), CH-1015 Lausanne (Switzerland); Flueckiger, Philippe [Center of MicroNanoTechnology (CMI), Ecole Polytechnique Federale de Lausanne (EPFL), CH-1015 Lausanne (Switzerland); Gasser, Philippe [EMPA, Swiss Federal Laboratories for Materials Testing and Research, Electronics/Metrology Laboratory, Ueberlandstrasse 129, CH-8600 Duebendorf (Switzerland); Hecht, Bert [Nano-Optics Group, National Center of Competence for Research in Nanoscale Science, Institute of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel (Switzerland)

    2007-03-28

    A method for the fabrication of bow-tie optical antennas at the apex of pyramidal Si{sub 3}N{sub 4} atomic force microscopy tips is described. We demonstrate that these novel optical probes are capable of sub-wavelength imaging of single quantum dots at room temperature. The enhanced and confined optical near-field at the antenna feed gap leads to locally enhanced photoluminescence (PL) of single quantum dots. Photoluminescence quenching due to the proximity of metal is found to be insignificant. The method holds promise for single quantum emitter imaging and spectroscopy at spatial resolution limited by the engineered antenna gap width exclusively.

  2. Optical measurement techniques - A push for digitization

    Science.gov (United States)

    Kulkarni, Rishikesh; Rastogi, Pramod

    2016-12-01

    Over the years, optical measurement techniques have been the problem-solving backbone of many engineering applications such as nondestructive testing of materials, measurement of various material properties, structural analysis and experimental mechanics [1-3]. Probably the most important advantage associated with any optical measurement system over other systems is its non-contact type of measurement capability. Apart from their non-contact nature, the optical measurement systems are capable of providing full-field measurements at scales ranging from milli-meters to nano-meters.

  3. Doppler optical coherence microscopy and tomography applied to inner ear mechanics

    Energy Technology Data Exchange (ETDEWEB)

    Page, Scott; Freeman, Dennis M. [Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Ghaffari, Roozbeh [Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States)

    2015-12-31

    While it is clear that cochlear traveling waves underlie the extraordinary sensitivity, frequency selectivity, and dynamic range of mammalian hearing, the underlying micromechanical mechanisms remain unresolved. Recent advances in low coherence measurement techniques show promise over traditional laser Doppler vibrometry and video microscopy, which are limited by low reflectivities of cochlear structures and restricted optical access. Doppler optical coherence tomography (DOCT) and Doppler optical coherence microscopy (DOCM) both utilize a broadband source to limit constructive interference of scattered light to a small axial depth called a coherence gate. The coherence gate can be swept axially to image and measure sub-nanometer motions of cochlear structures throughout the cochlear partition. The coherence gate of DOCT is generally narrower than the confocal gate of the focusing optics, enabling increased axial resolution (typically 15 μm) within optical sections of the cochlear partition. DOCM, frequently implemented in the time domain, centers the coherence gate on the focal plane, achieving enhanced lateral and axial resolution when the confocal gate is narrower than the coherence gate. We compare these two complementary systems and demonstrate their utility in studying cellular and micromechanical mechanisms involved in mammalian hearing.

  4. Structured Illumination-Based Super-Resolution Optical Microscopy for Hemato- and Cyto-Pathology Applications

    Directory of Open Access Journals (Sweden)

    Tieqiao Zhang

    2013-01-01

    Full Text Available Structured illumination fluorescence microscopy utilizes interfering light and the moiré effect to enhance spatial resolution to about a half of that of conventional light microscopy, i.e. approximately 90 nm. In addition to the enhancement in the x and y directions, it also allows enhancement of resolution in the z- direction by the same factor of two (to approximately 220 nm, making it a powerful tool for 3-D morphology studies of fluorescently labeled cells or thin tissue sections. In this report, we applied this technique to several types of blood cells that are commonly seen in hematopathology. Compared with standard brightfield and ordinary fluorescence microscopy images, the 3-D morphology results clearly reveal the morphological features of different types of normal blood cells. We have also used this technique to evaluate morphologies of abnormal erythrocytes and compare them with those recorded on normal cells. The results give a very intuitive presentation of morphological structures of erythrocytes with great details. This research illustrates the potential of this technique to be used in hematology and cyto-pathology studies aimed at identifying nanometer-sized features that cannot be distinguished otherwise with conventional optical microscopy.

  5. Structured illumination-based super-resolution optical microscopy for hemato- and cyto-pathology applications.

    Science.gov (United States)

    Zhang, Tieqiao; Osborn, Samantha; Brandow, Chloe; Dwyre, Denis; Green, Ralph; Lane, Stephen; Wachsmann-Hogiu, Sebastian

    2013-01-01

    Structured illumination fluorescence microscopy utilizes interfering light and the moiré effect to enhance spatial resolution to about a half of that of conventional light microscopy, i.e. approximately 90 nm. In addition to the enhancement in the x and y directions, it also allows enhancement of resolution in the z- direction by the same factor of two (to approximately 220 nm), making it a powerful tool for 3-D morphology studies of fluorescently labeled cells or thin tissue sections. In this report, we applied this technique to several types of blood cells that are commonly seen in hematopathology. Compared with standard brightfield and ordinary fluorescence microscopy images, the 3-D morphology results clearly reveal the morphological features of different types of normal blood cells. We have also used this technique to evaluate morphologies of abnormal erythrocytes and compare them with those recorded on normal cells. The results give a very intuitive presentation of morphological structures of erythrocytes with great details. This research illustrates the potential of this technique to be used in hematology and cyto-pathology studies aimed at identifying nanometer-sized features that cannot be distinguished otherwise with conventional optical microscopy.

  6. 3D automatic quantification applied to optically sectioned images to improve microscopy analysis

    Directory of Open Access Journals (Sweden)

    JE Diaz-Zamboni

    2009-08-01

    Full Text Available New fluorescence microscopy techniques, such as confocal or digital deconvolution microscopy, allow to easily obtain three-dimensional (3D information from specimens. However, there are few 3D quantification tools that allow extracting information of these volumes. Therefore, the amount of information acquired by these techniques is difficult to manipulate and analyze manually. The present study describes a model-based method, which for the first time shows 3D visualization and quantification of fluorescent apoptotic body signals, from optical serial sections of porcine hepatocyte spheroids correlating them to their morphological structures. The method consists on an algorithm that counts apoptotic bodies in a spheroid structure and extracts information from them, such as their centroids in cartesian and radial coordinates, relative to the spheroid centre, and their integrated intensity. 3D visualization of the extracted information, allowed us to quantify the distribution of apoptotic bodies in three different zones of the spheroid.

  7. Near-field scanning optical microscopy in cell biology and cytogenetics.

    Science.gov (United States)

    Hausmann, Michael; Perner, Birgit; Rapp, Alexander; Wollweber, Leo; Scherthan, Harry; Greulich, Karl-Otto

    2006-01-01

    Light microscopy has proven to be one of the most versatile analytical tools in cell biology and cytogenetics. The growing spectrum of scientific knowledge demands a continuous improvement of the optical resolution of the instruments. In far-field light microscopy, the attainable resolution is dictated by the limit of diffraction, which, in practice, is about 250 nm for high-numerical-aperture objective lenses. Near-field scanning optical microscopy (NSOM) was the first technique that has overcome this limit up to about one order of magnitude. Typically, the resolution range below 100 nm is accessed for biological applications. Using appropriately designed scanning probes allows for obtaining an extremely small near-field light excitation volume (some tens of nanometers in diameter). Because of the reduction of background illumination, high contrast imaging becomes feasible for light transmission and fluorescence microscopy. The height of the scanning probe is controlled by atomic force interactions between the specimen surface and the probe tip. The control signal can be used for the production of a topographic (nonoptical) image that can be acquired simultaneously. In this chapter, the principle of NSOM is described with respect to biological applications. A brief overview of some requirements in biology and applications described in the literature are given. Practical advice is focused on instruments with aperture-type illumination probes. Preparation protocols focussing on NSOM of cell surfaces and chromosomes are presented.

  8. Combining calcium imaging with other optical techniques.

    Science.gov (United States)

    Canepari, Marco; Zecevic, Dejan; Vogt, Kaspar E; Ogden, David; De Waard, Michel

    2013-12-01

    Ca(2+) imaging is a commonly used approach for measuring Ca(2+) signals at high spatial resolution. The method is often combined with electrode recordings to correlate electrical and chemical signals or to investigate Ca(2+) signals following an electrical stimulation. To obtain information on electrical activity at the same spatial resolution, Ca(2+) imaging must be combined with membrane potential imaging. Similarly, stimulation of subcellular compartments requires photostimulation. Thus, combining Ca(2+) imaging with an additional optical technique facilitates the study of a number of physiological questions. The aim of this article is to introduce some basic principles regarding the combination of Ca(2+) imaging with other optical techniques. We discuss the design of the optics, the design of experimental protocols, the optical characteristics of Ca(2+) indicators used in combination with an optical probe, and the affinity of the Ca(2+) indicator in relation to the type of measurement. This information will enable the reader to devise an optimal strategy for combined optical experiments.

  9. Dental diagnostics using optical coherence techniques

    Energy Technology Data Exchange (ETDEWEB)

    Nathel, H. [Lawrence Livermore National Lab., CA (United States); Colston, B. [Univ. of California, San Francisco, CA (United States); Armitage, G. [Univ. of California, Davis, CA (United States)] [and others

    1994-11-15

    Optical radiation can be used for diagnostic purposes in oral medicine. However, due to the turbid, amorphous, and inhomogeneous nature of dental tissue conventional techniques used to transilluminate materials are not well suited to dental tissues. Optical coherence techniques either in the time- of frequency-domain offer the capabilities of discriminating scattered from unscattered light, thus allowing for imaging through turbid tissue. Currently, using optical time-domain reflectometry we are able to discriminate specular from diffuse reflections occurring at tissue boundaries. We have determined the specular reflectivity of enamel and dentin to be approximately 6.6 x 10{sup -5} and 1.3 x 10{sup -6}, respectively. Implications to periodontal imaging will be discussed.

  10. Electro-optical techniques for signal conditioning

    Science.gov (United States)

    Helfrich, R. W.

    1981-01-01

    Electro-optical (EO) processing is discussed as a potential alternative to the all-digital approach to signal processing. Nonuniformity compensation can be done by normalizing all the single element detectors outputs in a staring array for both gain and level. Distortion correction can be accomplished with blackbodies, scene statistics or defocused optics. An algorithm used in digital signal conditioning that can be closely approximated by EO techniques is Local Area Brightness Control (LABC). In a digital processor, LABC is performed on a pixel-by-pixel basis, resulting in an enormous amount of calculation. A partially defocused optical system can be used in an EO analog to the digital system. For both nonuniformity compensation and LABC, the EO technique can result in great simplification.

  11. Optical Management Techniques for Organic Solar Cells

    CERN Document Server

    Rajagopal, Adharsh

    2016-01-01

    In this thesis, two different optical management techniques for organics based solar cells are explored. The first part is focused on the development of a textured rear reflector for OPVs. The use of textured reflector (TR) facilitates an increase in the optical path length along with light trapping within the active layer. TR was fabricated through a relatively simpler technique by depositing metal films over a microlens array (MLA). Zinc oxide nanoparticles were used to minimize the shadowing effect. Using TR, enhancements in short-circuit current density and power conversion efficiencies up to 10-25% were demonstrated for a polymer based organic solar cell. The second part is focused on improving the effectiveness of MLA incorporation in OPVs. The increase in path length achieved using MLA can be improved by increasing the refractive index of MLA and incorporating MLA directly on the transparent electrode instead of glass substrate. This approach could avoid the optical losses occurring at the interface be...

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

    Science.gov (United States)

    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.

  13. Perspective of remote optical measurement techniques

    OpenAIRE

    Gregorio, Eduard; Rocadenbosch Burillo, Francisco

    2007-01-01

    This article presents an intercomparison between four different ROMTs: differential optical absorption spectroscopy (DOAS), differential absorption LIDAR (DIAL), Fourier transform infrared spectroscopy (FTIR), and tunable diode laser absorption spectroscopy (TDLAS). The main focus is on the TDLAS technique, where the main laser-diode typologies and modulation schemes, namely, wavelength modulation spectroscopy (WMS) and frequency modulation spectroscopy (FMS), are reviewed. At present, new pr...

  14. Scanning microwave microscopy technique for nanoscale characterization of magnetic materials

    Science.gov (United States)

    Joseph, C. H.; Sardi, G. M.; Tuca, S. S.; Gramse, G.; Lucibello, A.; Proietti, E.; Kienberger, F.; Marcelli, R.

    2016-12-01

    In this work, microwave characterization of magnetic materials using the scanning microwave microscopy (SMM) technique is presented. The capabilities of the SMM are employed for analyzing and imaging local magnetic properties of the materials under test at the nanoscale. The analyses are performed by acquiring both amplitude and phase of the reflected microwave signal. The changes in the reflection coefficient S11 are related to the local properties of the material under investigation, and the changes in its magnetic properties have been studied as a function of an external DC magnetic bias. Yttrium iron garnet (YIG) films deposited by RF sputtering and grown by liquid phase epitaxial (LPE) on gadolinium gallium garnet (GGG) substrates and permalloy samples have been characterized. An equivalent electromagnetic transmission line model is discussed for the quantitative analysis of the local magnetic properties. We also observed the hysteretic behavior of the reflection coefficient S11 with an external bias field. The imaging and spectroscopy analysis on the experimental results are evidently indicating the possibilities of measuring local changes in the intrinsic magnetic properties on the surface of the material.

  15. Second harmonic generation microscopy is a novel technique for differential diagnosis of breast fibroepithelial lesions.

    Science.gov (United States)

    Tan, Wai Jin; Yan, Jie; Xu, Shuoyu; Thike, Aye Aye; Bay, Boon Huat; Yu, Hanry; Tan, Min-Han; Tan, Puay Hoon

    2015-12-01

    Breast fibroepithelial lesions, including fibroadenomas and phyllodes tumours, are commonly encountered in clinical practice. As histological differences between these two related entities may be subtle, resulting in a challenging differential diagnosis, pathological techniques to assist the differential diagnosis of these two entities are of high interest. An accurate diagnosis at biopsy is important given corresponding implications for clinical decision-making including surgical extent and monitoring. Second harmonic generation (SHG) microscopy is a recently developed optical imaging technique capable of robust, powerful and unbiased label-free direct detection of collagen fibril structure in tissue without the use of antibodies. We constructed tissue microarrays emulating limited materials on biopsy to investigate quantitative collagen signal in fibroepithelial lesions using SHG microscopy. Archived formalin-fixed paraffin-embedded materials of 47 fibroepithelial lesions (14 fibroadenomas and 33 phyllodes tumours) were evaluated. Higher collagen signal on SHG microscopy was observed in fibroadenomas than phyllodes tumours on SHG imaging (pmicroscopy for fibroadenoma classification was 71.4% and 84.4%, respectively. To corroborate these findings, we performed immunohistochemistry on tissue array sections using collagen I and III primary antibodies. Both collagen I and III immunohistochemical expressions were also significantly higher in fibroadenomas than in phyllodes tumours (pmicroscopy is a novel imaging approach that can aid the differential diagnosis of fibroepithelial lesions.

  16. Scanning Techniques For Optical Data Storage

    Science.gov (United States)

    Towner, David K.

    1987-01-01

    It seems almost paradoxical that beams of light can be moved and steered at very high speeds using a variety of scanning methods, yet the optical disk drives now being designed and marketed for data storage applications have comparatively long access times. Knowing that optical data storage has unrealized potential is of interest, but of more immediate concern is the recognition that poor access performance is a serious design issue. Magnetic disk drives offer average seek times in the 15-25ms range, compared to about 80-500ms (or more, for CD ROMS) for current optical drives. This performance disparity exists, in part, because the relatively massive "optical heads" in use today cannot be transported across the radius of a disk as quickly as a stack of much lighter magnetic heads. Any potential distance advantage that the optical drive might have, due to its substantially higher track density, is offset by the magnetic drive's use of a multi-disk stack. As a result, the drive must achieve similar radial accelerations during seeks if it is to have similar average access times. The inability of current optical drives to approach the access speeds of comparable magnetic drives significantly reduces the competitiveness of optical products in major segments of the very large data storage market. This shortcoming is especially disturbing when we know that opto-mechanical scanners typically operate in the 1-10ms range and that non-mechanical scanning techniques can be substantially faster than that.

  17. Spectral interferometric microscopy reveals absorption by individual optical nanoantennas from extinction phase.

    Science.gov (United States)

    Gennaro, Sylvain D; Sonnefraud, Yannick; Verellen, Niels; Van Dorpe, Pol; Moshchalkov, Victor V; Maier, Stefan A; Oulton, Rupert F

    2014-04-30

    Optical antennas transform light from freely propagating waves into highly localized excitations that interact strongly with matter. Unlike their radio frequency counterparts, optical antennas are nanoscopic and high frequency, making amplitude and phase measurements challenging and leaving some information hidden. Here we report a novel spectral interferometric microscopy technique to expose the amplitude and phase response of individual optical antennas across an octave of the visible to near-infrared spectrum. Although it is a far-field technique, we show that knowledge of the extinction phase allows quantitative estimation of nanoantenna absorption, which is a near-field quantity. To verify our method we characterize gold ring-disk dimers exhibiting Fano interference. Our results reveal that Fano interference only cancels a bright mode's scattering, leaving residual extinction dominated by absorption. Spectral interference microscopy has the potential for real-time and single-shot phase and amplitude investigations of isolated quantum and classical antennas with applications across the physical and life sciences.

  18. New optical technique for bulk magnetostriction measurement

    CERN Document Server

    Samata, H; Uchida, T; Abe, S

    2000-01-01

    A new optical technique was applied to the measurement of magnetostriction in bulk samples. This technique utilizes an optical fiber bundle, AC-modulated light and lock-in detection. Deformation of the sample is determined from the ratio of the incident and reflected light intensities. Noise due to the instability of the light source is eliminated by obtaining the ratio of the incident and reflected light intensities, and the noise caused in the detector circuit can be reduced by lock-in detection. The performance of this method was characterized with a series of measurements using a gold film and crystal disks of pure iron and nickel. This technique offers a resolution of 0.5 nm and is sensitive enough to measure magnetostriction as small as 5x10 sup - sup 7 in 1 mm thick samples.

  19. Combined Use of Electron and Light Microscopy Techniques Reveals False Secondary Shell Units in Megaloolithidae Eggshells.

    Directory of Open Access Journals (Sweden)

    Miguel Moreno-Azanza

    Full Text Available Abnormalities in the histo- and ultrastructure of the amniote eggshell are often related to diverse factors, such as ambient stress during egg formation, pathologies altering the physiology of the egg-laying females, or evolutionarily selected modifications of the eggshell structure that vary the physical properties of the egg, for example increasing its strength so as to avoid fracture during incubation. When dealing with fossil materials, all the above hypotheses are plausible, but a detailed taphonomical study has to be performed to rule out the possibility that secondary processes of recrystallization have occurred during fossilization. Traditional analyses, such as optical microscopy inspection and cathodoluminescence, have proven not to be enough to understand the taphonomic story of some eggshells. Recently, electron backscatter diffraction has been used, in combination with other techniques, to better understand the alteration of fossil eggshells. Here we present a combined study using scanning electron microscopy, optical microscopy, cathodoluminescence and electron backscatter diffraction of eggshell fragments assigned to Megaloolithus cf. siruguei from the Upper Cretaceous outcrops of the Cameros Basin. We focus our study on the presence of secondary shell units that mimic most aspects of the ultrastructure of the eggshell mammillae, but grow far from the inner surface of the eggshell. We call these structures extra-spherulites, describe their crystal structure and demonstrate their secondary origin. Our study has important implications for the interpretation of secondary shell units as biological or pathological structures. Thus, electron backscatter diffraction complements other microscope techniques as a useful tool for understanding taphonomical alterations in fossil eggshells.

  20. Optical and digital techniques for information security

    CERN Document Server

    2005-01-01

    Optical and Digital Techniques for Information Security is the first book in a series focusing on Advanced Sciences and Technologies for Security Applications. This book encompases the results of research investigation and technologies used to secure, verify, recognize, track, and authenticate objects and information from theft, counterfeiting, and manipulation by unauthorized persons and agencies. This Information Security book will draw on the diverse expertise in optical sciences and engineering, digital image processing, imaging systems, information processing, computer based information systems, sensors, detectors, and biometrics to report innovative technologies that can be applied to information security issues. The Advanced Sciences and Technologies for Security Applications series focuses on research monographs in the areas of: -Recognition and identification (including optical imaging, biometrics, authentication, verification, and smart surveillance systems) -Biological and chemical threat detection...

  1. All-optical photoacoustic microscopy using a MEMS scanning mirror

    Science.gov (United States)

    Chen, Sung-Liang; Xie, Zhixing; Ling, Tao; Wei, Xunbin; Guo, L. Jay; Wang, Xueding

    2013-03-01

    It has been studied that a potential marker to obtain prognostic information about bladder cancer is tumor neoangiogenesis, which can be quantified by morphometric characteristics such as microvascular density. Photoacoustic microscopy (PAM) can render sensitive three-dimensional (3D) mapping of microvasculature, providing promise to evaluate the neoangiogenesis that is closely related to the diagnosis of bladder cancer. To ensure good image quality, it is desired to acquire bladder PAM images from its inside via the urethra, like conventional cystoscope. Previously, we demonstrated all-optical PAM systems using polymer microring resonators to detect photoacoustic signals and galvanometer mirrors for laser scanning. In this work, we build a miniature PAM system using a microelectromechanical systems (MEMS) scanning mirror, demonstrating a prototype of an endoscopic PAM head capable of high imaging quality of the bladder. The system has high resolutions of 17.5 μm in lateral direction and 19 μm in the axial direction at a distance of 5.4 mm. Images of printed grids and the 3D structure of microvasculature in animal bladders ex vivo by the system are demonstrated.

  2. Surface Wear Measurement Using Optical Correlation Technique

    Science.gov (United States)

    Acinger, Kresimir

    1983-12-01

    The coherent optical correlation technique was applied for measuring the surface wear of a tappet (part of car engine), worn by friction with the camshaft. It was found that maximum correlation intensity decays exponentially with the number of wear cycles (i.e. camshaft revolutions). Tappets of the same make have an identical rate of correlation decay. Tappets of different makes have different rates of correlation decay which are in agreement with observed long term wear.

  3. Computational Techniques for LED Optical Microcavities

    OpenAIRE

    García Santiago, Xavier

    2015-01-01

    The project consist on the development of numerical methods and computational techniques to model the processes of light extraction in power LED (Light-Emitting Diodes) devices. We aim at the use of complex corrugated microstructures to boost the efficiency of our current LUXEON LED products. In order to study extraction efficiency in these devices a 3D optics model of thin film micro-structures must be developed and tested. In this project we develop a numerical model for computing and st...

  4. Noninvasive label-free monitoring of cosmetics and pharmaceuticals in human skin using nonlinear optical microscopy (Conference Presentation)

    Science.gov (United States)

    Osseiran, Sam; Wang, Hequn; Evans, Conor L.

    2017-02-01

    Over the past decade, nonlinear optical microscopy has seen a dramatic rise in its use in research settings due to its noninvasiveness, enhanced penetration depth, intrinsic optical sectioning, and the ability to probe chemical compounds with molecular specificity without exogenous contrast agents. Nonlinear optical techniques including two-photon excitation fluorescence (2PEF), fluorescence lifetime imaging microscopy (FLIM), second harmonic generation (SHG), coherent anti-Stokes and stimulated Raman scattering (CARS and SRS, respectively), as well as transient and sum frequency absorption (TA and SFA, respectively), have been widely used to explore the physiology and microanatomy of skin. Recently, these modalities have shed light on dermal processes that could not have otherwise been observed, including the spatiotemporal monitoring of cosmetics and pharmaceuticals. However, a challenge quickly arises when studying such chemicals in a dermatological context: many exogenous compounds have optical signatures that can interfere with the signals that would otherwise be acquired from intact skin. For example, oily solvents exhibit strong signals when probing CH2 vibrations with CARS/SRS; chemical sun filters appear bright in 2PEF microscopy; and darkly colored compounds readily absorb light across a broad spectrum, producing strong TA/SFA signals. Thus, this discussion will first focus on the molecular contrast in skin that can be probed using the aforementioned nonlinear optical techniques. This will be followed by an overview of strategies that take advantage of the exogenous compounds' optical signatures to probe spatiotemporal dynamics while preserving endogenous information from skin.

  5. Advanced magneto-optical microscopy: Imaging from picoseconds to centimeters - imaging spin waves and temperature distributions (invited

    Directory of Open Access Journals (Sweden)

    Necdet Onur Urs

    2016-05-01

    Full Text Available Recent developments in the observation of magnetic domains and domain walls by wide-field optical microscopy based on the magneto-optical Kerr, Faraday, Voigt, and Gradient effect are reviewed. Emphasis is given to the existence of higher order magneto-optical effects for advanced magnetic imaging. Fundamental concepts and advances in methodology are discussed that allow for imaging of magnetic domains on various length and time scales. Time-resolved imaging of electric field induced domain wall rotation is shown. Visualization of magnetization dynamics down to picosecond temporal resolution for the imaging of spin-waves and magneto-optical multi-effect domain imaging techniques for obtaining vectorial information are demonstrated. Beyond conventional domain imaging, the use of a magneto-optical indicator technique for local temperature sensing is shown.

  6. En face speckle reduction in optical coherence microscopy by frequency compounding

    Science.gov (United States)

    Magnain, Caroline; Wang, Hui; Sakadžić, Sava; Fischl, Bruce; Boas, David A.

    2017-01-01

    We report the use of frequency compounding to significantly reduce speckle noise in optical coherence microscopy, more specifically on the en face images. This method relies on the fact that the speckle patterns recorded from different wavelengths simultaneously are independent; hence their summation yields significant reduction in noise, with only a single acquisition. The results of our experiments with microbeads show that the narrow confocal parameter, due to a high numerical aperture objective, restricts the axial resolution loss that would otherwise theoretically broaden linearly with the number of optical frequency bands used. This speckle reduction scheme preserves the lateral resolution since it is performed on individual A-scans. Finally, we apply this technique to images of fixed human brain tissue, showing significant improvements in contrast-to-noise ratio with only moderate loss of axial resolution, in an effort to improve automatic three-dimensional detection of cells and fibers in the cortex. PMID:27128040

  7. Computational modeling of optical projection tomographic microscopy using the finite difference time domain method.

    Science.gov (United States)

    Coe, Ryan L; Seibel, Eric J

    2012-12-01

    We present a method for modeling image formation in optical projection tomographic microscopy (OPTM) using high numerical aperture (NA) condensers and objectives. Similar to techniques used in computed tomography, OPTM produces three-dimensional, reconstructed images of single cells from two-dimensional projections. The model is capable of simulating axial scanning of a microscope objective to produce projections, which are reconstructed using filtered backprojection. Simulation of optical scattering in transmission optical microscopy is designed to analyze all aspects of OPTM image formation, such as degree of specimen staining, refractive-index matching, and objective scanning. In this preliminary work, a set of simulations is performed to examine the effect of changing the condenser NA, objective scan range, and complex refractive index on the final reconstruction of a microshell with an outer radius of 1.5 μm and an inner radius of 0.9 μm. The model lays the groundwork for optimizing OPTM imaging parameters and triaging efforts to further improve the overall system design. As the model is expanded in the future, it will be used to simulate a more realistic cell, which could lead to even greater impact.

  8. Numerical study of super-resolved optical microscopy with partly staggered beams

    Science.gov (United States)

    He, Jinping; Wang, Nan; Kobayashi, Takayoshi

    2016-12-01

    The resolving power of optical microscopy involving two or even more beams, such as pump-probe microscopy and nonlinear optical microscopy, can be enhanced both laterally and longitudinally with partly staggered beams. A numerical study of the new super-resolution imaging technology is performed with vector diffraction theory. The influence of polarization is discussed. A resolving power of sub-100 nm and sub-300 nm in the lateral and longitudinal directions, respectively, is achievable.

  9. DMD-based LED-illumination Super-resolution and optical sectioning microscopy

    OpenAIRE

    Dan, Dan; Ming LEI; 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 ...

  10. Polishing techniques for MEGARA pupil elements optics

    Science.gov (United States)

    Izazaga, R.; Carrasco, E.; Aguirre, D.; Salas, A.; Gil de Paz, A.; Gallego, J.; Iglesias, J.; Arroyo, J. M.; Hernández, M.; López, N.; López, V.; Quechol, J. T.; Salazar, M. F.; Carballo, C.; Cruz, E.; Arriaga, J.; De la Luz, J. A.; Huepa, A.; Jaimes, G. L.; Reyes, J.

    2016-07-01

    MEGARA (Multi-Espectrógrafo en GTC de Alta Resolución para Astronomía) is the new integral-field and multi-object optical spectrograph for the 10.4m Gran Telescopio Canarias.. It will offer RFWHM 6,000, 12,000 and 18,700 for the low- , mid- and high-resolution, respectively in the wavelength range 3650-9700Å. .The dispersive elements are volume phase holographic (VPH) gratings, sandwiched between two flat Fused Silica windows of high optical precision in large apertures. The design, based in VPHs in combination with Ohara PBM2Y prisms allows to keep the collimator and camera angle fixed. Seventy three optical elements are being built in Mexico at INAOE and CIO. For the low resolution modes, the VPHs windows specifications in irregularity is 1 fringe in 210mm x 170mm and 0.5 fringe in 190mm x 160mm. for a window thickness of 25 mm. For the medium and high resolution modes the irregularity specification is 2 fringes in 220mm x 180mm and 1 fringe in 205mm x 160mm, for a window thickness of 20mm. In this work we present a description of the polishing techniques developed at INAOE optical workshop to fabricate the 36 Fused Silica windows and 24 PBM2Y prisms that allows us to achieve such demanding specifications. We include the processes of mounting, cutting, blocking, polishing and testing.

  11. Nonlinear optical microscopy and ultrasound imaging of human cervical structure

    Science.gov (United States)

    Reusch, Lisa M.; Feltovich, Helen; Carlson, Lindsey C.; Hall, Gunnsteinn; Campagnola, Paul J.; Eliceiri, Kevin W.; Hall, Timothy J.

    2013-03-01

    The cervix softens and shortens as its collagen microstructure rearranges in preparation for birth, but premature change may lead to premature birth. The global preterm birth rate has not decreased despite decades of research, likely because cervical microstructure is poorly understood. Our group has developed a multilevel approach to evaluating the human cervix. We are developing quantitative ultrasound (QUS) techniques for noninvasive interrogation of cervical microstructure and corroborating those results with high-resolution images of microstructure from second harmonic generation imaging (SHG) microscopy. We obtain ultrasound measurements from hysterectomy specimens, prepare the tissue for SHG, and stitch together several hundred images to create a comprehensive view of large areas of cervix. The images are analyzed for collagen orientation and alignment with curvelet transform, and registered with QUS data, facilitating multiscale analysis in which the micron-scale SHG images and millimeter-scale ultrasound data interpretation inform each other. This novel combination of modalities allows comprehensive characterization of cervical microstructure in high resolution. Through a detailed comparative study, we demonstrate that SHG imaging both corroborates the quantitative ultrasound measurements and provides further insight. Ultimately, a comprehensive understanding of specific microstructural cervical change in pregnancy should lead to novel approaches to the prevention of preterm birth.

  12. Optical multichannel analyzer techniques for high resolution optical spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Chao, J.L.

    1980-06-01

    The development of optical multichannel analyzer techniques for UV/VIS spectroscopy is presented. The research focuses on the development of spectroscopic techniques for measuring high resolution spectral lineshape functions from the exciton phosphorescence in H/sub 2/-1,2,4,5-tetrachlorobenzene. It is found that the temperature dependent frequency shifts and widths confirm a theoretical model based on an exchange theory. The exchange of low energy phonon modes which couple with excited state exciton transitions is shown to display the proper temperature dependent behavior. In addition to the techniques for using the optical multichannel analyzer (OMA) to perform low light level target integration, the use of the OMA for capturing spectral information in transient pulsed laser applications is discussed. An OMP data acquisition system developed for real-time signal processng is described. Both hardware and software interfacing considerations for control and data acquisition by a microcomputer are described. The OMA detector is described in terms of the principles behind its photoelectron detection capabilities and its design is compared with other optoelectronic devices.

  13. Scanning near-field optical microscopy on rough surfaces: applications in chemistry, biology, and medicine

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available Shear-force apertureless scanning near-field optical microscopy (SNOM with very sharp uncoated tapered waveguides relies on the unexpected enhancement of reflection in the shear-force gap. It is the technique for obtaining chemical (materials contrast in the optical image of “real world” surfaces that are rough and very rough without topographical artifacts, and it is by far less complicated than other SNOM techniques that can only be used for very flat surfaces. The experimental use of the new photophysical effect is described. The applications of the new technique are manifold. Important mechanistic questions in solid-state chemistry (oxidation, diazotization, photodimerization, surface hydration, hydrolysis are answered with respect to simultaneous AFM (atomic force microscopy and detailed crystal packing. Prehistoric petrified bacteria and concomitant pyrite inclusions are also investigated with local RAMAN SNOM. Polymer beads and unstained biological objects (rabbit heart, shrimp eye allow for nanoscopic analysis of cell organelles. Similarly, human teeth and a cancerous tissue are analyzed. Bladder cancer tissue is clearly differentiated from healthy tissue without staining and this opens a new highly promising diagnostic tool for precancer diagnosis. Industrial applications are demonstrated at the corrosion behavior of dental alloys (withdrawal of a widely used alloy, harmless substitutes, improvement of paper glazing, behavior of blood bags upon storage, quality assessment of metal particle preparations for surface enhanced RAMAN spectroscopy, and determination of diffusion coefficient and light fastness in textile fiber dyeing. The latter applications include fluorescence SNOM. Local fluorescence SNOM is also used in the study of partly aggregating dye nanoparticles within resin/varnish preparations. Unexpected new insights are obtained in all of the various fields that cannot be obtained by other techniques.

  14. Innovations of wide-field optical-sectioning fluorescence microscopy: toward high-speed volumetric bio-imaging with simplicity

    Science.gov (United States)

    Yu, Jiun-Yann

    Optical microscopy has become an indispensable tool for biological researches since its invention, mostly owing to its sub-cellular spatial resolutions, non-invasiveness, instrumental simplicity, and the intuitive observations it provides. Nonetheless, obtaining reliable, quantitative spatial information from conventional wide-field optical microscopy is not always intuitive as it appears to be. This is because in the acquired images of optical microscopy the information about out-of-focus regions is spatially blurred and mixed with in-focus information. In other words, conventional wide-field optical microscopy transforms the three-dimensional spatial information, or volumetric information about the objects into a two-dimensional form in each acquired image, and therefore distorts the spatial information about the object. Several fluorescence holography-based methods have demonstrated the ability to obtain three-dimensional information about the objects, but these methods generally rely on decomposing stereoscopic visualizations to extract volumetric information and are unable to resolve complex 3-dimensional structures such as a multi-layer sphere. The concept of optical-sectioning techniques, on the other hand, is to detect only two-dimensional information about an object at each acquisition. Specifically, each image obtained by optical-sectioning techniques contains mainly the information about an optically thin layer inside the object, as if only a thin histological section is being observed at a time. Using such a methodology, obtaining undistorted volumetric information about the object simply requires taking images of the object at sequential depths. Among existing methods of obtaining volumetric information, the practicability of optical sectioning has made it the most commonly used and most powerful one in biological science. However, when applied to imaging living biological systems, conventional single-point-scanning optical-sectioning techniques often

  15. Low-cost multimodal light sheet microscopy for optically cleared tissues and living specimens.

    Science.gov (United States)

    Rouger, Vincent; Alchini, Ricardo; Kazarine, Alexei; Gopal, Angelica A; Girouard, Marie-Pier; Fournier, Alyson E; Wiseman, Paul W

    2016-12-01

    Light sheet microscopy techniques have expanded with designs to address many new applications. Due to rapid advancements in computing power, camera/detector technologies, and tissue clearing techniques, light sheet methods are becoming increasingly popular for biomedical imaging applications at the cellular and tissue levels. Light sheet imaging modalities couple rapid imaging rates, low-levels of phototoxicity, and excellent signal to noise ratios, contributing to their popularity for experimental biology. However, the current major limitation of light sheet microscopy arises from optical aberrations, with the main drawback being the defocusing introduced by refractive index variations that accompany clearing techniques. Here, we propose an inexpensive and easy to build light sheet based instrumentation to overcome this limitation by optomechanically decoupling the sample scanning movement from the detection step. Our solution is relatively simple to implement and also provides increased modularity by using a swappable excitation arm. This expands the range of samples we can image on a single system, from high resolution for single cells at ? m spatial resolution, to tissues with mm spatial resolution. We demonstrate our approach, using the system to image iDISCO cleared embryos and sciatic nerves, and provide the full three-dimensional reconstruction of these objects in minutes.

  16. Near-Field Optical Microscopy and Spectroscopy with Pointed Probes

    Science.gov (United States)

    2006-01-01

    metal nanostructure can be viewed as an optical antenna . Of course, the efficiency depends on the material composition and the geometry of the...nanostructure. A simple form of optical antenna is a single ellipsoidal particle. This particle ex- hibits a distinct resonance for which the field...Grober RD, Schoelkopf RJ, Prober DE. 1997. Optical antenna : towards a unity efficiency near-field optical probe. Appl. Phys. Lett. 70:1354 54. Farahani

  17. Optical characterication of probes for photon scanning tunnelling microscopy

    DEFF Research Database (Denmark)

    Vohnsen, Brian; Bozhevolnyi, Sergey I.

    1999-01-01

    The photon scanning tunnelling microscope is a well-established member of the family of scanning near-field optical microscopes used for optical imaging at the sub-wavelength scale. The quality of the probes, typically pointed uncoated optical fibres, used is however difficult to evaluate...

  18. Second-harmonic scanning optical microscopy of semiconductor quantum dots

    DEFF Research Database (Denmark)

    Vohnsen, B.; Bozhevolnyi, S.I.; Pedersen, K.;

    2001-01-01

    Second-harmonic (SH) optical imaging of self-assembled InAlGaAs quantum dots (QD's) grown on a GaAs(0 0 1) substrate has been accomplished at room temperature by use of respectively a scanning far-field optical microscope in reflection mode and a scanning near-field optical microscope...

  19. Nanoscale chromatin structure characterization for optical applications: a transmission electron microscopy study (Conference Presentation)

    Science.gov (United States)

    Li, Yue; Cherkezyan, Lusik; Zhang, Di; Almassalha, Luay; Roth, Eric; Chandler, John; Bleher, Reiner; Subramanian, Hariharan; Dravid, Vinayak P.; Backman, Vadim

    2017-02-01

    Structural and biological origins of light scattering in cells and tissue are still poorly understood. We demonstrate how this problem might be addressed through the use of transmission electron microscopy (TEM). For biological samples, TEM image intensity is proportional to mass-density, and thus proportional to refractive index (RI). By calculating the autocorrelation function (ACF) of TEM image intensity of a thin-section of cells, we essentially maintain the nanoscale ACF of the 3D cellular RI distribution, given that the RI distribution is statistically isotropic. Using this nanoscale 3D RI ACF, we can simulate light scattering through biological samples, and thus guiding many optical techniques to quantify specific structures. In this work, we chose to use Partial Wave Spectroscopy (PWS) microscopy as a one of the nanoscale-sensitive optical techniques. Hela cells were prepared using standard protocol to preserve nanoscale ultrastructure, and a 50-nm slice was sectioned for TEM imaging at 6 nm resolution. The ACF was calculated for chromatin, and the PWS mean sigma was calculated by summing over the power spectral density in the visible light frequency of a random medium generated to match the ACF. A 1-µm slice adjacent to the 50-nm slice was sectioned for PWS measurement to guarantee identical chromatin structure. For 33 cells, we compared the calculated PWS mean sigma from TEM and the value measured directly, and obtained a strong correlation of 0.69. This example indicates the great potential of using TEM measured RI distribution to better understand the quantification of cellular nanostructure by optical methods.

  20. Nanoparticles and pop-off technique for electron microscopy: a known technique for a new purpose.

    Science.gov (United States)

    Lehmbecker, Annika; Rittinghausen, Susanne; Rohn, Kerstin; Baumgärtner, Wolfgang; Schaudien, Dirk

    2014-08-01

    Because of the size of the nanoparticles, their detection and exact anatomical localization in tissue samples are very difficult. Consequently, suitable methods are needed to prove their presence, especially co-localized to histological lesions. Therefore, the aim of this study was to investigate whether nanoparticles were detectable in specimens after reprocessing samples from glass slides using the pop-off technique. Tissue slides containing agglomerates of titanium dioxide nanoparticles already visible on a light microscopic level and amorphous silicium dioxide (SiO2) particles not observable in tissue slides were reprocessed. Furthermore, cytospots of bronchoalveolar lavage acquired from rats that previously inhaled carbon nanotubes were used. After reprocessing the samples, they were investigated using transmission electron microscopy. In all the reprocessed samples, the respective nanoparticles were detectable. Even the light microscopically invisible amorphous SiO2 particles were observed as electron dense structures. Titanium and silicium were additionally confirmed in the respective nanoparticles by energy-dispersive X-ray spectroscopy (EDX). In summary, the pop-off technique represents a fast and easy way to detect nanoparticles in histological sections. This enables further characterization of these particles by additional techniques such as EDX, and their direct correlation with light microscopic lesions at exactly the same location is investigated.

  1. Investigation into spiral phase plate contrast in optical and electron microscopy

    CERN Document Server

    Juchtmans, Roeland; Lubk, Axel; Verbeeck, Jo

    2016-01-01

    The use of phase plates in the back focal plane of a microscope is a well established technique in optical microscopy to increase the contrast of weakly interacting samples and is gaining interest in electron microscopy as well. In this paper we study the spiral phase plate (SPP), also called helical, vortex, or two-dimensional Hilbert phase plate, that adds an angularly dependent phase of the form $e^{i\\ell\\phi}$ to the exit wave in Fourier space. In the limit of large collection angles, we analytically calculate that the average of a pair of $\\ell=\\pm1$ SPP images is directly proportional to the gradient squared of the exit wave, explaining the edge contrast previously seen in optical SPP work. The difference between a clockwise-anticlockwise pair of SPP images and conditions where this difference vanishes and the gradient of the exit wave can be seen from one single SPP image, are discussed. Finally, we demonstrate how with three images, one without and one with each of an $\\ell=\\pm1$ SPP, may give enough ...

  2. An integrated optical coherence microscopy imaging and optical stimulation system for optogenetic pacing in Drosophila melanogaster (Conference Presentation)

    Science.gov (United States)

    Alex, Aneesh; Li, Airong; Men, Jing; Jerwick, Jason; Tanzi, Rudolph E.; Zhou, Chao

    2016-03-01

    Electrical stimulation is the clinical standard for cardiac pacing. Although highly effective in controlling cardiac rhythm, the invasive nature, non-specificity to cardiac tissues and possible tissue damage limits its applications. Optogenetic pacing of the heart is a promising alternative, which is non-invasive and more specific, has high spatial and temporal precision, and avoids the shortcomings in electrical stimulation. Drosophila melanogaster, which is a powerful model organism with orthologs of nearly 75% of human disease genes, has not been studied for optogenetic pacing in the heart. Here, we developed a non-invasive integrated optical pacing and optical coherence microscopy (OCM) imaging system to control the heart rhythm of Drosophila at different developmental stages using light. The OCM system is capable of providing high imaging speed (130 frames/s) and ultrahigh imaging resolutions (1.5 μm and 3.9 μm for axial and transverse resolutions, respectively). A light-sensitive pacemaker was developed in Drosophila by specifically expressing the light-gated cation channel, channelrhodopsin-2 (ChR2) in transgenic Drosophila heart. We achieved non-invasive and specific optical control of the Drosophila heart rhythm throughout the fly's life cycle (larva, pupa, and adult) by stimulating the heart with 475 nm pulsed laser light. Heart response to stimulation pulses was monitored non-invasively with OCM. This integrated non-invasive optogenetic control and in vivo imaging technique provides a novel platform for performing research studies in developmental cardiology.

  3. Fluorescent dyes with large Stokes shifts for super-resolution optical microscopy of biological objects: a review

    Science.gov (United States)

    Sednev, Maksim V.; Belov, Vladimir N.; Hell, Stefan W.

    2015-12-01

    The review deals with commercially available organic dyes possessing large Stokes shifts and their applications as fluorescent labels in optical microscopy based on stimulated emission depletion (STED). STED microscopy breaks Abbe’s diffraction barrier and provides optical resolution beyond the diffraction limit. STED microscopy is non-invasive and requires photostable fluorescent markers attached to biomolecules or other objects of interest. Up to now, in most biology-related STED experiments, bright and photoresistant dyes with small Stokes shifts of 20-40 nm were used. The rapid progress in STED microscopy showed that organic fluorophores possessing large Stokes shifts are indispensable in multi-color super-resolution techniques. The ultimate result of the imaging relies on the optimal combination of a dye, the bio-conjugation procedure and the performance of the optical microscope. Modern bioconjugation methods, basics of STED microscopy, as well as structures and spectral properties of the presently available fluorescent markers are reviewed and discussed. In particular, the spectral properties of the commercial dyes are tabulated and correlated with the available depletion wavelengths found in STED microscopes produced by LEICA Microsytems, Abberior Instruments and Picoquant GmbH.

  4. Tools and techniques to measure mitophagy using fluorescence microscopy.

    Science.gov (United States)

    Dolman, Nick J; Chambers, Kevin M; Mandavilli, Bhaskar; Batchelor, Robert H; Janes, Michael S

    2013-11-01

    Mitophagy is a specialized form of autophagy that removes damaged mitochondria, thereby maintaining efficient cellular metabolism and reducing cellular stress caused by aberrant oxidative bursts. Deficits in mitophagy underlie several diseases, and a substantial body of research has elucidated key steps in the pathways that lead to and execute autophagic clearance of mitochondria. Many of these studies employ fluorescence microscopy to visualize mitochondrial morphology, mass, and functional state. Studies in this area also examine colocalization/recruitment of accessory factors, components of the autophagic machinery and signaling molecules to mitochondria. In this review, we provide a brief summary of the current understanding about the processes involved in mitophagy followed by a discussion of probes commonly employed and important considerations of the methodologies to study and analyze mitophagy using fluorescence microscopy. Representative data, where appropriate, are provided to highlight the use of key probes to monitor mitophagy. The review will conclude with a consideration of new possibilities for mitophagy research and a discussion of recently developed technologies for this emerging area of cell biology.

  5. Comparison of mouse mammary gland imaging techniques and applications: Reflectance confocal microscopy, GFP Imaging, and ultrasound

    Directory of Open Access Journals (Sweden)

    Cotarla Ion

    2008-01-01

    Full Text Available Abstract Background Genetically engineered mouse models of mammary gland cancer enable the in vivo study of molecular mechanisms and signaling during development and cancer pathophysiology. However, traditional whole mount and histological imaging modalities are only applicable to non-viable tissue. Methods We evaluated three techniques that can be quickly applied to living tissue for imaging normal and cancerous mammary gland: reflectance confocal microscopy, green fluorescent protein imaging, and ultrasound imaging. Results In the current study, reflectance confocal imaging offered the highest resolution and was used to optically section mammary ductal structures in the whole mammary gland. Glands remained viable in mammary gland whole organ culture when 1% acetic acid was used as a contrast agent. Our application of using green fluorescent protein expressing transgenic mice in our study allowed for whole mammary gland ductal structures imaging and enabled straightforward serial imaging of mammary gland ducts in whole organ culture to visualize the growth and differentiation process. Ultrasound imaging showed the lowest resolution. However, ultrasound was able to detect mammary preneoplastic lesions 0.2 mm in size and was used to follow cancer growth with serial imaging in living mice. Conclusion In conclusion, each technique enabled serial imaging of living mammary tissue and visualization of growth and development, quickly and with minimal tissue preparation. The use of the higher resolution reflectance confocal and green fluorescent protein imaging techniques and lower resolution ultrasound were complementary.

  6. Simulation, Analysis, and Fabrication of Miniaturized Components with Applications in Optical Interconnects and Parallel Microscopy

    OpenAIRE

    Wohlfeld, Denis

    2009-01-01

    Optics and miniaturization of components are both key technologies supporting the progress in many multidisciplinary areas and enabling products with more functionality at reduced size and costs. Two applications of microscoptic integrations in the field of optical communications and parallel microscopy are investigated. This thesis also deals with light propagation in geometrical optics and scalar wave optics, both in homogeneous and inhomogeneous media. The fabrication and optimization of ...

  7. Bases for time-resolved probing of transient carrier dynamics by optical pump-probe scanning tunneling microscopy.

    Science.gov (United States)

    Yokota, Munenori; Yoshida, Shoji; Mera, Yutaka; Takeuchi, Osamu; Oigawa, Haruhiro; Shigekawa, Hidemi

    2013-10-07

    The tangled mechanism that produces optical pump-probe scanning tunneling microscopy spectra from semiconductors was analyzed by comparing model simulation data with experimental data. The nonlinearities reflected in the spectra, namely, the excitations generated by paired laser pulses with a delay time, the logarithmic relationship between carrier density and surface photovoltage (SPV), and the effect of the change in tunneling barrier height depending on SPV, were examined along with the delay-time-dependent integration process used in measurement. The optimum conditions required to realize reliable measurement, as well as the validity of the microscopy technique, were demonstrated for the first time.

  8. Dimensional metrology of lab-on-a-chip internal structures: a comparison of optical coherence tomography with confocal fluorescence microscopy.

    Science.gov (United States)

    Reyes, D R; Halter, M; Hwang, J

    2015-07-01

    The characterization of internal structures in a polymeric microfluidic device, especially of a final product, will require a different set of optical metrology tools than those traditionally used for microelectronic devices. We demonstrate that optical coherence tomography (OCT) imaging is a promising technique to characterize the internal structures of poly(methyl methacrylate) devices where the subsurface structures often cannot be imaged by conventional wide field optical microscopy. The structural details of channels in the devices were imaged with OCT and analyzed with an in-house written ImageJ macro in an effort to identify the structural details of the channel. The dimensional values obtained with OCT were compared with laser-scanning confocal microscopy images of channels filled with a fluorophore solution. Attempts were also made using confocal reflectance and interferometry microscopy to measure the channel dimensions, but artefacts present in the images precluded quantitative analysis. OCT provided the most accurate estimates for the channel height based on an analysis of optical micrographs obtained after destructively slicing the channel with a microtome. OCT may be a promising technique for the future of three-dimensional metrology of critical internal structures in lab-on-a-chip devices because scans can be performed rapidly and noninvasively prior to their use.

  9. Modeling the Effect of Wave-front Aberrations in Fiber-based Scanning Optical Microscopy

    NARCIS (Netherlands)

    Verstraete, H.R.G.W.; Verhaegen, M.H.G.; Kalkman, J.

    2013-01-01

    In scanning microscopy and optical coherence tomography, aberrations of the wave-front cause a loss in intensity and resolution. Intensity and resolution are quantified using Fresnel propagation, Fraunhofer diffraction, and the calculation of overlap integrals.

  10. Possibilities and Challenges of Scanning Hard X-ray Spectro-microscopy Techniques in Material Sciences

    Directory of Open Access Journals (Sweden)

    Andrea Somogyi

    2015-06-01

    Full Text Available Scanning hard X-ray spectro-microscopic imaging opens unprecedented possibilities in the study of inhomogeneous samples at different length-scales. It gives insight into the spatial variation of the major and minor components, impurities and dopants of the sample, and their chemical and electronic states at micro- and nano-meter scales. Measuring, modelling and understanding novel properties of laterally confined structures are now attainable. The large penetration depth of hard X-rays (several keV to several 10 keV beam energy makes the study of layered and buried structures possible also in in situ and in operando conditions. The combination of different X-ray analytical techniques complementary to scanning spectro-microscopy, such as X-ray diffraction, X-ray excited optical luminescence, secondary ion mass spectrometry (SIMS and nano-SIMS, provides access to optical characteristics and strain and stress distributions. Complex sample environments (temperature, pressure, controlled atmosphere/vacuum, chemical environment are also possible and were demonstrated, and allow as well the combination with other analysis techniques (Raman spectroscopy, infrared imaging, mechanical tensile devices, etc. on precisely the very same area of the sample. The use of the coherence properties of X-rays from synchrotron sources is triggering emerging experimental imaging approaches with nanometer lateral resolution. New fast analytical possibilities pave the way towards statistically significant studies at multi- length-scales and three dimensional tomographic investigations. This paper gives an overview of these techniques and their recent achievements in the field of material sciences.

  11. Study on metal nanoparticles induced third-order optical nonlinearity in phenylhydrazone derivatives with DFWM technique

    Science.gov (United States)

    Sudheesh, P.; Rao, D. Mallikharjuna; Chandrasekharan, K.

    2014-01-01

    The third-order nonlinear optical properties of newly synthesized phenylhydrazone derivatives and the influence of noble metal nanoparticles (Ag & Au) on their nonlinear optical responses were investigated by employing Degenerate Four wave Mixing (DFWM) technique with a 7 nanosecond, 10Hz Nd: YAG laser pulses at 532nm. Metal nanoparticles were prepared by laser ablation and the particle formation was confirmed using UV-Visible spectroscopy, Transmission Electron Microscopy (TEM). The nonlinear optical susceptibility were measured and found to be of the order 10-13esu. The results are encouraging and conclude that the materials are promising candidate for future optical device applications.

  12. Nonlinear photoacoustic microscopy via a loss modulation technique: from detection to imaging.

    Science.gov (United States)

    Lai, Yu-Hung; Lee, Szu-Yu; Chang, Chieh-Feng; Cheng, Yu-Hsiang; Sun, Chi-Kuang

    2014-01-13

    In order to achieve high-resolution deep-tissue imaging, multi-photon fluorescence microscopy and photoacoustic tomography had been proposed in the past two decades. However, combining the advantages of these two imaging systems to achieve optical-spatial resolution with an ultrasonic-penetration depth is still a field with challenges. In this paper, we investigate the detection of the two-photon photoacoustic ultrasound, and first demonstrate background-free two-photon photoacoustic imaging in a phantom sample. To generate the background-free two-photon photoacoustic signals, we used a high-repetition rate femtosecond laser to induce narrowband excitation. Combining a loss modulation technique, we successfully created a beating on the light intensity, which not only provides pure sinusoidal modulation, but also ensures the spectrum sensitivity and frequency selectivity. By using the lock-in detection, the power dependency experiment validates our methodology to frequency-select the source of the nonlinearity. This ensures our capability of measuring the background-free two-photon photoacoustic waves by detecting the 2nd order beating signal directly. Furthermore, by mixing the nanoparticles and fluorescence dyes as contrast agents, the two-photon photoacoustic signal was found to be enhanced and detected. In the end, we demonstrate subsurface two-photon photoacoustic bio-imaging based on the optical scanning mechanism inside phantom samples.

  13. Investigating the transverse optical structure of spider silk micro-fibers using quantitative optical microscopy

    Science.gov (United States)

    Little, Douglas J.; Kane, Deb M.

    2017-01-01

    The transverse optical structure of two orb-weaver (family Araneidae) spider dragline silks was investigated using a variant of the inverse-scattering technique. Immersing the silks in a closely refractive index-matched liquid, the minimum achievable image contrast was greater than expected for an optically homogeneous silk, given what is currently known about the optical absorption of these silks. This "excess contrast" indicated the presence of transverse optical structure within the spider silk. Applying electromagnetic scattering theory to a transparent double cylinder, the minimum achievable irradiance contrast for the Plebs eburnus and Argiope keyserlingi dragline silks was determined to be consistent with step index refractive index contrasts of 1-4×10-4 and 6-7×10-4, respectively, supposing outer-layer thicknesses consistent with previous TEM studies (50 nm and 100 nm, respectively). The possibility of graded index refractive index contrasts within the spider silks is also discussed. This is the strongest evidence, to date, that there is a refractive index contrast associated with the layered morphology of spider silks and/or variation of proportion of nanocrystalline components within the spider silk structure. The method is more generally applicable to optical micro-fibers, including those with refractive index variations on a sub-wavelength scale.

  14. Investigating the transverse optical structure of spider silk micro-fibers using quantitative optical microscopy

    Directory of Open Access Journals (Sweden)

    Little Douglas J.

    2016-10-01

    Full Text Available The transverse optical structure of two orb-weaver (family Araneidae spider dragline silks was investigated using a variant of the inverse-scattering technique. Immersing the silks in a closely refractive index-matched liquid, the minimum achievable image contrast was greater than expected for an optically homogeneous silk, given what is currently known about the optical absorption of these silks. This “excess contrast” indicated the presence of transverse optical structure within the spider silk. Applying electromagnetic scattering theory to a transparent double cylinder, the minimum achievable irradiance contrast for the Plebs eburnus and Argiope keyserlingi dragline silks was determined to be consistent with step index refractive index contrasts of 1−4×10−4 and 6–7×10−4, respectively, supposing outer-layer thicknesses consistent with previous TEM studies (50 nm and 100 nm, respectively. The possibility of graded index refractive index contrasts within the spider silks is also discussed. This is the strongest evidence, to date, that there is a refractive index contrast associated with the layered morphology of spider silks and/or variation of proportion of nanocrystalline components within the spider silk structure. The method is more generally applicable to optical micro-fibers, including those with refractive index variations on a sub-wavelength scale.

  15. Near-Field Optical Microscopy of Fractal Structures

    DEFF Research Database (Denmark)

    Coello, Victor; Bozhevolnyi, Sergey I.

    1999-01-01

    Using a photon scanning tunnelling microscope combined with a shear-force feedback system, we image both topographical and near-field optical images (at the wavelengths of 633 and 594 nm) of silver colloid fractals. Near-field optical imaging is calibrated with a standing evanescent wave pattern...

  16. Acousto-optic tuneable filters: advances and applications to microscopy

    Science.gov (United States)

    Pannell, C. N.; Wachman, E. S.; Farkas, D. L.; Ward, J.; Seale, W.

    2006-02-01

    The acousto-optic tunable filter (AOTF) is one example of a small number of commercially available optical filter technologies that lend themselves to imaging applications. In recent years the demand for high specification devices has increased significantly, and diffraction limited performance is being achieved.

  17. Differentiation of black gel inks using optical and chemical techniques.

    Science.gov (United States)

    Wilson, Jeffrey D; LaPorte, Gerald M; Cantu, Antonio A

    2004-03-01

    Gel ink pens have become a common writing instrument in the United States. Questioned document examiners often attempt to optically differentiate gel inks from each other and from other non-ballpoint ink writings (e.g., those from roller-ball pens). Since early formulations were primarily pigment-based, they do not elute when analyzed by thin-layer chromatography. However, recent gel ink formulations (i.e., within the past five years) include dye-based inks that can be easily separated. This study differentiates black gel inks using optical and chemical techniques. The techniques include: microscopy, visible and near infrared reflectance, near infrared luminescence, thin-layer chromatography (TLC), spot tests, and gas chromatography/mass spectrometry (GC/MS). As a result of this study a flow chart has been developed allowing for a systematic determination of a questioned ink. In addition, an analysis of volatile compounds found in gel inks revealed that there are some unique ingredients that may be found in gel inks that are not typically found in other non-ballpoint inks.

  18. Multiphoton microscopy, fluorescence lifetime imaging and optical spectroscopy for the diagnosis of neoplasia

    Science.gov (United States)

    Skala, Melissa Caroline

    2007-12-01

    Cancer morbidity and mortality is greatly reduced when the disease is diagnosed and treated early in its development. Tissue biopsies are the gold standard for cancer diagnosis, and an accurate diagnosis requires a biopsy from the malignant portion of an organ. Light, guided through a fiber optic probe, could be used to inspect regions of interest and provide real-time feedback to determine the optimal tissue site for biopsy. This approach could increase the diagnostic accuracy of current biopsy procedures. The studies in this thesis have characterized changes in tissue optical signals with carcinogenesis, increasing our understanding of the sensitivity of optical techniques for cancer detection. All in vivo studies were conducted on the dimethylbenz[alpha]anthracene treated hamster cheek pouch model of epithelial carcinogenesis. Multiphoton microscopy studies in the near infrared wavelength region quantified changes in tissue morphology and fluorescence with carcinogenesis in vivo. Statistically significant morphological changes with precancer included increased epithelial thickness, loss of stratification in the epithelium, and increased nuclear diameter. Fluorescence changes included a statistically significant decrease in the epithelial fluorescence intensity per voxel at 780 nm excitation, a decrease in the fluorescence lifetime of protein-bound nicotinamide adenine dinucleotide (NADH, an electron donor in oxidative phosphorylation), and an increase in the fluorescence lifetime of protein-bound flavin adenine dinucleotide (FAD, an electron acceptor in oxidative phosphorylation) with precancer. The redox ratio (fluorescence intensity of FAD/NADH, a measure of the cellular oxidation-reduction state) did not significantly change with precancer. Cell culture experiments (MCF10A cells) indicated that the decrease in protein-bound NADH with precancer could be due to increased levels of glycolysis. Point measurements of diffuse reflectance and fluorescence spectra in

  19. Three-dimensional microscopy and sectional image reconstruction using optical scanning holography.

    Science.gov (United States)

    Lam, Edmund Y; Zhang, Xin; Vo, Huy; Poon, Ting-Chung; Indebetouw, Guy

    2009-12-01

    Fast acquisition and high axial resolution are two primary requirements for three-dimensional microscopy. However, they are sometimes conflicting: imaging modalities such as confocal imaging can deliver superior resolution at the expense of sequential acquisition at different axial planes, which is a time-consuming process. Optical scanning holography (OSH) promises to deliver a good trade-off between these two goals. With just a single scan, we can capture the entire three-dimensional volume in a digital hologram; the data can then be processed to obtain the individual sections. An accurate modeling of the imaging system is key to devising an appropriate image reconstruction algorithm, especially for real data where random noise and other imaging imperfections must be taken into account. In this paper we demonstrate sectional image reconstruction by applying an inverse imaging sectioning technique to experimental OSH data of biological specimens and visualizing the sections using the OSA Interactive Science Publishing software.

  20. Super-resolution optical microscopy study of telomere structure

    Science.gov (United States)

    Phipps, Mary Lisa; Goodwin, Peter M.; Martinez, Jennifer S.; Goodwin, Edwin H.

    2016-09-01

    Chromosome ends are shielded from exonucleolytic attack and inappropriate end-joining by terminal structures called telomeres; these structures are potential targets for anticancer drugs. Telomeres are composed of a simple DNA sequence (5‧-TTAGGG-3‧ in humans) repeated more than a thousand times, a short 3‧ single-stranded overhang, and numerous proteins. Electron microscopy has shown that the 3‧ overhang pairs with the complementary strand at an internal site creating a small displacement loop and a large double-stranded "t-loop." Our goal is to determine whether all telomeres adopt the t-loop configuration, or whether there are two or more distinct configurations. Progress in optimizing super-resolution (SR) microscopy for this ongoing investigation is reported here. Results suggest that under certain conditions sample preparation procedures may disrupt chromatin by causing loss of nucleosomes. This finding may limit the use of SR microscopy in telomere studies.

  1. Programmable Colored Illumination Microscopy (PCIM): A practical and flexible optical staining approach for microscopic contrast enhancement

    Science.gov (United States)

    Zuo, Chao; Sun, Jiasong; Feng, Shijie; Hu, Yan; Chen, Qian

    2016-03-01

    Programmable colored illumination microscopy (PCIM) has been proposed as a flexible optical staining technique for microscopic contrast enhancement. In this method, we replace the condenser diaphragm of a conventional microscope with a programmable thin film transistor-liquid crystal display (TFT-LCD). By displaying different patterns on the LCD, numerous established imaging modalities can be realized, such as bright field, dark field, phase contrast, oblique illumination, and Rheinberg illuminations, which conventionally rely on intricate alterations in the respective microscope setups. Furthermore, the ease of modulating both the color and the intensity distribution at the aperture of the condenser opens the possibility to combine multiple microscopic techniques, or even realize completely new methods for optical color contrast staining, such as iridescent dark-field and iridescent phase-contrast imaging. The versatility and effectiveness of PCIM is demonstrated by imaging of several transparent colorless specimens, such as unstained lung cancer cells, diatom, textile fibers, and a cryosection of mouse kidney. Finally, the potentialities of PCIM for RGB-splitting imaging with stained samples are also explored by imaging stained red blood cells and a histological section.

  2. Simultaneous topographical, electrical and optical microscopy of optoelectronic devices at the nanoscale

    KAUST Repository

    Kumar, Naresh

    2017-01-12

    Novel optoelectronic devices rely on complex nanomaterial systems where the nanoscale morphology and local chemical composition are critical to performance. However, the lack of analytical techniques that can directly probe these structure-property relationships at the nanoscale presents a major obstacle to device development. In this work, we present a novel method for non-destructive, simultaneous mapping of the morphology, chemical composition and photoelectrical properties with <20 nm spatial resolution by combining plasmonic optical signal enhancement with electrical-mode scanning probe microscopy. We demonstrate that this combined approach offers subsurface sensitivity that can be exploited to provide molecular information with a nanoscale resolution in all three spatial dimensions. By applying the technique to an organic solar cell device, we show that the inferred surface and subsurface composition distribution correlates strongly with the local photocurrent generation and explains macroscopic device performance. For instance, the direct measurement of fullerene phase purity can distinguish between high purity aggregates that lead to poor performance and lower purity aggregates (fullerene intercalated with polymer) that result in strong photocurrent generation and collection. We show that the reliable determination of the structure-property relationship at the nanoscale can remove ambiguity from macroscopic device data and support the identification of the best routes for device optimisation. The multi-parameter measurement approach demonstrated herein is expected to play a significant role in guiding the rational design of nanomaterial-based optoelectronic devices, by opening a new realm of possibilities for advanced investigation via the combination of nanoscale optical spectroscopy with a whole range of scanning probe microscopy modes.

  3. All-optical histology using two photon laser scanning microscopy and ablation with ultrashort pulses

    Science.gov (United States)

    Tsai, Philbert S.

    This dissertation discusses the use of ultrashort laser pulses to image and manipulate tissue for the purpose of three-dimensional histological reconstruction of extended brain structures. Two photon laser scanning microscopy (TPLSM) and ultrashort pulsed laser ablation are used to provide in situ three-dimensional imaging through thick preparations of fixed tissue. Surface regions of fixed tissue are first imaged using TPLSM. The imaged regions are then removed by ablation with amplified, ultrashort laser pulses, thereby exposing a previously underlying tissue region for imaging. This process of imaging and ablation proceeds iteratively until the desired tissue volume has been processed. First, the principles, design, and construction of a two photon laser scanning microscope are discussed, followed by a discussion of the physical mechanisms of tissue ablation with ultrashort laser pulses. The compatibility of tissue ablation using ultrashort pulses with subsequent histological analysis, particularly with fluorescent microscopy, is evaluated. Tissue ablation with ultrashort laser pulses is found to produce ablated tissue surfaces that are smooth to within a micrometer. Intrinsic fluorescence as well as immunoreactivity are found to be resilient to the ablation process. The all-optical histological technique is demonstrated on brain tissue from rats and mice, including tissue from embryonic mouse as early at E15. The ablation process is shown to preserve both macroscopic and microscopic structures within tissue. To facilitate the all-optical histological analysis of neuronal vasculature and its relative distribution to surrounding neuronal tissue, a fluorescent gel perfusion technique is developed that provides a temperature-stabilized fluorescent label of the neuronal vasculature. The use of immunohistochemistry to label specific cell populations throughout an 800 micrometer-thick tissue section is demonstrated. Additionally, the immersion of fixed tissue in high

  4. The crocidolite fibres interaction with human mesothelial cells as investigated by combining electron microscopy, atomic force and scanning near-field optical microscopy.

    Science.gov (United States)

    Andolfi, Laura; Trevisan, Elisa; Zweyer, Marina; Prato, Stefano; Troian, Barbara; Vita, Francesca; Borelli, Violetta; Soranzo, Maria Rosa; Melato, Mauro; Zabucchi, Giuliano

    2013-03-01

    In this study, we have performed a morphological analysis of crocidolite fibres interaction with mesothelial cells (MET5A) by combining conventional electron microscopy with atomic force (AFM) and scanning near-field optical microscopy (SNOM). After 6-h exposure at a crocidolite dose of 5 μg cm(-2), 90% of MET5A cells interact with fibres that under these conditions have a low cytotoxic effect. SEM images point out that fibres can be either engulfed by the cells that lose their typical morphology or they can accumulate over or partially inside the cells, which preserve their typical spread morphology. By using AFM we are able to directly visualize the entry-site of nanometric-sized fibres at the plasma membrane of the spread mesothelial cells. More importantly, the crocidolite fibres that are observed to penetrate the plasma membrane in SNOM topography can be simultaneously followed beneath the cell surface in the SNOM optical images. The analysis of SNOM data demonstrates the entrance of crocidolite fibres in proximity of nuclear compartment, as observed also in the TEM images. Our findings indicate that the combination of conventional electron microscopy with novel nanoscopic techniques can be considered a promising approach to achieve a comprehensive morphological description of the interaction between asbestos fibres and mesothelial cells that represents the early event in fibre pathogenesis.

  5. Microscopic techniques bridging between nanoscale and microscale with an atomically sharpened tip - field ion microscopy/scanning probe microscopy/ scanning electron microscopy.

    Science.gov (United States)

    Tomitori, Masahiko; Sasahara, Akira

    2014-11-01

    Over a hundred years an atomistic point of view has been indispensable to explore fascinating properties of various materials and to develop novel functional materials. High-resolution microscopies, rapidly developed during the period, have taken central roles in promoting materials science and related techniques to observe and analyze the materials. As microscopies with the capability of atom-imaging, field ion microscopy (FIM), scanning tunneling microscopy (STM), atomic force microscopy (AFM) and transmission electron microscopy (TEM) can be cited, which have been highly evaluated as methods to ultimately bring forward the viewpoint of reductionism in materials science. On one hand, there have been difficulties to derive useful and practical information on large (micro) scale unique properties of materials using these excellent microscopies and to directly advance the engineering for practical materials. To make bridges over the gap between an atomic scale and an industrial engineering scale, we have to develop emergence science step-by-step as a discipline having hierarchical structures for future prospects by combining nanoscale and microscale techniques; as promising ways, the combined microscopic instruments covering the scale gap and the extremely sophisticated methods for sample preparation seem to be required. In addition, it is noted that spectroscopic and theoretical methods should implement the emergence science.Fundamentally, the function of microscope is to determine the spatial positions of a finite piece of material, that is, ultimately individual atoms, at an extremely high resolution with a high stability. To define and control the atomic positions, the STM and AFM as scanning probe microscopy (SPM) have successfully demonstrated their power; the technological heart of SPM lies in an atomically sharpened tip, which can be observed by FIM and TEM. For emergence science we would like to set sail using the tip as a base. Meanwhile, it is significant

  6. Improved 3D Superresolution Localization Microscopy Using Adaptive Optics

    CERN Document Server

    Piro, Nicolas; Olivier, Nicolas; Manley, Suliana

    2014-01-01

    We demonstrate a new versatile method for 3D super-resolution microscopy by using a deformable mirror to shape the point spread function of our microscope in a continuous and controllable way. We apply this for 3D STORM imaging of microtubules.

  7. DAPI staining and fluorescence microscopy techniques for phytoplasmas.

    Science.gov (United States)

    Andrade, Nancy M; Arismendi, Nolberto L

    2013-01-01

    The 4',6-diamidino-2-phenylindole (DAPI) stain technique is a simple method that was developed for confirming the presence of phytoplasmas in hand-cut or freezing microtome sections of infected tissues. DAPI binds AT-rich DNA preferentially, so that phytoplasmas, localized among phloem cells, can be visualized in a fluorescence microscope. The procedure is quick, easy to use, inexpensive, and can be used as a preliminary or quantitative method to detect or quantify phytoplasma-like bodies in infected plants.

  8. Spectroelectrochemistry: The Combination of Optical and Electrochemical Techniques.

    Science.gov (United States)

    Heineman, William R.

    1983-01-01

    Two different techniques, electrochemistry and spectroscopy, can be combined for studying the redox chemistry of inorganic, organic, and biological molecules. Several commonly used spectroelectrochemical methods and their applications are described. Includes discussions of optically transparent electrodes, optical absorption/fluorescence…

  9. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    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.

  10. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    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.

  11. Lasers, lenses and light curves : adaptive optics microscopy and peculiar transiting exoplanets

    NARCIS (Netherlands)

    Werkhoven, Theodorus Isaak Mattheus van

    2014-01-01

    In the first part of this thesis, we present an adaptive optics implementation for multi-photon microscopy correcting sample-induced wavefront aberrations using either direct wavefront sensing to run a close-loop adaptive optics system (Chapter 3), or use a model-based sensorless approach to iterati

  12. Heterodyne method of apertureless near-field scanning optical microscopy on periodic gold nanowells.

    Energy Technology Data Exchange (ETDEWEB)

    Hall, J. E.; Wiederrecht, G. P.; Gray, S. K.; Chang, S.-H.; Jeon, S.; Rogers, J. A.; Bachelot, R.; Royer, P.; Univ. of Illinois; Univ. of Technology at Troyes; Inst. of Electro-Optical Science and Engineering

    2007-04-02

    Heterodyne detection for apertureless near-field scanning optical microscopy was used to study periodic gold nanowell arrays. Optical near-field amplitude and phase signals were obtained simultaneously with the topography of the gold nanowells and with different polarizations. Theoretical calculations of the near-fields were consistent with the experiments; in particular, the calculated amplitudes were in especially good agreement. The heterodyne method is shown to be particularly effective for these types of periodic photonic structures and other highly scattering media, which can overwhelm the near-field scattered signal when conventional apertureless near-field scanning optical microscopy is used.

  13. All-optical thermal microscopy of laser-excited waveguides

    OpenAIRE

    He, R.; De Aldana, J.R.V.; Pedrola, G.L.; Chen, F.; JAQUE, D.

    2016-01-01

    We report on a unique combination of high-resolution confocal microscopy and ratiometric luminescence thermometry to obtain thermal images of 800 nm pumped ultrafast laser-inscribed waveguides in a Nd:YAG crystal. Thermal images evidence a strong localization of thermal load in the waveguide active volume. Comparison between experimental data and numerical simulations reveals that ultrafast laser-inscribed damage tracks in Nd:YAG crystals behave both as low-index and low-thermal conductivity ...

  14. Optical far-field super-resolution microscopy using nitrogen vacancy center ensemble in bulk diamond

    OpenAIRE

    Li, Shen; Chen, Xiang-Dong; Zhao, Bo-Wen; Dong, Yang; Zou, Chong-Wen; Guo, Guang-Can; Sun, Fang-Wen

    2016-01-01

    We demonstrate an optical far-field super-resolution microscopy using array of nitrogen vacancy centers in bulk diamond as near-field optical probes. The local optical field, which transmits through the nanostructures on the diamond surface, is measured by detecting the charge state conversion of nitrogen vacancy center. And the locating of nitrogen vacancy center with spatial resolution of 6.1 nm is realized with the charge state depletion nanoscopy. The nanostructures on the surface of diam...

  15. Ultra-precise measurement of optical aberrations for sub-Aangstroem transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Barthel, J.

    2008-06-15

    Quantitative investigations of material structures on an atomic scale by means of highresolution transmission electron microscopy (HRTEM) impose not only extreme demands on the mechanic and electromagnetic stability of the applied instruments but require also their precise electron-optical adjustment. Today a physical resolution well below one Aangstroem can be achieved with commercially available microscopes on a daily basis. However, the achieved resolution can often not be reliably exploited for the interpretation of the resulting microscopical data due to the presence of so-called higher-order lens aberrations. At the starting time of this work, a sufficiently accurate procedure to measure higher-order aberrations was urgently missing. Since aberration measurement is a mandatory prerequisite for any technique of aberration control enabling quantitative high-resolution microscopy, the goal of this work is to develop such a measurement procedure for the Sub-Aangstroem regime. The measurement procedures developed in the course of this work are based on the numerical evaluation of a series of images taken from an amorphous object under electron-beam illumination with varying tilt. New techniques have been developed for the evaluation of single images as well as for the optimised evaluation of the whole series. These procedures allow microscope users to perform quantitative HRTEM even at a resolution of 0.5 Aangstroem. The precision reached with the newly developed measurement procedures is unprecedented and surpasses existing solutions by at least one order of magnitude in any respect. All the concepts and procedures for aberration measurement developed in this work have been implemented in a software package which satisfies professional demands with respect to robustness, precision, speed and user-friendliness. The new automatic aberrationmeasurement procedures are suitable to establish HRTEM as a quantitative technique for material science investigations in the

  16. Measurement of intracellular calcium gradients in single living cells using optical sectioning microscopy

    Science.gov (United States)

    Yelamarty, Rao V.; Cheung, Joseph Y.

    1992-06-01

    Intracellular free calcium has been recognized as a regulator of many cellular processes and plays a key role in mediating actions of many drugs. To elucidate subcellular spatial calcium changes throughout the cell in three dimensions (3-D), optical sectioning microscopy was applied using digital imaging coupled fluorescence microscopy. The cell was loaded with a fluorescent indicator, fura-2, and a stack of sectional fluorescent images were acquired, digitized and finally stored on-line for post image analysis. Each sectional image was then deconvolved, to remove contaminating light signals from adjacent planes, using the Nearest Neighboring Deconvolution Algorithm (NNDA) and the overall imaging system's empirical Point Spread Function (PSF) that is measured with a 0.25 micrometers fluorescent bead. Using this technique, we measured that the addition of growth factors caused a 2 - 3 fold increase (1) in nuclear calcium compared to cytosolic calcium in blood cells and (2) in both nuclear and cytosolic calcium in liver cells. Such spatial information, which is important in understanding subcellular processes, would not be possible to measure with other methods.

  17. Surface diffusion studies by optical diffraction techniques

    Energy Technology Data Exchange (ETDEWEB)

    Xiao, X.D.

    1992-11-01

    The newly developed optical techniques have been combined with either second harmonic (SH) diffraction or linear diffraction off a monolayer adsorbate grating for surface diffusion measurement. Anisotropy of surface diffusion of CO on Ni(l10) was used as a demonstration for the second harmonic dim reaction method. The linear diffraction method, which possesses a much higher sensitivity than the SH diffraction method, was employed to study the effect of adsorbate-adsorbate interaction on CO diffusion on Ni(l10) surface. Results showed that only the short range direct CO-CO orbital overlapping interaction influences CO diffusion but not the long range dipole-dipole and CO-NI-CO interactions. Effects of impurities and defects on surface diffusion were further explored by using linear diffraction method on CO/Ni(110) system. It was found that a few percent S impurity can alter the CO diffusion barrier height to a much higher value through changing the Ni(110) surface. The point defects of Ni(l10) surface seem to speed up CO diffusion significantly. A mechanism with long jumps over multiple lattice distance initiated by CO filled vacancy is proposed to explain the observed defect effect.

  18. Removing lateral chromatic aberration in bright field optical microscopy.

    Science.gov (United States)

    Guzmán-Altamirano, Miguel; Gutiérrez-Medina, Braulio

    2015-06-01

    We present an efficient alternative to remove lateral chromatic aberration (LCA) in bright field light microscopy images. Our procedure is based on error calibration using time-sequential acquisition at different wavelengths, and error correction through digital image warping. Measurement of the displacements of fiducial marks in the red and green images relative to blue provide calibration factors that are subsequently used in test images to realign color channels digitally. We demonstrate quantitative improvement in the position and boundaries of objects in target slides and in the color content and morphology of specimens in stained biological samples. Our results show a reduction of LCA content below the 0.1% level.

  19. Characterization of nodular and thermal defects in hafnia/silica multilayer coatings using optical, photothermal, and atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Stolz, C.J.; Yoshiyama, J.M.; Salleo, A.; Wu, Z.L.; Green, J.; Krupka, R.

    1997-12-24

    Multilayer coatings manufactured from metallic hafnium and silica sources by reactive electron beam deposition, are being developed for high fluence optics in a fusion laser with a wavelength of 1053 nm and a 3 ns pulse length. Damage threshold studies have revealed a correlation between laser damage and nodular defects, but interestingly laser damage is also present in nodule-free regions. Photothermal studies of optical coatings reveal the existence of defects with strong optical absorption in nodule-free regions of the coating. A variety of microscopic techniques were employed to characterize the effects for a better understanding of the thermal properties of nodular defects and role of thermal defects in laser damage. Photothermal microscopy, utilizing the surface thermal lensing technique, was used to map the thermal characteristics of 3 mm x 3 mm areas of the coatings. High resolution subaperture scans, with a 1 pm step size and a 3 um pump beam diameter, W= conducted on the defects to characterize their photothermal properties. Optical and atomic force microscopy was used to visually identify defects and characterize their topography. The defects were then irradiated to determine the role of nodular and thermal defects in limiting the damage threshold of the multilayer.

  20. A minimal optical trapping and imaging microscopy system.

    Directory of Open Access Journals (Sweden)

    Carmen Noemí Hernández Candia

    Full Text Available We report the construction and testing of a simple and versatile optical trapping apparatus, suitable for visualizing individual microtubules (∼25 nm in diameter and performing single-molecule studies, using a minimal set of components. This design is based on a conventional, inverted microscope, operating under plain bright field illumination. A single laser beam enables standard optical trapping and the measurement of molecular displacements and forces, whereas digital image processing affords real-time sample visualization with reduced noise and enhanced contrast. We have tested our trapping and imaging instrument by measuring the persistence length of individual double-stranded DNA molecules, and by following the stepping of single kinesin motor proteins along clearly imaged microtubules. The approach presented here provides a straightforward alternative for studies of biomaterials and individual biomolecules.

  1. Depth-Encoded Spectral Domain Phase Microscopy for Simultaneous Multi-Site Nanoscale Optical Measurements.

    Science.gov (United States)

    Hendargo, Hansford C; Bower, Bradley A; Reinstein, Alex S; Shepherd, Neal; Tao, Yuankai K; Izatt, Joseph A

    2011-09-01

    Spectral domain phase microscopy (SDPM) is an extension of spectral domain optical coherence tomography (SDOCT) that exploits the extraordinary phase stability of spectrometer-based systems with common-path geometry to resolve sub-wavelength displacements within a sample volume. This technique has been implemented for high resolution axial displacement and velocity measurements in biological samples, but since axial displacement information is acquired serially along the lateral dimension, it has been unable to measure fast temporal dynamics in extended samples. Depth-Encoded SDPM (DESDPM) uses multiple sample arms with unevenly spaced common path reference reflectors to multiplex independent SDPM signals from separate lateral positions on a sample simultaneously using a single interferometer, thereby reducing the time required to detect unique optical events to the integration period of the detector. Here, we introduce DESDPM and demonstrate the ability to acquire useful phase data concurrently at two laterally separated locations in a phantom sample as well as a biological preparation of spontaneously beating chick cardiomyocytes. DESDPM may be a useful tool for imaging fast cellular phenomena such as nervous conduction velocity or contractile motion.

  2. Refractometry of melanocyte cell nuclei using optical scatter images recorded by digital Fourier microscopy.

    Science.gov (United States)

    Seet, Katrina Y T; Nieminen, Timo A; Zvyagin, Andrei V

    2009-01-01

    The cell nucleus is the dominant optical scatterer in the cell. Neoplastic cells are characterized by cell nucleus polymorphism and polychromism-i.e., the nuclei exhibits an increase in the distribution of both size and refractive index. The relative size parameter, and its distribution, is proportional to the product of the nucleus size and its relative refractive index and is a useful discriminant between normal and abnormal (cancerous) cells. We demonstrate a recently introduced holographic technique, digital Fourier microscopy (DFM), to provide a sensitive measure of this relative size parameter. Fourier holograms were recorded and optical scatter of individual scatterers were extracted and modeled with Mie theory to determine the relative size parameter. The relative size parameter of individual melanocyte cell nuclei were found to be 16.5+/-0.2, which gives a cell nucleus refractive index of 1.38+/-0.01 and is in good agreement with previously reported data. The relative size parameters of individual malignant melanocyte cell nuclei are expected to be greater than 16.5.

  3. Improving the visualization of electron-microscopy data through optical flow interpolation

    KAUST Repository

    Carata, Lucian

    2013-01-01

    Technical developments in neurobiology have reached a point where the acquisition of high resolution images representing individual neurons and synapses becomes possible. For this, the brain tissue samples are sliced using a diamond knife and imaged with electron-microscopy (EM). However, the technique achieves a low resolution in the cutting direction, due to limitations of the mechanical process, making a direct visualization of a dataset difficult. We aim to increase the depth resolution of the volume by adding new image slices interpolated from the existing ones, without requiring modifications to the EM image-capturing method. As classical interpolation methods do not provide satisfactory results on this type of data, the current paper proposes a re-framing of the problem in terms of motion volumes, considering the depth axis as a temporal axis. An optical flow method is adapted to estimate the motion vectors of pixels in the EM images, and this information is used to compute and insert multiple new images at certain depths in the volume. We evaluate the visualization results in comparison with interpolation methods currently used on EM data, transforming the highly anisotropic original dataset into a dataset with a larger depth resolution. The interpolation based on optical flow better reveals neurite structures with realistic undistorted shapes, and helps to easier map neuronal connections. © 2011 ACM.

  4. Single myelin fiber imaging in living rodents without labeling by deep optical coherence microscopy

    Science.gov (United States)

    Ben Arous, Juliette; Binding, Jonas; Léger, Jean-François; Casado, Mariano; Topilko, Piotr; Gigan, Sylvain; Claude Boccara, A.; Bourdieu, Laurent

    2011-11-01

    Myelin sheath disruption is responsible for multiple neuropathies in the central and peripheral nervous system. Myelin imaging has thus become an important diagnosis tool. However, in vivo imaging has been limited to either low-resolution techniques unable to resolve individual fibers or to low-penetration imaging of single fibers, which cannot provide quantitative information about large volumes of tissue, as required for diagnostic purposes. Here, we perform myelin imaging without labeling and at micron-scale resolution with >300-μm penetration depth on living rodents. This was achieved with a prototype [termed deep optical coherence microscopy (deep-OCM)] of a high-numerical aperture infrared full-field optical coherence microscope, which includes aberration correction for the compensation of refractive index mismatch and high-frame-rate interferometric measurements. We were able to measure the density of individual myelinated fibers in the rat cortex over a large volume of gray matter. In the peripheral nervous system, deep-OCM allows, after minor surgery, in situ imaging of single myelinated fibers over a large fraction of the sciatic nerve. This allows quantitative comparison of normal and Krox20 mutant mice, in which myelination in the peripheral nervous system is impaired. This opens promising perspectives for myelin chronic imaging in demyelinating diseases and for minimally invasive medical diagnosis.

  5. Dye-enhanced reflectance and fluorescence confocal microscopy as an optical pathology tool

    Science.gov (United States)

    Yaroslavsky, Anna N.; Salomatina, Elena; Novak, John; Amat-Roldan, Ivan; Castano, Ana; Hamblin, Michael

    2006-02-01

    Early detection and precise excision of neoplasms are imperative requirements for successful cancer treatment. In this study we evaluated the use of dye-enhanced confocal microscopy as an optical pathology tool in the ex vivo trial with fresh thick non-melanoma skin cancer excisions and in vivo trial with B16F10 melanoma cancer in mice. For the experiments the tumors were rapidly stained using aqueous solutions of either toluidine blue or methylene blue and imaged using multimodal confocal microscope. Reflectance images were acquired at the wavelengths of 630nm and 650 nm. Fluorescence was excited at 630 nm and 650 nm. Fluorescence emission was registered in the range between 680 nm and 710 nm. The images were compared to the corresponding en face frozen H&E sections. The results of the study indicate confocal images of stained cancerous tissue closely resemble corresponding H&E sections both in vivo and in vitro. This remarkable similarity enables interpretation of confocal images in a manner similar to that of histopathology. The developed technique may provide an efficient real-time optical tool for detecting skin pathology.

  6. Single myelin fiber imaging in living rodents without labeling by deep optical coherence microscopy.

    Science.gov (United States)

    Ben Arous, Juliette; Binding, Jonas; Léger, Jean-François; Casado, Mariano; Topilko, Piotr; Gigan, Sylvain; Boccara, A Claude; Bourdieu, Laurent

    2011-11-01

    Myelin sheath disruption is responsible for multiple neuropathies in the central and peripheral nervous system. Myelin imaging has thus become an important diagnosis tool. However, in vivo imaging has been limited to either low-resolution techniques unable to resolve individual fibers or to low-penetration imaging of single fibers, which cannot provide quantitative information about large volumes of tissue, as required for diagnostic purposes. Here, we perform myelin imaging without labeling and at micron-scale resolution with >300-μm penetration depth on living rodents. This was achieved with a prototype [termed deep optical coherence microscopy (deep-OCM)] of a high-numerical aperture infrared full-field optical coherence microscope, which includes aberration correction for the compensation of refractive index mismatch and high-frame-rate interferometric measurements. We were able to measure the density of individual myelinated fibers in the rat cortex over a large volume of gray matter. In the peripheral nervous system, deep-OCM allows, after minor surgery, in situ imaging of single myelinated fibers over a large fraction of the sciatic nerve. This allows quantitative comparison of normal and Krox20 mutant mice, in which myelination in the peripheral nervous system is impaired. This opens promising perspectives for myelin chronic imaging in demyelinating diseases and for minimally invasive medical diagnosis.

  7. Optical far-field super-resolution microscopy using nitrogen vacancy center ensemble in bulk diamond

    CERN Document Server

    Li, Shen; Zhao, Bo-Wen; Dong, Yang; Zou, Chong-Wen; Guo, Guang-Can; Sun, Fang-Wen

    2016-01-01

    We demonstrate an optical far-field super-resolution microscopy using array of nitrogen vacancy centers in bulk diamond as near-field optical probes. The local optical field, which transmits through the nanostructures on the diamond surface, is measured by detecting the charge state conversion of nitrogen vacancy center. And the locating of nitrogen vacancy center with spatial resolution of 6.1 nm is realized with the charge state depletion nanoscopy. The nanostructures on the surface of diamond are then imaged with resolution below optical diffraction limit. The results offer an approach to built a general-purpose optical super-resolution microscopy and a convenient platform for high spatial resolution quantum sensing with nitrogen vacancy center.

  8. Depth-profiling by confocal Raman microscopy (CRM): data correction by numerical techniques.

    Science.gov (United States)

    Tomba, J Pablo; Eliçabe, Guillermo E; Miguel, María de la Paz; Perez, Claudio J

    2011-03-01

    The data obtained in confocal Raman microscopy (CRM) depth profiling experiments with dry optics are subjected to significant distortions, including an artificial compression of the depth scale, due to the combined influence of diffraction, refraction, and instrumental effects that operate on the measurement. This work explores the use of (1) regularized deconvolution and (2) the application of simple rescaling of the depth scale as methodologies to obtain an improved, more precise, confocal response. The deconvolution scheme is based on a simple predictive model for depth resolution and the use of regularization techniques to minimize the dramatic oscillations in the recovered response typical of problem inversion. That scheme is first evaluated using computer simulations on situations that reproduce smooth and sharp sample transitions between two materials and finally it is applied to correct genuine experimental data, obtained in this case from a sharp transition (planar interface) between two polymeric materials. It is shown that the methodology recovers very well most of the lost profile features in all the analyzed situations. The use of simple rescaling appears to be only useful for correcting smooth transitions, particularly those extended over distances larger than those spanned by the operative depth resolution, which limits the strategy to the study of profiles near the sample surface. However, through computer simulations, it is shown that the use of water immersion objectives may help to reduce optical distortions and to expand the application window of this simple methodology, which could be useful, for instance, to safely monitor Fickean sorption/desorption of penetrants in polymer films/coatings in a nearly noninvasive way.

  9. Cognitive Heterogeneous Reconfigurable Optical Networks (CHRON): Enabling Technologies and Techniques

    DEFF Research Database (Denmark)

    Tafur Monroy, Idelfonso; Zibar, Darko; Guerrero Gonzalez, Neil;

    2011-01-01

    We present the approach of cognition applied to heterogeneous optical networks developed in the framework of the EU project CHRON: Cognitive Heterogeneous Reconfigurable Optical Network. We introduce and discuss in particular the technologies and techniques that will enable a cognitive optical ne...

  10. Optical Fourier techniques for medical image processing and phase contrast imaging.

    Science.gov (United States)

    Yelleswarapu, Chandra S; Kothapalli, Sri-Rajasekhar; Rao, D V G L N

    2008-04-01

    This paper briefly reviews the basics of optical Fourier techniques (OFT) and applications for medical image processing as well as phase contrast imaging of live biological specimens. Enhancement of microcalcifications in a mammogram for early diagnosis of breast cancer is the main focus. Various spatial filtering techniques such as conventional 4f filtering using a spatial mask, photoinduced polarization rotation in photosensitive materials, Fourier holography, and nonlinear transmission characteristics of optical materials are discussed for processing mammograms. We also reviewed how the intensity dependent refractive index can be exploited as a phase filter for phase contrast imaging with a coherent source. This novel approach represents a significant advance in phase contrast microscopy.

  11. Optical Measurement Techniques for Optical Fiber and Waveguide Devices

    Institute of Scientific and Technical Information of China (English)

    D.Y.; Kim; Y.; Park; N.H.; Seong; Y.C.Youk; J.Y.; Lee; S.; Moon; I.H.; Shin; H.S.; Ryu

    2003-01-01

    We describe three major optical characterization methods for fiber and fiber devices. A simple servo controlled scanning fiber-optic confocal microscope is proposed for determining the refractive index profile of an optical fiber. To measure the chromatic dispersion of a short length fiber a Mach-Zehnder fiber interferometer with a novel interferometric distance meter is introduced. At the end, a tomographic method is demonstrated for determining the 2-D stress profile of a fiber.

  12. Second-order nonlinear optical microscopy of spider silk

    Science.gov (United States)

    Zhao, Yue; Hien, Khuat Thi Thu; Mizutani, Goro; Rutt, Harvey N.

    2017-06-01

    Asymmetric β-sheet protein structures in spider silk should induce nonlinear optical interaction such as second harmonic generation (SHG) which is experimentally observed for a radial line and dragline spider silk using an imaging femtosecond laser SHG microscope. By comparing different spider silks, we found that the SHG signal correlates with the existence of the protein β-sheets. Measurements of the polarization dependence of SHG from the dragline indicated that the β-sheet has a nonlinear response depending on the direction of the incident electric field. We propose a model of what orientation the β-sheet takes in spider silk.

  13. Optical-resolution photoacoustic microscopy by use of a multimode fiber

    CERN Document Server

    Papadopoulos, Ioannis N; Farahi, Salma; Huignard, Jean Pierre; Bossy, Emmanuel; Psaltis, Demetri; Moser, Christophe

    2013-01-01

    We demonstrate Optical-Resolution Photoacoustic Microscopy (OR-PAM), where the optical field is focused and scanned using Digital Phase Conjugation (DPC) through a multimode fiber. The focus is scanned across the field of view using digital means, and the acoustic signal induced is collected by a transducer. Optical-resolution photoacoustic images of a knot made by two absorptive wires are obtained and we report on resolution smaller than 1.5{\\mu}m across a 201{\\mu}m by 201{\\mu}m field of view. The use of a multimode optical fiber for the optical excitation part can pave the way for miniature endoscopes that can provide optical-resolution photoacoustic images at large optical depth.

  14. Digital processing optical transmission and coherent receiving techniques

    CERN Document Server

    Binh, Le Nguyen

    2013-01-01

    With coherent mixing in the optical domain and processing in the digital domain, advanced receiving techniques employing ultra-high speed sampling rates have progressed tremendously over the last few years. These advances have brought coherent reception systems for lightwave-carried information to the next stage, resulting in ultra-high capacity global internetworking. Digital Processing: Optical Transmission and Coherent Receiving Techniques describes modern coherent receiving techniques for optical transmission and aspects of modern digital optical communications in the most basic lines. The

  15. Light propagation studies on laser modified waveguides using scanning near-field optical microscopy

    DEFF Research Database (Denmark)

    Borrise, X.; Berini, Abadal Gabriel; Jimenez, D.

    2001-01-01

    By means of direct laser writing on Al, a new method to locally modify optical waveguides is proposed. This technique has been applied to silicon nitride waveguides, allowing modifications of the optical propagation along the guide. To study the formed structures, a scanning near-held optical...

  16. All Optical Signal-Processing Techniques Utilizing Four Wave Mixing

    Directory of Open Access Journals (Sweden)

    Refat Kibria

    2015-02-01

    Full Text Available Four Wave Mixing (FWM based optical signal-processing techniques are reviewed. The use of FWM in arithmetical operation like subtraction, wavelength conversion and pattern recognition are three key parts discussed in this paper after a brief introduction on FWM and its comparison with other nonlinear mixings. Two different approaches to achieve correlation are discussed, as well as a novel technique to realize all optical subtraction of two optical signals.

  17. Antifouling leaching technique for optical lenses

    Science.gov (United States)

    Strahle, William J.; Perez, C. L.; Martini, Marinna A.

    1994-01-01

    The effectiveness of optical lenses deployed in water less than 100 m deep is significantly reduced by biofouling caused by the settlement of macrofauna, such as barnacles, hydroids, and tunicates. However, machineable porous plastic rings can be used to dispense antifoulant into the water in front of the lens to retard macrofaunal growth without obstructing the light path. Unlike coatings which can degrade the optical performance, antifouling rings do not interfere with the instrument optics. The authors have designed plastic, reusable cup-like antifouling rings to slip over the optical lenses of a transmissometer. These rings have been used for several deployments on shallow moorings in Massachusetts Bay, MA and have increased the time before fouling degrades optical characteristics

  18. Optical Microscopy Characterization for Borehole U-15n#12 in Support of NCNS Source Physics Experiment

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, Jennifer E. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Sussman, Aviva Joy [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2015-05-22

    Optical microscopy characterization of thin sections from corehole U-15n#12 is part of a larger material characterization effort for the Source Physics Experiment (SPE). The SPE program was conducted in Nevada with a series of explosive tests designed to study the generation and propagation of seismic waves inside Stock quartz monzonite. Optical microscopy analysis includes the following: 1) imaging of full thin sections (scans and mosaic maps); 2) high magnification imaging of petrographic texture (grain size, foliations, fractures, etc.); and 3) measurement of microfracture density.

  19. Imaging of membrane proteins using antenna-based optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Hoeppener, Christiane; Novotny, Lukas [Institute of Optics and Department of Biomedical Engineering, University of Rochester, Rochester, NY 14627 (United States)], E-mail: novotny@optics.rochester.edu

    2008-09-24

    The localization and identification of individual proteins is of key importance for the understanding of biological processes on the molecular scale. Here, we demonstrate near-field fluorescence imaging of single proteins in their native cell membrane. Incident laser radiation is localized and enhanced with an optical antenna in the form of a spherical gold particle attached to a pointed dielectric tip. Individual proteins can be identified with a diffraction-unlimited spatial resolution of {approx}50 nm. Besides determining the concentration and distribution of specific membrane proteins, this approach makes it possible to study the colocalization of different membrane proteins. Moreover, it enables a simultaneous recording of the membrane topology. Protein distributions can be correlated with the local membrane topology, thereby providing important information on the chemical and structural organization of cellular membranes.

  20. Neural imaging in songbirds using fiber optic fluorescence microscopy

    Science.gov (United States)

    Nooshabadi, Fatemeh; Hearn, Gentry; Lints, Thierry; Maitland, Kristen C.

    2012-02-01

    The song control system of juvenile songbirds is an important model for studying the developmental acquisition and generation of complex learned vocal motor sequences, two processes that are fundamental to human speech and language. To understand the neural mechanisms underlying song production, it is critical to characterize the activity of identified neurons in the song control system when the bird is singing. Neural imaging in unrestrained singing birds, although technically challenging, will advance our understanding of neural ensemble coding mechanisms in this system. We are exploring the use of a fiber optic microscope for functional imaging in the brain of behaving and singing birds in order to better understand the contribution of a key brain nucleus (high vocal center nucleus; HVC) to temporal aspects of song motor control. We have constructed a fluorescence microscope with LED illumination, a fiber bundle for transmission of fluorescence excitation and emission light, a ~2x GRIN lens, and a CCD for image acquisition. The system has 2 μm resolution, 375 μm field of view, 200 μm working distance, and 1 mm outer diameter. As an initial characterization of this setup, neurons in HVC were imaged using the fiber optic microscope after injection of quantum dots or fluorescent retrograde tracers into different song nuclei. A Lucid Vivascope confocal microscope was used to confirm the imaging results. Long-term imaging of the activity of these neurons in juvenile birds during singing may lead us to a better understanding of the central motor codes for song and the central mechanism by which auditory experience modifies song motor commands to enable vocal learning and imitation.

  1. Follow-up review: recent progress in the development of super-resolution optical microscopy.

    Science.gov (United States)

    Fujita, Katsumasa

    2016-08-01

    The advent of super-resolution microscopy brought a huge impact to various research fields ranging from the fundamental science to medical and industrial applications. The technological development is still ongoing with involving different scientific disciplines and often changing the standard of optical imaging. In this review, I would like to introduce the recent research progress in super-resolution microscopy as a follow-up for the featured issue in Microscopy (Vol. 64, No. 4, 2015) with discussions especially on the current trends and new directions in the technological development.

  2. Dispersion effect in optical microscopy systems with a supersphere solid immersion lens

    Institute of Scientific and Technical Information of China (English)

    Zhang Yao-Ju; Zhuang You-Yi

    2009-01-01

    This paper studies the dispersion effect of the supersphere solid immersion lens (SIL) on a near field optical microscopy system by using the vector diffraction theory. Results show that when a real non-monochromatic beam illuminates a supersphere SIL microscopy, the dispersion effect of the SIL has an important influence on the image quality. As the wavelength bandwidth of the non-monochromatic beam increases, the size of the focused spot increases and its intensity decreases in near-field microscopy systems with a supersphere SIL.

  3. Multicolor 3D super-resolution imaging by quantum dot stochastic optical reconstruction microscopy.

    Science.gov (United States)

    Xu, Jianquan; Tehrani, Kayvan F; Kner, Peter

    2015-03-24

    We demonstrate multicolor three-dimensional super-resolution imaging with quantum dots (QSTORM). By combining quantum dot asynchronous spectral blueing with stochastic optical reconstruction microscopy and adaptive optics, we achieve three-dimensional imaging with 24 nm lateral and 37 nm axial resolution. By pairing two short-pass filters with two appropriate quantum dots, we are able to image single blueing quantum dots on two channels simultaneously, enabling multicolor imaging with high photon counts.

  4. Super-resolution optical microscopy by using dielectric microwires

    Science.gov (United States)

    Darafsheh, Arash; Wu, Gaoxiang; Yang, Shu; Finlay, Jarod C.

    2016-03-01

    We demonstrate that super-resolution imaging of specimens containing sub-diffraction-limited features is feasible by using dielectric microwires fabricated through capillary force lithography followed by photopatterning. As supplementary micron scale cylindrical lenses, we fabricated uniform-sized microwires with and 5 and 10 μm diameters and refractive index ~1.3-1.6. The microwires are placed in contact with the specimen to collect the information of the sub-wavelength features of the specimen and transmit them to the far-field with magnification enabling imaging with two-fold resolution improvement. Potential applications of our imaging technique include biological imaging, microfluidics, and nanophotonics applications.

  5. Generalized spectral method for near-field optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, B.-Y.; Zhang, L. M.; Basov, D. N.; Fogler, M. M. [Department of Physics, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093 (United States); Castro Neto, A. H. [Department of Physics, Boston University, 590 Commonwealth Avenue, Boston, Massachusetts 02215 (United States); Centre for Advanced 2D Materials and Graphene Research Centre, National University of Singapore, Singapore, Singapore 117542 (Singapore)

    2016-02-07

    Electromagnetic interaction between a sub-wavelength particle (the “probe”) and a material surface (the “sample”) is studied theoretically. The interaction is shown to be governed by a series of resonances corresponding to surface polariton modes localized near the probe. The resonance parameters depend on the dielectric function and geometry of the probe as well as on the surface reflectivity of the material. Calculation of such resonances is carried out for several types of axisymmetric probes: spherical, spheroidal, and pear-shaped. For spheroids, an efficient numerical method is developed, capable of handling cases of large or strongly momentum-dependent surface reflectivity. Application of the method to highly resonant materials, such as aluminum oxide (by itself or covered with graphene), reveals a rich structure of multi-peak spectra and nonmonotonic approach curves, i.e., the probe-sample distance dependence. These features also strongly depend on the probe shape and optical constants of the model. For less resonant materials such as silicon oxide, the dependence is weak, so that the spheroidal model is reliable. The calculations are done within the quasistatic approximation with radiative damping included perturbatively.

  6. Ultra-sensitive Magnetic Microscopy with an Optically Pumped Magnetometer

    Science.gov (United States)

    Kim, Young Jin; Savukov, Igor

    2016-04-01

    Optically pumped magnetometers (OPMs) based on lasers and alkali-metal vapor cells are currently the most sensitive non-cryogenic magnetic field sensors. Many applications in neuroscience and other fields require high-resolution, high-sensitivity magnetic microscopic measurements. In order to meet this demand we combined a cm-size spin-exchange relaxation-free (SERF) OPM and flux guides (FGs) to realize an ultra-sensitive FG-OPM magnetic microscope. The FGs serve to transmit the target magnetic flux to the OPM thus improving both the resolution and sensitivity to small magnetic objects. We investigated the performance of the FG-OPM device using experimental and numerical methods, and demonstrated that an optimized device can achieve a unique combination of high resolution (80 μm) and high sensitivity (8.1 pT/). In addition, we also performed numerical calculations of the magnetic field distribution in the FGs to estimate the magnetic noise originating from the domain fluctuations in the material of the FGs. We anticipate many applications of the FG-OPM device such as the detection of micro-biological magnetic fields; the detection of magnetic nano-particles; and non-destructive testing. From our theoretical estimate, an FG-OPM could detect the magnetic field of a single neuron, which would be an important milestone in neuroscience.

  7. Ultra-sensitive Magnetic Microscopy with an Optically Pumped Magnetometer.

    Science.gov (United States)

    Kim, Young Jin; Savukov, Igor

    2016-04-22

    Optically pumped magnetometers (OPMs) based on lasers and alkali-metal vapor cells are currently the most sensitive non-cryogenic magnetic field sensors. Many applications in neuroscience and other fields require high-resolution, high-sensitivity magnetic microscopic measurements. In order to meet this demand we combined a cm-size spin-exchange relaxation-free (SERF) OPM and flux guides (FGs) to realize an ultra-sensitive FG-OPM magnetic microscope. The FGs serve to transmit the target magnetic flux to the OPM thus improving both the resolution and sensitivity to small magnetic objects. We investigated the performance of the FG-OPM device using experimental and numerical methods, and demonstrated that an optimized device can achieve a unique combination of high resolution (80 μm) and high sensitivity (8.1 pT/). In addition, we also performed numerical calculations of the magnetic field distribution in the FGs to estimate the magnetic noise originating from the domain fluctuations in the material of the FGs. We anticipate many applications of the FG-OPM device such as the detection of micro-biological magnetic fields; the detection of magnetic nano-particles; and non-destructive testing. From our theoretical estimate, an FG-OPM could detect the magnetic field of a single neuron, which would be an important milestone in neuroscience.

  8. Magnetic and optical properties of the nickel thin film deposited by GLAD technique

    Directory of Open Access Journals (Sweden)

    Potočnik Jelena M.

    2014-01-01

    Full Text Available In this work, nickel thin film was deposited on glass sample using Glancing Angle Deposition (GLAD technique, to a thickness of 1 μm. Glass sample was positioned 15 degrees with respect to the nickel vapor flux. The nickel thin film was characterized by Atomic Force Microscopy (AFM, Magneto- Optical Kerr effect Microscopy and Spectroscopic Ellipsometry. According to an AFM cross-section imaging, it was found that the nickel thin film has a columnar structure. The values of the coercively, obtained from the magnetic hysteresis loops, were analyzed as a function of the sample rotation in the magnetic field. It was found that the direction of magnetization easy axis lies toward the structure growth. Optical properties of the nickel thin film were studied at the wavelength of 455 nm. From the shape of the refractive index and the extinction coefficient diagrams could be concluded that the nickel thin film has an optical anisotropy.

  9. Investigation into local cell mechanics by atomic force microscopy mapping and optical tweezer vertical indentation

    Science.gov (United States)

    Coceano, G.; Yousafzai, M. S.; Ma, W.; Ndoye, F.; Venturelli, L.; Hussain, I.; Bonin, S.; Niemela, J.; Scoles, G.; Cojoc, D.; Ferrari, E.

    2016-02-01

    Investigating the mechanical properties of cells could reveal a potential source of label-free markers of cancer progression, based on measurable viscoelastic parameters. The Young’s modulus has proved to be the most thoroughly studied so far, however, even for the same cell type, the elastic modulus reported in different studies spans a wide range of values, mainly due to the application of different experimental conditions. This complicates the reliable use of elasticity for the mechanical phenotyping of cells. Here we combine two complementary techniques, atomic force microscopy (AFM) and optical tweezer microscopy (OTM), providing a comprehensive mechanical comparison of three human breast cell lines: normal myoepithelial (HBL-100), luminal breast cancer (MCF-7) and basal breast cancer (MDA-MB-231) cells. The elastic modulus was measured locally by AFM and OTM on single cells, using similar indentation approaches but different measurement parameters. Peak force tapping AFM was employed at nanonewton forces and high loading rates to draw a viscoelastic map of each cell and the results indicated that the region on top of the nucleus provided the most meaningful results. OTM was employed at those locations at piconewton forces and low loading rates, to measure the elastic modulus in a real elastic regime and rule out the contribution of viscous forces typical of AFM. When measured by either AFM or OTM, the cell lines’ elasticity trend was similar for the aggressive MDA-MB-231 cells, which were found to be significantly softer than the other two cell types in both measurements. However, when comparing HBL-100 and MCF-7 cells, we found significant differences only when using OTM.

  10. Chemical Silver Coating of Fiber Tips in Near-Field Scanning Optical Microscopy

    Science.gov (United States)

    Vikram, Chandra S.; Witherow, William K.

    1998-01-01

    We report what is believed to be the first experimental demonstration of silver coating by a wet chemical process on tapered fiber tips used in near-field scanning optical microscopy. The process is at room temperature and pressure and takes only a few minutes to complete. Many tips can be simultaneously coated.

  11. Fast Calcium Imaging with Optical Sectioning via HiLo Microscopy.

    Science.gov (United States)

    Lauterbach, Marcel A; Ronzitti, Emiliano; Sternberg, Jenna R; Wyart, Claire; Emiliani, Valentina

    2015-01-01

    Imaging intracellular calcium concentration via reporters that change their fluorescence properties upon binding of calcium, referred to as calcium imaging, has revolutionized our way to probe neuronal activity non-invasively. To reach neurons densely located deep in the tissue, optical sectioning at high rate of acquisition is necessary but difficult to achieve in a cost effective manner. Here we implement an accessible solution relying on HiLo microscopy to provide robust optical sectioning with a high frame rate in vivo. We show that large calcium signals can be recorded from dense neuronal populations at high acquisition rates. We quantify the optical sectioning capabilities and demonstrate the benefits of HiLo microscopy compared to wide-field microscopy for calcium imaging and 3D reconstruction. We apply HiLo microscopy to functional calcium imaging at 100 frames per second deep in biological tissues. This approach enables us to discriminate neuronal activity of motor neurons from different depths in the spinal cord of zebrafish embryos. We observe distinct time courses of calcium signals in somata and axons. We show that our method enables to remove large fluctuations of the background fluorescence. All together our setup can be implemented to provide efficient optical sectioning in vivo at low cost on a wide range of existing microscopes.

  12. Single molecule mapping of the optical field distribution of probes for near-field microscopy

    NARCIS (Netherlands)

    Veerman, J.A.; Garcia Parajo, M.F.; Kuipers, L.; van Hulst, N.F.

    1999-01-01

    The most difficult task in near-field scanning optical microscopy (NSOM) is to make a high quality subwavelength aperture probe, Recently we have developed high definition NSOM probes by focused ion beam (FIB) milling. These probes have a higher brightness, better polarization characteristics,

  13. Correlative nonlinear optical microscopy and infrared nanoscopy reveals collagen degradation in altered parchments

    OpenAIRE

    Gaël Latour; Laurianne Robinet; Alexandre Dazzi; François Portier; Ariane Deniset-Besseau; Marie-Claire Schanne-Klein

    2016-01-01

    International audience; This paper presents the correlative imaging of collagen denaturation by nonlinear optical microscopy (NLO) and nanoscale infrared (IR) spectroscopy to obtain morphological and chemical information at different length scales. Such multiscale correlated measurements are applied to the investigation of ancient parchments, which are mainly composed of dermal fibrillar collagen. The main issue is to characterize gelatinization, the ultimate and irreversible alteration corre...

  14. Single molecule mapping of the optical field distribution of probes for near-field microscopy

    NARCIS (Netherlands)

    Veerman, J.A.; Garcia-Parajo, M.F.; Kuipers, L.; Hulst, van N.F.

    1999-01-01

    The most difficult task in near-field scanning optical microscopy (NSOM) is to make a high quality subwavelength aperture probe, Recently we have developed high definition NSOM probes by focused ion beam (FIB) milling. These probes have a higher brightness, better polarization characteristics, bette

  15. Multimodal super-resolution optical microscopy visualizes the close connection between membrane and the cytoskeleton in liver sinusoidal endothelial cell fenestrations

    Science.gov (United States)

    Mönkemöller, Viola; Øie, Cristina; Hübner, Wolfgang; Huser, Thomas; McCourt, Peter

    2015-11-01

    Liver sinusoidal endothelial cells (LSECs) act as a filter between blood and the hepatocytes. LSECs are highly fenestrated cells; they contain transcellular pores with diameters between 50 to 200 nm. The small sizes of the fenestrae have so far prohibited any functional analysis with standard and advanced light microscopy techniques. Only the advent of super-resolution optical fluorescence microscopy now permits the recording of such small cellular structures. Here, we demonstrate the complementary use of two different super-resolution optical microscopy modalities, 3D structured illumination microscopy (3D-SIM) and single molecule localization microscopy in a common optical platform to obtain new insights into the association between the cytoskeleton and the plasma membrane that supports the formation of fenestrations. We applied 3D-SIM to multi-color stained LSECs to acquire highly resolved overviews of large sample areas. We then further increased the spatial resolution for imaging fenestrations by single molecule localization microscopy applied to select small locations of interest in the same sample on the same microscope setup. We optimized the use of fluorescent membrane stains for these imaging conditions. The combination of these techniques offers a unique opportunity to significantly improve studies of subcellular ultrastructures such as LSEC fenestrations.

  16. Automatic forensic analysis of automotive paints using optical microscopy.

    Science.gov (United States)

    Thoonen, Guy; Nys, Bart; Vander Haeghen, Yves; De Roy, Gilbert; Scheunders, Paul

    2016-02-01

    The timely identification of vehicles involved in an accident, such as a hit-and-run situation, bears great importance in forensics. To this end, procedures have been defined for analyzing car paint samples that combine techniques such as visual analysis and Fourier transform infrared spectroscopy. This work proposes a new methodology in order to automate the visual analysis using image retrieval. Specifically, color and texture information is extracted from a microscopic image of a recovered paint sample, and this information is then compared with the same features for a database of paint types, resulting in a shortlist of candidate paints. In order to demonstrate the operation of the methodology, a test database has been set up and two retrieval experiments have been performed. The first experiment quantifies the performance of the procedure for retrieving exact matches, while the second experiment emulates the real-life situation of paint samples that experience changes in color and texture over time. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  17. A Study of Synchronization Techniques for Optical Communication Systems

    Science.gov (United States)

    Gagliardi, R. M.

    1975-01-01

    The study of synchronization techniques and related topics in the design of high data rate, deep space, optical communication systems was reported. Data cover: (1) effects of timing errors in narrow pulsed digital optical systems, (2) accuracy of microwave timing systems operating in low powered optical systems, (3) development of improved tracking systems for the optical channel and determination of their tracking performance, (4) development of usable photodetector mathematical models for application to analysis and performance design in communication receivers, and (5) study application of multi-level block encoding to optical transmission of digital data.

  18. Super-resolution spinning-disk confocal microscopy using optical photon reassignment.

    Science.gov (United States)

    Azuma, Takuya; Kei, Takayuki

    2015-06-01

    Spinning-disk confocal microscopy is a proven technology for investigating 3D structures of biological specimens. Here we report a super-resolution method based on spinning-disk confocal microscopy that optically improves lateral resolution by a factor of 1.37 with a single exposure. Moreover, deconvolution yields twofold improvement over the diffraction limit. With the help of newly modified Nipkow disk which comprises pinholes and micro-lenses on the front and back respectively, emitted photons from specimen can be optically reassigned to the most probable locations they originate from. Consequently, the improvement in resolution is achieved preserving inherent sectioning capabilities of confocal microscopy. This extremely simple implementation will enable reliable observations at super high resolution in biomedical routine research.

  19. Cell tracking with gadophrin-2: a bifunctional contrast agent for MR imaging, optical imaging, and fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Daldrup-Link, Heike E. [Department of Radiology, UCSF Medical Center, University of California in San Francisco, 513 Parnassus Ave, CA 94143, San Francisco (United States); Rudelius, Martina; Piontek, Guido; Schlegel, Juergen [Institute of Pathology, Technical University, Munich (Germany); Metz, Stephan; Settles, Marcus; Rummeny, Ernst J. [Department of Radiology, Technical University, Munich (Germany); Pichler, Bernd [Department of Biomedical Engineering, University of California Davis, Davis (United States); Heinzmann, Ulrich [National Research Center for Environment and Health, Technical University, Munich (Germany); Oostendorp, Robert A.J. [3. Clinic of Internal Medicine, Laboratory of Stem Cell Physiology, Technical University, Munich (Germany)

    2004-09-01

    The purpose of this study was to assess the feasibility of use of gadophrin-2 to trace intravenously injected human hematopoietic cells in athymic mice, employing magnetic resonance (MR) imaging, optical imaging (OI), and fluorescence microscopy. Mononuclear peripheral blood cells from GCSF-primed patients were labeled with gadophrin-2 (Schering AG, Berlin, Germany), a paramagnetic and fluorescent metalloporphyrin, using established transfection techniques with cationic liposomes. The labeled cells were evaluated in vitro with electron microscopy and inductively coupled plasma atomic emission spectrometry. Then, 1 x 10{sup 6}-3 x 10{sup 8} labeled cells were injected into 14 nude Balb/c mice and the in vivo cell distribution was evaluated with MR imaging and OI before and 4, 24, and 48 h after intravenous injection (p.i.). Five additional mice served as controls: three mice were untreated controls and two mice were investigated after injection of unlabeled cells. The contrast agent effect was determined quantitatively for MR imaging by calculating signal-to-noise-ratio (SNR) data. After completion of in vivo imaging studies, fluorescence microscopy of excised organs was performed. Intracellular cytoplasmatic uptake of gadophrin-2 was confirmed by electron microscopy. Spectrometry determined an uptake of 31.56 nmol Gd per 10{sup 6} cells. After intravenous injection, the distribution of gadophrin-2 labeled cells in nude mice could be visualized by MR, OI, and fluorescence microscopy. At 4 h p.i., the transplanted cells mainly distributed to lung, liver, and spleen, and 24 h p.i. they also distributed to the bone marrow. Fluorescence microscopy confirmed the distribution of gadophrin-2 labeled cells to these target organs. Gadophrin-2 is suited as a bifunctional contrast agent for MR imaging, OI, and fluorescence microscopy and may be used to combine the advantages of each individual imaging modality for in vivo tracking of intravenously injected hematopoietic cells

  20. Visualisation of biopolymer mixtures using confocal scanning laser microscopy (CSLM) and covalent labelling techniques

    NARCIS (Netherlands)

    Velde, van de F.; Weinbreck, F.; Edelman, M.W.; Linden, van der E.; Tromp, R.H.

    2003-01-01

    Confocal scanning laser microscopy (CSLM) has been used to study the behaviour of mixtures of proteins, gelatine, whey proteins and ß-lactoglobulin, and polysaccharides, dextran, gellan gum, carrageenan, gum Arabic, and starch. CSLM proved to be a suitable technique to visualise the microstructure o

  1. Optically stimulated luminescence techniques in retrospective dosimetry

    DEFF Research Database (Denmark)

    Bøtter-Jensen, L.; Murray, A.S.

    2001-01-01

    Optically stimulated luminescence signals from natural quartz and feldspar are now used routinely in dating geological and archaeological materials. More recently they have also been employed in accident dosimetry, i.e. the retrospective assessment of doses received as a result of a nuclear...

  2. Line-scan Raman microscopy complements optical coherence tomography for tumor boundary detection

    Science.gov (United States)

    Sudheendran, Narendran; Qi, Ji; Young, Eric D.; Lazar, Alexander J.; Lev, Dina C.; Pollock, Raphael E.; Larin, Kirill V.; Shih, Wei-Chuan

    2014-10-01

    Current technique for tumor resection requires biopsy of the tumor region and histological confirmation before the surgeon can be certain that the entire tumor has been resected. This confirmation process is time consuming both for the surgeon and the patient and also requires sacrifice of healthy tissue, motivating the development of novel technologies which can enable real-time detection of tumor-healthy tissue boundary for faster and more efficient surgeries. In this study, the potential of combining structural information from optical coherence tomography (OCT) and molecular information from line-scan Raman microscopy (LSRM) for such an application is presented. The results show a clear presence of boundary between myxoid liposarcoma and normal fat which is easily identifiable both from structural and molecular information. In cases where structural images are indistinguishable, for example, in normal fat and well differentiated liposarcoma (WDLS) or gastrointestinal sarcoma tumor (GIST) and myxoma, distinct molecular spectra have been obtained. The results suggest LSRM can effectively complement OCT to tumor boundary demarcation with high specificity.

  3. Surface diffusion of Sb on Ge(111) monitored quantitatively with optical second harmonic microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Schultz, K.A.; Seebauer, E.G. (Department of Chemical Engineering, University of Illinois, Urbana, Illinois 61801 (United States))

    1992-11-01

    Surface diffusion of Sb on Ge(111) has been measured with the newly developed technique of optical second harmonic microscopy. In this method, concentration profiles at submonolayer coverage are imaged directly by surface second harmonic generation with 5 {mu} spatial resolution. A Boltzmann--Matano analysis yields the coverage dependence of the diffusivity {ital D} without parametrization. Experiments were performed at roughly 70% of the bulk melting temperature {ital T}{sub {ital m}}. In the coverage range 0{le}{theta}{le}0.6, the activation energy {ital E}{sub diff} remains constant at 47.5{plus minus}1.5 kcal/mol, but the pre-exponential factor {ital D}{sub 0} decreases from 8.7{times}10{sup 3{plus minus}0.4} to 1.6{times}10{sup 2{plus minus}0.4} cm{sup 2}/s. Both {ital E}{sub diff} and {ital D}{sub 0} are quite large, which is consistent with high-temperature measurements in other systems. The inadequacies of current theories for high-temperature surface diffusion are outlined, and a new vacancy model is proposed for low-coverage diffusion. The model accounts semiquantitatively for the large values of {ital E}{sub diff} and {ital D}{sub 0}, and suggests that these quantities may be manipulated using doping levels and photon illumination. An islanding mechanism is proposed to explain the decrease in {ital D}{sub 0} with {theta}.

  4. Simultaneous topographical, electrical and optical microscopy of optoelectronic devices at the nanoscale.

    Science.gov (United States)

    Kumar, Naresh; Zoladek-Lemanczyk, Alina; Guilbert, Anne A Y; Su, Weitao; Tuladhar, Sachetan M; Kirchartz, Thomas; Schroeder, Bob C; McCulloch, Iain; Nelson, Jenny; Roy, Debdulal; Castro, Fernando A

    2017-02-23

    Novel optoelectronic devices rely on complex nanomaterial systems where the nanoscale morphology and local chemical composition are critical to performance. However, the lack of analytical techniques that can directly probe these structure-property relationships at the nanoscale presents a major obstacle to device development. In this work, we present a novel method for non-destructive, simultaneous mapping of the morphology, chemical composition and photoelectrical properties with performance. For instance, the direct measurement of fullerene phase purity can distinguish between high purity aggregates that lead to poor performance and lower purity aggregates (fullerene intercalated with polymer) that result in strong photocurrent generation and collection. We show that the reliable determination of the structure-property relationship at the nanoscale can remove ambiguity from macroscopic device data and support the identification of the best routes for device optimisation. The multi-parameter measurement approach demonstrated herein is expected to play a significant role in guiding the rational design of nanomaterial-based optoelectronic devices, by opening a new realm of possibilities for advanced investigation via the combination of nanoscale optical spectroscopy with a whole range of scanning probe microscopy modes.

  5. Towards automated segmentation of cells and cell nuclei in nonlinear optical microscopy.

    Science.gov (United States)

    Medyukhina, Anna; Meyer, Tobias; Schmitt, Michael; Romeike, Bernd F M; Dietzek, Benjamin; Popp, Jürgen

    2012-11-01

    Nonlinear optical (NLO) imaging techniques based e.g. on coherent anti-Stokes Raman scattering (CARS) or two photon excited fluorescence (TPEF) show great potential for biomedical imaging. In order to facilitate the diagnostic process based on NLO imaging, there is need for an automated calculation of quantitative values such as cell density, nucleus-to-cytoplasm ratio, average nuclear size. Extraction of these parameters is helpful for the histological assessment in general and specifically e.g. for the determination of tumor grades. This requires an accurate image segmentation and detection of locations and boundaries of cells and nuclei. Here we present an image processing approach for the detection of nuclei and cells in co-registered TPEF and CARS images. The algorithm developed utilizes the gray-scale information for the detection of the nuclei locations and the gradient information for the delineation of the nuclear and cellular boundaries. The approach reported is capable for an automated segmentation of cells and nuclei in multimodal TPEF-CARS images of human brain tumor samples. The results are important for the development of NLO microscopy into a clinically relevant diagnostic tool.

  6. Automated centreline extraction of neuronal dendrite from optical microscopy image stacks

    Science.gov (United States)

    Xiao, Liang; Zhang, Fanbiao

    2010-11-01

    In this work we present a novel vision-based pipeline for automated skeleton detection and centreline extraction of neuronal dendrite from optical microscopy image stacks. The proposed pipeline is an integrated solution that merges image stacks pre-processing, the seed points detection, ridge traversal procedure, minimum spanning tree optimization and tree trimming into to a unified framework to deal with the challenge problem. In image stacks preprocessing, we first apply a curvelet transform based shrinkage and cycle spinning technique to remove the noise. This is followed by the adaptive threshold method to compute the result of neuronal object segmentation, and the 3D distance transformation is performed to get the distance map. According to the eigenvalues and eigenvectors of the Hessian matrix, the skeleton seed points are detected. Staring from the seed points, the initial centrelines are obtained using ridge traversal procedure. After that, we use minimum spanning tree to organize the geometrical structure of the skeleton points, and then we use graph trimming post-processing to compute the final centreline. Experimental results on different datasets demonstrate that our approach has high reliability, good robustness and requires less user interaction.

  7. Multimodal nonlinear optical microscopy improves the accuracy of early diagnosis of squamous intraepithelial neoplasia

    Science.gov (United States)

    Teh, Seng Khoon; Zheng, Wei; Li, Shuxia; Li, Dong; Zeng, Yan; Yang, Yanqi; Qu, Jianan Y.

    2013-03-01

    We explore diagnostic utility of a multicolor excitation multimodal nonlinear optical (NLO) microscopy for noninvasive detection of squamous epithelial precancer in vivo. The 7,12-dimenthylbenz(a)anthracene treated hamster cheek pouch was used as an animal model of carcinogenesis. The NLO microscope system employed was equipped with the ability to collect multiple tissue endogenous NLO signals such as two-photon excited fluorescence of keratin, nicotinamide adenine dinucleotide, collagen, and tryptophan, and second harmonic generation of collagen in spectral and time domains simultaneously. A total of 34 (11 controlled and 23 treated) Golden Syrian hamsters with 62 in vivo spatially distinct measurement sites were assessed in this study. High-resolution label-free NLO images were acquired from stratum corneum, stratum granulosum-stratum basale, and stroma for all tissue measurement sites. A total of nine and eight features from 745 and 600 nm excitation wavelengths, respectively, involving tissue structural and intrinsic biochemical properties were found to contain significant diagnostic information for precancers detection (p<0.05). Particularly, 600 nm excited tryptophan fluorescence signals emanating from stratum corneum was revealed to provide remarkable diagnostic utility. Multivariate statistical techniques confirmed the integration of diagnostically significant features from multicolor excitation wavelengths yielded improved diagnostic accuracy as compared to using the individual wavelength alone.

  8. Combining scanning haptic microscopy and fibre optic Raman spectroscopy for tissue characterization.

    Science.gov (United States)

    Candefjord, S; Murayama, Y; Nyberg, M; Hallberg, J; Ramser, K; Ljungberg, B; Bergh, A; Lindahl, O A

    2012-08-01

    The tactile resonance method (TRM) and Raman spectroscopy (RS) are promising for tissue characterization in vivo. Our goal is to combine these techniques into one instrument, to use TRM for swift scanning, and RS for increasing the diagnostic power. The aim of this study was to determine the classification accuracy, using support vector machines, for measurements on porcine tissue and also produce preliminary data on human prostate tissue. This was done by developing a new experimental set-up combining micro-scale TRM-scanning haptic microscopy (SHM)-for assessing stiffness on a micro-scale, with fibre optic RS measurements for assessing biochemical content. We compared the accuracy using SHM alone versus SHM combined with RS, for different degrees of tissue homogeneity. The cross-validation classification accuracy for healthy porcine tissue types using SHM alone was 65-81%, and when RS was added it increased to 81-87%. The accuracy for healthy and cancerous human tissue was 67-70% when only SHM was used, and increased to 72-77% for the combined measurements. This shows that the potential for swift and accurate classification of healthy and cancerous prostate tissue is high. This is promising for developing a tool for probing the surgical margins during prostate cancer surgery.

  9. Characterization of arbitrary fiber taper profiles with optical microscopy and image processing algorithms

    Science.gov (United States)

    Farias, Heric D.; Sebem, Renan; Paterno, Aleksander S.

    2014-08-01

    This work reports results from the development of a software to process the parameters involved in the characterization of fiber taper profiles, while using optical microscopy, a high-definition camera and a high- precision translation stage as the moveable base on which the taper is positioned. In addition to this procedure, image processing algorithms were customized to process the acquired images. With edge detection algorithms in the stitched image, one would be able to characterize the given taper radius curve that represents the taper profile when the camera has a sufficient resolution. As a consequence, the proposed fiber taper characterization procedure is a first step towards a high-resolution characterization of fiber taper diameters with arbitrary profiles, specially this case, in which tapers are fabricated with the stepwise technique that allows the production of non- biconical profiles. The parameters of the stitched images depends on the used microscope objective and the length of the characterized tapers. A non-biconical arbitrary taper is measured as an example for the illustration of the developed software and procedure.

  10. Quantifying three-dimensional rodent retina vascular development using optical tissue clearing and light-sheet microscopy

    Science.gov (United States)

    Singh, Jasmine N.; Nowlin, Taylor M.; Seedorf, Gregory J.; Abman, Steven H.; Shepherd, Douglas P.

    2017-07-01

    Retinal vasculature develops in a highly orchestrated three-dimensional (3-D) sequence. The stages of retinal vascularization are highly susceptible to oxygen perturbations. We demonstrate that optical tissue clearing of intact rat retinas and light-sheet microscopy provides rapid 3-D characterization of vascular complexity during retinal development. Compared with flat mount preparations that dissect the retina and primarily image the outermost vascular layers, intact cleared retinas imaged using light-sheet fluorescence microscopy display changes in the 3-D retinal vasculature rapidly without the need for point scanning techniques. Using a severe model of retinal vascular disruption, we demonstrate that a simple metric based on Sholl analysis captures the vascular changes observed during retinal development in 3-D. Taken together, these results provide a methodology for rapidly quantifying the 3-D development of the entire rodent retinal vasculature.

  11. The thin layer technique and its application to electron microscopy; La technique des couches minces et son application a la microscopie electronique

    Energy Technology Data Exchange (ETDEWEB)

    Ranc, G. [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

    1957-10-15

    This work deals with the technique of thin layers obtained by evaporation under vacuum, in the thickness range extending from a few monoatomic layers to several hundred angstroms. The great theoretical and practical interest of these layers has, it is well known, given rise to many investigations from Faraday onwards. Within the necessarily restricted limits of this study, we shall approach the problem more particularly from the point of view of: - their production; - their use in electron microscopy. A critical appraisal is made, in the light of present-day knowledge, based on our personal experience and on an extensive bibliography which we have collected on the subject. (author) [French] Le present travail concerne la technique des couches minces obtenues par evaporation sous vide, dans le domaine d'epaisseur qui s'etend de quelques couches monoatomiques a plusieurs centaines d'angstroms. L'interet theorique et pratique considerable de ces couches a suscite, comme on sait, de nombreux travaux depuis Faraday. Dans le cadre necessairement restreint de cette these, nous aborderons plus particulierement le point de vue de: - leur obtention; - leur utilisation en microscopie electronique. Il s'agit d'une mise au point critique, a la lumiere des connaissances actuelles, appuyee sur notre experience personnelle et sur une importante bibliographie, qu'il nous a ete donne de reunir a ce sujet. (auteur)

  12. Measurement of time-varying displacement fields in cell culture for traction force optical coherence microscopy (Conference Presentation)

    Science.gov (United States)

    Mulligan, Jeffrey A.; Adie, Steven G.

    2017-02-01

    Mechanobiology is an emerging field which seeks to link mechanical forces and properties to the behaviors of cells and tissues in cancer, stem cell growth, and other processes. Traction force microscopy (TFM) is an imaging technique that enables the study of traction forces exerted by cells on their environment to migrate as well as sense and manipulate their surroundings. To date, TFM research has been performed using incoherent imaging modalities and, until recently, has been largely confined to the study of cell-induced tractions within two-dimensions using highly artificial and controlled environments. As the field of mechanobiology advances, and demand grows for research in physiologically relevant 3D culture and in vivo models, TFM will require imaging modalities that support such settings. Optical coherence microscopy (OCM) is an interferometric imaging modality which enables 3D cellular resolution imaging in highly scattering environments. Moreover, optical coherence elastography (OCE) enables the measurement of tissue mechanical properties. OCE relies on the principle of measuring material deformations in response to artificially applied stress. By extension, similar techniques can enable the measurement of cell-induced deformations, imaged with OCM. We propose traction force optical coherence microscopy (TF-OCM) as a natural extension and partner to existing OCM and OCE methods. We report the first use of OCM data and digital image correlation to track temporally varying displacement fields exhibited within a 3D culture setting. These results mark the first steps toward the realization of TF-OCM in 2D and 3D settings, bolstering OCM as a platform for advancing research in mechanobiology.

  13. Interferometric and nonlinear-optical spectral-imaging techniques for outer space and live cells

    Science.gov (United States)

    Itoh, Kazuyoshi

    2015-12-01

    Multidimensional signals such as the spectral images allow us to have deeper insights into the natures of objects. In this paper the spectral imaging techniques that are based on optical interferometry and nonlinear optics are presented. The interferometric imaging technique is based on the unified theory of Van Cittert-Zernike and Wiener-Khintchine theorems and allows us to retrieve a spectral image of an object in the far zone from the 3D spatial coherence function. The retrieval principle is explained using a very simple object. The promising applications to space interferometers for astronomy that are currently in progress will also be briefly touched on. An interesting extension of interferometric spectral imaging is a 3D and spectral imaging technique that records 4D information of objects where the 3D and spectral information is retrieved from the cross-spectral density function of optical field. The 3D imaging is realized via the numerical inverse propagation of the cross-spectral density. A few techniques suggested recently are introduced. The nonlinear optical technique that utilizes stimulated Raman scattering (SRS) for spectral imaging of biomedical targets is presented lastly. The strong signals of SRS permit us to get vibrational information of molecules in the live cell or tissue in real time. The vibrational information of unstained or unlabeled molecules is crucial especially for medical applications. The 3D information due to the optical nonlinearity is also the attractive feature of SRS spectral microscopy.

  14. Machine learning techniques in optical communication

    DEFF Research Database (Denmark)

    Zibar, Darko; Piels, Molly; Jones, Rasmus Thomas

    2015-01-01

    Techniques from the machine learning community are reviewed and employed for laser characterization, signal detection in the presence of nonlinear phase noise, and nonlinearity mitigation. Bayesian filtering and expectation maximization are employed within nonlinear state-space framework...

  15. Machine learning techniques in optical communication

    DEFF Research Database (Denmark)

    Zibar, Darko; Piels, Molly; Jones, Rasmus Thomas

    2016-01-01

    Machine learning techniques relevant for nonlinearity mitigation, carrier recovery, and nanoscale device characterization are reviewed and employed. Markov Chain Monte Carlo in combination with Bayesian filtering is employed within the nonlinear state-space framework and demonstrated for parameter...

  16. Optical skin friction measurement technique in hypersonic wind tunnel

    Science.gov (United States)

    Chen, Xing; Yao, Dapeng; Wen, Shuai; Pan, Junjie

    2016-10-01

    Shear-sensitive liquid-crystal coatings (SSLCCs) have an optical characteristic that they are sensitive to the applied shear stress. Based on this, a novel technique is developed to measure the applied shear stress of the model surface regarding both its magnitude and direction in hypersonic flow. The system of optical skin friction measurement are built in China Academy of Aerospace Aerodynamics (CAAA). A series of experiments of hypersonic vehicle is performed in wind tunnel of CAAA. Global skin friction distribution of the model which shows complicated flow structures is discussed, and a brief mechanism analysis and an evaluation on optical measurement technique have been made.

  17. Optical Frequency Upconversion Technique for Transmission of Wireless MIMO-Type Signals over Optical Fiber

    OpenAIRE

    R. Q. Shaddad; Mohammad, A. B.; S. A. Al-Gailani; Al-Hetar, A. M.

    2014-01-01

    The optical fiber is well adapted to pass multiple wireless signals having different carrier frequencies by using radio-over-fiber (ROF) technique. However, multiple wireless signals which have the same carrier frequency cannot propagate over a single optical fiber, such as wireless multi-input multi-output (MIMO) signals feeding multiple antennas in the fiber wireless (FiWi) system. A novel optical frequency upconversion (OFU) technique is proposed to solve this problem. In this paper, the n...

  18. Optical correlator techniques applied to robotic vision

    Science.gov (United States)

    Hine, Butler P., III; Reid, Max B.; Downie, John D.

    1991-01-01

    Vision processing is one of the most computationally intensive tasks required of an autonomous robot. The data flow from a single typical imaging sensor is roughly 60 Mbits/sec, which can easily overload current on-board processors. Optical correlator-based processing can be used to perform many of the functions required of a general robotic vision system, such as object recognition, tracking, and orientation determination, and can perform these functions fast enough to keep pace with the incoming sensor data. We describe a hybrid digital electronic/analog optical robotic vision processing system developed at Ames Research Center to test concepts and algorithms for autonomous construction, inspection, and maintenance of space-based habitats. We discuss the system architecture design and implementation, its performance characteristics, and our future plans. In particular, we compare the performance of the system to a more conventional all digital electronic system developed concurrently. The hybrid system consistently outperforms the digital electronic one in both speed and robustness.

  19. Tear film measurement by optical reflectometry technique.

    Science.gov (United States)

    Lu, Hui; Wang, Michael R; Wang, Jianhua; Shen, Meixiao

    2014-02-01

    Evaluation of tear film is performed by an optical reflectometer system with alignment guided by a galvanometer scanner. The reflectometer system utilizes optical fibers to deliver illumination light to the tear film and collect the film reflectance as a function of wavelength. Film thickness is determined by best fitting the reflectance-wavelength curve. The spectral reflectance acquisition time is 15 ms, fast enough for detecting film thickness changes. Fast beam alignment of 1 s is achieved by the galvanometer scanner. The reflectometer was first used to evaluate artificial tear film on a model eye with and without a contact lens. The film thickness and thinning rate have been successfully quantified with the minimum measured thickness of about 0.3 μm. Tear films in human eyes, with and without a contact lens, have also been evaluated. A high-contrast spectral reflectance signal from the precontact lens tear film is clearly observed, and the thinning dynamics have been easily recorded from 3.69 to 1.31 μm with lipid layer thickness variation in the range of 41 to 67 nm. The accuracy of the measurement is better than ±0.58% of the film thickness at an estimated tear film refractive index error of ±0.001. The fiber-based reflectometer system is compact and easy to handle.

  20. Activated sludge characterization through microscopy: A review on quantitative image analysis and chemometric techniques

    Energy Technology Data Exchange (ETDEWEB)

    Mesquita, Daniela P. [IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal); Amaral, A. Luís [IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal); Instituto Politécnico de Coimbra, ISEC, DEQB, Rua Pedro Nunes, Quinta da Nora, 3030-199 Coimbra (Portugal); Ferreira, Eugénio C., E-mail: ecferreira@deb.uminho.pt [IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal)

    2013-11-13

    Graphical abstract: -- Highlights: •Quantitative image analysis shows potential to monitor activated sludge systems. •Staining techniques increase the potential for detection of operational problems. •Chemometrics combined with quantitative image analysis is valuable for process monitoring. -- Abstract: In wastewater treatment processes, and particularly in activated sludge systems, efficiency is quite dependent on the operating conditions, and a number of problems may arise due to sludge structure and proliferation of specific microorganisms. In fact, bacterial communities and protozoa identification by microscopy inspection is already routinely employed in a considerable number of cases. Furthermore, quantitative image analysis techniques have been increasingly used throughout the years for the assessment of aggregates and filamentous bacteria properties. These procedures are able to provide an ever growing amount of data for wastewater treatment processes in which chemometric techniques can be a valuable tool. However, the determination of microbial communities’ properties remains a current challenge in spite of the great diversity of microscopy techniques applied. In this review, activated sludge characterization is discussed highlighting the aggregates structure and filamentous bacteria determination by image analysis on bright-field, phase-contrast, and fluorescence microscopy. An in-depth analysis is performed to summarize the many new findings that have been obtained, and future developments for these biological processes are further discussed.

  1. Two-photon microscopy with diffractive optical elements and spatial light modulators

    Directory of Open Access Journals (Sweden)

    Brendon O Watson

    2010-09-01

    Full Text Available Two-photon microscopy is often performed at slow frame rates, due to the need to serially scan all points in a field of view with a single laser beam. To overcome this problem, we have developed two optical methods that split and multiplex a laser beam across the sample. In the first method a diffractive optical element (DOE generates a fixed number of beamlets that are scanned in parallel, resulting in a corresponding increase in speed, or in signal-to-noise ratio, in time-lapse measurements. The second method uses a computer-controlled spatial light modulator (SLM, to generate any arbitrary spatio-temporal light pattern. With an SLM one can image or photostimulate any predefined region of the image, such as neurons or dendritic spines. In addition, SLMs can be used to mimic a large number of optical transfer functions, including light path corrections or as adaptive optical devices.

  2. Optical Super-Resolution Imaging of β-Amyloid Aggregation In Vitro and In Vivo: Method and Techniques.

    Science.gov (United States)

    Pinotsi, Dorothea; Kaminski Schierle, Gabriele S; Kaminski, Clemens F

    2016-01-01

    Super-resolution microscopy has emerged as a powerful and non-invasive tool for the study of molecular processes both in vitro and in live cells. In particular, super-resolution microscopy has proven valuable for research studies in protein aggregation. In this chapter we present details of recent advances in this method and the specific techniques, enabling the study of amyloid beta aggregation optically, both in vitro and in cells. First, we show that variants of optical super-resolution microscopy provide a capability to visualize oligomeric and fibrillar structures directly, providing detailed information on species morphology in vitro and even in situ, in the cellular environment. We focus on direct Stochastic Optical Reconstruction Microscopy, dSTORM, which provides morphological detail on spatial scales below 20 nm, and provide detailed protocols for its implementation in the context of amyloid beta research. Secondly, we present a range of optical techniques that offer super-resolution indirectly, which we call multi-parametric microscopy. The latter offers molecular scale information on self-assembly reactions via changes in protein or fluorophore spectral signatures. These techniques are empowered by our recent discovery that disease related amyloid proteins adopt intrinsic energy states upon fibrilisation. We show that fluorescence lifetime imaging provides a particularly sensitive readout to report on the aggregation state, which is robustly quantifiable for experiments performed either in vitro or in vivo.

  3. Phase retrieval techniques for adaptive optics

    Energy Technology Data Exchange (ETDEWEB)

    Carrano, C. J., LLNL

    1998-03-01

    We have developed and tested a method for minimizing static aberrations in adaptive optics systems. In order to correct the static phase aberrations, we need to measure the aberrations through the entire system. We have employed various phase retrieval algorithms to detect these aberrations. We have performed simulations of our experimental setup demonstrating that phase retrieval can improve the static aberrations to below the 20 nm rms level, with the limiting factor being local turbulence in the A0 system. Experimentally thus far, we have improved the static aberrations down to the 50 nm level, with the limiting factor being the ability to adjust the deformable mirror. This should be improved with better control algorithms now being implemented.

  4. Optical image segmentation using wavelet filtering techniques

    Science.gov (United States)

    Veronin, Christopher P.

    1990-12-01

    This research effort successfully implemented an automatic, optically based image segmentation scheme for locating potential targets in a cluttered FLIR image. Such a design is critical to achieve real-time segmentation and classification for machine vision applications. The segmentation scheme used in this research was based on texture discrimination and employs orientation specific, bandpass spatial filters as its main component. The orientation specific, bandpass spatial filters designed during this research include symmetrically located circular apertures implemented on heavy, black aluminum foil; cosine and sine Gabor filters implemented with detour-phase computer generated holography photoreduced onto glass slides; and symmetrically located circular apertures implemented on a liquid crystal television (LCTV) for real-time filter selection. The most successful design was the circular aperture pairs implemented on the aluminum foil. Segmentation was illustrated for simple and complex texture slides, glass template slides, and static and real-time FLIR imagery displayed on an LCTV.

  5. Field results of antifouling techniques for optical instruments

    Science.gov (United States)

    Strahle, W.J.; Hotchkiss, F.S.; Martini, M.A.

    1998-01-01

    An anti-fouling technique is developed for the protection of optical instruments from biofouling which leaches a bromide compound into a sample chamber and pumps new water into the chamber prior to measurement. The primary advantage of using bromide is that it is less toxic than the metal-based antifoulants. The drawback of the bromide technique is also discussed.

  6. Revealing T-Tubules in Striated Muscle with New Optical Super-Resolution Microscopy Techniquess.

    Science.gov (United States)

    Jayasinghe, Isuru D; Clowsley, Alexander H; Munro, Michelle; Hou, Yufeng; Crossman, David J; Soeller, Christian

    2015-01-07

    The t-tubular system plays a central role in the synchronisation of calcium signalling and excitation-contraction coupling in most striated muscle cells. Light microscopy has been used for imaging t-tubules for well over 100 years and together with electron microscopy (EM), has revealed the three-dimensional complexities of the t-system topology within cardiomyocytes and skeletal muscle fibres from a range of species. The emerging super-resolution single molecule localisation microscopy (SMLM) techniques are offering a near 10-fold improvement over the resolution of conventional fluorescence light microscopy methods, with the ability to spectrally resolve nanometre scale distributions of multiple molecular targets. In conjunction with the next generation of electron microscopy, SMLM has allowed the visualisation and quantification of intricate t-tubule morphologies within large areas of muscle cells at an unprecedented level of detail. In this paper, we review recent advancements in the t-tubule structural biology with the utility of various microscopy techniques. We outline the technical considerations in adapting SMLM to study t-tubules and its potential to further our understanding of the molecular processes that underlie the sub-micron scale structural alterations observed in a range of muscle pathologies.

  7. Optical and digital microscopic imaging techniques and applications in pathology

    OpenAIRE

    Xiaodong Chen; Bin Zheng; Hong Liu

    2011-01-01

    The conventional optical microscope has been the primary tool in assisting pathological examinations. The modern digital pathology combines the power of microscopy, electronic detection, and computerized analysis. It enables cellular-, molecular-, and genetic-imaging at high efficiency and accuracy to facilitate clinical screening and diagnosis. This paper first reviews the fundamental concepts of microscopic imaging and introduces the technical features and associated clinical applications o...

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

    Science.gov (United States)

    Kettel, Johannes; Reinecke, Holger; Müller, Claas

    2016-04-01

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

  9. Biophotonic endoscopy: a review of clinical research techniques for optical imaging and sensing of early gastrointestinal cancer.

    Science.gov (United States)

    Coda, Sergio; Siersema, Peter D; Stamp, Gordon W H; Thillainayagam, Andrew V

    2015-10-01

    Detection, characterization, and staging constitute the fundamental elements in the endoscopic diagnosis of gastrointestinal diseases, but histology still remains the diagnostic gold standard. New developments in endoscopic techniques may challenge histopathology in the near future. An ideal endoscopic technique should combine a wide-field, "red flag" screening technique with an optical contrast or microscopy method for characterization and staging, all simultaneously available during the procedure. In theory, biophotonic advances have the potential to unite these elements to allow in vivo "optical biopsy." These techniques may ultimately offer the potential to increase the rates of detection of high risk lesions and the ability to target biopsies and resections, and so reduce the need for biopsy, costs, and uncertainty for patients. However, their utility and sensitivity in clinical practice must be evaluated against those of conventional histopathology. This review describes some of the most recent applications of biophotonics in endoscopic optical imaging and metrology, along with their fundamental principles and the clinical experience that has been acquired in their deployment as tools for the endoscopist. Particular emphasis has been placed on translational label-free optical techniques, such as fluorescence spectroscopy, fluorescence lifetime imaging microscopy (FLIM), two-photon and multi-photon microscopy, second harmonic generation (SHG) and third harmonic generation (THG) imaging, optical coherence tomography (OCT), diffuse reflectance, Raman spectroscopy, and molecular imaging.

  10. Mono-Cycle Photonics and Optical Scanning Tunneling Microscopy Route to Femtosecond Ångstrom Technology

    CERN Document Server

    Yamashita, Mikio; Morita, Ryuji

    2005-01-01

    "Mono-Cycle Photonics and Optical Scanning Tunneling Microscopy" deals with both the ultrashort laser-pulse technology in the few- to mono-cycle region and the laser-surface-controlled scanning-tunneling microscopy (STM) extending into the spatiotemporal extreme technology. The former covers the theory of nonlinear pulse propagation beyond the slowly-varing-envelope approximation, the generation and active chirp compensation of ultrabroadband optical pulses, the amplitude and phase characterization of few- to mono-cycle pulses, and the feedback field control for the mono-cycle-like pulse generation. In addition, the wavelength-multiplex shaping of ultrabroadband pulse is described. The latter covers the CW-laser-excitation STM, the femtosecond-time-resolved STM and atomic-level surface phenomena controlled by femtosecond pulses.

  11. Image edge-enhancement in optical microscopy with a phase mismatched spiral phase plate

    Institute of Scientific and Technical Information of China (English)

    Shibiao Wei; Jing Bu; Siwei Zhu; Xiaocong Yuan

    2011-01-01

    @@ We present a spiral phase filtering system with a large tolerance for edge enhancement of both phase and amplitude objects in optical microscopy.The method is based on a Fourier 4-f spatial filtering system.A phase mismatched spiral phase plate (SPP) fabricated by electron beam lithography is employed as the radial Hilbert transform for image edge enhancement.Compared with holography, SPP is simple,economical, reliable, and easy to integrate.%We present a spiral phase filtering system with a large tolerance for edge enhancement of both phase and amplitude objects in optical microscopy. The method is based on a Fourier 4-f spatial filtering system.A phase mismatched spiral phase plate (SPP) fabricated by electron beam lithography is employed as the radial Hilbert transform for image edge enhancement. Compared with holography, SPP is simple,economical, reliable, and easy to integrate.

  12. Performance Characterization of a Switchable Acoustic Resolution and Optical Resolution Photoacoustic Microscopy System

    Science.gov (United States)

    Moothanchery, Mohesh; Pramanik, Manojit

    2017-01-01

    Photoacoustic microscopy (PAM) is a scalable bioimaging modality; one can choose low acoustic resolution with deep penetration depth or high optical resolution with shallow imaging depth. High spatial resolution and deep penetration depth is rather difficult to achieve using a single system. Here we report a switchable acoustic resolution and optical resolution photoacoustic microscopy (AR-OR-PAM) system in a single imaging system capable of both high resolution and low resolution on the same sample. Lateral resolution of 4.2 µm (with ~1.4 mm imaging depth) and lateral resolution of 45 μm (with ~7.6 mm imaging depth) was successfully demonstrated using a switchable system. In vivo blood vasculature imaging was also performed for its biological application. PMID:28208676

  13. Imaging interferometric microscopy-approaching the linear systems limits of optical resolution.

    Science.gov (United States)

    Kuznetsova, Yuliya; Neumann, Alexander; Brueck, S R

    2007-05-28

    The linear systems optical resolution limit is a dense grating pattern at a lambda/2 pitch or a critical dimension (resolution) of lambda/4. However, conventional microscopy provides a (Rayleigh) resolution of only ~ 0.6lambda/NA, approaching lambda/1.67 as NA ?lambda1. A synthetic aperture approach to reaching the lambda/4 linear-systems limit, extending previous developments in imaginginterferometric microscopy, is presented. Resolution of non-periodic 180-nm features using 633-nm illumination (lambda/3.52) and of a 170-nm grating (lambda/3.72) is demonstrated. These results are achieved with a 0.4-NA optical system and retain the working distance, field-of-view, and depth-of-field advantages of low-NA systems while approaching ultimate resolution limits.

  14. Optical characters and texture maps of skin and the aging mechanism by use of multiphoton microscopy and optical coherence tomography

    Science.gov (United States)

    Wu, Shulian; Li, Hui; Zhang, Xiaoman; Huang, Yudian; Xu, Xiaohui

    2012-03-01

    Cutaneous aging is a complicated biological process affecting different constituents of skin, which can be divided into two types: the chronological aging and the photo-aging. The two cutaneous aging processes often co-exist accompanying with each other. The effects are often overlapped including changes in epithelium and dermis. The degeneration of collagen is a major factor in dermal alteration with aging. In this study, multiphoton microscopy (MPM) with its high resolution imaging and optical coherence tomography (OCT) with its depth resolved imaging were used to study the anti-aging dermatology in vivo. It was attempted to make the optical parameter and texture feature to evaluate the process of aging skin using mathematical image processing. The links among optical parameter, spectrum and texture feature in collagen with aging process were established to uncover mechanism of aging skin.

  15. Nano-scale measurement of biomolecules by optical microscopy and semiconductor nanoparticles

    Directory of Open Access Journals (Sweden)

    Taro eIchimura

    2014-07-01

    Full Text Available Over the past decade, great developments in optical microscopy have made this technology increasingly compatible with biological studies. Fluorescence microscopy has especially contributed to investigating the dynamic behaviors of live specimens and can now resolve objects with nanometer precision and resolution due to super-resolution imaging. Additionally, single particle tracking provides information on the dynamics of individual proteins at the nanometr scale both in vitro and in cells. Complementing advances in microscopy technologies has been the development of fluorescent probes. The quantum dot, a semi-conductor fluorescent nanoparticle, is particularly suitable for single particle tracking and super-resolution imaging. This article overviews the principles of single particle tracking and super resolution along with describing their application to the nanometer measurement/observation of biological systems when combined with quantum dot technologies.

  16. Optical-resolution photoacoustic microscopy of amyloid-β deposits in vivo

    Science.gov (United States)

    Hu, Song; Yan, Ping; Maslov, Konstantin; Lee, Jin-Moo; Wang, Lihong V.

    2010-02-01

    Advances in high-resolution imaging have permitted microscopic observations within the brains of living animals. Applied to Alzheimer's disease (AD) mouse models, multiphoton microscopy has opened a new window to study the real-time appearance and growth of amyloid plaques. Here, we report an alternative technology-optical-resolution photoacoustic microscopy (OR-PAM)-for in vivo imaging of amyloid plaques in a transgenic AD mouse model. In vivo validation using multiphoton microscopy shows that OR-PAM has sufficient sensitivity and spatial resolution to identify amyloid plaques in living brains. In addition, with dual-wavelength OR-PAM, the three-dimensional morphology of amyloid plaques and the surrounding microvasculature are imaged simultaneously through a cranial window. In vivo transcranial OR-PAM imaging of amyloid plaques is highly likely once the imaging parameters are optimized.

  17. Resolution of Internal Total Reflection Scanning Near-field Optical Microscopy

    Institute of Scientific and Technical Information of China (English)

    GE Huayong; GUO Qizhi; TAN Weihan

    2002-01-01

    In this paper, the probe-sample interaction equation based on Mie′s scattering theory is derived, and the resolution of scanning near field optical microscopy is calculated numerically. The results show that the offset of far-field component to near-field component in total field plays an important role in the resolution and the size of samples also has influence on resolution.

  18. Optical tweezers and multiphoton microscopies integrated photonic tool for mechanical and biochemical cell processes studies

    Science.gov (United States)

    de Thomaz, A. A.; Faustino, W. M.; Fontes, A.; Fernandes, H. P.; Barjas-Castro, M. d. L.; Metze, K.; Giorgio, S.; Barbosa, L. C.; Cesar, C. L.

    2007-09-01

    The research in biomedical photonics is clearly evolving in the direction of the understanding of biological processes at the cell level. The spatial resolution to accomplish this task practically requires photonics tools. However, an integration of different photonic tools and a multimodal and functional approach will be necessary to access the mechanical and biochemical cell processes. This way we can observe mechanicaly triggered biochemical events or biochemicaly triggered mechanical events, or even observe simultaneously mechanical and biochemical events triggered by other means, e.g. electricaly. One great advantage of the photonic tools is its easiness for integration. Therefore, we developed such integrated tool by incorporating single and double Optical Tweezers with Confocal Single and Multiphoton Microscopies. This system can perform 2-photon excited fluorescence and Second Harmonic Generation microscopies together with optical manipulations. It also can acquire Fluorescence and SHG spectra of specific spots. Force, elasticity and viscosity measurements of stretched membranes can be followed by real time confocal microscopies. Also opticaly trapped living protozoas, such as leishmania amazonensis. Integration with CARS microscopy is under way. We will show several examples of the use of such integrated instrument and its potential to observe mechanical and biochemical processes at cell level.

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

    Science.gov (United States)

    Harrell, Shelley; Zaretsky, Erwin V.

    1961-01-01

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

  20. Tumor margin detection using optical biopsy techniques

    Science.gov (United States)

    Zhou, Yan; Liu, Cheng-hui; Li, Jiyou; Li, Zhongwu; Zhou, Lixin; Chen, Ke; Pu, Yang; He, Yong; Zhu, Ke; Li, Qingbo; Alfano, Robert R.

    2014-03-01

    The aim of this study is to use the Resonance Raman (RR) and fluorescence spectroscopic technique for tumor margin detection with high accuracy based on native molecular fingerprints of breast and gastrointestinal (GI) tissues. This tumor margins detection method utilizes advantages of RR spectroscopic technique in situ and in real-time to diagnose tumor changes providing powerful tools for clinical guiding intraoperative margin assessments and postoperative treatments. The tumor margin detection procedures by RR spectroscopy were taken by scanning lesion from center or around tumor region in ex-vivo to find the changes in cancerous tissues with the rim of normal tissues using the native molecular fingerprints. The specimens used to analyze tumor margins include breast and GI carcinoma and normal tissues. The sharp margin of the tumor was found by the changes of RR spectral peaks within 2 mm distance. The result was verified using fluorescence spectra with 300 nm, 320 nm and 340 nm excitation, in a typical specimen of gastric cancerous tissue within a positive margin in comparison with normal gastric tissues. This study demonstrates the potential of RR and fluorescence spectroscopy as new approaches with labeling free to determine the intraoperative margin assessment.

  1. New Technique for Successful Thermal Barrier Coating Specimen Preparation for Transmission Electron Microscopy.

    Science.gov (United States)

    Brickey; Lee

    2000-05-01

    Reliability of thermal barrier coatings (TBC) hinges on the adhesion of a thermally grown oxide scale to an insulative ceramic topcoat and an underlying metallic bondcoat. The width of the scale and its interfaces makes transmission electron microscopy (TEM) an appropriate tool for its analysis. However, specimen preparation has proven to be a challenging obstacle leading to a dearth of TEM research on TBCs. A new approach to cross-section TBC TEM specimen preparation is described. The principal advantages of this technique are reproducibility, reduced specimen damage, and time savings resulting from decreased ion milling. This technique has been successfully applied to numerous TBC specimens with various thermal histories.

  2. Visualizing and Calculating Tip-Substrate Distance in Nanoscale Scanning Electrochemical Microscopy Using 3-Dimensional Super-Resolution Optical Imaging.

    Science.gov (United States)

    Sundaresan, Vignesh; Marchuk, Kyle; Yu, Yun; Titus, Eric J; Wilson, Andrew J; Armstrong, Chadd M; Zhang, Bo; Willets, Katherine A

    2017-01-03

    We report a strategy for the optical determination of tip-substrate distance in nanoscale scanning electrochemical microscopy (SECM) using three-dimensional super-resolution fluorescence imaging. A phase mask is placed in the emission path of our dual SECM/optical microscope, generating a double helix point spread function at the image plane, which allows us to measure the height of emitting objects relative to the focus of the microscope. By exciting both a fluorogenic reaction at the nanoscale electrode tip as well as fluorescent nanoparticles at the substrate, we are able to calculate the tip-substrate distance as the tip approaches the surface with precision better than 25 nm. Attachment of a fluorescent particle to the insulating sheath of the SECM tip extends this technique to nonfluorogenic electrochemical reactions. Correlated electrochemical and optical determination of tip-substrate distance yielded excellent agreement between the two techniques. Not only does super-resolution imaging offer a secondary feedback mechanism for measuring the tip-sample gap during SECM experiments, it also enables facile tip alignment and a strategy for accounting for electrode tilt relative to the substrate.

  3. Surface diffusion of In on Ge(111) studied by optical second harmonic microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Suni, I.I.; Seebauer, E.G. (Department of Chemical Engineering, University of Illinois, Urbana, Illinois 61801 (United States))

    1994-05-01

    Surface diffusion of In on Ge(111) has been measured by optical second harmonic microscopy. This technique employs surface second harmonic generation to directly image submonolayer surface concentration profiles. The coverage dependence of the diffusivity [ital D] can then be obtained from a Boltzmann--Matano analysis. In the coverage range 0.1[lt][theta][lt]0.48, the activation energy [ital E][sub diff] decreased with increasing coverage, ranging from 31 kcal/mol at [theta]=0.1 to 23 kcal/mol at [theta]=0.48. Over the same coverage range, the pre-exponential factor [ital D][sub 0] decreased from 5[times]10[sup 2] to 1[times]10[sup [minus]1] cm[sup 2]/s. This gradual change reflects a change in diffusion mechanism arising from the disordered nature of the Ge(111) surface. At low coverages, In adatoms sink into the top layer of Ge, and diffusion is dominated by thermal formation of adatom-vacancy pairs. At high coverages, diffusion occurs by normal site-to-site hopping. The gradual change in diffusion parameters with coverage was interrupted by an apparent phase transition at [theta]=0.16. At this point, both [ital E][sub diff] and [ital D][sub 0] peaked sharply at 41 kcal/mol and 6[times]10[sup 5] cm[sup 2]/s, respectively. The desorption energy [ital E][sub des] was measured by temperature programmed desorption. [ital E][sub des] decreased from 60 kcal/mol at submonolayer coverages to 55 kcal/mol at multilayer coverages.

  4. Developing new optical imaging techniques for single particle and molecule tracking in live cells

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Wei [Iowa State Univ., Ames, IA (United States)

    2010-01-01

    Differential interference contrast (DIC) microscopy is a far-field as well as wide-field optical imaging technique. Since it is non-invasive and requires no sample staining, DIC microscopy is suitable for tracking the motion of target molecules in live cells without interfering their functions. In addition, high numerical aperture objectives and condensers can be used in DIC microscopy. The depth of focus of DIC is shallow, which gives DIC much better optical sectioning ability than those of phase contrast and dark field microscopies. In this work, DIC was utilized to study dynamic biological processes including endocytosis and intracellular transport in live cells. The suitability of DIC microscopy for single particle tracking in live cells was first demonstrated by using DIC to monitor the entire endocytosis process of one mesoporous silica nanoparticle (MSN) into a live mammalian cell. By taking advantage of the optical sectioning ability of DIC, we recorded the depth profile of the MSN during the endocytosis process. The shape change around the nanoparticle due to the formation of a vesicle was also captured. DIC microscopy was further modified that the sample can be illuminated and imaged at two wavelengths simultaneously. By using the new technique, noble metal nanoparticles with different shapes and sizes were selectively imaged. Among all the examined metal nanoparticles, gold nanoparticles in rod shapes were found to be especially useful. Due to their anisotropic optical properties, gold nanorods showed as diffraction-limited spots with disproportionate bright and dark parts that are strongly dependent on their orientation in the 3D space. Gold nanorods were developed as orientation nanoprobes and were successfully used to report the self-rotation of gliding microtubules on kinesin coated substrates. Gold nanorods were further used to study the rotational motions of cargoes during the endocytosis and intracellular transport processes in live mammalian

  5. Photothermal and thermoelastic microscopies: two alternative techniques for the non-destructive testing of materials; Microscopies photothermiques et thermoelastiques: deux techniques alternatives pour le CND des materiaux

    Energy Technology Data Exchange (ETDEWEB)

    Fouaidy, M. [Institut de Physique Nucleaire, IN2P3/CNRS 91 - Orsay (France); Ridouane, H. [Faculte des Sciences Ben M' sik, LPPPC, Casablanca (Morocco)

    2002-07-01

    This work aims at evaluating the possibility of application of photothermal and thermoelastic microscopies to the non-destructive testing of materials, such as niobium used in the fabrication of superconductive RF cavities. The theoretical results obtained in this study show the diagnostic potentialities of these techniques when applied to niobium sheets or directly to cavities. The microscopes that use an intensity modulated laser as excitation source have a lateral resolution comprised between 1 {mu}m for f{sub mod} = 10 MHz and 30 to 50 {mu}m for f{sub mod} = 10 kHz with a 1 {mu}m diameter beam. These techniques allow the detection, localization, and sometimes the characterization, of subsurface and deep defects and inclusions. In far field regime the resolution of the method depends on the thermal diffusion depth. Thanks to the strong dependence between the laser induced stress and the thickness of the target, the photothermal and thermoelastic microscopes can be used also for the measurement of cavities thickness and internal profile. (J.S.)

  6. Endoscopic Microscopy

    Science.gov (United States)

    Sokolov, Konstantin; Sung, Kung-Bin; Collier, Tom; Clark, Anne; Arifler, Dizem; Lacy, Alicia; Descour, Michael; Richards-Kortum, Rebecca

    2002-01-01

    In vivo endoscopic optical microscopy provides a tool to assess tissue architecture and morphology with contrast and resolution similar to that provided by standard histopathology – without need for physical tissue removal. In this article, we focus on optical imaging technologies that have the potential to dramatically improve the detection, prevention, and therapy of epithelial cancers. Epithelial pre-cancers and cancers are associated with a variety of morphologic, architectural, and molecular changes, which currently can be assessed only through invasive, painful biopsy. Optical imaging is ideally suited to detecting cancer-related alterations because it can detect biochemical and morphologic alterations with sub-cellular resolution throughout the entire epithelial thickness. Optical techniques can be implemented non-invasively, in real time, and at low cost to survey the tissue surface at risk. Our manuscript focuses primarily on modalities that currently are the most developed: reflectance confocal microscopy (RCM) and optical coherence tomography (OCT). However, recent advances in fluorescence-based endoscopic microscopy also are reviewed briefly. We discuss the basic principles of these emerging technologies and their current and potential applications in early cancer detection. We also present research activities focused on development of exogenous contrast agents that can enhance the morphological features important for cancer detection and that have the potential to allow vital molecular imaging of cancer-related biomarkers. In conclusion, we discuss future improvements to the technology needed to develop robust clinical devices. PMID:14646041

  7. Advanced Time-Resolved Fluorescence Microscopy Techniques for the Investigation of Peptide Self-Assembly

    Science.gov (United States)

    Anthony, Neil R.

    The ubiquitous cross beta sheet peptide motif is implicated in numerous neurodegenerative diseases while at the same time offers remarkable potential for constructing isomorphic high-performance bionanomaterials. Despite an emerging understanding of the complex folding landscape of cross beta structures in determining disease etiology and final structure, we lack knowledge of the critical initial stages of nucleation and growth. In this dissertation, I advance our understanding of these key stages in the cross-beta nucleation and growth pathways using cutting-edge microscopy techniques. In addition, I present a new combined time-resolved fluorescence analysis technique with the potential to advance our current understanding of subtle molecular level interactions that play a pivotal role in peptide self-assembly. Using the central nucleating core of Alzheimer's Amyloid-beta protein, Abeta(16 22), as a model system, utilizing electron, time-resolved, and non-linear microscopy, I capture the initial and transient nucleation stages of peptide assembly into the cross beta motif. In addition, I have characterized the nucleation pathway, from monomer to paracrystalline nanotubes in terms of morphology and fluorescence lifetime, corroborating the predicted desolvation process that occurs prior to cross-beta nucleation. Concurrently, I have identified unique heterogeneous cross beta domains contained within individual nanotube structures, which have potential bionanomaterials applications. Finally, I describe a combined fluorescence theory and analysis technique that dramatically increases the sensitivity of current time-resolved techniques. Together these studies demonstrate the potential for advanced microscopy techniques in the identification and characterization of the cross-beta folding pathway, which will further our understanding of both amyloidogenesis and bionanomaterials.

  8. Fossilised microcenoses and microdebris in Cretaceous amber Alava (Spain) explored using several microscopy techniques

    Science.gov (United States)

    Ascaso, C.; Wierzchos, J.; Corral, J. C.; López, R.; Alonso, J.

    2003-04-01

    It is obvious that Mars return missions are not likely to provide us with fossil-bearing amber resins. Nonetheless, there is much to learn from the study of the biodiversity of fossilised microorganisms, their fossilization processes and detection strategies. In particular, if we are able to determine the endurance of biomolecules in this ancient material, this might contribute to existing knowledge on the persistence of dormant forms over millennia. Amber is a superb medium for the fossilization of organisms. Besides light microscopy techniques, this report describes the use of scanning electron microscopy both in backscattered electron (SEM-BSE) and low temperature (LTSEM) modes, and confocal laser scanning microscopy to examine microorganisms and microdebris (remains of pluricellular organisms). These novel techniques were applied to inclusions in amber (dated as Allaian: Early Cretaceous) from Álava (N Spain). Confocal microscopy provides a 3D image of microcenoses showing very well preserved biomolecules; fungal hyphae and protozoan cells emitting a strong autofluorescence signal. The huge potential of SEM-BSE was demonstrated by high resolution images, in which the relationship between fossilized (mineralized) protozoa and fungal hyphae could be observed. Moreover, this technique enabled the observation and description of further ultrastructural details of the cytoplasm of protozoa and fungal hyphae. Energy dispersive X-ray spectroscopy revealed that protozoan and fungal cells were transformed by mineralisation process into fossils composed of iron sulphide with highly preserved ultrastructural details. LTSEM performed on protozoan inclusions generated images showing many vacuoles. Finally, SEM in secondary electron detection mode was found to provide micromorphological information on mummified (not mineralized) bacteria-like microbiota trapped in gas bubbles

  9. Scattering-type scanning near-field optical microscopy with reconstruction of vertical interaction

    Science.gov (United States)

    Wang, Le; Xu, Xiaoji G.

    2015-11-01

    Scattering-type scanning near-field optical microscopy provides access to super-resolution spectroscopic imaging of the surfaces of a variety of materials and nanostructures. In addition to chemical identification, it enables observations of nano-optical phenomena, such as mid-infrared plasmons in graphene and phonon polaritons in boron nitride. Despite the high lateral spatial resolution, scattering-type near-field optical microscopy is not able to provide characteristics of near-field responses in the vertical dimension, normal to the sample surface. Here, we present an accurate and fast reconstruction method to obtain vertical characteristics of near-field interactions. For its first application, we investigated the bound electromagnetic field component of surface phonon polaritons on the surface of boron nitride nanotubes and found that it decays within 20 nm with a considerable phase change in the near-field signal. The method is expected to provide characterization of the vertical field distribution of a wide range of nano-optical materials and structures.

  10. Remote optical sensing on the nanometer scale with a bowtie aperture nano-antenna on a fiber tip of scanning near-field optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Atie, Elie M.; Xie, Zhihua; El Eter, Ali; Salut, Roland; Baida, Fadi I.; Grosjean, Thierry, E-mail: thierry.grosjean@univ-fcomte.fr [Institut FEMTO-ST, UMR CNRS 6174, Université de Franche-Comté, Département d' Optique P.M. Duffieux, 15B avenue des Montboucons, 25030 Besançon cedex (France); Nedeljkovic, Dusan [Lovalite s.a.s., 7 rue Xavier Marmier, 25000 Besançon (France); Tannous, Tony [Department of Physics, University of Balamand, P.O. Box 100 Tripoli (Lebanon)

    2015-04-13

    Plasmonic nano-antennas have proven the outstanding ability of sensing chemical and physical processes down to the nanometer scale. Sensing is usually achieved within the highly confined optical fields generated resonantly by the nano-antennas, i.e., in contact to the nanostructures. In this paper, we demonstrate the sensing capability of nano-antennas to their larger scale environment, well beyond their plasmonic confinement volume, leading to the concept of “remote” (non contact) sensing on the nanometer scale. On the basis of a bowtie-aperture nano-antenna (BNA) integrated at the apex of a SNOM (Scanning Near-field Optical Microscopy) fiber tip, we introduce an ultra-compact, moveable, and background-free optical nanosensor for the remote sensing of a silicon surface (up to distance of 300 nm). Sensitivity of the BNA to its large scale environment is high enough to expect the monitoring and control of the spacing between the nano-antenna and a silicon surface with sub-nanometer accuracy. This work paves the way towards an alternative class of nanopositioning techniques, based on the monitoring of diffraction-free plasmon resonance, that are alternative to nanomechanical and diffraction-limited optical interference-based devices.

  11. Remote optical sensing on the nanometer scale with a bowtie aperture nano-antenna on a fiber tip of scanning near-field optical microscopy

    Science.gov (United States)

    Atie, Elie M.; Xie, Zhihua; El Eter, Ali; Salut, Roland; Nedeljkovic, Dusan; Tannous, Tony; Baida, Fadi I.; Grosjean, Thierry

    2015-04-01

    Plasmonic nano-antennas have proven the outstanding ability of sensing chemical and physical processes down to the nanometer scale. Sensing is usually achieved within the highly confined optical fields generated resonantly by the nano-antennas, i.e., in contact to the nanostructures. In this paper, we demonstrate the sensing capability of nano-antennas to their larger scale environment, well beyond their plasmonic confinement volume, leading to the concept of "remote" (non contact) sensing on the nanometer scale. On the basis of a bowtie-aperture nano-antenna (BNA) integrated at the apex of a SNOM (Scanning Near-field Optical Microscopy) fiber tip, we introduce an ultra-compact, moveable, and background-free optical nanosensor for the remote sensing of a silicon surface (up to distance of 300 nm). Sensitivity of the BNA to its large scale environment is high enough to expect the monitoring and control of the spacing between the nano-antenna and a silicon surface with sub-nanometer accuracy. This work paves the way towards an alternative class of nanopositioning techniques, based on the monitoring of diffraction-free plasmon resonance, that are alternative to nanomechanical and diffraction-limited optical interference-based devices.

  12. Study on application of optical clearing technique in skin diseases

    Science.gov (United States)

    Shan, Hao; Liang, Yanmei; Wang, Jingyi; Li, Yan

    2012-11-01

    So far, the study of the optical clearing is almost always about healthy tissue. However, the ultimate goal is to detect diseases for clinical application. Optical clearing on diseased skins is explored. The effect is evaluated by applying a combined liquid paraffin and glycerol mixed solution on several kinds of diseased skins in vitro. Scanning experiments from optical coherence tomography show that it has different effects among fibroma, pigmented nevus, and seborrheic keratosis. Based on the results, we conclude that different skin diseases have different compositions and structures, and their optical parameters and biological characteristics should be different, which implies that the optical clearing technique may have selectivity and may not be suitable for all kinds of skin diseases.

  13. Advanced Equalization Techniques for Digital Coherent Optical Receivers

    DEFF Research Database (Denmark)

    Arlunno, Valeria

    This PhD thesis addresses the design and performance evaluation of advanced Digital Signal Processing (DSP) algorithms for coherent optical fiber transmission systems. The research results presented in this thesis report on transmission of highly spectrally efficient optical communication systems...... employing multiplexing techniques with polarization multiplexing and multi-level modulations format. Advanced digital signal processing techniques offer robustness and flexibility for next generation high capacity optical fibre networks and are therefore considered as key building blocks in next generation...... format detection. Feedback equalization structure have been investigated in high order modulation formats transmission, when combined with coding techniques, and for closed spaced multiplexing scenario. Highlight results presented in this PhD thesis include evaluation and implementation of a novel...

  14. Nanohybrids Near-Field Optical Microscopy: From Image Shift to Biosensor Application

    Directory of Open Access Journals (Sweden)

    Nayla El-Kork

    2016-01-01

    Full Text Available Near-Field Optical Microscopy is a valuable tool for the optical and topographic study of objects at a nanometric scale. Nanoparticles constitute important candidates for such type of investigations, as they bear an important weight for medical, biomedical, and biosensing applications. One, however, has to be careful as artifacts can be easily reproduced. In this study, we examined hybrid nanoparticles (or nanohybrids in the near-field, while in solution and attached to gold nanoplots. We found out that they can be used for wavelength modulable near-field biosensors within conditions of artifact free imaging. In detail, we refer to the use of topographic/optical image shift and the imaging of Local Surface Plasmon hot spots to validate the genuineness of the obtained images. In summary, this study demonstrates a new way of using simple easily achievable comparative methods to prove the authenticity of near-field images and presents nanohybrid biosensors as an application.

  15. Congestion estimation technique in the optical network unit registration process.

    Science.gov (United States)

    Kim, Geunyong; Yoo, Hark; Lee, Dongsoo; Kim, Youngsun; Lim, Hyuk

    2016-07-01

    We present a congestion estimation technique (CET) to estimate the optical network unit (ONU) registration success ratio for the ONU registration process in passive optical networks. An optical line terminal (OLT) estimates the number of collided ONUs via the proposed scheme during the serial number state. The OLT can obtain congestion level among ONUs to be registered such that this information may be exploited to change the size of a quiet window to decrease the collision probability. We verified the efficiency of the proposed method through simulation and experimental results.

  16. Techniques for labeling of optical signals in bust switched networks

    DEFF Research Database (Denmark)

    Tafur Monroy, Idelfonso; Koonen, A. M. J.; Zhang, Jianfeng

    2003-01-01

    multiplexed (WDM) networks due to its single forwarding algorithm, thus yielding low latency, and it enables scaling to terabit rates. Moreover, LOBS is compatible with the general multiprotocol label switching (GMPLS) framework for a unified control plane. We present a review on techniques for labeling......We present a review of significant issues related to labeled optical burst switched (LOBS) networks and technologies enabling future optical internet networks. Labeled optical burst switching provides a quick and efficient forwarding mechanism of IP packets/bursts over wavelength division...

  17. Optical design and multi-length-scale scanning spectro-microscopy possibilities at the Nanoscopium beamline of Synchrotron Soleil.

    Science.gov (United States)

    Somogyi, Andrea; Medjoubi, Kadda; Baranton, Gil; Le Roux, Vincent; Ribbens, Marc; Polack, François; Philippot, Pascal; Samama, Jean Pierre

    2015-07-01

    The Nanoscopium 155 m-long beamline of Synchrotron Soleil is dedicated to scanning hard X-ray nanoprobe techniques. Nanoscopium aims to reach ≤100 nm resolution in the 5-20 keV energy range for routine user experiments. The beamline design tackles the tight stability requirements of such a scanning nanoprobe by creating an overfilled secondary source, implementing all horizontally reflecting main beamline optics, applying high mechanical stability equipment and constructing a dedicated high-stability building envelope. Multi-technique scanning imaging and tomography including X-ray fluorescence spectrometry and spectro-microscopy, absorption, differential phase and dark-field contrasts are implemented at the beamline in order to provide simultaneous information on the elemental distribution, speciation and sample morphology. This paper describes the optical concept and the first measured performance of the Nanoscopium beamline followed by the hierarchical length-scale multi-technique imaging experiments performed with dwell times down to 3 ms per pixel.

  18. Measurement of dynamic cell-induced 3D displacement fields in vitro for traction force optical coherence microscopy.

    Science.gov (United States)

    Mulligan, Jeffrey A; Bordeleau, François; Reinhart-King, Cynthia A; Adie, Steven G

    2017-02-01

    Traction force microscopy (TFM) is a method used to study the forces exerted by cells as they sense and interact with their environment. Cell forces play a role in processes that take place over a wide range of spatiotemporal scales, and so it is desirable that TFM makes use of imaging modalities that can effectively capture the dynamics associated with these processes. To date, confocal microscopy has been the imaging modality of choice to perform TFM in 3D settings, although multiple factors limit its spatiotemporal coverage. We propose traction force optical coherence microscopy (TF-OCM) as a novel technique that may offer enhanced spatial coverage and temporal sampling compared to current methods used for volumetric TFM studies. Reconstructed volumetric OCM data sets were used to compute time-lapse extracellular matrix deformations resulting from cell forces in 3D culture. These matrix deformations revealed clear differences that can be attributed to the dynamic forces exerted by normal versus contractility-inhibited NIH-3T3 fibroblasts embedded within 3D Matrigel matrices. Our results are the first step toward the realization of 3D TF-OCM, and they highlight the potential use of OCM as a platform for advancing cell mechanics research.

  19. Evaluation of agave fiber delignification by means of microscopy techniques and image analysis.

    Science.gov (United States)

    Hernández-Hernández, Hilda M; Chanona-Pérez, Jorge J; Calderón-Domínguez, Georgina; Perea-Flores, María J; Mendoza-Pérez, Jorge A; Vega, Alberto; Ligero, Pablo; Palacios-González, Eduardo; Farrera-Rebollo, Reynold R

    2014-10-01

    Recently, the use of different types of natural fibers to produce paper and textiles from agave plants has been proposed. Agave atrovirens can be a good source of cellulose and lignin; nevertheless, the microstructural changes that happen during delignification have scarcely been studied. The aim of this work was to study the microstructural changes that occur during the delignification of agave fibers by means of microscopy techniques and image analysis. The fibers of A. atrovirens were obtained from leaves using convective drying, milling, and sieving. Fibers were processed using the Acetosolv pulping method at different concentrations of acetic acid; increasing acid concentration promoted higher levels of delignification, structural damage, and the breakdown of fiber clumps. Delignification followed by spectrometric analysis and microstructural studies were carried out by light, confocal laser scanning and scanning electron microscopy and showed that the delignification process follows three stages: initial, bulk, and residual. Microscopy techniques and image analysis were efficient tools for microstructural characterization during delignification of agave fibers, allowing quantitative evaluation of the process and the development of linear prediction models. The data obtained integrated numerical and microstructural information that could be valuable for the study of pulping of lignocellulosic materials.

  20. Coherent light microscopy

    CERN Document Server

    Ferraro, Pietro; Zalevsky, Zeev

    2011-01-01

    This book deals with the latest achievements in the field of optical coherent microscopy. While many other books exist on microscopy and imaging, this book provides a unique resource dedicated solely to this subject. Similarly, many books describe applications of holography, interferometry and speckle to metrology but do not focus on their use for microscopy. The coherent light microscopy reference provided here does not focus on the experimental mechanics of such techniques but instead is meant to provide a users manual to illustrate the strengths and capabilities of developing techniques. Th

  1. Two-axis water-immersible microscanning mirror for scanning optics and acoustic microscopy

    Science.gov (United States)

    Xu, Song; Zou, Jun

    2016-10-01

    Fast multiaxis scanning is useful for not only optical but also acoustic microscopic imaging. Although they have been used for optical scanning, the application of (MEMS) scanning mirrors in acoustic microscopy is still very limited due to their small mirror plate size, and more importantly, inability to operate in liquids (as ultrasound coupling media). A microfabricated two-axis water-immersible scanning mirror for optical and acoustic microscopy is reported. It has an optical and acoustically reflective mirror plate (6 mm×4 mm) to provide numerical aperture for ultrasound beam steering. Electromagnetic and mechanical analysis and simulation were conducted to estimate the mechanical tilting angle and resonance frequency of both fast and slow axes, which matches well with the measurement results. The fast axis has a resonant frequency of 320 Hz in air and 220 Hz in water, which is more than 10 times higher than that of the slow axis (24 Hz in air and 14 Hz in water). Under a 100-mA driving current, the scanning angles of the fast axis reached ±9.5 deg in both air and water at the resonance frequency, respectively. The scanning angles of the slow axis reached ±15 deg in air and ±12.5 deg in water at resonant frequencies, respectively. Raster scanning of a collimated laser beam was achieved by driving both axes simultaneously close to their own resonance frequencies. The feasibility of using the two-axis water-immersible scanning mirror in scanning acoustic microscopy was also demonstrated.

  2. Optical characteristics of transparent samarium oxide thin films deposited by the radio-frequency sputtering technique

    Indian Academy of Sciences (India)

    A A ATTA; M M EL-NAHASS; KHALED M ELSABAWY; M M ABD EL-RAHEEM; A M HASSANIEN; A ALHUTHALI; ALI BADAWI; AMAR MERAZGA

    2016-11-01

    Transparent metal oxide thin films of samarium oxide (Sm$_2$O$_3$) were prepared on pre-cleaned fused optically flat quartz substrates by radio-frequency (RF) sputtering technique. The as-deposited thin films were annealed at different temperatures (873, 973 and 1073 K) for 4 h in air under normal atmospheric pressure. The topological morphology of the film surface was characterized by using atomic force microscopy (AFM). The optical properties of the as-prepared and annealed thin films were studied using their reflectance and transmittance spectra at nearly normal incident light. The estimated direct optical band gap energy (E$^{d}_{g}$ ) values were found to increase by increasing the annealing temperatures. The dispersion curves of the refractive index of Sm$_2$O$_3$ thin films were found to obey the single oscillator model.

  3. Study of fossil bones by synchrotron radiation micro-spectroscopic techniques and scanning electron microscopy.

    Science.gov (United States)

    Zougrou, I M; Katsikini, M; Pinakidou, F; Paloura, E C; Papadopoulou, L; Tsoukala, E

    2014-01-01

    Earlymost Villafranchian fossil bones of an artiodactyl and a perissodactyl from the Milia excavation site in Grevena, Greece, were studied in order to evaluate diagenetic effects. Optical microscopy revealed the different bone types (fibro-lamellar and Haversian, respectively) of the two fragments and their good preservation state. The spatial distribution of bone apatite and soil-originating elements was studied using micro-X-ray fluorescence (µ-XRF) mapping and scanning electron microscopy. The approximate value of the Ca/P ratio was 2.2, as determined from scanning electron microscopy measurements. Bacterial boring was detected close to the periosteal region and Fe bearing oxides were found to fill bone cavities, e.g. Haversian canals and osteocyte lacunae. In the perissodactyl bone considerable amounts of Mn were detected close to cracks (the Mn/Fe weight ratio takes values up to 3.5). Goethite and pyrite were detected in both samples by means of metallographic microscopy. The local Ca/P ratio determined with µ-XRF varied significantly in metal-poor spots indicating spatial inhomogeneities in the ionic substitutions. XRF line scans that span the bone cross sections revealed that Fe and Mn contaminate the bones from both the periosteum and medullar cavity and aggregate around local maxima. The formation of goethite, irrespective of the local Fe concentration, was verified by the Fe K-edge X-ray absorption fine structure (XAFS) spectra. Finally, Sr K-edge extended XAFS (EXAFS) revealed that Sr substitutes for Ca in bone apatite without obvious preference to the Ca1 or Ca2 unit-cell site occupation.

  4. Transfer doping of single isolated nanodiamonds, studied by scanning probe microscopy techniques.

    Science.gov (United States)

    Bolker, Asaf; Saguy, Cecile; Kalish, Rafi

    2014-09-26

    The transfer doping of diamond surfaces has been applied in various novel two-dimensional electronic devices. Its extension to nanodiamonds (ND) is essential for ND-based applications in many fields. In particular, understanding the influence of the crystallite size on transfer doping is desirable. Here, we report the results of a detailed study of the electronic energetic band structure of single, isolated transfer-doped nanodiamonds with nanometric resolution using a combination of scanning tunneling spectroscopy and Kelvin force microscopy measurements. The results show how the band gap, the valence band maximum, the electron affinity and the work function all depend on the ND's size and nanoparticle surface properties. The present analysis, which combines information from both scanning tunneling spectroscopy and Kelvin force microscopy, should be applicable to any nanoparticle or surface that can be measured with scanning probe techniques.

  5. Transfer doping of single isolated nanodiamonds, studied by scanning probe microscopy techniques

    Science.gov (United States)

    Bolker, Asaf; Saguy, Cecile; Kalish, Rafi

    2014-09-01

    The transfer doping of diamond surfaces has been applied in various novel two-dimensional electronic devices. Its extension to nanodiamonds (ND) is essential for ND-based applications in many fields. In particular, understanding the influence of the crystallite size on transfer doping is desirable. Here, we report the results of a detailed study of the electronic energetic band structure of single, isolated transfer-doped nanodiamonds with nanometric resolution using a combination of scanning tunneling spectroscopy and Kelvin force microscopy measurements. The results show how the band gap, the valence band maximum, the electron affinity and the work function all depend on the ND’s size and nanoparticle surface properties. The present analysis, which combines information from both scanning tunneling spectroscopy and Kelvin force microscopy, should be applicable to any nanoparticle or surface that can be measured with scanning probe techniques.

  6. Optical Frequency Upconversion Technique for Transmission of Wireless MIMO-Type Signals over Optical Fiber

    Directory of Open Access Journals (Sweden)

    R. Q. Shaddad

    2014-01-01

    Full Text Available The optical fiber is well adapted to pass multiple wireless signals having different carrier frequencies by using radio-over-fiber (ROF technique. However, multiple wireless signals which have the same carrier frequency cannot propagate over a single optical fiber, such as wireless multi-input multi-output (MIMO signals feeding multiple antennas in the fiber wireless (FiWi system. A novel optical frequency upconversion (OFU technique is proposed to solve this problem. In this paper, the novel OFU approach is used to transmit three wireless MIMO signals over a 20 km standard single mode fiber (SMF. The OFU technique exploits one optical source to produce multiple wavelengths by delivering it to a LiNbO3 external optical modulator. The wireless MIMO signals are then modulated by LiNbO3 optical intensity modulators separately using the generated optical carriers from the OFU process. These modulators use the optical single-sideband with carrier (OSSB+C modulation scheme to optimize the system performance against the fiber dispersion effect. Each wireless MIMO signal is with a 2.4 GHz or 5 GHz carrier frequency, 1 Gb/s data rate, and 16-quadrature amplitude modulation (QAM. The crosstalk between the wireless MIMO signals is highly suppressed, since each wireless MIMO signal is carried on a specific optical wavelength.

  7. Optical frequency upconversion technique for transmission of wireless MIMO-type signals over optical fiber.

    Science.gov (United States)

    Shaddad, R Q; Mohammad, A B; Al-Gailani, S A; Al-Hetar, A M

    2014-01-01

    The optical fiber is well adapted to pass multiple wireless signals having different carrier frequencies by using radio-over-fiber (ROF) technique. However, multiple wireless signals which have the same carrier frequency cannot propagate over a single optical fiber, such as wireless multi-input multi-output (MIMO) signals feeding multiple antennas in the fiber wireless (FiWi) system. A novel optical frequency upconversion (OFU) technique is proposed to solve this problem. In this paper, the novel OFU approach is used to transmit three wireless MIMO signals over a 20 km standard single mode fiber (SMF). The OFU technique exploits one optical source to produce multiple wavelengths by delivering it to a LiNbO3 external optical modulator. The wireless MIMO signals are then modulated by LiNbO3 optical intensity modulators separately using the generated optical carriers from the OFU process. These modulators use the optical single-sideband with carrier (OSSB+C) modulation scheme to optimize the system performance against the fiber dispersion effect. Each wireless MIMO signal is with a 2.4 GHz or 5 GHz carrier frequency, 1 Gb/s data rate, and 16-quadrature amplitude modulation (QAM). The crosstalk between the wireless MIMO signals is highly suppressed, since each wireless MIMO signal is carried on a specific optical wavelength.

  8. Reciprocity theory of apertureless scanning near-field optical microscopy with point-dipole probes.

    Science.gov (United States)

    Esslinger, Moritz; Vogelgesang, Ralf

    2012-09-25

    Near-field microscopy offers the opportunity to reveal optical contrast at deep subwavelength scales. In scanning near-field optical microscopy (SNOM), the diffraction limit is overcome by a nanoscopic probe in close proximity to the sample. The interaction of the probe with the sample fields necessarily perturbs the bare sample response, and a critical issue is the interpretation of recorded signals. For a few specific SNOM configurations, individual descriptions have been modeled, but a general and intuitive framework is still lacking. Here, we give an exact formulation of the measurable signals in SNOM which is easily applicable to experimental configurations. Our results are in close analogy with the description Tersoff and Hamann have derived for the tunneling currents in scanning tunneling microscopy. For point-like scattering probe tips, such as used in apertureless SNOM, the theory simplifies dramatically to a single scalar relation. We find that the measured signal is directly proportional to the field of the coupled tip-sample system at the position of the tip. For weakly interacting probes, the model thus verifies the empirical findings that the recorded signal is proportional to the unperturbed field of the bare sample. In the more general case, it provides guidance to an intuitive and faithful interpretation of recorded images, facilitating the characterization of tip-related distortions and the evaluation of novel SNOM configurations, both for aperture-based and apertureless SNOM.

  9. Characterization of bacterial spore germination using integrated phase contrast microscopy, Raman spectroscopy, and optical tweezers.

    Science.gov (United States)

    Kong, Lingbo; Zhang, Pengfei; Setlow, Peter; Li, Yong-Qing

    2010-05-01

    We present a methodology that combines external phase contrast microscopy, Raman spectroscopy, and optical tweezers to monitor a variety of changes during the germination of single Bacillus cereus spores in both nutrient (l-alanine) and non-nutrient (Ca-dipicolinic acid (DPA)) germinants with a temporal resolution of approximately 2 s. Phase contrast microscopy assesses changes in refractility of individual spores during germination, while Raman spectroscopy gives information on changes in spore-specific molecules. The results obtained include (1) the brightness of the phase contrast image of an individual dormant spore is proportional to the level of CaDPA in that spore; (2) the end of the first Stage of germination, revealed as the end of the rapid drop in spore refractility by phase contrast microscopy, precisely corresponds to the completion of the release of CaDPA as revealed by Raman spectroscopy; and (3) the correspondence between the rapid drop in spore refractility and complete CaDPA release was observed not only for spores germinating in the well-controlled environment of an optical trap but also for spores germinating when adhered on a microscope coverslip. Using this latter method, we also simultaneously characterized the distribution of the time-to-complete-CaDPA release (T(release)) of hundreds of individual B. cereus spores germinating with both saturating and subsaturating concentrations of l-alanine and with CaDPA.

  10. Optical and atomic force microscopy of an explanted AcrySof intraocular lens with glistenings.

    Science.gov (United States)

    Dogru, M; Tetsumoto, K; Tagami, Y; Kato, K; Nakamae, K

    2000-04-01

    To assess the surface morphology and cause of glistenings in an explanted AcrySof intraocular lens (IOL). Shakai Hoken Kobe Central Hospital, Kobe, Japan. A 63-year-old Japanese man had implantation of an AcrySof IOL in the capsular bag. One month postoperatively, he had a neodymium:YAG laser capsulotomy for posterior capsule opacification, which changed the IOL's position in the capsular bag. A few months later, the patient developed disabling night glare from intralenticular glistenings and progressive hyperopic refractive error. The IOL was explanted and then analyzed by optical microscopy and atomic force microscopy (AFM). Laboratory analysis of control AcrySof IOLs kept in a balanced salt solution at steady room and body temperature for 2 months was also performed to evaluate the cause of the glistenings observed clinically. Optical microscopy showed that the explanted AcrySof IOL had several microvacuoles; no abnormalities were observed in the control AcrySof IOLs before or after folding at the room and body temperatures. The AFM analysis showed a significant change in the surface morphology of the explanted IOL, including vacuolar formations in the posterior surface as well as numerous anterior surface irregularities. No microvacuoles or surface morphology alterations were observed in the control AcrySof IOLs by AFM analysis. The glistenings in the explanted AcrySof IOL were likely caused by temperature changes and not mechanical stress from folding.

  11. Laser speckle reduction techniques for mid-infrared microscopy and stand-off spectroscopy

    Science.gov (United States)

    Furstenberg, Robert; Kendziora, Christopher A.; Breshike, Christopher J.; Nguyen, Viet; McGill, R. Andrew

    2017-05-01

    Due to their high brightness, infrared (IR) lasers (such as tunable quantum cascade lasers, QCLLs) are very attractive illumination sources in both stand-off spectroscopy and micro-spectroscopy. In fact, they are the enabling device for trace-level spectroscopy. However, due to their high coherence as laser beams, QCLLs can cause speckle, especially when illuminating a rough surface. This is highly detrimental to the signal-to-noise ratio (SNR) of thee collected spectra and can easily negate the gains from using aa high brightness source. In most cases, speckle reduction is performed at the expense of optical power. In this paper, we examine several speckle reduction approaches and evaluate them for their ability to reduce speckle contrast while at the same time preserving aa high optical throughput. We analyze multi-mode fibers, integrating spheres, and stationary and moving diffusers for their speckle reduction potential. Speckle-contrast is measured directly by acquiring beam profiles of the illumination beam or, indirectly, by observing speckle formation from illuminating a rough surface (e.g. Infragold® coated surface) with an IR micro-bolometer camera. We also report on a novel speckle-reducing device with increased optical throughput. We characterize speckle contrast reduction from spatial, temporal and wavelength averaging for both CWW and pulsed QCLs. Examples of effect of speckle-reduction on hyperspectral images in both standoff and microscopy configurations are given.

  12. Multi-color quantum dot stochastic optical reconstruction microscopy (qSTORM)

    Science.gov (United States)

    Tehrani, Kayvan F.; Xu, Jianquan; Kner, Peter A.

    2015-03-01

    Although Single Molecule Localization (SML) techniques have pushed the resolution of fluorescence microscopy beyond the diffraction limit, the accuracy of SML has been limited by the brightness of the fluorophores. The introduction of Quantum Dots (QD) for SML promises to overcome this barrier, and the QD Blueing technique provides a novel approach to SML microscopy. QDs have a higher quantum yield and absorption cross-section, making them brighter, thereby providing a higher accuracy of localization. The blueing technique is also faster and more quantitative than other SML techniques such as dSTORM. The initial bleaching step required by dSTORM is not necessary and each QD is imaged only once as its emission spectrum moves through the blueing window in contrast to dSTORM where the same molecule might be imaged multiple times. Single color QD Blueing has been demonstrated. However in biological imaging, multi-color imaging is essential for understanding the samples under study. Here we introduce two color superresolution microscopy using QD Blueing on biological samples. We demonstrate simultaneous imaging of microtubules and mitochondria in HepG2 cells with a localization accuracy of 40nm. We further show how QD Blueing can be optimized through the control of the sample mounting medium.

  13. Dynamic light scattering and atomic force microscopy techniques for size determination of polyurethane nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Giehl Zanetti-Ramos, Betina [Laboratorio de Bioenergetica e Bioquimica de Macromoleculas, Departamento de Ciencias Farmaceuticas (Brazil)], E-mail: betinagzramos@pq.cnpq.br; Beddin Fritzen-Garcia, Mauricia [Laboratorio de Bioenergetica e Bioquimica de Macromoleculas, Departamento de Ciencias Farmaceuticas (Brazil); Schweitzer de Oliveira, Cristian; Avelino Pasa, Andre [Laboratorio de Filmes Finos e Superficie, Departamento de Fisica (Brazil); Soldi, Valdir [Grupo de Estudos em Materiais Polimericos, Departamento de Quimica, Universidade Federal de Santa Catarina, 88040-900, Florianopolis, SC (Brazil); Borsali, Redouane [Centre de Recherche sur les Macromolecules Vegetales CERMAV/CNRS, 38041 - Grenoble (France); Creczynski-Pasa, Tania Beatriz [Laboratorio de Bioenergetica e Bioquimica de Macromoleculas, Departamento de Ciencias Farmaceuticas (Brazil)

    2009-03-01

    Nanoparticles have applications in various industrial fields principally in drug delivery. Nowadays, there are several processes for manufacturing colloidal polymeric systems and methods of preparation as well as of characterization. In this work, Dynamic Light Scattering and Atomic Force Microscopy techniques were used to characterize polyurethane nanoparticles. The nanoparticles were prepared by miniemulsion technique. The lipophilic monomers, isophorone diisocyanate (IPDI) and natural triol, were emulsified in water containing surfactant. In some formulations the poly(ethylene glycol) was used as co-monomer to obtain the hydrophilic and pegylated nanoparticles. Polyurethane nanoparticles observed by atomic force microscopy (AFM) were spherical with diameter around 209 nm for nanoparticles prepared without PEG. From AFM imaging two populations of nanoparticles were observed in the formulation prepared with PEG (218 and 127 nm) while dynamic light scattering (DLS) measurements showed a monodisperse size distribution around 250 nm of diameters for both formulations. The polydispersity index of the formulations and the experimental procedures could influence the particle size determination with these techniques.

  14. An Optical Cryostat for Use in Microscopy Cooled by Stirling-Type Pulse Tube Cryocooler

    Science.gov (United States)

    Liubiao, Chen; Qiang, Zhou; Xiaoshuang, Zhu; Yuan, Zhou; Junjie, Wang

    The few products of an optical cryostat for use in microscopy in commercialapplications are generally cooled by liquid nitrogen, liquid helium or cryocoolers such as G-M cryocooler or G-M type pulse tube cryocooler (PTC). Sometimes it is not convenient to use G-M cryocooler or G-M type PTC because of its noise and big size; and in some places, liquid nitrogen, especially liquid helium, is not easily available. To overcome this limitation, an optical cryostat for use in microscopy cooled by a Stirling-type pulse tube cryocooler (SPTC) has been designed, built and tested. The refrigerator system SPTC is an important component of the optical cryostat; it has the advantages of compactness, high efficiency, and low vibration. For simplification and compactness, single-stage configuration with coaxial arrangement was employed in the developed SPTC. In order to lower the vibration, the separated configuration was adopted; its compressor and pulse tube are connected with a flexible connecting tube. At present, a lowest temperature of 20 K could be achieved. The temperature fluctuation can be controlled at ±10 mK by adjusting the input electric power to the compressor; and some considerations for further improvement will also be described in this paper.

  15. Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells

    Science.gov (United States)

    Hofemeier, Arne D.; Hachmeister, Henning; Pilger, Christian; Schürmann, Matthias; Greiner, Johannes F. W.; Nolte, Lena; Sudhoff, Holger; Kaltschmidt, Christian; Huser, Thomas; Kaltschmidt, Barbara

    2016-05-01

    Tissue engineering by stem cell differentiation is a novel treatment option for bone regeneration. Most approaches for the detection of osteogenic differentiation are invasive or destructive and not compatible with live cell analysis. Here, non-destructive and label-free approaches of Raman spectroscopy, coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) microscopy were used to detect and image osteogenic differentiation of human neural crest-derived inferior turbinate stem cells (ITSCs). Combined CARS and SHG microscopy was able to detect markers of osteogenesis within 14 days after osteogenic induction. This process increased during continued differentiation. Furthermore, Raman spectroscopy showed significant increases of the PO43‑ symmetric stretch vibrations at 959 cm‑1 assigned to calcium hydroxyapatite between days 14 and 21. Additionally, CARS microscopy was able to image calcium hydroxyapatite deposits within 14 days following osteogenic induction, which was confirmed by Alizarin Red-Staining and RT- PCR. Taken together, the multimodal label-free analysis methods Raman spectroscopy, CARS and SHG microscopy can monitor osteogenic differentiation of adult human stem cells into osteoblasts with high sensitivity and spatial resolution in three dimensions. Our findings suggest a great potential of these optical detection methods for clinical applications including in vivo observation of bone tissue–implant-interfaces or disease diagnosis.

  16. 3D imaging of optically cleared tissue using a simplified CLARITY method and on-chip microscopy

    KAUST Repository

    Zhang, Yibo

    2017-08-12

    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.

  17. Characterization of a circular optical nanoantenna by nonlinear photoemission electron microscopy

    CERN Document Server

    Kaiser, Thomas; Qi, Jing; Klein, Angela; Steinert, Michael; Menzel, Christoph; Rockstuhl, Carsten; Pertsch, Thomas

    2015-01-01

    We report on the investigation of an advanced circular plasmonic nanoantenna under ultrafast excitation using nonlinear photoemission electron microscopy (PEEM) under near-normal incidence. The circular nanoantenna is enhanced in its performance by a supporting grating and milled out from a gold film. The considered antenna shows a sophisticated physical resonance behavior that is ideal to demonstrate the possibilities of PEEM for the experimental investigations of plasmonic effects on the nanoscale. Field profiles of the antenna resonance for both possible linear polarizations of the incident field are measured with high spatial resolution. In addition, outward propagating Hankel plasmons, which are also excited by the structure, are measured and analyzed. We compare our findings to measurements of an isolated plasmonic nanodisc resonator and scanning near-field optical microscopy (SNOM) measurements of both structures. All results are in very good agreement with numerical simulations as well as analytial mo...

  18. Photoemission Electron Microscopy as a tool for the investigation of optical near fields

    CERN Document Server

    Cinchetti, M; Nepjiko, S A; Sch"onhense, G; Rochholz, H; Kreiter, M

    2005-01-01

    Photoemission electron microscopy was used to image the electrons photoemitted from specially tailored Ag nanoparticles deposited on a Si substrate (with its native oxide SiO$_{x}$). Photoemission was induced by illumination with a Hg UV-lamp (photon energy cutoff $\\hbar\\omega_{UV}=5.0$ eV, wavelength $\\lambda_{UV}=250$ nm) and with a Ti:Sapphire femtosecond laser ($\\hbar\\omega_{l}=3.1$ eV, $\\lambda_{l}=400$ nm, pulse width below 200 fs), respectively. While homogeneous photoelectron emission from the metal is observed upon illumination at energies above the silver plasmon frequency, at lower photon energies the emission is localized at tips of the structure. This is interpreted as a signature of the local electrical field therefore providing a tool to map the optical near field with the resolution of emission electron microscopy.

  19. Nanostructured PLD-grown gadolinia doped ceria: Chemical and structural characterization by transmission electron microscopy techniques

    DEFF Research Database (Denmark)

    Rodrigo, Katarzyna Agnieszka; Wang, Hsiang-Jen; Heiroth, Sebastian

    2011-01-01

    The morphology as well as the spatially resolved elemental and chemical characterization of 10 mol% gadolinia doped ceria (CGO10) structures prepared by pulsed laser deposition (PLD) technique are investigated by scanning transmission electron microscopy accompanied with electron energy loss...... spectroscopy and energy dispersive X-ray spectroscopy. A dense, columnar and structurally inhomogeneous CGO10 film, i.e. exhibiting grain size refinement across the film thickness, is obtained in the deposition process. The cerium M4,5 edges, used to monitor the local electronic structure of the grains...

  20. An Optical Fiber Displacement Sensor Using RF Interrogation Technique.

    Science.gov (United States)

    Kim, Hyeon-Ho; Choi, Sang-Jin; Jeon, Keum Soo; Pan, Jae-Kyung

    2016-02-24

    We propose a novel non-contact optical fiber displacement sensor. It uses a radio frequency (RF) interrogation technique which is based on bidirectional modulation of a Mach-Zehnder electro-optical modulator (MZ-EOM). The displacement is measured from the free spectral range (FSR) which is determined by the dip frequencies of the modulated MZ-EOM transfer function. In experiments, the proposed sensor showed a sensitivity of 456 kHz/mm or 1.043 kHz/V in a measurement range of 7 mm. The displacement resolution of the proposed sensor depends on the linewidth and the power of the optical source. Resolution better than 0.05 μm would be achieved if an optical source which has a linewidth narrower than 1.5 nm and a received power larger than -36 dBm is used. Also, the multiplexing characteristic of the proposed sensor was experimentally validated.

  1. Combining digital holographic microscopy and optical tweezers: a new route in microfluidic

    Science.gov (United States)

    Miccio, L.; Memmolo, P.; Merola, F.; Paturzo, M.; Finizio, A.; Grilli, S.; Ferraro, P.

    2012-04-01

    An optical configuration is realized to obtain quantitative phase-contrast maps able to characterize particles floating in a microfluidic chamber by interference microscopy. The novelty is the possibility to drive the sample and measure it thorough the same light path. That is realized by an optical setup made of two light beams coming from the same laser source. One beam provides the optical forces for driving the particle along the desired path and, at same time, it works as object beam in the digital holographic microscope (DHM). The second one acts as reference beam, allowing recording of an interference fringe pattern (i.e., the digital hologram) in an out-of-focus image plane. This work finds application in the field of micromanipulation as, the devise developed allows to operate in microfluidic chambers driving samples flowing in very small volumes. Recently, the field of optical particle micro-manipulation has had rapid growth, due to Optical Tweezers development. A particle is trapped or moved along certain trajectories according to the intensity and phase distribution of the laser beam used. Here, particles freely floating are driven by optical forces along preferential directions and then analyzed by a DHM to numerically calculate their phase-contrast signature. The improvement is that one laser source is employed for making two jobs: driving and analyze the sample. We use two slightly off-axis laser beams coming from a single laser source. The interference between them gives the possibility to record in real-time a sequence of digital holograms, while one of the beam creates the driving force. By this method, a great amount of particles can be analyzed by a real-time recording of DH movies. This allows one to examine each particle at time and characterize it. The optical configuration and the working method are illustrated. Experimental results are shown for polymeric particles and in-vitro.

  2. Nonlinear optical microscopy for histology of fresh normal and cancerous pancreatic tissues.

    Directory of Open Access Journals (Sweden)

    Wenyan Hu

    Full Text Available BACKGROUND: Pancreatic cancer is a lethal disease with a 5-year survival rate of only 1-5%. The acceleration of intraoperative histological examination would be beneficial for better management of pancreatic cancer, suggesting an improved survival. Nonlinear optical methods based on two-photon excited fluorescence (TPEF and second harmonic generation (SHG of intrinsic optical biomarkers show the ability to visualize the morphology of fresh tissues associated with histology, which is promising for real-time intraoperative evaluation of pancreatic cancer. METHODOLOGY/PRINCIPAL FINDINGS: In order to investigate whether the nonlinear optical imaging methods have the ability to characterize pancreatic histology at cellular resolution, we studied different types of pancreatic tissues by using label-free TPEF and SHG. Compared with other routine methods for the preparation of specimens, fresh tissues without processing were found to be most suitable for nonlinear optical imaging of pancreatic tissues. The detailed morphology of the normal rat pancreas was observed and related with the standard histological images. Comparatively speaking, the preliminary images of a small number of chemical-induced pancreatic cancer tissues showed visible neoplastic differences in the morphology of cells and extracellular matrix. The subcutaneous pancreatic tumor xenografts were further observed using the nonlinear optical microscopy, showing that most cells are leucocytes at 5 days after implantation, the tumor cells begin to proliferate at 10 days after implantation, and the extracellular collagen fibers become disordered as the xenografts grow. CONCLUSIONS/SIGNIFICANCE: In this study, nonlinear optical imaging was used to characterize the morphological details of fresh pancreatic tissues for the first time. We demonstrate that it is possible to provide real-time histological evaluation of pancreatic cancer by the nonlinear optical methods, which present an

  3. Transmission line resonance technique for eccentric core optical fibers

    Science.gov (United States)

    Georgantzos, E.; Boucouvalas, A. C.

    2016-12-01

    In several cases optical fibers in telecommunications have cores of non circular geometry. Fibre optic deformations appear in optical fibres for many reasons. Optical fibre core ellipticity for example where the fibre optic core is not perfectly circular due to fibre optic manufacturing tolerances, is measured and often is a problem. Optical fibre core eccentricity, where the fibre core is not on the axis of the fibre, but it is offset by a small length. This is another issue and very important for ensuring performance low loss splices and connector losses. Both of ellipticity and eccentricity are specified in accordance to international standards for fibre optic manufacturing telecommunications grade fibres. The present paper studies ellipticity and core eccentricity specifically and presents a new method for analysing their effect. We present an extension of the transmission line technique as a means of studying such fibers and deriving necessary parameters. Conformal mapping on the other hand is a simple mathematical tool by which we can generate sets of orthogonal two-dimensional coordinate systems. Shortly a conformal map of Cartesian two-dimensional space is defined by any analytical function W(z) where z, w, are: z = x + jy, W = θ + j φ The function deriving by the conformal mapping transformation h(θ ,φ )=| ∂w/∂z | = 1/|∂z/∂w|, can be used in order to define ∇A → and ∇×A → where A → is the magnetic or electric field in the derived orthogonal coordinate system. Useful conformal maps for fiber optics applications should have the property that the equation θ(x, y) = constant, is forming closed curves in a Cartesian two-dimensional space (x,y). If θ(x, y) = constant represents a set of co-eccentric circles, we obtain the normal case of conventional fibers with circular cores. If θ(x, y) = constant represents a set of eclipses, we are have the formation of elliptic core optical fibers. If θ(x, y) = constant represents a set of

  4. Massively parallel data processing for quantitative total flow imaging with optical coherence microscopy and tomography

    Science.gov (United States)

    Sylwestrzak, Marcin; Szlag, Daniel; Marchand, Paul J.; Kumar, Ashwin S.; Lasser, Theo

    2017-08-01

    We present an application of massively parallel processing of quantitative flow measurements data acquired using spectral optical coherence microscopy (SOCM). The need for massive signal processing of these particular datasets has been a major hurdle for many applications based on SOCM. In view of this difficulty, we implemented and adapted quantitative total flow estimation algorithms on graphics processing units (GPU) and achieved a 150 fold reduction in processing time when compared to a former CPU implementation. As SOCM constitutes the microscopy counterpart to spectral optical coherence tomography (SOCT), the developed processing procedure can be applied to both imaging modalities. We present the developed DLL library integrated in MATLAB (with an example) and have included the source code for adaptations and future improvements. Catalogue identifier: AFBT_v1_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/AFBT_v1_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: GNU GPLv3 No. of lines in distributed program, including test data, etc.: 913552 No. of bytes in distributed program, including test data, etc.: 270876249 Distribution format: tar.gz Programming language: CUDA/C, MATLAB. Computer: Intel x64 CPU, GPU supporting CUDA technology. Operating system: 64-bit Windows 7 Professional. Has the code been vectorized or parallelized?: Yes, CPU code has been vectorized in MATLAB, CUDA code has been parallelized. RAM: Dependent on users parameters, typically between several gigabytes and several tens of gigabytes Classification: 6.5, 18. Nature of problem: Speed up of data processing in optical coherence microscopy Solution method: Utilization of GPU for massively parallel data processing Additional comments: Compiled DLL library with source code and documentation, example of utilization (MATLAB script with raw data) Running time: 1,8 s for one B-scan (150 × faster in comparison to the CPU

  5. Approximate Bayesian computation for estimating number concentrations of monodisperse nanoparticles in suspension by optical microscopy

    Science.gov (United States)

    Röding, Magnus; Zagato, Elisa; Remaut, Katrien; Braeckmans, Kevin

    2016-06-01

    We present an approximate Bayesian computation scheme for estimating number concentrations of monodisperse diffusing nanoparticles in suspension by optical particle tracking microscopy. The method is based on the probability distribution of the time spent by a particle inside a detection region. We validate the method on suspensions of well-controlled reference particles. We illustrate its usefulness with an application in gene therapy, applying the method to estimate number concentrations of plasmid DNA molecules and the average number of DNA molecules complexed with liposomal drug delivery particles.

  6. Optical parametric oscillator-based light source for coherent Raman scattering microscopy: practical overview

    Science.gov (United States)

    Brustlein, Sophie; Ferrand, Patrick; Walther, Nico; Brasselet, Sophie; Billaudeau, Cyrille; Marguet, Didier; Rigneault, Hervé

    2011-02-01

    We present the assets and constraints of using optical parametric oscillators (OPOs) to perform point scanning nonlinear microscopy and spectroscopy with special emphasis on coherent Raman spectroscopy. The difterent possible configurations starting with one OPO and two OPOs are described in detail and with comments that are intended to be practically useful for the user. Explicit examples on test samples such as nonlinear organic crystal, polystyrene beads, and fresh mouse tissues are given. Special emphasis is given to background-free coherent Raman anti-Stokes scattering (CARS) imaging, including CARS hyperspectral imaging in a fully automated mode with commercial OPOs.

  7. What advances in microscopy are required for combined MRI and optical functional brain imaging? (Conference Presentation)

    Science.gov (United States)

    Kleinfeld, David

    2016-03-01

    This overview talk will focus on forward-looking scientific needs and physical limits to images of neuronal processes. The challenge in nervous systems is that the basic unit for "switching" events in the nervous system occurs on the one micrometer scale of synaptic spines, while computations involve communication between individual neurons across the full expanse of cortex, which is ten millimeters for mouse cortex. I will address hoped-for advances in optical microscopy, within the context of existing and proposed contrast mechanisms of neuronal function, that span the four orders of magnitude of length scales for neuronal processing

  8. Analysis of the efficiency of hair removal by different optical methods: comparison of Trichoscan, reflectance confocal microscopy, and optical coherence tomography

    Science.gov (United States)

    Kuck, Monika; Schanzer, Sabine; Ulrich, Martina; Bartels, Natalie Garcia; Meinke, Martina C.; Fluhr, Joachim; Krah, Martin; Blume-Peytavi, Ulrike; Stockfleth, Eggert; Lademann, Jürgen

    2012-10-01

    Noninvasive diagnostic tools, such as Trichoscan, reflectance confocal microscopy (RCM), and optical coherence tomography (OCT), are efficient methods of hair shaft and growth evaluation. The aim of this study was to carry out a comparative assessment of these three medical procedures by measuring the hair shaft and hair growth after hair removal for a defined period of five days. The application of these techniques was demonstrated by measuring hair growth on the lower leg of six female volunteers. After removal of the hair shaft with a shaving system, the hair follicle infundibula and the length of the growing hairs were measured with the Trichoscan, RCM, and OCT method. All three methods are reliable hair measuring tools after hair removal. Trichoscan is best suited in the implementation of hair growth measurement and RCM in the analysis of hair follicles, whereas the OCT system can be consulted as an additional measurement for the evaluation of the hair follicle and length.

  9. Electro-optic techniques in electron beam diagnostics

    Energy Technology Data Exchange (ETDEWEB)

    van Tilborg, Jeroen; Toth, Csaba; Matlis, Nicholas; Plateau, Guillaume; Leemans, Wim

    2011-06-17

    Electron accelerators such as laser wakefield accelerators, linear accelerators driving free electron lasers, or femto-sliced synchrotrons, are capable of producing femtosecond-long electron bunches. Single-shot characterization of the temporal charge profile is crucial for operation, optimization, and application of such accelerators. A variety of electro-optic sampling (EOS) techniques exists for the temporal analysis. In EOS, the field profile from the electron bunch (or the field profile from its coherent radiation) will be transferred onto a laser pulse co-propagating through an electro-optic crystal. This paper will address the most common EOS schemes and will list their advantages and limitations. Strong points that all techniques share are the ultra-short time resolution (tens of femtoseconds) and the single-shot capabilities. Besides introducing the theory behind EOS, data from various research groups is presented for each technique.

  10. Automated cantilever exchange and optical alignment for High-throughput, parallel atomic force microscopy

    CERN Document Server

    Bijnagte, Tom; Kramer, Lukas; Dekker, Bert; Herfst, Rodolf; Sadeghian, Hamed

    2016-01-01

    In atomic force microscopy (AFM), the exchange and alignment of the AFM cantilever with respect to the optical beam and position-sensitive detector (PSD) are often performed manually. This process is tedious and time-consuming and sometimes damages the cantilever or tip. To increase the throughput of AFM in industrial applications, the ability to automatically exchange and align the cantilever in a very short time with sufficient accuracy is required. In this paper, we present the development of an automated cantilever exchange and optical alignment instrument. We present an experimental proof of principle by exchanging various types of AFM cantilevers in 6 seconds with an accuracy better than 2 um. The exchange and alignment unit is miniaturized to allow for integration in a parallel AFM. The reliability of the demonstrator has also been evaluated. Ten thousand continuous exchange and alignment cycles were performed without failure. The automated exchange and alignment of the AFM cantilever overcome a large ...

  11. Near-field optical microscopy and spectroscopy of few-layer black phosphorous

    Science.gov (United States)

    Frenzel, A. J.; Tran, S.; Hinton, J. P.; Sternbach, A. J.; Yang, J.; Gillgren, N.; Lau, C. N.; Basov, D. N.

    Few-layer black phosphorous is a recent addition to the family of two-dimensional (2D) materials which exhibits strongly anisotropic transport and optical properties due to its puckered honeycomb structure. It was recently predicted that this intrinsic anisotropy should manifest in the plasmon dispersion. Additionally, tuning layer number and carrier density can control the dispersion of these collective modes. Scanning near-field optical microscopy (SNOM) has been demonstrated as a powerful method to probe electronic properties, including propagating collective modes, in layered 2D materials. We used SNOM to investigate anisotropic carrier response in few-layer black phosphorous encapsulated by hexagonal boron nitride. In addition to exploring gate-voltage tunability of the electronic response, we demonstrate effective modulation of the near-field signal by ultrafast photoexcitation.

  12. Recent advancements in nanoelectrodes and nanopipettes used in combined scanning electrochemical microscopy techniques.

    Science.gov (United States)

    Kranz, Christine

    2014-01-21

    In recent years, major developments in scanning electrochemical microscopy (SECM) have significantly broadened the application range of this electroanalytical technique from high-resolution electrochemical imaging via nanoscale probes to large scale mapping using arrays of microelectrodes. A major driving force in advancing the SECM methodology is based on developing more sophisticated probes beyond conventional micro-disc electrodes usually based on noble metals or carbon microwires. This critical review focuses on the design and development of advanced electrochemical probes particularly enabling combinations of SECM with other analytical measurement techniques to provide information beyond exclusively measuring electrochemical sample properties. Consequently, this critical review will focus on recent progress and new developments towards multifunctional imaging.

  13. Innovative, Inexpensive Etching Technique Developed for Polymer Electro- Optical Structures

    Science.gov (United States)

    Nguyen, Hung D.

    1999-01-01

    Electro-optic, polymer-based integrated optic devices for high-speed communication and computing applications offer potentially significant advantages over conventional inorganic electro-optic crystals. One key area of integrated optical technology--primary processing and fabrication--may particularly benefit from the use of polymer materials. However, as efforts concentrate on the miniaturization of electro-integrated circuit pattern geometries, the ability to etch fine features and smoothly sloped sidewalls is essential to make polymers useful for electro-integrated circuit applications. There are many existing processes available to etch polymer materials, but they all yield nearly vertical sidewalls. Vertical sidewalls are too difficult to reliably cover with a metal layer, and incomplete metalization degrades microwave performance, particularly at high frequency. However, obtaining a very sloped sidewall greatly improves the deposition of metal on the sidewall, leading to low-loss characteristics, which are essential to integrating these devices in highspeed electro-optic modulators. The NASA Lewis Research Center has developed in-house an inexpensive etching technique that uses a photolithography method followed by a simple, wet chemical etching process to etch through polymer layers. In addition to being simpler and inexpensive, this process can be used to fabricate smoothly sloped sidewalls by using a commercial none rodible mask: Spin-On-Glass. A commercial transparent material, Spin-On-Glass, uses processes and equipment similar to that for photoresist techniques.

  14. Integrated atomic force microscopy techniques for analysis of biomaterials: Study of membrane proteins

    Science.gov (United States)

    Connelly, Laura S.

    Atomic Force Microscopy (AFM) is the prominent techniques for structural studies of biological materials in physiological relevant fluidic environments. AFM has been used to resolve the three-dimensional (3D) surface structure of cells, membranes, and proteins structures. Ion channels, formed by membrane proteins, are the key structures that control the activity of all living systems. This dissertation focuses on the structural evaluation of membrane proteins through atomic force microscopy. In Part I, AFM is utilized to study one of the most prominent medical issues facing our society, Alzheimer's Disease (AD). AD is a misfolded protein disease characterized by the accumulation of beta-amyloid (Abeta) peptide as senile plaques, progressive neurodegeneration, and memory loss. Recent evidence suggests that AD pathology is linked to the destabilization of cellular ionic homeostasis mediated by toxic channel structures composed of Abeta peptides. Selectively engineered sequences of Abeta were examined by AFM to elucidate the substructures and thus activity Abeta channels. Key residues were evaluated with the intent better understand the exact nature by which these pores conduct electrical and molecular signals, which could aid in identifying potential therapeutic targets for the prevention/treatment of AD. Additionally, AFM was used to analyze brain derived Abeta and newly developed pharmacological agents to study membranes and Abeta. Part II, presents a novel technology that incorporates electrophysiology into the AFM interface, enabling simultaneous imaging and complementary conductance measurements. The activity of ion channels is studied by various techniques, including patch clamp, free standing lipid bilayers, droplet interface bilayers, and supported lipid bilayers. However, direct correlation with channel structures has remained a challenge. The integrated atomic force microscopy system presented offers a solution to this challenge. The functionality of the

  15. Imaging of Scleral Collagen Deformation Using Combined Confocal Raman Microspectroscopy and Polarized Light Microscopy Techniques.

    Science.gov (United States)

    Chakraborty, Nilay; Wang, Mian; Solocinski, Jason; Kim, Wonsuk; Argento, Alan

    2016-01-01

    This work presents an optospectroscopic characterization technique for soft tissue microstructure using site-matched confocal Raman microspectroscopy and polarized light microscopy. Using the technique, the microstructure of soft tissue samples is directly observed by polarized light microscopy during loading while spatially correlated spectroscopic information is extracted from the same plane, verifying the orientation and arrangement of the collagen fibers. Results show the response and orientation of the collagen fiber arrangement in its native state as well as during tensile and compressive loadings in a porcine sclera model. An example is also given showing how the data can be used with a finite element program to estimate the strain in individual collagen fibers. The measurements demonstrate features that indicate microstructural reorganization and damage of the sclera's collagen fiber arrangement under loading. The site-matched confocal Raman microspectroscopic characterization of the tissue provides a qualitative measure to relate the change in fibrillar arrangement with possible chemical damage to the collagen microstructure. Tests and analyses presented here can potentially be used to determine the stress-strain behavior, and fiber reorganization of the collagen microstructure in soft tissue during viscoelastic response.

  16. An ultra-low noise optical head for liquid environment atomic force microscopy.

    Science.gov (United States)

    Schlesinger, I; Kuchuk, K; Sivan, U

    2015-08-01

    The design considerations and eventual performance of a new, ultra-low noise optical head for dynamic atomic force microscopy (AFM) are presented. The head, designed specifically for the study of hydration layers and ion organization next to solid surfaces and biomolecules, displays an integrated tip-sample distance noise below 3 pm. The sensitivity of the optical beam deflection sensor, operating at frequencies up to 8.6 MHz (3 dB roll-off), is typically below 10 fm/√Hz, enabling utilization of high frequency cantilevers of low thermal noise for fundamental and higher mode imaging. Exceptional signal stability and low optical noise are achieved by replacing the commonly used laser diode with a helium-neon laser. An integral photothermal excitation of the cantilever produces pure harmonic oscillations, minimizing the generation of higher cantilever modes and deleterious sound waves characterizing the commonly used excitation by a piezoelectric crystal. The optical head is designed to fit on top of the widespread Multimode(®) (Bruker) piezo-tube and accommodate its commercial liquid cell. The performance of the new AFM head is demonstrated by atomic resolution imaging of a muscovite mica surface in aqueous solution.

  17. Confocal Raman microscopy supported by optical clearing treatment of the skin—influence on collagen hydration

    Science.gov (United States)

    Sdobnov, Anton Yu; Tuchin, Valery V.; Lademann, Juergen; E Darvin, Maxim

    2017-07-01

    Confocal Raman microscopy (CRM) is employed to study the skin physiology, drug permeation and skin disease monitoring. In order to increase the depth of investigations, the effect of optical clearing was observed on porcine ear skin ex vivo. The optical clearing agents (OCAs) glycerol and iohexol (Omnipaque™) were applied to the porcine ear skin and investigated by CRM after 30 and 60 min of treatment. The extent of optical clearing by utilizing concentrations of 70% glycerol and 100% Omnipaque™ was evaluated. The intensity of the skin-related Raman peaks significantly increased starting from the depth 160 µm for Omnipaque™ and 40 µm for glycerol (p  ⩽  0.05) after 60 min of treatment. The OCAs’ influence on the collagen hydration in the deep-located dermis was investigated. Both OCAs induce skin dehydration, but the effect of glycerol treatment (30 min and 60 min) is stronger. The obtained results demonstrate that with increasing the treatment time, both glycerol and Omnipaque™ solutions improve the optical clearing of porcine skin making the deep-located dermal regions able for investigations. At the used concentrations and time intervals, glycerol is more effective than Omnipaque™. However, Omnipaque™ is more promising than glycerol for future in vivo applications as it is an already approved pharmaceutic substance without any known impact on the skin structure.

  18. An ultra-low noise optical head for liquid environment atomic force microscopy

    Science.gov (United States)

    Schlesinger, I.; Kuchuk, K.; Sivan, U.

    2015-08-01

    The design considerations and eventual performance of a new, ultra-low noise optical head for dynamic atomic force microscopy (AFM) are presented. The head, designed specifically for the study of hydration layers and ion organization next to solid surfaces and biomolecules, displays an integrated tip-sample distance noise below 3 pm. The sensitivity of the optical beam deflection sensor, operating at frequencies up to 8.6 MHz (3 dB roll-off), is typically below 10 fm / √{ Hz } , enabling utilization of high frequency cantilevers of low thermal noise for fundamental and higher mode imaging. Exceptional signal stability and low optical noise are achieved by replacing the commonly used laser diode with a helium-neon laser. An integral photothermal excitation of the cantilever produces pure harmonic oscillations, minimizing the generation of higher cantilever modes and deleterious sound waves characterizing the commonly used excitation by a piezoelectric crystal. The optical head is designed to fit on top of the widespread Multimode® (Bruker) piezo-tube and accommodate its commercial liquid cell. The performance of the new AFM head is demonstrated by atomic resolution imaging of a muscovite mica surface in aqueous solution.

  19. Evaluation of the Chondromalacia Patella Using a Microscopy Coil: Comparison of the Two-Dimensional Fast Spin Echo Techniques and the Three-Dimensional Fast Field Echo Techniques

    OpenAIRE

    2011-01-01

    Objective We wanted to compare the two-dimensional (2D) fast spin echo (FSE) techniques and the three-dimensional (3D) fast field echo techniques for the evaluation of the chondromalacia patella using a microscopy coil. Materials and Methods Twenty five patients who underwent total knee arthroplasty were included in this study. Preoperative MRI evaluation of the patella was performed using a microscopy coil (47 mm). The proton density-weighted fast spin echo images (PD), the fat-suppressed PD...

  20. Micro-anatomical and functional assessment of ciliated epithelium in mouse trachea using optical coherence phase microscopy.

    Science.gov (United States)

    Ansari, Rehman; Buj, Christian; Pieper, Mario; König, Peter; Schweikard, Achim; Hüttmann, Gereon

    2015-09-07

    Motile cilia perform a range of important mechanosensory and chemosensory functions, along with expulsion of mucus and inhaled pathogens from the lungs. Here we demonstrate that spectral domain optical coherence phase microscopy (SD-OCPM), which combines the principles of optical coherence tomography (OCT) and confocal microscopy, is particularly well-suited for characterization of both morphology and the ciliary dynamics of mouse trachea. We present micro-anatomical images of mouse trachea, where different cell types can be clearly visualized. The phase contrast, which measures the sub-nanometer changes in axial optical pathlength is used to determine the frequency and direction of cilia beatings.

  1. A new method of assessing the surgical margin in rectal carcinoma—using nonlinear optical microscopy

    Science.gov (United States)

    Li, Lianhuang; Chen, Zhifen; Kang, Deyong; Deng, Tongxin; Jiang, Liwei; Zhou, Yi; Liu, Xing; Jiang, Weizhong; Zhuo, Shuangmu; Guan, Guoxian; Chi, Pan; Chen, Jianxin

    2016-06-01

    Nowadays, surgical resection is still the most effective treatment strategy for rectal carcinoma and one of the most important factors affecting whether the operation is successful or not is the surgical margin determination, especially in the distal rectal carcinoma which should take the sphincter-preserving issue into consideration. However, until recently no reliable evaluation method has been developed for this purpose. There are some shortcomings in intraoperative negative surgical margin assessment such as either lack of enough detailed information of biological tissues or the fact that it is time-consuming. Multiphoton microscopy (MPM)—nonlinear optical microscopy, which is based on the nonlinear optical process two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), has the ability to label freely and noninvasively visualize tissue micro-architecture at the sub-cellular level. The advantage of providing high contrast and high resolution biomedical image in real time makes MPM have a wide range of applications in life sciences. In this study, we introduced MPM to identify the boundary between normal and abnormal rectal tissues. MPM images clearly exhibit biological tissue microstructure and its morphological changes in the regions of our interest, which enable it to determine the surgical margin in rectal carcinoma. It can be foreseen that once MPM imaging system is used in clinical examination, it will greatly improve the accuracy of surgical resection.

  2. Membrane distribution of the glycine receptor α3 studied by optical super-resolution microscopy.

    Science.gov (United States)

    Notelaers, Kristof; Rocha, Susana; Paesen, Rik; Swinnen, Nina; Vangindertael, Jeroen; Meier, Jochen C; Rigo, Jean-Michel; Ameloot, Marcel; Hofkens, Johan

    2014-07-01

    In this study, the effect of glycine receptor (GlyR) α3 alternative RNA splicing on the distribution of receptors in the membrane of human embryonic kidney 293 cells is investigated using optical super-resolution microscopy. Direct stochastic optical reconstruction microscopy is used to image both α3K and α3L splice variants individually and together using single- and dual-color imaging. Pair correlation analysis is used to extract quantitative measures from the resulting images. Autocorrelation analysis of the individually expressed variants reveals clustering of both variants, yet with differing properties. The cluster size is increased for α3L compared to α3K (mean radius 92 ± 4 and 56 ± 3 nm, respectively), yet an even bigger difference is found in the cluster density (9,870 ± 1,433 and 1,747 ± 200 μm(-2), respectively). Furthermore, cross-correlation analysis revealed that upon co-expression, clusters colocalize on the same spatial scales as for individually expressed receptors (mean co-cluster radius 94 ± 6 nm). These results demonstrate that RNA splicing determines GlyR α3 membrane distribution, which has consequences for neuronal GlyR physiology and function.

  3. In vivo optical virtual biopsy of human oral mucosa with harmonic generation microscopy

    Science.gov (United States)

    Tsai, Ming-Rung; Chen, Szu-Yu; Shieh, Dar-Bin; Lou, Pei-Jen; Sun, Chi-Kuang

    2011-01-01

    Recent clinical studies on human skin indicated that in vivo multi-harmonic generation microscopy (HGM) can achieve sub-micron resolution for histopathological analysis with a high penetration depth and leave no energy or photodamages in the interacted tissues. It is thus highly desired to apply HGM for in vivo mucosa histopathological diagnosis. In this paper, the first in vivo optical virtual biopsy of human oral mucosa by using epi-HGM is demonstrated. We modified an upright microscope to rotate the angle of objective for in vivo observation. Our clinical study reveals the capability of HGM to in vivo image cell distributions in human oral mucosa, including epithelium and lamina propria with a high penetration depth greater than 280 μm and a high spatial resolution better than 500 nm. We also found that the third-harmonic-generation (THG) contrast on nucleus depends strongly on its thicknesses, in agreement with a numerical simulation. Besides, 4% acetic acid was found to be able to enhance the THG contrast of nucleus in oral mucosa, while such enhancement was found to decay due to the metabolic clearance of the contrast enhancer by the oral mucosa. Our clinical study indicated that, the combined epi-THG and epi-second-harmonic-generation (SHG) microscopy is a promising imaging tool for in vivo noninvasive optical virtual biopsy and disease diagnosis in human mucosa. PMID:21833368

  4. Imaging arterial cells, atherosclerosis, and restenosis by multimodal nonlinear optical microscopy

    Science.gov (United States)

    Wang, Han-Wei; Simianu, Vlad; Locker, Matthew J.; Sturek, Michael; Cheng, Ji-Xin

    2008-02-01

    By integrating sum-frequency generation (SFG), and two-photon excitation fluorescence (TPEF) on a coherent anti-Stokes Raman scattering (CARS) microscope platform, multimodal nonlinear optical (NLO) imaging of arteries and atherosclerotic lesions was demonstrated. CARS signals arising from CH II-rich membranes allowed visualization of endothelial cells and smooth muscle cells in a carotid artery. Additionally, CARS microscopy allowed vibrational imaging of elastin and collagen fibrils which are rich in CH II bonds in their cross-linking residues. The extracellular matrix organization was further confirmed by TPEF signals arising from elastin's autofluorescence and SFG signals arising from collagen fibrils' non-centrosymmetric structure. The system is capable of identifying different atherosclerotic lesion stages with sub-cellular resolution. The stages of atherosclerosis, such as macrophage infiltration, lipid-laden foam cell accumulation, extracellular lipid distribution, fibrous tissue deposition, plaque establishment, and formation of other complicated lesions could be viewed by our multimodal CARS microscope. Collagen percentages in the region adjacent to coronary artery stents were resolved. High correlation between NLO and histology imaging evidenced the validity of the NLO imaging. The capability of imaging significant components of an arterial wall and distinctive stages of atherosclerosis in a label-free manner suggests the potential application of multimodal nonlinear optical microscopy to monitor the onset and progression of arterial diseases.

  5. In Vivo Confocal Microscopy and Anterior Segment Optic Coherence Tomography Findings in Ocular Ochronosis

    Directory of Open Access Journals (Sweden)

    Elif Demirkilinc Biler

    2015-01-01

    Full Text Available Purpose. To report clinical and in vivo confocal microscopy (IVCM findings of two patients with ocular ochronosis secondary due to alkaptonuria. Materials and Methods. Complete ophthalmologic examinations, including IVCM (HRT II/Rostock Cornea Module, Heidelberg, Germany, anterior segment optical coherence tomography (AS-OCT (Topcon 3D spectral-domain OCT 2000, Topcon Medical Systems, Paramus, NJ, USA, corneal topography (Pentacam, OCULUS Optikgeräte GmbH, Wetzlar, Germany, and anterior segment photography, were performed. Results. Biomicroscopic examination showed bilateral darkly pigmented lesions of the nasal and temporal conjunctiva and episclera in both patients. In vivo confocal microscopy of the lesions revealed prominent degenerative changes, including vacuoles and fragmentation of collagen fibers in the affected conjunctival lamina propria and episclera. Hyperreflective pigment granules in different shapes were demonstrated in the substantia propria beneath the basement membrane. AS-OCT of Case 1 demonstrated hyporeflective areas. Fundus examination was within normal limits in both patients, except tilted optic discs with peripapillary atrophy in one of the patients. Corneal topography, thickness, and macular OCT were normal bilaterally in both cases. Conclusion. The degenerative and anatomic changes due to ochronotic pigment deposition in alkaptonuria can be demonstrated in detail with IVCM and AS-OCT. Confocal microscopic analysis in ocular ochronosis may serve as a useful adjunct in diagnosis and monitoring of the disease progression.

  6. Probing single processive molecular motors with high-speed optical tweezers and fluorescence microscopy

    Science.gov (United States)

    Gardini, L.; Pavone, F. S.; Capitanio, M.

    2017-02-01

    Here we present development of optical techniques for the study of single processive myosin motors based on the combination of high-speed optical tweezers force spectroscopy and single molecule fluorescence imaging. Ultrafast force-clamp spectroscopy1 is applied to study the dependence of single chemo-mechanical steps of processive myosin motors on the applied load. On the other hand, single molecule localization through FIONA (Fluorescence Imaging with One Nanometer Accuracy)2, 3 is applied to in vitro motility assay to measure parameters such as the runlength, velocity and step size of single myosin V motors, labeled with Quantum Dots, under unloaded conditions.

  7. Development of a Fiber Laser with Independently Adjustable Properties for Optical Resolution Photoacoustic Microscopy.

    Science.gov (United States)

    Aytac-Kipergil, Esra; Demirkiran, Aytac; Uluc, Nasire; Yavas, Seydi; Kayikcioglu, Tunc; Salman, Sarper; Karamuk, Sohret Gorkem; Ilday, Fatih Omer; Unlu, Mehmet Burcin

    2016-12-08

    Photoacoustic imaging is based on the detection of generated acoustic waves through thermal expansion of tissue illuminated by short laser pulses. Fiber lasers as an excitation source for photoacoustic imaging have recently been preferred for their high repetition frequencies. Here, we report a unique fiber laser developed specifically for multiwavelength photoacoustic microscopy system. The laser is custom-made for maximum flexibility in adjustment of its parameters; pulse duration (5-10 ns), pulse energy (up to 10 μJ) and repetition frequency (up to 1 MHz) independently from each other and covers a broad spectral region from 450 to 1100 nm and also can emit wavelengths of 532, 355, and 266 nm. The laser system consists of a master oscillator power amplifier, seeding two stages; supercontinuum and harmonic generation units. The laser is outstanding since the oscillator, amplifier and supercontinuum generation parts are all-fiber integrated with custom-developed electronics and software. To demonstrate the feasibility of the system, the images of several elements of standardized resolution test chart are acquired at multiple wavelengths. The lateral resolution of optical resolution photoacoustic microscopy system is determined as 2.68 μm. The developed system may pave the way for spectroscopic photoacoustic microscopy applications via widely tunable fiber laser technologies.

  8. Development of a Fiber Laser with Independently Adjustable Properties for Optical Resolution Photoacoustic Microscopy

    Science.gov (United States)

    Aytac-Kipergil, Esra; Demirkiran, Aytac; Uluc, Nasire; Yavas, Seydi; Kayikcioglu, Tunc; Salman, Sarper; Karamuk, Sohret Gorkem; Ilday, Fatih Omer; Unlu, Mehmet Burcin

    2016-12-01

    Photoacoustic imaging is based on the detection of generated acoustic waves through thermal expansion of tissue illuminated by short laser pulses. Fiber lasers as an excitation source for photoacoustic imaging have recently been preferred for their high repetition frequencies. Here, we report a unique fiber laser developed specifically for multiwavelength photoacoustic microscopy system. The laser is custom-made for maximum flexibility in adjustment of its parameters; pulse duration (5-10 ns), pulse energy (up to 10 μJ) and repetition frequency (up to 1 MHz) independently from each other and covers a broad spectral region from 450 to 1100 nm and also can emit wavelengths of 532, 355, and 266 nm. The laser system consists of a master oscillator power amplifier, seeding two stages; supercontinuum and harmonic generation units. The laser is outstanding since the oscillator, amplifier and supercontinuum generation parts are all-fiber integrated with custom-developed electronics and software. To demonstrate the feasibility of the system, the images of several elements of standardized resolution test chart are acquired at multiple wavelengths. The lateral resolution of optical resolution photoacoustic microscopy system is determined as 2.68 μm. The developed system may pave the way for spectroscopic photoacoustic microscopy applications via widely tunable fiber laser technologies.

  9. Towards a more realistic picture of in situ biocide actions: Combining physiological and microscopy techniques

    Energy Technology Data Exchange (ETDEWEB)

    Speranza, M., E-mail: speranzamariela@gmail.com [Museo Nacional de Ciencias Naturales, MNCN-CSIC, Serrano 115 bis, 28006 Madrid (Spain); Wierzchos, J.; De Los Rios, A.; Perez-Ortega, S. [Museo Nacional de Ciencias Naturales, MNCN-CSIC, Serrano 115 bis, 28006 Madrid (Spain); Souza-Egipsy, V. [Instituto de Ciencias Agrarias, ICA-CSIC, Serrano 115 bis, 28006 Madrid (Spain); Ascaso, C., E-mail: ascaso@mncn.csic.es [Museo Nacional de Ciencias Naturales, MNCN-CSIC, Serrano 115 bis, 28006 Madrid (Spain)

    2012-11-15

    In this study, we combined chlorophyll a fluorescence (ChlaF) measurements, using pulse-amplitude-modulate (PAM) equipment, with scanning electron microscopy in backscattered electron mode (SEM-BSE) and transmission electron microscopy (TEM) images to evaluate the actions of Koretrel at lower concentrations on Verrucaria nigrescens colonising a dolostone. ChlaF measurements are good indicators of the damaging effects of biocides. However, these indicators only provide an incomplete view of the mechanism of biocides used to control biodeterioration agents. The death of the V. nigrescens photobiont at two biocide concentrations was revealed by PAM, SEM-BSE and TEM. Once Koretrel was applied, the Fv/Fm ratios markedly fell in the first few hours after the 1.5% treatment, and ratios for the 3% dilution remained close to zero throughout the study. The algal zone shows the plasmolysed appearance of the photobiont cells, and important aspects related to the action of the biocide on free and lichenised fungi were also detected using SEM-BSE. Many of the mycobiont cells had only their cell walls preserved; although, some fungal hyphae in lichen thalli and some microorganisms in endolithic clusters maintained lipid storage in their cytoplasm. These results indicated that the combination of physiological and microscopy techniques improves the assessment of biocide action in situ and this will help to optimize protocols in order to reduce the emission of these compounds to the environment. -- Highlights: Black-Right-Pointing-Pointer We combined ChlaF measurements with EM images to analyses the biocides action on stone biodeterioration agents. Black-Right-Pointing-Pointer At lower biocide concentrations damage to photobiont and mycobiont cells integrity, ultrastructure and vitality were observed. Black-Right-Pointing-Pointer The limited action of biocides on fungi and algae were detected using SEM-BSE. Black-Right-Pointing-Pointer The combination of physiological and microscopy

  10. Sachse optical urethrotomy, a modified technique: 6 years of experience.

    Science.gov (United States)

    Sandozi, S; Ghazali, S

    1988-11-01

    Urethrotomy with the Sachse optical urethrotome has radically modified our management of strictures of the male urethra. A total of 143 patients underwent 210 procedures during 6 years. The strictures resolved in 85 per cent of the patients and they required no further treatment, including 52 patients followed for more than 6 years. A modified surgical technique using a pediatric cystoscope is described to inspect, traverse and cannulate the urethral stricture before optical urethrotomy, thus, enabling successful management of even the most difficult strictures. Our results confirm that the best prospects of cure were short and single urethral strictures, and that optical urethrotomy is the treatment of choice for strictures of the male urethra.

  11. Optical resilient packet ring (O-RPR) based on all-optical buffering techniques

    Science.gov (United States)

    Wu, Chongqing; Sheng, Xingzhi; Fu, Songnian; Wei, Bin; Li, Yajie; Liu, Aiming

    2006-01-01

    This paper reports the progress of the 863 high-technology project of China "Optical Resilient Packet Ring (O-RPR) Based on All-optical Buffering Techniques". In this ring network, for the packet through an intermediate node the conversion of O/E/O is not needed in order to overcome the bottleneck of O/E/O. In all-optical node a Dual Loop Optical Buffer (DLOB) is used to revolve the collision between the packet, which pass through the node, and add packet from local user to ring. The principle of DLOB is introduced. The bit-rate of head of optical frame is lower than the bit-rate of payload in a packet, in order to increase the efficiency of transmission link. This paper will introduce the network topology, layers and the structure of optical node. It includes an optical splitter, optical delay line as input buffer, a SOA as optical switch, which switch the packet dropping down form the ring or pass through the node, a DLOB and an electric buffer. An ARM is used for regulation of different buffers. The experiment results of a demonstrate network including 3 nodes are given.

  12. Integrating Nanostructured Artificial Receptors with Whispering Gallery Mode Optical Microresonators via Inorganic Molecular Imprinting Techniques.

    Science.gov (United States)

    Hammond, G Denise; Vojta, Adam L; Grant, Sheila A; Hunt, Heather K

    2016-06-15

    The creation of label-free biosensors capable of accurately detecting trace contaminants, particularly small organic molecules, is of significant interest for applications in environmental monitoring. This is achieved by pairing a high-sensitivity signal transducer with a biorecognition element that imparts selectivity towards the compound of interest. However, many environmental pollutants do not have corresponding biorecognition elements. Fortunately, biomimetic chemistries, such as molecular imprinting, allow for the design of artificial receptors with very high selectivity for the target. Here, we perform a proof-of-concept study to show how artificial receptors may be created from inorganic silanes using the molecular imprinting technique and paired with high-sensitivity transducers without loss of device performance. Silica microsphere Whispering Gallery Mode optical microresonators are coated with a silica thin film templated by a small fluorescent dye, fluorescein isothiocyanate, which serves as our model target. Oxygen plasma degradation and solvent extraction of the template are compared. Extracted optical devices are interacted with the template molecule to confirm successful sorption of the template. Surface characterization is accomplished via fluorescence and optical microscopy, ellipsometry, optical profilometry, and contact angle measurements. The quality factors of the devices are measured to evaluate the impact of the coating on device sensitivity. The resulting devices show uniform surface coating with no microstructural damage with Q factors above 10⁶. This is the first report demonstrating the integration of these devices with molecular imprinting techniques, and could lead to new routes to biosensor creation for environmental monitoring.

  13. Integrating Nanostructured Artificial Receptors with Whispering Gallery Mode Optical Microresonators via Inorganic Molecular Imprinting Techniques

    Directory of Open Access Journals (Sweden)

    G. Denise Hammond

    2016-06-01

    Full Text Available The creation of label-free biosensors capable of accurately detecting trace contaminants, particularly small organic molecules, is of significant interest for applications in environmental monitoring. This is achieved by pairing a high-sensitivity signal transducer with a biorecognition element that imparts selectivity towards the compound of interest. However, many environmental pollutants do not have corresponding biorecognition elements. Fortunately, biomimetic chemistries, such as molecular imprinting, allow for the design of artificial receptors with very high selectivity for the target. Here, we perform a proof-of-concept study to show how artificial receptors may be created from inorganic silanes using the molecular imprinting technique and paired with high-sensitivity transducers without loss of device performance. Silica microsphere Whispering Gallery Mode optical microresonators are coated with a silica thin film templated by a small fluorescent dye, fluorescein isothiocyanate, which serves as our model target. Oxygen plasma degradation and solvent extraction of the template are compared. Extracted optical devices are interacted with the template molecule to confirm successful sorption of the template. Surface characterization is accomplished via fluorescence and optical microscopy, ellipsometry, optical profilometry, and contact angle measurements. The quality factors of the devices are measured to evaluate the impact of the coating on device sensitivity. The resulting devices show uniform surface coating with no microstructural damage with Q factors above 106. This is the first report demonstrating the integration of these devices with molecular imprinting techniques, and could lead to new routes to biosensor creation for environmental monitoring.

  14. Sensitivity and Specificity of Cardiac Tissue Discrimination Using Fiber-Optics Confocal Microscopy.

    Science.gov (United States)

    Huang, Chao; Sachse, Frank B; Hitchcock, Robert W; Kaza, Aditya K

    2016-01-01

    Disturbances of the cardiac conduction system constitute a major risk after surgical repair of complex cases of congenital heart disease. Intraoperative identification of the conduction system may reduce the incidence of these disturbances. We previously developed an approach to identify cardiac tissue types using fiber-optics confocal microscopy and extracellular fluorophores. Here, we applied this approach to investigate sensitivity and specificity of human and automated classification in discriminating images of atrial working myocardium and specialized tissue of the conduction system. Two-dimensional image sequences from atrial working myocardium and nodal tissue of isolated perfused rodent hearts were acquired using a fiber-optics confocal microscope (Leica FCM1000). We compared two methods for local application of extracellular fluorophores: topical via pipette and with a dye carrier. Eight blinded examiners evaluated 162 randomly selected images of atrial working myocardium (n = 81) and nodal tissue (n = 81). In addition, we evaluated the images using automated classification. Blinded examiners achieved a sensitivity and specificity of 99.2 ± 0.3% and 98.0 ± 0.7%, respectively, with the dye carrier method of dye application. Sensitivity and specificity was similar for dye application via a pipette (99.2 ± 0.3% and 94.0 ± 2.4%, respectively). Sensitivity and specificity for automated methods of tissue discrimination were similarly high. Human and automated classification achieved high sensitivity and specificity in discriminating atrial working myocardium and nodal tissue. We suggest that our findings facilitate clinical translation of fiber-optics confocal microscopy as an intraoperative imaging modality to reduce the incidence of conduction disturbances during surgical correction of congenital heart disease.

  15. Sensitivity and Specificity of Cardiac Tissue Discrimination Using Fiber-Optics Confocal Microscopy

    Science.gov (United States)

    Huang, Chao; Sachse, Frank B.; Hitchcock, Robert W.; Kaza, Aditya K.

    2016-01-01

    Disturbances of the cardiac conduction system constitute a major risk after surgical repair of complex cases of congenital heart disease. Intraoperative identification of the conduction system may reduce the incidence of these disturbances. We previously developed an approach to identify cardiac tissue types using fiber-optics confocal microscopy and extracellular fluorophores. Here, we applied this approach to investigate sensitivity and specificity of human and automated classification in discriminating images of atrial working myocardium and specialized tissue of the conduction system. Two-dimensional image sequences from atrial working myocardium and nodal tissue of isolated perfused rodent hearts were acquired using a fiber-optics confocal microscope (Leica FCM1000). We compared two methods for local application of extracellular fluorophores: topical via pipette and with a dye carrier. Eight blinded examiners evaluated 162 randomly selected images of atrial working myocardium (n = 81) and nodal tissue (n = 81). In addition, we evaluated the images using automated classification. Blinded examiners achieved a sensitivity and specificity of 99.2±0.3% and 98.0±0.7%, respectively, with the dye carrier method of dye application. Sensitivity and specificity was similar for dye application via a pipette (99.2±0.3% and 94.0±2.4%, respectively). Sensitivity and specificity for automated methods of tissue discrimination were similarly high. Human and automated classification achieved high sensitivity and specificity in discriminating atrial working myocardium and nodal tissue. We suggest that our findings facilitate clinical translation of fiber-optics confocal microscopy as an intraoperative imaging modality to reduce the incidence of conduction disturbances during surgical correction of congenital heart disease. PMID:26808149

  16. Sensitivity and Specificity of Cardiac Tissue Discrimination Using Fiber-Optics Confocal Microscopy.

    Directory of Open Access Journals (Sweden)

    Chao Huang

    Full Text Available Disturbances of the cardiac conduction system constitute a major risk after surgical repair of complex cases of congenital heart disease. Intraoperative identification of the conduction system may reduce the incidence of these disturbances. We previously developed an approach to identify cardiac tissue types using fiber-optics confocal microscopy and extracellular fluorophores. Here, we applied this approach to investigate sensitivity and specificity of human and automated classification in discriminating images of atrial working myocardium and specialized tissue of the conduction system. Two-dimensional image sequences from atrial working myocardium and nodal tissue of isolated perfused rodent hearts were acquired using a fiber-optics confocal microscope (Leica FCM1000. We compared two methods for local application of extracellular fluorophores: topical via pipette and with a dye carrier. Eight blinded examiners evaluated 162 randomly selected images of atrial working myocardium (n = 81 and nodal tissue (n = 81. In addition, we evaluated the images using automated classification. Blinded examiners achieved a sensitivity and specificity of 99.2 ± 0.3% and 98.0 ± 0.7%, respectively, with the dye carrier method of dye application. Sensitivity and specificity was similar for dye application via a pipette (99.2 ± 0.3% and 94.0 ± 2.4%, respectively. Sensitivity and specificity for automated methods of tissue discrimination were similarly high. Human and automated classification achieved high sensitivity and specificity in discriminating atrial working myocardium and nodal tissue. We suggest that our findings facilitate clinical translation of fiber-optics confocal microscopy as an intraoperative imaging modality to reduce the incidence of conduction disturbances during surgical correction of congenital heart disease.

  17. Full-field illumination approach with multiple speckle for optical-resolution photoacoustic microscopy (Conference Presentation)

    Science.gov (United States)

    Poisson, Florian; Bossy, Emmanuel

    2016-03-01

    Optical-resolution photoacoustic endomicroscopy (OR-PAE) allows going beyond the limited penetration depth of conventional optical-resolution photoacoustic systems. Recently, it has been shown that OR-PAE may be performed through minimally invasive multimode fibers, by raster scanning a focus spot with optical wavefront shaping [1]. Here we introduce for the first time an approach to perform OR-PAE through a multimode fiber with a full-field illumination approach. By using multiple known speckle patterns, we show that it is possible to obtain optical-diffraction limited photoacoustic images, with the same resolution as that obtained by raster scanning a focus spot, i.e that of the speckle grain size. The fluctuations patterns of the photoacoustic amplitude at each pixel in the sample plane with the series of multiple speckle illumination were used to encode each pixel. This approach with known speckle illumination requires an initial calibration stage, that consists in learn a set of fluctuation patterns pixel per pixel, which will encode patterns each pixel of the scanned area. A point-like absorber was scanned across the filed-of-view during the calibration stage to acquire the reference patterns. Image reconstruction may be carried out by cross-correlating the series of photoacoustic amplitude measured with the sample to the reference patterns obtained during the calibration stage. In this work, the approach above was carried out both theoretically with Monte-carlo simulations and experimentally through a multi-mode fiber with samples made of absorbing spheres. [1] Papadopoulos et al., " Optical-resolution photoacoustic microscopy by use of a multimode fiber", Appl. Phys. Lett., 102(21), 2013

  18. Optical, infrared and radio astronomy from techniques to observation

    CERN Document Server

    Poggiani, Rosa

    2017-01-01

    This textbook presents the established sciences of optical, infrared, and radio astronomy as distinct research areas, focusing on the science targets and the constraints that they place on instrumentation in the different domains. It aims to bridge the gap between specialized books and practical texts, presenting the state of the art in different techniques. For each type of astronomy, the discussion proceeds from the orders of magnitude for observable quantities that drive the building of instrumentation and the development of advanced techniques. The specific telescopes and detectors are then presented, together with the techniques used to measure fluxes and spectra. Finally, the instruments and their limits are discussed to assist readers in choice of setup, planning and execution of observations, and data reduction. The volume also includes worked examples and problem sets to improve student understanding; tables and figures in chapters summarize the state of the art of instrumentation and techniques.

  19. Optical Microstructures Fabricated with Direct Laser Writing Technique

    Directory of Open Access Journals (Sweden)

    Kowalczyk M.

    2014-12-01

    Full Text Available Three-dimensional photolitography, also known as Direct Laser Writing (DLW, is a powerful technique for fabrication of photonic microstructures. In this paper we present the basics of the relevant technology and discuss some features of the fabrication process. We also describe the experimental setup designed for making colour filters based on diffraction gratings, fibre-tip-integrated lens and anti-reflective coating designed for telecom wavelength (1550 nm. The results obtained demonstrate the DLW technique to be a promising fast prototyping fabrication method that may allow manipulating the properties of optical materials.

  20. Optical fiber sensors fabricated by the focused ion beam technique

    DEFF Research Database (Denmark)

    Yuan, Scott Wu; Wang, Fei; Bang, Ole

    2012-01-01

    crystal fiber (PCF). Using this technique we fabricate a highly compact fiber-optic Fabry-Pérot (FP) refractive index sensor near the tip of fiber taper, and a highly sensitive in-line temperature sensor in PCF. We also demonstrate the potential of using FIB to selectively fill functional fluid......Focused ion beam (FIB) is a highly versatile technique which helps to enable next generation of lab-on-fiber sensor technologies. In this paper, we demonstrate the use application of FIB to precisely mill the fiber taper and end facet of both conventional single mode fiber (SMF) and photonic...

  1. Ultrabroadband Phased-Array Receivers Based on Optical Techniques

    Science.gov (United States)

    2016-02-26

    AFRL-AFOSR-VA-TR-2016-0121 Ultrabroadband Phased-array Receivers Based on Optical Techniques Christopher Schuetz UNIVERSITY OF DELAWARE Final Report...NUMBER 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) University of Delaware 210 Hullihen Hall Newark, DE 19716 8. PERFORMING ORGANIZATION...Rev. 8/98) Prescribed by ANSI Std . Z39.18 Adobe Professional 7.0 Reset INSTRUCTIONS FOR COMPLETING SF 298 1. REPORT DATE. Full publication date

  2. Structural and optical properties of CdO thin films deposited by RF magnetron sputtering technique

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, G. Anil, E-mail: anilhcu@gmail.com; Reddy, M. V. Ramana, E-mail: anilhcu@gmail.com [Department of Physics, Osmania University, Hyderabad-500007 (India); Reddy, Katta Narasimha, E-mail: anilhcu@gmail.com [Department of Physics, Mahatma Gandhi University, Nalgonda-508003 (India)

    2014-04-24

    Cadmium oxide (CdO) thin films were deposited on glass substrate by r.f. magnetron sputtering technique using a high purity (99.99%) Cd target of 2-inch diameter and 3 mm thickness in an Argon and oxygen mixed atmosphere with sputtering power of 50W and sputtering pressure of 2×10{sup −2} mbar. The prepared films were characterized by X-ray diffraction (XRD), optical spectroscopy and scanning electron microscopy (SEM). The XRD analysis reveals that the films were polycrystalline with cubic structure. The visible range transmittance was found to be over 70%. The optical band gap increased from 2.7 eV to2.84 eV with decrease of film thickness.

  3. Optical probe design with extended depth-of-focus for optical coherence microscopy and optical coherence tomography

    Science.gov (United States)

    Lee, Seungwan; Choi, Minseog; Lee, Eunsung; Jung, Kyu-Dong; Chang, Jong-hyeon; Kim, Woonbae

    2013-03-01

    In this report, Optical probe system for modality, optical coherence tomography (OCT) and optical coherence microscope (OCM), is presented. In order to control the back focal length from 2.2 mm to 27 mm, optical probe is designed using two liquid lenses and several lenses. The narrow depth of focus (DOF) in microscope is extended by phase filter such as cubic filter. The filter is modified so that DOF is extended only In the OCM mode. The section for the extended DOF of probe is controlled by iris. Therefore in OCT mode, the phase filter does not affect on the DOF of lens. In OCM mode, the Gaussian light and modified light will affect the DOF. The probe dimension is less than 4 mm diameter and less than 60 mm long. The scan range of system is 0.88 mm wide, 1 mm deep in the OCT and 510 μm wide, 1 mm deep in the OCM mode. The lens curvature and iris aperture are operated by digital microelectrofluidic lens and iris.

  4. Optical resolution photoacoustic microscopy using a Blu-ray DVD pickup head

    Science.gov (United States)

    Li, Meng-Lin; Wang, Po-Hsun

    2014-03-01

    Optical resolution photoacoustic microscopy (OR-PAM) has been shown as a promising tool for label-free microvascular and single-cell imaging in clinical and bioscientific applications. However, most OR-PAM systems are realized by using a bulky laser for photoacoustic excitation. The large volume and high price of the laser may restrain the popularity of OR-PAM. In this study, we attempt to develop a compact, portable, and low cost OR-PAM based on a consumer Blu-ray (405 nm) DVD pickup head for label-free micro-vascular imaging and red-blood-cell related blood examination. According to the high optical absorption of the hemoglobin at 405 nm, the proposed OR-PAM has potential to be an alternative for the conventional optical microscopy in the examinations of hematological morphology for blood routine. We showed that the Blu-ray DVD pickup head owns the required laser energy and focusing optics for OR-PAM. The firmware of a Blu-ray DVD drive was modified to allow its pickup head to generate nano-second laser pulses with a tunable pulse repetition rate of >30 kHz and a tunable pulse width ranging from 10 to 30 ns. The laser beam was focused onto the target after passing through a transparent cover slide, and then aligned to be confocal with a 50-MHz focused ultrasonic transducer in forward mode. To keep the target on focus, a scan involving auto-tracking procedure was performed. The measured maximum achievable lateral resolution was 1 μm which was mainly limited by the minimum step size of the used motorized stage. A blood smear was imaged without any staining. The red blood cells were well resolved and the biconcave structure could be clearly visualized. In addition, to verify the in vivo imaging capability of the proposed OR-PAM, the micro-vasculature of a mouse ear was imaged without any contrast agent. The results showed that it performed better than a 200x digital optical microscope in terms of image contrast and vascular morphology. In summaries, the proposed OR

  5. Cavity-aided magnetic-resonance microscopy of atoms in optical lattices

    CERN Document Server

    Purdy, Tom P; Brooks, Daniel W C; Botter, Thierry; Stamper-Kurn, Dan M

    2010-01-01

    Magnetic resonance imaging (MRI) is a powerful technique for investigating the microscopic properties and dynamics of physical systems. In this work we demonstrate state-sensitive MRI of ultracold atoms in an optical lattice. Single-shot spatial resolution is 120 nm, well below the lattice spacing, and number sensitivity is +/-2.4 for 150 atoms on a single site, well below Poissonian atom-number fluctuations. We achieve this by combining high-spatial-resolution control over the atomic spin using an atom chip, together with nearly quantum-limited spin measurement, obtained by dispersively coupling the atoms to light in a high-finesse optical cavity. The MRI is minimally disruptive of the atoms' internal state, preserving the magnetisation of the gas for subsequent experiments. Using this technique, we observe the nonequilibrium transport dynamics of the atoms among individual lattice sites. We see the atom cloud initially expand ballistically, followed by the onset of interaction-inhibited transport.

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

    KAUST Repository

    Adhikari, Aniruddha

    2016-12-15

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

  7. Microscopy and calorimetry as complementary techniques to analyze sugar crystallization from amorphous systems.

    Science.gov (United States)

    Mazzobre, María F; Aguilera, José M; Buera, María P

    2003-03-14

    A comparison of microscopic and macroscopic techniques to evaluate sugar crystallization kinetics is presented using amorphous lactose and lactose-trehalose mixtures. Polarized light video microscopy (PLV) and differential scanning calorimetry (DSC) were applied to measure crystallization kinetics, induction times and time for complete sugar crystallization at different storage temperatures (60-95 degrees C). DSC was also employed to measure the glass transition temperature (T(ag)) of the systems. PLV permitted direct observation, in real time, of growth of individual crystals and morphological aspects at a scale not detected by DSC. Taking the average of several microscopic observations, the results for temperature dependence of crystallization rate and time to complete lactose crystallization were similar to those obtained by DSC. Both PLV and DSC techniques showed that the presence of trehalose delayed lactose crystallization, without affecting the T(ag) value. For the analysis of sugar crystallization in amorphous systems, PLV and DSC proved to be complementary techniques. Validation of results obtained by PLV with results from DSC opens a new area of microstructural analysis of crystallizing systems.

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

    Science.gov (United States)

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

    2017-01-11

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

  9. Ultrathin forward-imaging short multimode fiber probe for full-field optical coherence microscopy

    Science.gov (United States)

    Sato, Manabu; Saito, Daisuke; Shouji, Kou; Kurotani, Reiko; Abe, Hiroyuki; Nishidate, Izumi

    2016-12-01

    To extend the applications of optical coherence tomography (OCT) to the fields of physiology and clinical medicine, less invasive, robust, and reliable optical probes are required. Thus, we demonstrate an ultrathin forward-imaging short multimode fiber (SMMF) optical coherence microscopy (OCM) probe with a 50 μm core diameter, 125 μm total diameter, and 5.12 mm length. Imaging conditions and magnification were analyzed, and they correspond closely to the measured results. The dispersion of the SMMF was investigated, and the modal dispersion coefficient was found to be 2.3% of the material dispersion coefficient. The axial resolution was minimized at 2.15 μm using a 0.885-mm-thick dispersion compensator. The lateral resolution was evaluated to be 4.38 μm using a test pattern. The contrast of the OCM images was 5.7 times higher than that of the signal images owing to the coherence gate. The depth of focus and diameter of the field of view were measured to be 60 μm and 40-50 μm, respectively. OCM images of the dried fins of small fish (Medaka) were measured and internal structures could be recognized.

  10. In vivo optical virtual biopsy of human oral cavity with harmonic generation microscopy

    Science.gov (United States)

    Tsai, M.-R.; Chen, S.-Y.; Shieh, D.-B.; Lou, P.-J.; Sun, C.-K.

    2010-02-01

    Oral cancer ranked number four in both cancer incident and mortality in Taiwanese male population. Early disease diagnosis and staging is essential for its clinical success. However, most patients were diagnosed in their late disease stage as ideal prescreening procedures are yet to be developed especially when dealing with a large surface of precancerous lesions. Therefore, how to detect and confirm the diagnosis of these early stage lesions are of significant clinical value. Harmonic generation process naturally occurred in biological molecules and requires no energy deposition to the target molecule. Thus harmonic generation microscopy (HGM) could potentially serve as a noninvasive tool for screening of human oral mucosal diseases. The in vivo optical biopsy of human oral cavity with HGM could be achieved with high spatial resolution to resolve dynamic physiological process in the oral mucosal tissue with equal or superior quality but devoid of complicated physical biopsy procedures. The second harmonic generation (SHG) provide significant image contrast for biomolecules with repetitive structures such as the collagen fibers in the lamina propria and the mitotic spindles in dividing cells. The cell morphology in the epithelial layer, blood vessels and blood cells flow through the capillaries can be revealed by third harmonic generation (THG) signals. Tissue transparent technology was used to increase the optical penetration of the tissue. In conclusion, this report demonstrates the first in vivo optical virtual biopsy of human oral mucosa using HGM and revealed a promising future for its clinical application for noninvasive in vivo diseases diagnosis.

  11. Calcium effect on membrane of an optically trapped erythrocyte studied by digital holographic microscopy

    Science.gov (United States)

    Farzam Rad, Vahideh; Tavakkoli, Rahim; Moradi, Ali-Reza; Anand, Arun; Javidi, Bahram

    2017-08-01

    The calcium level in blood affects the morphological and rheological properties of red blood cell (RBC) membranes. In this paper, we present an integrated optical system for a single cell study of hypercalcemia. The system consists of holographic optical tweezers and blinking optical tweezers, for photo-damage-free immobilization of the cells, combined with digital holographic microscopy, for quantitative analysis and live visualization of the cells. Digital holograms were recorded live, while the concentration of calcium ions in the buffer is gradually increased. Full morphometric data of RBCs were obtained by numerical reconstruction of the holograms. Morphological changes are expressed in terms of various parameters such as root mean square, skewness, and kurtosis of the cell membrane thickness distribution. We have observed dramatic changes of the cell morphology, which are attributed to the formation of calcium-induced hydrophobic aggregates of phospholipid molecules in the RBC membrane, resulting in a net change in membrane rigidity. Our experimental results are in agreement with previous biological studies of RBCs under the Ca2+ influence.

  12. All-optical delay technique for supporting multiple antennas in a hybrid optical - wireless transmission system

    DEFF Research Database (Denmark)

    Prince, Kamau; Chiuchiarelli, A; Presi, M

    2008-01-01

    We introduce a novel continuously-variable optical delay technique to support beam-forming wireless communications systems using antenna arrays. We demonstrate delay with 64-QAM modulated signals at a rate of 15 Msymbol/sec with 2.5 GHz carrier frequency.......We introduce a novel continuously-variable optical delay technique to support beam-forming wireless communications systems using antenna arrays. We demonstrate delay with 64-QAM modulated signals at a rate of 15 Msymbol/sec with 2.5 GHz carrier frequency....

  13. Surface nanobubbles studied by atomic force microscopy techniques: Facts, fiction, and open questions

    Science.gov (United States)

    Schönherr, Holger; Hain, Nicole; Walczyk, Wiktoria; Wesner, Daniel; Druzhinin, Sergey I.

    2016-08-01

    In this review surface nanobubbles, which are presumably gas-filled enclosures found at the solid-liquid interface, are introduced and discussed together with key experimental findings that suggest that these nanoscale features indeed exist and are filled with gas. The most prominent technique used thus far has been atomic force microscopy (AFM). However, due to its potentially invasive nature, AFM data must be interpreted with great care. Owing to their curved interface, the Laplace internal pressure of surface nanobubbles exceeds substantially the outside ambient pressure, and the experimentally observed long term stability is in conflict with estimates of gas transport rates and predicted surface nanobubble lifetimes. Despite recent explanations of both the stability and the unusual nanoscopic contact angles, the development of new co-localization approaches and the adequate analysis of AFM data of surface nanobubbles are important as a means to confirm the gaseous nature and correctly estimate the interfacial curvature.

  14. Electron Correlation Microscopy: A New Technique for Studying Local Atom Dynamics Applied to a Supercooled Liquid.

    Science.gov (United States)

    He, Li; Zhang, Pei; Besser, Matthew F; Kramer, Matthew Joseph; Voyles, Paul M

    2015-08-01

    Electron correlation microscopy (ECM) is a new technique that utilizes time-resolved coherent electron nanodiffraction to study dynamic atomic rearrangements in materials. It is the electron scattering equivalent of photon correlation spectroscopy with the added advantage of nanometer-scale spatial resolution. We have applied ECM to a Pd40Ni40P20 metallic glass, heated inside a scanning transmission electron microscope into a supercooled liquid to measure the structural relaxation time τ between the glass transition temperature T g and the crystallization temperature, T x . τ determined from the mean diffraction intensity autocorrelation function g 2(t) decreases with temperature following an Arrhenius relationship between T g and T g +25 K, and then increases as temperature approaches T x . The distribution of τ determined from the g 2(t) of single speckles is broad and changes significantly with temperature.

  15. Stroboscobic near-field scanning optical microscopy for 3D mapping of mode profiles of plasmonic nanostructures (Conference Presentation)

    Science.gov (United States)

    Dana, Aykutlu; Ozgur, Erol; Torunoglu, Gamze

    2016-09-01

    We present a dynamic approach to scanning near field optical microscopy that extends the measurement technique to the third dimension, by strobing the illumination in sync with the cantilever oscillation. Nitrogen vacancy (NV) centers in nanodiamonds placed on cantilever tips are used as stable emitters for emission enhancement. Local field enhancement and modulation of optical density states are mapped in three dimensions based on fluorescence intensity and spectrum changes as the tip is scanned over plasmonic nanostructures. The excitation of NV centers is done using a total internal reflection setup. Using a digital phase locked loop to pulse the excitation in various tip sample separations, 2D slices of fluorescence enhancement can be recorded. Alternatively, a conventional SNOM tip can be used to selectively couple wideband excitation to the collection path, with subdiffraction resolution of 60 nm in x and y and 10 nm in z directions. The approach solves the problem of tip-sample separation stabilization over extended periods of measurement time, required to collect data resolved in emission wavelength and three spatial dimensions. The method can provide a unique way of accessing the three dimensional field and mode profiles of nanophotonics structures.

  16. Developing a New Biophysical Tool to Combine Magneto-Optical Tweezers with Super-Resolution Fluorescence Microscopy

    Directory of Open Access Journals (Sweden)

    Zhaokun Zhou

    2015-06-01

    Full Text Available We present a novel experimental setup in which magnetic and optical tweezers are combined for torque and force transduction onto single filamentous molecules in a transverse configuration to allow simultaneous mechanical measurement and manipulation. Previously we have developed a super-resolution imaging module which, in conjunction with advanced imaging techniques such as Blinking assisted Localisation Microscopy (BaLM, achieves localisation precision of single fluorescent dye molecules bound to DNA of ~30 nm along the contour of the molecule; our work here describes developments in producing a system which combines tweezing and super-resolution fluorescence imaging. The instrument also features an acousto-optic deflector that temporally divides the laser beam to form multiple traps for high throughput statistics collection. Our motivation for developing the new tool is to enable direct observation of detailed molecular topological transformation and protein binding event localisation in a stretching/twisting mechanical assay that previously could hitherto only be deduced indirectly from the end-to-end length variation of DNA. Our approach is simple and robust enough for reproduction in the lab without the requirement of precise hardware engineering, yet is capable of unveiling the elastic and dynamic properties of filamentous molecules that have been hidden using traditional tools.

  17. Path radiance technique for retrieving aerosol optical thickness over land

    Energy Technology Data Exchange (ETDEWEB)

    Wen, Guoyong [Joint Center for Earth Systems Technology, University of Maryland, Baltimore County, Catonsville (United States); Tsay, Si-Chee [NASA Goddard Space Flight Center, Greenbelt, Maryland (United States); Cahalan, Robert F. [Joint Center for Earth Systems Technology, University of Maryland, Baltimore County, Catonsville (United States); NASA Goddard Space Flight Center, Greenbelt, Maryland (United States); Oreopoulos, Lazaros [Joint Center for Earth Systems Technology, University of Maryland, Baltimore County, Catonsville (United States)

    1999-12-27

    The key issue in retrieving aerosol optical thickness over land from shortwave satellite radiances is to identify and separate the signal due to scattering by a largely transparent aerosol layer from the noise due to reflection by the background surface, where the signal is relatively uniform compared to the highly inhomogeneous surface contribution. Sensitivity studies in aerosol optical thickness retrievals reveal that the apparent reflectance at the top of the atmosphere is very susceptible to the surface reflectance, especially when aerosol optical thickness is small. Uncertainties associated with surface reflectance estimation can greatly amplify the error of the aerosol optical thickness retrieval. To reduce these uncertainties, we have developed a ''path radiance'' method to retrieve aerosol optical thickness over land by extending the traditional technique that uses the ''dark object'' approach to extract the aerosol signal. This method uses the signature of the correlation of visible and middle-IR reflectance at the surface and couples the correlation with the atmospheric effect. We have applied this method to a Landsat TM (Thematic Mapper) image acquired over the Oklahoma southern Great Plains site of the Department of Energy Atmospheric Radiation Measurement (ARM) program on September 27, 1997, a very clear day (aerosol optical thickness of 0.07 at 0.5 {mu}m) during the first Landsat Intensive Observation Period. The retrieved mean aerosol optical thickness for TM band 1 at 0.49 {mu}m and band 3 at 0.66 {mu}m agree very well with the ground-based Sun photometer measurements at the ARM site. The ability to retrieve small aerosol optical thickness makes this path radiance technique promising. More importantly, the path radiance is relatively insensitive to surface inhomogeneity. The retrieved mean path radiances in reflectance units have very small standard deviations for both TM blue and red bands. This small

  18. Nonlinear optical techniques for imaging and manipulating the mouse central nervous system

    Science.gov (United States)

    Farrar, Matthew John

    The spinal cord of vertebrates serves as the conduit for somatosensory information and motor control, as well as being the locus of neural circuits that govern fast reflexes and patterned behaviors, such as walking in mammals or swimming in fish. Consequently, pathologies of the spinal cord -such as spinal cord injury (SCI)- lead to loss of motor control and sensory perception, with accompanying decline in life expectancy and quality of life. Despite the devastating effects of these diseases, few therapies exist to substantially ameliorate patient outcome. In part, studies of spinal cord pathology have been limited by the inability to perform in vivo imaging at the level of cellular processes. The focus of this thesis is to present the underlying theory for and demonstration of novel multi-photon microscopy (MPM) and optical manipulation techniques as they apply to studies the mouse central nervous system (CNS), with an emphasis on the spinal cord. The scientific findings which have resulted from the implementation of these techniques are also presented. In particular, we have demonstrated that third harmonic generation is a dye-free method of imaging CNS myelin, a fundamental constituent of the spinal cord that is difficult to label using exogenous dyes and/or transgenic constructs. Since gaining optical access to the spinal cord is a prerequisite for spinal cord imaging, we review our development of a novel spinal cord imaging chamber and surgical procedure which allowed us to image for multiple weeks following implantation without the need for repeated surgeries. We also have used MPM to characterize spinal venous blood flow before and after point occlusions. We review a novel nonlinear microscopy technique that may serve to show optical interfaces in three dimensions inside scattering tissue. Finally, we discuss a model and show results of optoporation, a means of transfecting cells with genetic constructs. Brief reviews of MPM and SCI are also presented.

  19. Three dimensional time lapse imaging of live cell mitochondria with photothermal optical lock-in optical coherence microscopy (Conference Presentation)

    Science.gov (United States)

    Sison, Miguel; Chakrabortty, Sabyasachi; Extermann, Jerome; Nahas, Amir; Pache, Christophe; Weil, Tanja; Lasser, Theo

    2016-03-01

    The photothermal optical lock-in optical coherence microscope (poli-OCM) introduced molecular specificity to OCM imaging, which is conventionally, a label-free technique. Here we achieve three-dimensional live cell and mitochondria specific imaging using ~4nm protein-functionalized gold nanoparticles (AuNPs). These nanoparticles do not photobleach and we demonstrate they're suitability for long-term time lapse imaging. We compare the accuracy of labelling with these AuNPs using classical fluorescence confocal imaging with a standard mitochondria specific marker. Furthermore, time lapse poli-OCM imaging every 5 minutes over 1.5 hours period was achieved, revealing the ability for three-dimensional monitoring of mitochondria dynamics.

  20. Closer to the native state. Critical evaluation of cryo-techniques for Transmission Electron Microscopy: preparation of biological samples.

    Science.gov (United States)

    Mielanczyk, Lukasz; Matysiak, Natalia; Michalski, Marek; Buldak, Rafal; Wojnicz, Romuald

    2014-01-01

    Over the years Transmission Electron Microscopy (TEM) has evolved into a powerful technique for the structural analysis of cells and tissues at various levels of resolution. However, optimal sample preservation is required to achieve results consistent with reality. During the last few decades, conventional preparation methods have provided most of the knowledge about the ultrastructure of organelles, cells and tissues. Nevertheless, some artefacts can be introduced at all stagesofstandard electron microscopy preparation technique. Instead, rapid freezing techniques preserve biological specimens as close as possible to the native state. Our review focuses on different cryo-preparation approaches, starting from vitrification methods dependent on sample size. Afterwards, we discuss Cryo-Electron Microscopy Of VItreous Sections (CEMOVIS) and the main difficulties associated with this technique. Cryo-Focused Ion Beam (cryo-FIB) is described as a potential alternative for CEMOVIS. Another post-processing route for vitrified samples is freeze substitution and embedding in resin for structural analysis or immunolocalization analysis. Cryo-sectioning according to Tokuyasu is a technique dedicated to high efficiency immunogold labelling. Finally, we introduce hybrid techniques, which combine advantages of primary techniques originally dedicated to different approaches. Hybrid approaches permit to perform the study of difficult-to-fix samples and antigens or help optimize the sample preparation protocol for the integrated Laser and Electron Microscopy (iLEM) technique.

  1. Imaging photonic crystals using Fourier plane imaging and Fourier ptychographic microscopy techniques implemented with a computer controlled hemispherical digital condenser

    Science.gov (United States)

    Sen, Sanchari; Desai, Darshan B.; Alsubaie, Meznh H.; Zhelyeznyakov, Maksym V.; Molina, L.; Sarraf, Hamed Sari; Bernussi, Ayrton A.; Peralta, Luis Grave de

    2017-01-01

    Fourier plane imaging (FPIM) and Fourier ptychographic (FPM) microscopy techniques were used to image photonic crystals. A computer-controlled hemispherical digital condenser provided required sample illumination with variable inclination. Notable improvement in image resolution was obtained with both methods. However, it was determined that the FPM technique cannot surpass the Rayleigh resolution limit when imaging photonic crystals.

  2. Fluorescence microscopy techniques for quantitative evaluation of organic biocide distribution in antifouling paint coatings: application to model antifouling coatings.

    Science.gov (United States)

    Goodes, L R; Dennington, S P; Schuppe, H; Wharton, J A; Bakker, M; Klijnstra, J W; Stokes, K R

    2012-01-01

    A test matrix of antifouling (AF) coatings including pMMA, an erodible binder and a novel trityl copolymer incorporating Cu₂O and a furan derivative (FD) natural product, were subjected to pontoon immersion and accelerated rotor tests. Fluorescence and optical microscopy techniques were applied to these coatings for quantification of organic biocide and pigment distribution. Total leaching of the biocide from the novel copolymer binder was observed within 6 months of rotor immersion, compared to 35% from the pMMA coating. In pontoon immersions, 61% of the additive was lost from the pMMA coating, and 53% from the erodible binder. Profiles of FD content in the binders revealed an accelerated loss of additive from the surface of the CDP resulting from rosin degradation, compared to even depletion from pMMA. In all samples, release of the biocide was inhibited beyond the Cu₂O front, corresponding to the leached layer in samples where Cu₂O release occurred.

  3. Techniques of advanced light microscopy and their applications to morphological analysis of human extra-embryonic membranes.

    Science.gov (United States)

    Ockleford, C D; Mongan, L C; Hubbard, A R

    The science of light microscopy has advanced dramatically in recent years through the introduction of new technology. A brief description of scanning light microscopes, laser illumination, the confocal principle, digital imaging, and image processing reveals a number of theoretical advantages which are particularly useful in improving epifluorescence microscope images. Examples of results from several studies of human extra-embryonic membranes conducted in our laboratory show how the application of these techniques has been used to describe structures such as microtrabeculae and rivets for the first time, to map the microscopic distribution of a wide range of proteins, and to observe the activity of placental villi at the microscopic level in an environmentally controlled microscope stage. High-sensitivity detectors have permitted the "super-resolution" detection of structures smaller than the theoretically calculated limits of light microscope resolution. Rendering images in false colour is demonstrably useful in detecting subtle variations in fluorescence intensity at different intracellular sites and at different sites within tissues of fetal membranes. Processing stacks of digital images using appropriate software allows the 3-D reconstruction of suitably sized extra-embryonic membrane components. These digital images created from optical sections through the tissue are obtained non-destructively, and the relationships in space of the components are well preserved.

  4. Near-Field Optical Microscopy of Defects in Cholesteric Oligomeric Liquid Crystal Films

    Energy Technology Data Exchange (ETDEWEB)

    Lukishova, S.G.; Schmid, A.W.

    2006-08-18

    This paper describes formation of 2-D hexagonal structures with a periodicity ~0.5-0.8 um in the defects of thin films of cholesteric oligomeric liquid crystals prepared by the evaporation of the solvent from the oligomer solution on the substrate. These regular arrays were observed by scanning near-field optical and concurrent atomic force microscopy. The mechanisms considered are both Benard-Marangoni and buoyancy conventions induced by solvent evaporation and air-bubble creation around the condensed water droplets from the air during evaporative cooling. Hexagonal structures prepared by this method can be used in photonic devices for emission enhancement, for instance, in liquid crystal lasers and single photon sources with oligomeric liquid crystal hosts.

  5. Single-beam photothermal microscopy - a new diagnostic tool for optical materials

    Energy Technology Data Exchange (ETDEWEB)

    Feit, M D; Kozlowski, M; Natoli, J Y; Rubenchik, A M; Sheehan L; Wu, Z L; Yan, M

    1998-12-22

    A novel photothermal microscopy (PTM) is developed which uses only one laser beam, working as both the pump and the probe. The principle of this single-beam PTM is based on the detection of the second harmonic component of the laser modulated scattering (LMS) signal. This component has a linear dependence on the optical absorptance of the tested area and a quadratic dependence on the pump laser power. Using a pump laser at the wavelengths of 514.5- and 532-nm high-resolution photothermal scans are performed for polished fused silica surfaces and a HfO{sub 2}/SiO{sub 2} multilayer coatings. The results are compared with those from the traditional two-beam PTM mapping. It is demonstrated that the single-beam PTM is more user-friendly (i.e. no alignment is needed) than conventional two-beam PTM and, offers a higher spatial resolution for defect detection.

  6. Stent-induced coronary artery stenosis characterized by multimodal nonlinear optical microscopy

    Science.gov (United States)

    Wang, Han-Wei; Simianu, Vlad; Locker, Mattew J.; Cheng, Ji-Xin; Sturek, Michael

    2011-02-01

    We demonstrate for the first time the applicability of multimodal nonlinear optical (NLO) microscopy to the interrogation of stented coronary arteries under different diet and stent deployment conditions. Bare metal stents and Taxus drug-eluting stents (DES) were placed in coronary arteries of Ossabaw pigs of control and atherogenic diet groups. Multimodal NLO imaging was performed to inspect changes in arterial structures and compositions after stenting. Sum frequency generation, one of the multimodalities, was used for the quantitative analysis of collagen content in the peristent and in-stent artery segments of both pig groups. Atherogenic diet increased lipid and collagen in peristent segments. In-stent segments showed decreased collagen expression in neointima compared to media. Deployment of DES in atheromatous arteries inhibited collagen expression in the arterial media.

  7. Label-free imaging of developing vasculature in zebrafish with phase variance optical coherence microscopy

    Science.gov (United States)

    Chen, Yu; Fingler, Jeff; Trinh, Le A.; Fraser, Scott E.

    2016-03-01

    A phase variance optical coherence microscope (pvOCM) has been created to visualize blood flow in the vasculature of zebrafish embryos, without using exogenous labels. The pvOCM imaging system has axial and lateral resolutions of 2 μm in tissue, and imaging depth of more than 100 μm. Imaging of 2-5 days post-fertilization zebrafish embryos identified the detailed structures of somites, spinal cord, gut and notochord based on intensity contrast. Visualization of the blood flow in the aorta, veins and intersegmental vessels was achieved with phase variance contrast. The pvOCM vasculature images were confirmed with corresponding fluorescence microscopy of a zebrafish transgene that labels the vasculature with green fluorescent protein. The pvOCM images also revealed functional information of the blood flow activities that is crucial for the study of vascular development.

  8. Crystallization kinetics of poly-(lactic acid) with and without talc: Optical microscopy and calorimetric analysis

    Science.gov (United States)

    Refaa, Z.; Boutaous, M.; Rousset, F.; Fulchiron, R.; Zinet, M.; Xin, S.; Bourgin, P.

    2014-05-01

    Poly-(lactic acid) or PLA is a biodegradable polymer synthesized from renewable resources. Recently, the discovery of new polymerization routes has allowed increasing the produced volumes. As a consequence, PLA is becoming of great interest for reducing the dependence on petroleum-based plastics. Because of its interesting mechanical properties, PLA is seen as a potential substitute for some usual polymers. However, its relatively slow crystallization kinetics can be a disadvantage with regard to industrial applications. The crystallization kinetics of PLA can be enhanced by adding nucleating agents, which also influences on crystalline morphology and rheological behavior. In the present work, the isothermal quiescent crystallization kinetics of both neat PLA and PLA/talc composite (5 wt% talc) are investigated. The effects of talc on the overall crystallization kinetics and on the crystalline morphology are analyzed using both optical microscopy measurements and thermal analysis by differential scanning calorimetry.

  9. Dual tree complex wavelet transform based denoising of optical microscopy images.

    Science.gov (United States)

    Bal, Ufuk

    2012-12-01

    Photon shot noise is the main noise source of optical microscopy images and can be modeled by a Poisson process. Several discrete wavelet transform based methods have been proposed in the literature for denoising images corrupted by Poisson noise. However, the discrete wavelet transform (DWT) has disadvantages such as shift variance, aliasing, and lack of directional selectivity. To overcome these problems, a dual tree complex wavelet transform is used in our proposed denoising algorithm. Our denoising algorithm is based on the assumption that for the Poisson noise case threshold values for wavelet coefficients can be estimated from the approximation coefficients. Our proposed method was compared with one of the state of the art denoising algorithms. Better results were obtained by using the proposed algorithm in terms of image quality metrics. Furthermore, the contrast enhancement effect of the proposed method on collagen fıber images is examined. Our method allows fast and efficient enhancement of images obtained under low light intensity conditions.

  10. Imaging immune and metabolic cells of visceral adipose tissues with multimodal nonlinear optical microscopy.

    Directory of Open Access Journals (Sweden)

    Yasuyo Urasaki

    Full Text Available Visceral adipose tissue (VAT inflammation is recognized as a mechanism by which obesity is associated with metabolic diseases. The communication between adipose tissue macrophages (ATMs and adipocytes is important to understanding the interaction between immunity and energy metabolism and its roles in obesity-induced diseases. Yet visualizing adipocytes and macrophages in complex tissues is challenging to standard imaging methods. Here, we describe the use of a multimodal nonlinear optical (NLO microscope to characterize the composition of VATs of lean and obese mice including adipocytes, macrophages, and collagen fibrils in a label-free manner. We show that lipid metabolism processes such as lipid droplet formation, lipid droplet microvesiculation, and free fatty acids trafficking can be dynamically monitored in macrophages and adipocytes. With its versatility, NLO microscopy should be a powerful imaging tool to complement molecular characterization of the immunity-metabolism interface.

  11. Near-field optical microscopy of localized excitations on rough surfaces: influence of a probe

    DEFF Research Database (Denmark)

    Bozhevolnyi, Sergey I.

    1999-01-01

    Starting from the general principles of near-field optical microscopy. I consider the influence of a probe when being used to image localized dipolar excitations and suggest a way of evaluating the perturbation thus introduced. Using the rigorous microscopic (electric) point-dipole description, I...... calculate the self consistent field intensity at the site of a probe dipole scanning over resonantly interacting object dipoles and show that the intensity distribution deviates from that existing in the absence of a probe. I demonstrate that this difference increases with an increase in the polarizability...... of the probe dipole, resulting eventually in a completely different intensity distribution, The calculations also show that the perturbation of the intensity distribution due to the presence of a probe decreases with an increase in the probe-sample distance. In order to evaluate the degree of perturbation, I...

  12. Nonlinear Optical Imaging of Individual Carbon Nanotubes with Four-Wave-Mixing Microscopy

    Science.gov (United States)

    Kim, Hyunmin; Sheps, Tatyana; Collins, Philip G.; Potma, Eric O.

    2014-01-01

    Dual color four-wave-mixing (FWM) microscopy is used to spatially resolve the third-order optical response from individual carbon nanotubes. Good signal-to-noise is obtained from single-walled carbon nanotubes (SWNT) sitting on substrates, when the excitation beams are resonant with electronic transitions of the nanotube, by detecting the FWM response at the anti-Stokes frequency. Whereas the coherent anti-Stokes (CAS) signal is sensitive to both electronic and vibrational resonances of the material, it is shown that the signal from individual SWNTs is dominated by the electronic response. The CAS signal is strongly polarization dependent, with the highest signals found parallel with the enhanced electronic polarizibility along the long axis of the SWNT. PMID:19637886

  13. Macro-optical trapping for sample confinement in light sheet microscopy.

    Science.gov (United States)

    Yang, Zhengyi; Piksarv, Peeter; Ferrier, David E K; Gunn-Moore, Frank J; Dholakia, Kishan

    2015-08-01

    Light sheet microscopy is a powerful approach to construct three-dimensional images of large specimens with minimal photo-damage and photo-bleaching. To date, the specimens are usually mounted in agents such as agarose, potentially restricting the development of live samples, and also highly mobile specimens need to be anaesthetized before imaging. To overcome these problems, here we demonstrate an integrated light sheet microscope which solely uses optical forces to trap and hold the sample using a counter-propagating laser beam geometry. Specifically, tobacco plant cells and living Spirobranchus lamarcki larvae were successfully trapped and sectional images acquired. This novel approach has the potential to significantly expand the range of applications for light sheet imaging.

  14. PEO-LiClO4-ZSM5 composite polymer electrolyte (IV): Polarized optical microscopy study

    Institute of Scientific and Technical Information of China (English)

    XI Jingyu; QIU Xinping; ZHU Wentao; CHEN Liquan

    2005-01-01

    Polarized optical microscopy (POM) results show that ZSM5 has great influence on both the nucleation stage and the growth stage of PEO spherulites. Part of ZSM5 particles can act as the nucleus of PEO spherulites and thus increase the amount of PEO spherulites. On the other hand, ZSM5 can restrain the recrystallization tendency of PEO chains through Lewis acid-base interaction and hence decrease the growth speed of PEO spherulites. The increasing amount of PEO spherulites, decreasing size of PEO spherulites, and the incomplete crystallization are all beneficial for creating more continuous amorphous phases of PEO, which is very important for the transporting of Li+ ions. An adequate amount of ZSM5 can enhance the room temperature ionic conductivity of PEO-LiClO4 based polymer electrolyte for more than two magnitudes.

  15. A tiled CCD detector with 2x2 array and tapered fibre optics for electron microscopy

    CERN Document Server

    Faruqi, A R; Cattermole, D M; Stubbings, S

    2002-01-01

    Charge coupled devices (CCD)-based detectors have made a major impact on data collection in electron microscopy over the past few years. There have been a number of successful applications of CCDs in electron crystallography of two-dimensional protein crystal arrays but high-resolution imaging has been hampered by the relatively poor spatial resolution (and fewer independent pixels) compared to film. A partial solution to this problem, presented in this paper, are to design detectors with larger effective pixel sizes and with more pixels. A CCD detector with a much greater number of 'independent' pixels, achieved by tiling a 2x2 array of CCDs, each of which has 1242x1152 pixels is described here. The sensitive area of the detector, using fibre optics with a demagnification of 2.5 : 1, is 140x130 mm sup 2; the pixel size is 56 mu m square and there is a total of approx 2500x2300 pixels.

  16. New optical tomographic & topographic techniques for biomedical applications

    Science.gov (United States)

    Buytaert, Jan

    The mammalian middle ear contains the eardrum and the three auditory ossicles, and forms an impedance match between sound in air and pressure waves in the fluid of the inner ear. Without this intermediate system, with its unsurpassed efficiency and dynamic range, we would be practically deaf. Physics-based modeling of this extremely complex mechanical system is necessary to help our basic understanding of the functioning of hearing. Highly realistic models will make it possible to predict the outcome of surgical interventions and to optimize design of ossicle prostheses and active middle ear implants. To obtain such models and with realistic output, basic input data is still missing. In this dissertation I developed and used two new optical techniques to obtain two essential sets of data: accurate three-dimensional morphology of the middle ear structures, and elasticity parameters of the eardrum. The first technique is a new method for optical tomography of macroscopic biomedical objects, which makes it possible to measure the three-dimensional geometry of the middle ear ossicles and soft tissues which are connecting and suspending them. I made a new and high-resolution version of this orthogonal-plane fluorescence optical sectioning method, to obtain micrometer resolution in macroscopic specimens. The result is thus a complete 3-D model of the middle (and inner) ear of gerbil in unprecedented quality. On top of high-resolution morphological models of the middle ear structures, I applied the technique in other fields of research as well. The second device works according to a new optical profilometry technique which allows to measure shape and deformations of the eardrum and other membranes or objects. The approach is called projection moire profilometry, and creates moire interference fringes which contain the height information. I developed a setup which uses liquid crystal panels for grid projection and optical demodulation. Hence no moving parts are present and

  17. Development of fast two-dimensional standing wave microscopy using acousto-optic deflectors

    Science.gov (United States)

    Gliko, Olga; Reddy, Duemani G.; Brownell, William E.; Saggau, Peter

    2008-02-01

    A novel scheme for two-dimensional (2D) standing wave fluorescence microscopy (SWFM) using acousto-optic deflectors (AODs) is proposed. Two laser beams were coupled into an inverted microscope and focused at the back focal plane of the objective lens. The position of each of two beams at the back focal plane was controlled by a pair of AODs. This resulted in two collimated beams that interfered in the focal plane, creating a lateral periodic excitation pattern with variable spacing and orientation. The phase of the standing wave pattern was controlled by phase delay between two RF sinusoidal signals driving the AODs. Nine SW patterns of three different orientations about the optical axis and three different phases were generated. The excitation of the specimen using these patterns will result in a SWFM image with enhanced 2D lateral resolution with a nearly isotropic effective point-spread function. Rotation of the SW pattern relative to specimen and varying the SW phase do not involve any mechanical movements and are only limited by the time required for the acoustic wave to fill the aperture of AOD. The resulting total acquisition time can be as short as 100 µs and is only further limited by speed and sensitivity of the employed CCD camera. Therefore, this 2D SWFM can provide a real time imaging of subresolution processes such as docking and fusion of synaptic vesicles. In addition, the combination of 2D SWFM with variable angle total internal reflection (TIR) can extend this scheme to fast microscopy with enhanced three-dimensional (3D) resolution.

  18. Metrology Optical Power Budgeting in SIM Using Statistical Analysis Techniques

    Science.gov (United States)

    Kuan, Gary M

    2008-01-01

    The Space Interferometry Mission (SIM) is a space-based stellar interferometry instrument, consisting of up to three interferometers, which will be capable of micro-arc second resolution. Alignment knowledge of the three interferometer baselines requires a three-dimensional, 14-leg truss with each leg being monitored by an external metrology gauge. In addition, each of the three interferometers requires an internal metrology gauge to monitor the optical path length differences between the two sides. Both external and internal metrology gauges are interferometry based, operating at a wavelength of 1319 nanometers. Each gauge has fiber inputs delivering measurement and local oscillator (LO) power, split into probe-LO and reference-LO beam pairs. These beams experience power loss due to a variety of mechanisms including, but not restricted to, design efficiency, material attenuation, element misalignment, diffraction, and coupling efficiency. Since the attenuation due to these sources may degrade over time, an accounting of the range of expected attenuation is needed so an optical power margin can be book kept. A method of statistical optical power analysis and budgeting, based on a technique developed for deep space RF telecommunications, is described in this paper and provides a numerical confidence level for having sufficient optical power relative to mission metrology performance requirements.

  19. Rapid cost-effective silicon carbide optical component manufacturing technique

    Science.gov (United States)

    Casstevens, John M.; Plummer, Ronald; Jarocki, Jim

    1999-10-01

    Silicon carbide may well be the best known material for the manufacture of high performance optical components. A combination of extremely high specific stiffness (r/E), high thermal conductivity and outstanding dimensional stability make silicon carbide superior overall to beryllium and low- expansion glass ceramics. A major impediment to wide use of silicon carbide in optical systems has been the costs of preliminary pressing, casting, shaping and final finishing of silicon carbide. Diamond grinding of silicon carbide is a slow and expensive process even on machines specially designed for the task. The process described here begins by machining the component from a special type of graphite. This graphite is easily machined with multi-axis CNC machine tools to any level of complexity and lightweighting required. The graphite is then converted completely to silicon carbide with very small and very predictable dimensional change. After conversion to silicon carbide the optical surface is coated with very fine grain silicon carbide which is easily polished to extreme smoothness using conventional optical polishing techniques. The fabrication process and a 6 inch diameter development mirror is described.

  20. Fluorescent Probes and Fluorescence (Microscopy Techniques — Illuminating Biological and Biomedical Research

    Directory of Open Access Journals (Sweden)

    Gregor P. C. Drummen

    2012-11-01

    Full Text Available Fluorescence, the absorption and re-emission of photons with longer wavelengths, is one of those amazing phenomena of Nature. Its discovery and utilization had, and still has, a major impact on biological and biomedical research, since it enables researchers not just to visualize normal physiological processes with high temporal and spatial resolution, to detect multiple signals concomitantly, to track single molecules in vivo, to replace radioactive assays when possible, but also to shed light on many pathobiological processes underpinning disease states, which would otherwise not be possible. Compounds that exhibit fluorescence are commonly called fluorochromes or fluorophores and one of these fluorescent molecules in particular has significantly enabled life science research to gain new insights in virtually all its sub-disciplines: Green Fluorescent Protein. Because fluorescent proteins are synthesized in vivo, integration of fluorescent detection methods into the biological system via genetic techniques now became feasible. Currently fluorescent proteins are available that virtually span the whole electromagnetic spectrum. Concomitantly, fluorescence imaging techniques were developed, and often progress in one field fueled innovation in the other. Impressively, the properties of fluorescence were utilized to develop new assays and imaging modalities, ranging from energy transfer to image molecular interactions to imaging beyond the diffraction limit with super-resolution microscopy. Here, an overview is provided of recent developments in both fluorescence imaging and fluorochrome engineering, which together constitute the “fluorescence toolbox” in life science research.

  1. Development of a digital video-microscopy technique to study lactose crystallisation kinetics in situ.

    Science.gov (United States)

    Arellano, María Paz; Aguilera, José Miguel; Bouchon, Pedro

    2004-11-15

    Polarised light microscopy was employed non-invasively to monitor lactose crystallisation from non-seeded supersaturated solutions in real time. Images were continuously recorded, processed and characterised by image analysis, and the results were compared with those obtained by refractometry. Three crystallisation temperatures (10, 20 and 30 degrees C) and three different levels of initial relative supersaturation (C/C(s)=1.95; 2.34; 3.15) were investigated. Induction times using the imaging technique proved to be substantially lower than those determined using refractive index. Lactose crystals were isolated digitally to determine geometrical parameters of interest, such as perimeter, diameter, area, roundness and Feret mean, and to derive crystal growth rates. Mean growth rates obtained for single crystals were fitted to a combined mass transfer model (R(2)=0.9766). The model allowed the effects of temperature and supersaturation on crystallisation rate to be clearly identified. It also suggested that, in this set of experiments, surface integration seemed to be the rate controlling step. It is believed that a similar experimental set-up could be implemented in a real food system to characterise a particular process where crystallisation control is of interest and where traditional techniques are difficult to implement.

  2. Local delivery of fluorescent dye for fiber-optics confocal microscopy of the living heart.

    Science.gov (United States)

    Huang, Chao; Kaza, Aditya K; Hitchcock, Robert W; Sachse, Frank B

    2014-01-01

    Fiber-optics confocal microscopy (FCM) is an emerging imaging technology with various applications in basic research and clinical diagnosis. FCM allows for real-time in situ microscopy of tissue at sub-cellular scale. Recently FCM has been investigated for cardiac imaging, in particular, for discrimination of cardiac tissue during pediatric open-heart surgery. FCM relies on fluorescent dyes. The current clinical approach of dye delivery is based on systemic injection, which is associated with high dye consumption, and adverse clinical events. In this study, we investigated approaches for local dye delivery during FCM imaging based on dye carriers attached to the imaging probe. Using three-dimensional confocal microscopy, automated bench tests, and FCM imaging we quantitatively characterized dye release of carriers composed of open-pore foam only and foam loaded with agarose hydrogel. In addition, we compared local dye delivery with a model of systemic dye delivery in the isolated perfused rodent heart. We measured the signal-to-noise ratio (SNR) of images acquired in various regions of the heart. Our evaluations showed that foam-agarose dye carriers exhibited a prolonged dye release vs. foam-only carriers. Foam-agarose dye carriers allowed reliable imaging of 5-9 lines, which is comparable to 4-8 min of continuous dye release. Our study in the living heart revealed that the SNR of FCM images using local and systemic dye delivery is not different. However, we observed differences in the imaged tissue microstructure with the two approaches. Structural features characteristic of microvasculature were solely observed for systemic dye delivery. Our findings suggest that local dye delivery approach for FCM imaging constitutes an important alternative to systemic dye delivery. We suggest that the approach for local dye delivery will facilitate clinical translation of FCM, for instance, for FCM imaging during pediatric heart surgery.

  3. Correlation between polarization sensitive optical coherence tomography and SHG microscopy in articular cartilage

    Science.gov (United States)

    Zhou, Xin; Ju, Myeong Jin; Huang, Lin; Tang, Shuo

    2017-02-01

    Polarization-sensitive optical coherence tomography (PS-OCT) and second harmonic generation (SHG) microscopy are two imaging modalities with different resolutions, field-of-views (FOV), and contrasts, while they both have the capability of imaging collagen fibers in biological tissues. PS-OCT can measure the tissue birefringence which is induced by highly organized fibers while SHG can image the collagen fiber organization with high resolution. Articular cartilage, with abundant structural collagen fibers, is a suitable sample to study the correlation between PS-OCT and SHG microscopy. Qualitative conjecture has been made that the phase retardation measured by PS-OCT is affected by the relationship between the collagen fiber orientation and the illumination direction. Anatomical studies show that the multilayered architecture of articular cartilage can be divided into four zones from its natural surface to the subchondral bone: the superficial zone, the middle zone, the deep zone, and the calcified zone. The different zones have different collagen fiber orientations, which can be studied by the different slopes in the cumulative phase retardation in PS-OCT. An algorithm is developed based on the quantitative analysis of PS-OCT phase retardation images to analyze the microstructural features in swine articular cartilage tissues. This algorithm utilizes the depth-dependent slope changing of phase retardation A-lines to segment structural layers. The results show good consistency with the knowledge of cartilage morphology and correlation with the SHG images measured at selected depth locations. The correlation between PS-OCT and SHG microscopy shows that PS-OCT has the potential to analyze both the macro and micro characteristics of biological tissues with abundant collagen fibers and other materials that may cause birefringence.

  4. Local Delivery of Fluorescent Dye For Fiber-Optics Confocal Microscopy of the Living Heart

    Directory of Open Access Journals (Sweden)

    Chao eHuang

    2014-09-01

    Full Text Available Fiber-optics confocal microscopy (FCM is an emerging imaging technology with various applications in basic research and clinical diagnosis. FCM allows for real-time in situ microscopy of tissue at sub-cellular scale. Recently FCM has been investigated for cardiac imaging, in particular, for discrimination of cardiac tissue during pediatric open-heart surgery. FCM relies on fluorescent dyes. The current clinical approach of dye delivery is based on systemic injection, which is associated with high dye consumption and adverse clinical events. In this study, we investigated approaches for local dye delivery during FCM imaging based on dye carriers attached to the imaging probe. Using three-dimensional confocal microscopy, automated bench tests, and FCM imaging we quantitatively characterized dye release of carriers composed of open-pore foam only and foam loaded with agarose hydrogel. In addition, we compared local dye delivery with a model of systemic dye delivery in the isolated perfused rodent heart. We measured the signal-to-noise ratio of images acquired in various regions of the heart. Our evaluations showed that foam-agarose dye carriers exhibited a prolonged dye release versus foam-only carriers. Foam-agarose dye carriers allowed reliable imaging of 5-9 lines, which is comparable to 4-8 min of continuous dye release. Our study in the living heart revealed that the SNR of FCM images using local and systemic dye delivery is not different. However, we observed differences in the imaged tissue microstructure with the two approaches. Structural features characteristic of microvasculature were solely observed for systemic dye delivery. Our findings suggest that local dye delivery approach for FCM imaging constitutes an important alternative to systemic dye delivery. We suggest that the approach for local dye delivery will facilitate clinical translation of FCM, for instance, for FCM imaging during pediatric heart surgery.

  5. Optical tracking of embryonic vertebrates behavioural responses using automated time-resolved video-microscopy system

    Science.gov (United States)

    Walpitagama, Milanga; Kaslin, Jan; Nugegoda, Dayanthi; Wlodkowic, Donald

    2016-12-01

    The fish embryo toxicity (FET) biotest performed on embryos of zebrafish (Danio rerio) has gained significant popularity as a rapid and inexpensive alternative approach in chemical hazard and risk assessment. The FET was designed to evaluate acute toxicity on embryonic stages of fish exposed to the test chemical. The current standard, similar to most traditional methods for evaluating aquatic toxicity provides, however, little understanding of effects of environmentally relevant concentrations of chemical stressors. We postulate that significant environmental effects such as altered motor functions, physiological alterations reflected in heart rate, effects on development and reproduction can occur at sub-lethal concentrations well below than LC10. Behavioral studies can, therefore, provide a valuable integrative link between physiological and ecological effects. Despite the advantages of behavioral analysis development of behavioral toxicity, biotests is greatly hampered by the lack of dedicated laboratory automation, in particular, user-friendly and automated video microscopy systems. In this work we present a proof-of-concept development of an optical system capable of tracking embryonic vertebrates behavioral responses using automated and vastly miniaturized time-resolved video-microscopy. We have employed miniaturized CMOS cameras to perform high definition video recording and analysis of earliest vertebrate behavioral responses. The main objective was to develop a biocompatible embryo positioning structures that were suitable for high-throughput imaging as well as video capture and video analysis algorithms. This system should support the development of sub-lethal and behavioral markers for accelerated environmental monitoring.

  6. Correlative nonlinear optical microscopy and infrared nanoscopy reveals collagen degradation in altered parchments.

    Science.gov (United States)

    Latour, Gaël; Robinet, Laurianne; Dazzi, Alexandre; Portier, François; Deniset-Besseau, Ariane; Schanne-Klein, Marie-Claire

    2016-05-19

    This paper presents the correlative imaging of collagen denaturation by nonlinear optical microscopy (NLO) and nanoscale infrared (IR) spectroscopy to obtain morphological and chemical information at different length scales. Such multiscale correlated measurements are applied to the investigation of ancient parchments, which are mainly composed of dermal fibrillar collagen. The main issue is to characterize gelatinization, the ultimate and irreversible alteration corresponding to collagen denaturation to gelatin, which may also occur in biological tissues. Key information about collagen and gelatin signatures is obtained in parchments and assessed by characterizing the denaturation of pure collagen reference samples. A new absorbing band is observed near the amide I band in the IR spectra, correlated to the onset of fluorescence signals in NLO images. Meanwhile, a strong decrease is observed in Second Harmonic signals, which are a structural probe of the fibrillar organization of the collagen at the micrometer scale. NLO microscopy therefore appears as a powerful tool to reveal collagen degradation in a non-invasive way. It should provide a relevant method to assess or monitor the condition of collagen-based materials in museum and archival collections and opens avenues for a broad range of applications regarding this widespread biological material.

  7. Distinguishing between ultrafast optical harmonic generation and multi-photon-induced luminescence from ZnO thin films by interferometric frequency-resolved autocorrelation microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, Slawa; Mascheck, Manfred; Silies, Martin [Carl-von-Ossietzky-Universitaet, Oldenburg (Germany); Yatsui, Takashi; Kitamura, Kokoro; Ohtsu, Motoichi; Lienau, Christoph [University of Tokyo (Japan)

    2011-07-01

    The nonlinear optical properties of a thin ZnO film are studied using interferometric frequency-resolved autocorrelation (IFRAC) microscopy. By exciting the film with 6-fs, below-bandgap laser pulses at 800nm focused to a spot size of 1 {mu}m two emission bands in the blue and bluegreen spectral region with distinctly different coherence properties can be detected. We show that an analysis of the wavelength-dependence of the interference fringes in the IFRAC signal allows for an unambiguous assignment of these bands as coherent second harmonic emission and incoherent, multiphoton-induced photoluminescence, respectively. More generally our analysis shows that IFRAC allows for a complete characterization of the coherence properties of the nonlinear optical emission from nanostructures in a single-beam experiment. Since this technique combines a very high temporal and spatial resolution we anticipate broad applications in nonlinear nano-optics.

  8. Properties of grain boundary networks in the NEEM ice core analyzed by combined transmission and reflection optical microscopy

    Science.gov (United States)

    Binder, Tobias; Weikusat, Ilka; Garbe, Christoph; Svensson, Anders; Kipfstuhl, Sepp

    2014-05-01

    Microstructure analysis of ice cores is vital to understand the processes controlling the flow of ice on the microscale. To quantify the microstructural variability (and thus occurring processes) on centimeter, meter and kilometer scale along deep polar ice cores, a large number of sections has to be analyzed. In the last decade, two different methods have been applied: On the one hand, transmission optical microscopy of thin sections between crossed polarizers yields information on the distribution of crystal c-axes. On the other hand, reflection optical microscopy of polished and controlled sublimated section surfaces allows to characterize the high resolution properties of a single grain boundary, e.g. its length, shape or curvature (further developed by [1]). Along the entire NEEM ice core (North-West Greenland, 2537 m length) drilled in 2008-2011 we applied both methods to the same set of vertical sections. The data set comprises series of six consecutive 6 x 9 cm2 sections in steps of 20 m - in total about 800 images. A dedicated method for automatic processing and matching both image types has recently been developed [2]. The high resolution properties of the grain boundary network are analyzed. Furthermore, the automatic assignment of c-axis misorientations to visible sublimation grooves enables us to quantify the degree of similarity between the microstructure revealed by both analysis techniques. The reliability to extract grain boundaries from both image types as well as the appearance of sublimation groove patterns exhibiting low misorientations is investigated. X-ray Laue diffraction measurements (yielding full crystallographic orientation) have validated the sensitivity of the surface sublimation method for sub-grain boundaries [3]. We introduce an approach for automatic extraction of sub-grain structures from sublimation grooves. A systematic analysis of sub-grain boundary densities indicates a possible influence of high impurity contents (amongst

  9. Neurophotonics: non-invasive optical techniques for monitoring brain functions

    Science.gov (United States)

    Torricelli, Alessandro; Contini, Davide; Mora, Alberto Dalla; Pifferi, Antonio; Re, Rebecca; Zucchelli, Lucia; Caffini, Matteo; Farina, Andrea; Spinelli, Lorenzo

    2014-01-01

    Summary The aim of this review is to present the state of the art of neurophotonics, a recently founded discipline lying at the interface between optics and neuroscience. While neurophotonics also includes invasive techniques for animal studies, in this review we focus only on the non-invasive methods that use near infrared light to probe functional activity in the brain, namely the fast optical signal, diffuse correlation spectroscopy, and functional near infrared spectroscopy methods. We also present an overview of the physical principles of light propagation in biological tissues, and of the main physiological sources of signal. Finally, we discuss the open issues in models, instrumentation, data analysis and clinical approaches. PMID:25764252

  10. Dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy

    Science.gov (United States)

    Li, Yongbo; Shinohara, Ryosuke; Iwami, Kentaro; Ohta, Yoshihiro; Umeda, Norihiro

    2010-08-01

    A novel method to observe pH distribution by dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy (SNOM) is developed. In this method, in order to investigate not only the pH of mitochondrial membrane but also its distribution in the vicinity, a pH sensitive fluorescent reagent covers mitochondria instead of injecting it to mitochondria. This method utilizes a dual-emission pH sensitive dye and SNOM with a themally-pulled and metal-coated optical fiber to improve the spatial resolution. Time-dependence of Fluorescent intensity ratio (FIR) under acid addition is investigated. As the distances between the dropped point and the SNOM probe becomes closer, the time when FIR changes becomes earlier. The response of mitochondria under supplement of nutrition is studied by using this method. While the probe is near to mitochondria, the ratio quickly becomes to increase. In conclusion, it was confirmed that the temporal variation of pH can be detected by this method, and pH distribution in the vicinity of mitochondria is able to be measured by this method.

  11. Rat brain imaging using full field optical coherence microscopy with short multimode fiber probe

    Science.gov (United States)

    Sato, Manabu; Saito, Daisuke; Kurotani, Reiko; Abe, Hiroyuki; Kawauchi, Satoko; Sato, Shunichi; Nishidate, Izumi

    2017-02-01

    We demonstrated FF OCM(full field optical coherence microscopy) using an ultrathin forward-imaging SMMF (short multimode fiber) probe of 50 μm core diameter, 125 μm diameter, and 7.4 mm length, which is a typical graded-index multimode fiber for optical communications. The axial resolution was measured to be 2.20 μm, which is close to the calculated axial resolution of 2.06 μm. The lateral resolution was evaluated to be 4.38 μm using a test pattern. Assuming that the FWHM of the contrast is the DOF (depth of focus), the DOF of the signal is obtained at 36 μm and that of the OCM is 66 μm. The contrast of the OCT images was 6.1 times higher than that of the signal images due to the coherence gate. After an euthanasia the rat brain was resected and cut at 2.6mm tail from Bregma. Contacting SMMF to the primary somatosensory cortex and the agranular insular cortex of ex vivo brain, OCM images of the brain were measured 100 times with 2μm step. 3D OCM images of the brain were measured, and internal structure information was obtained. The feasibility of an SMMF as an ultrathin forward-imaging probe in full-field OCM has been demonstrated.

  12. Micron-scale resolution optical tomography of entire mouse brains with confocal light sheet microscopy.

    Science.gov (United States)

    Silvestri, Ludovico; Bria, Alessandro; Costantini, Irene; Sacconi, Leonardo; Peng, Hanchuan; Iannello, Giulio; Pavone, Francesco Saverio

    2013-10-08

    Understanding the architecture of mammalian brain at single-cell resolution is one of the key issues of neuroscience. However, mapping neuronal soma and projections throughout the whole brain is still challenging for imaging and data management technologies. Indeed, macroscopic volumes need to be reconstructed with high resolution and contrast in a reasonable time, producing datasets in the TeraByte range. We recently demonstrated an optical method (confocal light sheet microscopy, CLSM) capable of obtaining micron-scale reconstruction of entire mouse brains labeled with enhanced green fluorescent protein (EGFP). Combining light sheet illumination and confocal detection, CLSM allows deep imaging inside macroscopic cleared specimens with high contrast and speed. Here we describe the complete experimental pipeline to obtain comprehensive and human-readable images of entire mouse brains labeled with fluorescent proteins. The clearing and the mounting procedures are described, together with the steps to perform an optical tomography on its whole volume by acquiring many parallel adjacent stacks. We showed the usage of open-source custom-made software tools enabling stitching of the multiple stacks and multi-resolution data navigation. Finally, we illustrated some example of brain maps: the cerebellum from an L7-GFP transgenic mouse, in which all Purkinje cells are selectively labeled, and the whole brain from a thy1-GFP-M mouse, characterized by a random sparse neuronal labeling.

  13. Ultrafast random-access scanning in two-photon microscopy using acousto-optic deflectors.

    Science.gov (United States)

    Salomé, R; Kremer, Y; Dieudonné, S; Léger, J-F; Krichevsky, O; Wyart, C; Chatenay, D; Bourdieu, L

    2006-06-30

    Two-photon scanning microscopy (TPSM) is a powerful tool for imaging deep inside living tissues with sub-cellular resolution. The temporal resolution of TPSM is however strongly limited by the galvanometric mirrors used to steer the laser beam. Fast physiological events can therefore only be followed by scanning repeatedly a single line within the field of view. Because acousto-optic deflectors (AODs) are non-mechanical devices, they allow access at any point within the field of view on a microsecond time scale and are therefore excellent candidates to improve the temporal resolution of TPSM. However, the use of AOD-based scanners with femtosecond pulses raises several technical difficulties. In this paper, we describe an all-digital TPSM setup based on two crossed AODs. It includes in particular an acousto-optic modulator (AOM) placed at 45 degrees with respect to the AODs to pre-compensate for the large spatial distortions of femtosecond pulses occurring in the AODs, in order to optimize the spatial resolution and the fluorescence excitation. Our setup allows recording from freely selectable point-of-interest at high speed (1kHz). By maximizing the time spent on points of interest, random-access TPSM (RA-TPSM) constitutes a promising method for multiunit recordings with millisecond resolution in biological tissues.

  14. Determination of crystal grain orientations by optical microscopy at textured surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Lausch, D.; Gläser, M.; Hagendorf, C. [Team Mikrostrukturdiagnostik und Analytik, Fraunhofer-Center für Silizium-Photovoltaik CSP, Walter-Hülse-Straße 1 Halle (Saale), Sachsen-Anhalt D-06120 (Germany)

    2013-11-21

    In this contribution, a new method to determine the crystal orientation with the example of chemical treated silicon wafers by means of optical microscopy has been demonstrated. The introduced procedure represents an easy method to obtain all relevant parameters to describe the crystal structure of the investigated material, i.e., the crystal grain orientation and the grain boundary character. The chemical treatment is a standard mono-texture for solar cells, well known in the solar industry. In general, this concept can also be applied to other crystalline materials, i.e., GaAs, SiC, etc., the only thing that needs to be adjusted is the texturing method to reveal specific crystal planes and the calculation model. In conclusion, an application of this method is shown with the example of the defect classification of recombination active defects in mc-Si solar cell. The introduced method demonstrates a simple and quick opportunity to improve the crystallization process and the quality of electronic devices by means of an optical microscope and a chemical treatment of the material.

  15. Smart microscope: an adaptive optics learning system for aberration correction in multiphoton confocal microscopy.

    Science.gov (United States)

    Albert, O; Sherman, L; Mourou, G; Norris, T B; Vdovin, G

    2000-01-01

    Off-axis aberrations in a beam-scanning multiphoton confocal microscope are corrected with a deformable mirror. The optimal mirror shape for each pixel is determined by a genetic learning algorithm, in which the second-harmonic or two-photon fluorescence signal from a reference sample is maximized. The speed of the convergence is improved by use of a Zernike polynomial basis for the deformable mirror shape. This adaptive optical correction scheme is implemented in an all-reflective system by use of extremely short (10-fs) optical pulses, and it is shown that the scanning area of an f:1 off-axis parabola can be increased by nine times with this technique.

  16. High-Resolution Optical Tweezers Combined With Single-Molecule Confocal Microscopy.

    Science.gov (United States)

    Whitley, K D; Comstock, M J; Chemla, Y R

    2017-01-01

    We describe the design, construction, and application of an instrument combining dual-trap, high-resolution optical tweezers and a confocal microscope. This hybrid instrument allows nanomechanical manipulation and measurement simultaneously with single-molecule fluorescence detection. We present the general design principles that overcome the challenges of maximizing optical trap resolution while maintaining single-molecule fluorescence sensitivity, and provide details on the construction and alignment of the instrument. This powerful new tool is just beginning to be applied to biological problems. We present step-by-step instructions on an application of this technique that highlights the instrument's capabilities, detecting conformational dynamics in a nucleic acid-processing enzyme. © 2017 Elsevier Inc. All rights reserved.

  17. Energy Landscape of Alginate-Epimerase Interactions Assessed by Optical Tweezers and Atomic Force Microscopy.

    Directory of Open Access Journals (Sweden)

    Armend Gazmeno Håti

    Full Text Available Mannuronan C-5 epimerases are a family of enzymes that catalyze epimerization of alginates at the polymer level. This group of enzymes thus enables the tailor-making of various alginate residue sequences to attain various functional properties, e.g. viscosity, gelation and ion binding. Here, the interactions between epimerases AlgE4 and AlgE6 and alginate substrates as well as epimerization products were determined. The interactions of the various epimerase-polysaccharide pairs were determined over an extended range of force loading rates by the combined use of optical tweezers and atomic force microscopy. When studying systems that in nature are not subjected to external forces the access to observations obtained at low loading rates, as provided by optical tweezers, is a great advantage since the low loading rate region for these systems reflect the properties of the rate limiting energy barrier. The AlgE epimerases have a modular structure comprising both A and R modules, and the role of each of these modules in the epimerization process were examined through studies of the A- module of AlgE6, AlgE6A. Dynamic strength spectra obtained through combination of atomic force microscopy and the optical tweezers revealed the existence of two energy barriers in the alginate-epimerase complexes, of which one was not revealed in previous AFM based studies of these complexes. Furthermore, based on these spectra estimates of the locations of energy transition states (xβ, lifetimes in the absence of external perturbation (τ0 and free energies (ΔG# were determined for the different epimerase-alginate complexes. This is the first determination of ΔG# for these complexes. The values determined were up to 8 kBT for the outer barrier, and smaller values for the inner barriers. The size of the free energies determined are consistent with the interpretation that the enzyme and substrate are thus not tightly locked at all times but are able to relocate

  18. Energy Landscape of Alginate-Epimerase Interactions Assessed by Optical Tweezers and Atomic Force Microscopy.

    Science.gov (United States)

    Håti, Armend Gazmeno; Aachmann, Finn Lillelund; Stokke, Bjørn Torger; Skjåk-Bræk, Gudmund; Sletmoen, Marit

    2015-01-01

    Mannuronan C-5 epimerases are a family of enzymes that catalyze epimerization of alginates at the polymer level. This group of enzymes thus enables the tailor-making of various alginate residue sequences to attain various functional properties, e.g. viscosity, gelation and ion binding. Here, the interactions between epimerases AlgE4 and AlgE6 and alginate substrates as well as epimerization products were determined. The interactions of the various epimerase-polysaccharide pairs were determined over an extended range of force loading rates by the combined use of optical tweezers and atomic force microscopy. When studying systems that in nature are not subjected to external forces the access to observations obtained at low loading rates, as provided by optical tweezers, is a great advantage since the low loading rate region for these systems reflect the properties of the rate limiting energy barrier. The AlgE epimerases have a modular structure comprising both A and R modules, and the role of each of these modules in the epimerization process were examined through studies of the A- module of AlgE6, AlgE6A. Dynamic strength spectra obtained through combination of atomic force microscopy and the optical tweezers revealed the existence of two energy barriers in the alginate-epimerase complexes, of which one was not revealed in previous AFM based studies of these complexes. Furthermore, based on these spectra estimates of the locations of energy transition states (xβ), lifetimes in the absence of external perturbation (τ0) and free energies (ΔG#) were determined for the different epimerase-alginate complexes. This is the first determination of ΔG# for these complexes. The values determined were up to 8 kBT for the outer barrier, and smaller values for the inner barriers. The size of the free energies determined are consistent with the interpretation that the enzyme and substrate are thus not tightly locked at all times but are able to relocate. Together with the

  19. Energy Landscape of Alginate-Epimerase Interactions Assessed by Optical Tweezers and Atomic Force Microscopy

    Science.gov (United States)

    Håti, Armend Gazmeno; Aachmann, Finn Lillelund; Stokke, Bjørn Torger; Skjåk-Bræk, Gudmund; Sletmoen, Marit

    2015-01-01

    Mannuronan C-5 epimerases are a family of enzymes that catalyze epimerization of alginates at the polymer level. This group of enzymes thus enables the tailor-making of various alginate residue sequences to attain various functional properties, e.g. viscosity, gelation and ion binding. Here, the interactions between epimerases AlgE4 and AlgE6 and alginate substrates as well as epimerization products were determined. The interactions of the various epimerase–polysaccharide pairs were determined over an extended range of force loading rates by the combined use of optical tweezers and atomic force microscopy. When studying systems that in nature are not subjected to external forces the access to observations obtained at low loading rates, as provided by optical tweezers, is a great advantage since the low loading rate region for these systems reflect the properties of the rate limiting energy barrier. The AlgE epimerases have a modular structure comprising both A and R modules, and the role of each of these modules in the epimerization process were examined through studies of the A- module of AlgE6, AlgE6A. Dynamic strength spectra obtained through combination of atomic force microscopy and the optical tweezers revealed the existence of two energy barriers in the alginate-epimerase complexes, of which one was not revealed in previous AFM based studies of these complexes. Furthermore, based on these spectra estimates of the locations of energy transition states (xβ), lifetimes in the absence of external perturbation (τ0) and free energies (ΔG#) were determined for the different epimerase–alginate complexes. This is the first determination of ΔG# for these complexes. The values determined were up to 8 kBT for the outer barrier, and smaller values for the inner barriers. The size of the free energies determined are consistent with the interpretation that the enzyme and substrate are thus not tightly locked at all times but are able to relocate. Together with the

  20. Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

    Science.gov (United States)

    Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

    2014-08-01

    Primary biological aerosol particles (PBAPs) can contribute significantly to the coarse particle burden in many environments. PBAPs can thus influence climate and precipitation systems as cloud nuclei and can spread disease to humans, animals, and plants. Measurement data and techniques for PBAPs in natural environments at high time- and size resolution are, however, sparse, and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in southwestern Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of a waveband integrated bioaerosol sensor (WIBS-4) with a ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behavior, with increased fluorescent bioparticle concentrations at night, when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each was correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multimodal distributions turning into a broad featureless single mode after averaging, and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent

  1. In vivo fluorescence fiber-optic microscopy of superfast Ca^2+ transients in syrinx muscles

    Science.gov (United States)

    Dall'Asén, Analía; Méndez, Jorge; Goller, Franz

    2010-03-01

    Optical techniques in conjunction with fluorescent markers have revolutionized the investigation of dynamical cellular processes, such as studies of calcium ion (Ca^2+) dynamics in muscles. Recently, it was shown that songbirds have superfast syringeal muscles, which can modulate song acoustics up to 250 Hz. Such rapid contraction cycles most likely require very rapid Ca^2+ kinetics. We developed a technique to measure Ca^2+ transients in syringeal muscles of anesthetized birds with a custom-built endoscope. The fluorescence measurements were carried out before and after applying a calcium indicator dye and while muscles were stimulated electrically. The results show fast (˜50-ms FWHM) and superfast (˜7-ms FWHM) Ca^2+ transients. The strongest signals were observed 30-40 minutes after applying the dye. This study confirms that rapid Ca^2+ transients in syringeal muscles facilitate superfast contraction kinetics and demonstrates the feasibility of this optical technique as a biosensor for detecting fluorescence signals of muscular calcium activity on a ms timescale of living animals.

  2. Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM).

    Science.gov (United States)

    Tang, Anson H L; Lai, Queenie T K; Chung, Bob M F; Lee, Kelvin C M; Mok, Aaron T Y; Yip, G K; Shum, Anderson H C; Wong, Kenneth K Y; Tsia, Kevin K

    2017-06-28

    Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring. Incorporating imaging capabilities into flow cytometry compromises the assay throughput, primarily due to the limitations on speed and sensitivity in the camera technologies. To overcome this speed or throughput challenge facing imaging flow cytometry while preserving the image quality, asymmetric-detection time-stretch optical microscopy (ATOM) has been demonstrated to enable high-contrast, single-cell imaging with sub-cellular resolution, at an imaging throughput as high as 100,000 cells/s. Based on the imaging concept of conventional time-stretch imaging, which relies on all-optical image encoding and retrieval through the use of ultrafast broadband laser pulses, ATOM further advances imaging performance by enhancing the image contrast of unlabeled/unstained cells. This is achieved by accessing the phase-gradient information of the cells, which is spectrally encoded into single-shot broadband pulses. Hence, ATOM is particularly advantageous in high-throughput measurements of single-cell morphology and texture - information indicative of cell types, states, and even functions. Ultimately, this could become a powerful imaging flow cytometry platform for the biophysical phenotyping of cells, complementing the current state-of-the-art biochemical-marker-based cellular assay. This work describes a protocol to establish the key modules of an ATOM system (from optical frontend to data processing and visualization

  3. Optical modulation techniques for analog signal processing and CMOS compatible electro-optic modulation

    Science.gov (United States)

    Gill, Douglas M.; Rasras, Mahmoud; Tu, Kun-Yii; Chen, Young-Kai; White, Alice E.; Patel, Sanjay S.; Carothers, Daniel; Pomerene, Andrew; Kamocsai, Robert; Beattie, James; Kopa, Anthony; Apsel, Alyssa; Beals, Mark; Mitchel, Jurgen; Liu, Jifeng; Kimerling, Lionel C.

    2008-02-01

    Integrating electronic and photonic functions onto a single silicon-based chip using techniques compatible with mass-production CMOS electronics will enable new design paradigms for existing system architectures and open new opportunities for electro-optic applications with the potential to dramatically change the management, cost, footprint, weight, and power consumption of today's communication systems. While broadband analog system applications represent a smaller volume market than that for digital data transmission, there are significant deployments of analog electro-optic systems for commercial and military applications. Broadband linear modulation is a critical building block in optical analog signal processing and also could have significant applications in digital communication systems. Recently, broadband electro-optic modulators on a silicon platform have been demonstrated based on the plasma dispersion effect. The use of the plasma dispersion effect within a CMOS compatible waveguide creates new challenges and opportunities for analog signal processing since the index and propagation loss change within the waveguide during modulation. We will review the current status of silicon-based electrooptic modulators and also linearization techniques for optical modulation.

  4. Calibration-free absolute quantification of optical absorption coefficients using acoustic spectra in 3D photoacoustic microscopy of biological tissue.

    Science.gov (United States)

    Guo, Zijian; Hu, Song; Wang, Lihong V

    2010-06-15

    Optical absorption is closely associated with many physiological important parameters, such as the concentration and oxygen saturation of hemoglobin, and it can be used to quantify the concentrations of nonfluorescent molecules. We propose a method to use acoustic spectra of photoacoustic signals to quantify the absolute optical absorption. This method is self-calibrating and thus insensitive to variations in the optical fluence. Factors such as system bandwidth and acoustic attenuation can affect the quantification but can be canceled by dividing the acoustic spectra measured at two optical wavelengths. Using optical-resolution photoacoustic microscopy, we quantified the absolute optical absorption of black ink samples with various concentrations. We also quantified both the concentration and oxygen saturation of hemoglobin in a live mouse in absolute units.

  5. Optical waveguide behavior of Se-doped and undoped CdS one-dimensional nanostructures using near-field optical microscopy

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao; LIU Dan; PAN Anlian; FANG Zheyu; HUANG Shan; ZHU Xing

    2009-01-01

    The optical waveguide behaviors of CdS and CdSxSe1-x nanostructures are studied using near-field optical microscopy. Optical measurements demonstrate that light may be guided on sub-wavelength scales along CdS nanoribbons in straight or bent structures. The photoluminescence (PL) spectra from nanoribbon emission using scanning near-field optical microscopy are analyzed under different inci-dent laser intensities. The PL spectra along Se-doped and undoped CdS nanoribbons at different propagation distances are investigated. Both the guided PL spectra of Se-doped and undoped CdS nanoribbons show red-shifts because of the band-edge absorption. Our results are useful for the de-velopment of new kinds of functional nano devices.

  6. Optical waveguide behavior of Se-doped and undoped CdS one-dimensional nanostructures using near-field optical microscopy

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The optical waveguide behaviors of CdS and CdSxSe1?x nanostructures are studied using near-field optical microscopy. Optical measurements demonstrate that light may be guided on sub-wavelength scales along CdS nanoribbons in straight or bent structures. The photoluminescence (PL) spectra from nanoribbon emission using scanning near-field optical microscopy are analyzed under different incident laser intensities. The PL spectra along Se-doped and undoped CdS nanoribbons at different propagation distances are investigated. Both the guided PL spectra of Se-doped and undoped CdS nanoribbons show red-shifts because of the band-edge absorption. Our results are useful for the development of new kinds of functional nano devices.

  7. Effect of thickness on optical properties of nickel vertical posts deposited by GLAD technique

    Science.gov (United States)

    Potočnik, J.; Nenadović, M.; Bundaleski, N.; Popović, M.; Rakočević, Z.

    2016-12-01

    Nickel (Ni) thin films of different thicknesses (25 nm to 150 nm) were deposited on glass substrates using Glancing Angle Deposition technique. Characterization of obtained Ni films was performed by scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, spectroscopic ellipsometry and by four-point probe method. Variations in optical parameters with thickness correlated with structural, chemical and electrical properties of nanostructured nickel thin films were studied. The results showed that deposit is porous and consists of nano-scaled columns, which grow perpendicular to the substrate. It was found that the size of the columns and the surface roughness change with film thickness. Spectroscopic ellipsometry revealed that the refractive index and extinction coefficient varied with thickness, which can be correlated with changes in microstructure of Ni films. Additionally, the relationship between the film microstructure and its resistivity was also analyzed. It was found that the variations in Ni films resistivity could be attributed to the changes in the width of the columns. The increasing of layer thickness leads to overall decrease of optical resistivity of nickel thin films.

  8. Local thermal conductivity of polycrystalline AlN ceramics measured by scanning thermal microscopy and complementary scanning electron microscopy techniques

    Institute of Scientific and Technical Information of China (English)

    Zhang Yue-Fei; Wang Li; R. Heiderhoff; A. K. Geinzer; Wei Bin; Ji Yuan; Han Xiao-Dong; L. J. Balk; Zhang Ze

    2012-01-01

    The local thermal conductivity of polycrystalline aluminum nitride (AlN) ceramics is measured and imaged by using a scanning thermal microscope (SThM) and complementary scanning electron microscope (SEM) based techniques at room temperature.The quantitative thermal conductivity for the AlN sample is gained by using a SThM with a spatial resolution of sub-micrometer scale through using the 3w method.A thermal conductivity of 308 W/m·K withingrains corresponding to that of high-purity single crystal AlN is obtained.The slight differences in thermal conduction between the adjacent grains are found to result from crystallographic misorientations,as demonstrated in the electron backscattered diffraction.A much lower thermal conductivity at the grain boundary is due to impurities and defects enriched in these sites,as indicated by energy dispersive X-ray spectroscopy.

  9. Detecting Trichinella infections using inverse microscopy and an improved larval counting technique.

    Science.gov (United States)

    Makrutzki, G; Riehn, K; Hamedy, A; Petroff, D; Hasenclever, D; Meiler, H; Lücker, E

    2014-12-01

    Several methods for the detection of Trichinella in meat are legally prescribed in regulation (EC) No 2075/2005, which prescribes the magnetic stirrer method for pooled sample digestion (MSM) as the reference method. However, the MSM's multistage protocol requires several preparatory steps that seem to be accountable for the loss of larvae. Here we present a modified MSM (mMSM) based on: (1) an inversion of the optical path using inverse microscopy; and (2) a modified larval counting basin (mLCB, 'Trichoview'). This enables one to examine samples of up to 40 ml and reduces the examination area from 72 to 10.3 cm2. Preparatory steps that might cause the loss of Trichinella larvae are eliminated from the new protocol. Correspondingly, the overall analytical time is reduced. In a direct and blinded comparison using 60 digest samples containing spiked vital Trichinella larvae (1-90 L1), both methods performed well for both small and large numbers of L1. However, 1278 of 1285 L1 (99.4%) were detected using the mMSM, while MSM recovered only 1225 L1 (95.3%). The improvement stems largely from samples with small numbers of L1: in all samples spiked with fewer than 10 L1, the recovery rate of mMSM was 100% compared to only 93% with MSM. Our data suggest that the use of the mMSM can improve the recovery rate by about 4% and therefore reduce the chances of a false-negative result in a sample containing 5 larvae by a factor of about 4.

  10. Viability and antibacterial efficacy of four root canal disinfection techniques evaluated using confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    Joan Mathew

    2014-01-01

    Full Text Available Background: Several disinfection techniques have been recently introduced with the main objective of improving root canal disinfection in the inaccessible areas of the root canal system. This in vitro study was done to evaluate the antimicrobial effect and viability of Enterococcus faecalis biofilms using conventional irrigation, EndoActivator (Dentsply, Tulsa Dental, USA, diode laser irradiation and photon-initiated photoacoustic streaming (PIPS. Materials and Methods: Root canals of 130 single rooted mandibular premolars, standardized to a uniform length of 20 mm were instrumented until finishing file, F1 (Universal Protaper Rotary System, Dentsply, Tulsa Dental Specialties, USA. After smear layer removal and sterilization, five teeth were randomly selected to assure sterility before bacterial inoculation. The remaining 125 samples were contaminated with E. faecalis suspension, incubated for 21 days and divided into five groups (n = 25. In Group 1; untreated group (positive control, the root canals were not subjected to any disinfection procedure. Sampling was performed within the canals and the colony-forming unit count was evaluated for 20 samples. Five samples were selected to visualize the pattern of colonization at Level 1 (4 mm from the apex and Level 2 (1 mm from the apex by confocal laser scanning microscopy. Samples in Groups 2-5 namely conventional needle irrigation, EndoActivator, diode laser and PIPS were subjected to their respective disinfection procedures. Postdisinfection sample evaluation criteria was followed for all groups as same as that for Group 1. Results: Diode laser displayed the highest antibacterial efficacy and least viable bacteria than the other three disinfection techniques. Conclusion: Diode laser group showed better antibacterial efficacy and least viable bacteria when compared to conventional needle irrigation, PIPS and EndoActivator groups in minimally instrumented, experimentally infected root canals.

  11. A constant compliance force modulation technique for scanning force microscopy (SFM) imaging of polymer surface elasticity

    Science.gov (United States)

    Stroup, E.W.; Pungor, A/

    2012-01-01

    A new method of force modulation scanning force microscopy (SFM) imaging based on a constant compliance feedback loop is presented. The feedback adjusts the loading force applied by the SFM tip to the surface in order to maintain a constant compliance beneath the tip. The new method, constant compliance force modulation (CCFM), has the advantage of being able to quantify the loading force exerted by the tip onto the sample surface and thus to estimate the elastic modulus of the material probed by the SFM tip. Once the elastic modulus of one region is known, the elastic moduli of other surface regions can be estimated from the spatial map of loading forces using the Hertz model of deformation. Force vs. displacement measurements made on one surface locality could also be used to estimate the local modulus. Several model surfaces, including a rubber-toughened epoxy polymer blend which showed clearly resolved compliant rubber phases within the harder epoxy matrix, were analyzed with the CCFM technique to illustrate the method’s application. PMID:9195751

  12. Trade-offs between spatial and temporal resolutions in stochastic super-resolution microscopy techniques

    CERN Document Server

    Rupprecht, Jean-Francois; Tessier, Gilles

    2016-01-01

    Widefield stochastic microscopy techniques such as PALM or STORM rely on the progressive accumulation of a large number of frames, each containing a scarce number of super-resolved point images. We justify that the redundancy in the localization of detected events imposes a specific limit on the temporal resolution. Based on a theoretical model, we derive analytical predictions for the minimal time required to obtain a reliable image at a given spatial resolution, called image completion time. In contrast to standard assumptions, we find that the image completion time scales logarithmically with the ratio of the image size by the spatial resolution volume. We justify that this non-linear relation is the hallmark of a random coverage problem. We propose a method to estimate the risk that the image reconstruction is not complete, which we apply to an experimental data set. Our results provide a theoretical framework to quantify the pattern detection efficiency and to optimize the trade-off between image coverag...

  13. Drying techniques for the visualisation of agarose-based chromatography media by scanning electron microscopy.

    Science.gov (United States)

    Nweke, Mauryn C; Turmaine, Mark; McCartney, R Graham; Bracewell, Daniel G

    2017-03-01

    The drying of chromatography resins prior to scanning electron microscopy is critical to image resolution and hence understanding of the bead structure at sub-micron level. Achieving suitable drying conditions is especially important with agarose-based chromatography resins, as over-drying may cause artefact formation, bead damage and alterations to ultrastructural properties; and under-drying does not provide sufficient resolution for visualization under SEM. This paper compares and contrasts the effects of two drying techniques, critical point drying and freeze drying, on the morphology of two agarose based resins (MabSelect™/dw ≈85 µm and Capto™ Adhere/dw ≈75 µm) and provides a complete method for both. The results show that critical point drying provides better drying and subsequently clearer ultrastructural visualization of both resins under SEM. Under this protocol both the polymer fibers (thickness ≈20 nm) and the pore sizes (diameter ≈100 nm) are clearly visible. Freeze drying is shown to cause bead damage to both resins, but to different extents. MabSelect resin encounters extensive bead fragmentation, whilst Capto Adhere resin undergoes partial bead disintegration, corresponding with the greater extent of agarose crosslinking and strength of this resin. While freeze drying appears to be the less favorable option for ultrastructural visualization of chromatography resin, it should be noted that the extent of fracturing caused by the freeze drying process may provide some insight into the mechanical properties of agarose-based chromatography media.

  14. Acousto-optic multiphoton laser scanning microscopy and multiphoton photon counting spectroscopy: Applications and implications for optical neurobiology

    Science.gov (United States)

    Iyer, Vijay

    Multiphoton excitation of molecular probes has become an important tool in experimental neurobiology owing to the intrinsic optical sectioning and low light scattering it affords. Using molecular functional indicators, multiphoton excitation allows physiological signals within single neurons to be observed from within living brain tissue. Ideally, it would be possible to record from multiple sites located throughout the elaborately branching dendritic arbors, in order to study the correlations of structure and function both within and across experiments. However, existing multiphoton microscope systems based on scanning mirrors do not allow optical recordings to be obtained from more than a handful of sites simultaneously at the high rates required to capture the fast physiological signals of interest (>100Hz for Ca2+ signals, >1kHz for membrane potential transients). In order to overcome this limitation, two-dimensional acousto-optic deflection was employed, to allow an ultrafast laser beam suited for multiphoton excitation to be rapidly repositioned with low latency (˜15mus). This supports a random-access scanning mode in which the beam can repeatedly visit a succession of user-selected sites of interest within the microscope's field-of-view at high rates, with minimal sacrifice of pixel dwell time. This technique of acousto-optic multiphoton laser scanning microscope (AO-MPLSM) was demonstrated to allow the spatial profile of signals arising in response to physiological stimulation to be rapidly mapped. Means to compensate or avoid problems of dispersion which have hampered AO-MPLSM in the past are presented, with the latter being implemented. Separately, the combination of photon counting detection with multiphoton excitation, termed generally multiphoton photon counting spectroscopy (MP-PCS), was also considered, with particular emphasis on the technique of fluorescence correlation spectroscopy (FCS). MP-PCS was shown to allow information about molecular

  15. In-vivo multiphoton microscopy (MPM) of laser-induced optical breakdown (LIOB) in human skin (Conference Presentation)

    Science.gov (United States)

    Balu, Mihaela; Lentsch, Griffin; Korta, Dorota; Konig, Karsten; Kelly, Kristen M.; Tromberg, Bruce J.; Zachary, Christopher B.

    2017-02-01

    We use a multiphoton microscopy (MPM)-based clinical microscope (MPTflex, JenLab, Germany) to describe changes in human skin following treatment with a fractional non-ablative laser (PicoWay, Candela). The treatment was based on a fractionated picosecond Nd:YAG laser (1064 and 532nm, 3mJ and 1.5mJ (no attenuation), respectively maximum energy/pulse, 100 microbeams/6mmx6mm). Improvements in skin appearance resulting from treatment with this laser have been noted but optimizing the efficacy depends on a thorough understanding of the specific skin response to treatment. MPM is a nonlinear laser scanning microscopy technique that features sub-cellular resolution and label-free molecular contrast. MPM contrast in skin is derived from second-harmonic generation of collagen and two-photon excited fluorescence of NADH/FAD+, elastin, keratin, melanin. In this pilot study, two areas on the arm of a volunteer (skin type II) were treated with the picoWay laser (1064nm, 3mJ; 532nm, 1.5mJ; 1pass). The skin response to treatment was imaged in-vivo at 8 time points over the following 4 weeks. MPM revealed micro-injuries present in epidermis. Damaged individual cells were distinguished after 3h and 24h from treatment with both wavelengths. Pigmented cells were particularly damaged in the process, suggesting that melanin is the main absorber and the primary target for laser induced optical breakdown. At later time points, clusters of cellular necrotic debris were imaged across the treated epidermis. These results represent the groundwork for future longitudinal studies on expanded number of subjects to understand the response to treatment in different skin types at different laser parameters, critical factors in optimizing treatment outcomes.

  16. Characterization techniques for nano-electronics, with emphasis to electron microscopy. The role of the European Project ANNA

    Science.gov (United States)

    Armigliato, A.

    2008-07-01

    , however, European laboratories with high-level expertise in materials characterization still operate in a largely independent way; this adversely affects the competitivity of European science and industry at the international level. For this reason the European Commission has started an Integrated Infrastructure Initiative (I3) in the sixth Framework Programme (now continuing in FP7) and funded a project called ANNA (2006-2010). This acronym stands for European Integrated Activity of Excellence and Networking for Nano and Micro- Electronics Analysis. The consortium includes 12 partners from 7 European countries and is coordinated by the Fondazione B.Kessler (FBK) in Trento (Italy); CNR-IMM is one of the 12 partners. Aim of ANNA is the onset of strong, long-term collaboration among the partners, so to form an integrated multi-site analytical facility, able to offer to the European community a wide variety of top-level analytical expertise and services in the field of micro- and nano-electronics. They include X-ray diffraction and scattering, SIMS, electron microscopy, medium-energy ion scattering, optical and electrical techniques. The project will be focused on three main activities: Networking (standardization of samples and methodologies, establishment of accredited reference laboratories), Transnational Access to laboratories located in the partners' premises to perform specific analytical experiments (an example is given by the two STEM methodologies discussed above) and Joint Research activity, which is targeted at the improvement and extension of the methodologies through a continuous instrumental and technical development. It is planned that the European joint analytical laboratory will continue its activity beyond the end of the project in 2010.

  17. Application of optical spectroscopic techniques for disease diagnosis

    Science.gov (United States)

    Saha, Anushree

    Optical spectroscopy, a truly non-invasive tool for remote diagnostics, is capable of providing valuable information on the structure and function of molecules. However, most spectroscopic techniques suffer from drawbacks, which limit their application. As a part of my dissertation work, I have developed theoretical and experimental methods to address the above mentioned issues. I have successfully applied these methods for monitoring the physical, chemical and biochemical parameters of biomolecules involved in some specific life threatening diseases like lead poisoning and age-related macular degeneration (AMD). I presented optical studies of melanosomes, which are one of the vital organelles in the human eye, also known to be responsible for a disease called age-related macular degeneration (AMD), a condition of advanced degeneration which causes progressive blindness. I used Raman spectroscopy, to first chemically identify the composition of melanosome, and then monitor the changes in its functional and chemical behavior due to long term exposure to visible light. The above study, apart from explaining the role of melanosomes in AMD, also sets the threshold power for lasers used in surgeries and other clinical applications. In the second part of my dissertation, a battery of spectroscopic techniques was successfully applied to explore the different binding sites of lead ions with the most abundant carrier protein molecule in our circulatory system, human serum albumin. I applied optical spectroscopic tools for ultrasensitive detection of heavy metal ions in solution which can also be used for lead detection at a very early stage of lead poisoning. Apart from this, I used Raman microspectroscopy to study the chemical alteration occurring inside a prostate cancer cell as a result of a treatment with a low concentrated aqueous extract of a prospective drug, Nerium Oleander. The experimental methods used in this study has tremendous potential for clinical

  18. Optical analysis of nanomaterial-cell interactions: flow cytometry and digital holographic microscopy

    Science.gov (United States)

    Mues, Sarah; Antunovic, Jan; Ossig, Rainer; Kemper, Björn; Schnekenburger, Jürgen

    2015-05-01

    The in vitro cytotoxicity assessment of engineered nanoparticles commonly involves the measurement of different endpoints like the formation of reactive oxygen species, cell viability or cell death. Usually these parameters are determined by optical readouts of enzymatically converted substrates that often interfere with the tested nanomaterials. Using cell viability (WST-8) and cell death (LDH) as parameter we have initially investigated the toxic effects of spherical (NM 300) and rod shaped (NM 302) silver nanomaterials with a matrix of four cell lines representing different functions: lung and kidney epithelial cells, macrophages and fibroblasts. In addition, we have used a label-free flow cytometer configuration to investigate interactions of particles and macrophages by side scatter signal analysis. Finally, we explored digital holographic microscopy (DHM) for multimodal label-free analysis of nanomaterial toxicity. Quantitative DHM phase images were analyzed for cell thickness, volume, density, dry mass and refractive index. We could demonstrate that silver spheres lead to more cytotoxic effects than rods in all four examined cell lines and both assay. Exemplarily a dose dependent interaction increase of cells with NM 300 and NM 302 analyzed by flow cytometry is shown. Furthermore, we found that the refractive index of cells is influenced by incubation with NM 300 in a decreasing manner. A 24 hours time-lapse measurement revealed a dose dependent decrease of dry mass and surface area development indicating reduced cell viability and cell death. Our results demonstrate digital holographic microscopy and flow cytometry as valuable label-free tools for nanomaterial toxicity and cell interaction studies.

  19. Atomic Force and Optical Microscopy Characterization of the Deformation of Individual Carbon Nanotubes and Nanofibers

    Directory of Open Access Journals (Sweden)

    Terry P. Bigioni

    2008-01-01

    Full Text Available A popular technique for characterizing the mechanical properties of carbon nanotubes is to apply a one-dimension axial compression and measure its response to the compressive force. At some critical compression, a dramatic decrease in the force is observed. This has previously been attributed to Euler buckling, allowing the elastic modulus to be calculated from the Euler buckling force. We have attached individual plasma enhanced chemical vapor deposition (PECVD grown carbon nanofibers (CNFs and thermal chemical vapor deposition (CVD grown carbon nanotubes (CNTs to the apex of an atomic force microscope (AFM cantilever to examine this mechanical response. By combining the force measurements and simultaneous video microscopy, we are able to observe the mechanical deformation and correlate points in the force curve with phenomena such as slipping and bending. Analysis of the mechanical response must therefore be interpreted in terms of bending and/or slipping of a tube compressed by an off-normal force.

  20. Characterization of Si3N4/SiO2 optical channel waveguides by photon scanning tunneling microscopy

    Science.gov (United States)

    Wang, Yan; Chudgar, Mona H.; Jackson, Howard E.; Miller, Jeffrey S.; De Brabander, Gregory N.; Boyd, Joseph T.

    1993-01-01

    Photon scanning tunneling microscopy (PSTM) is used to characterize Si3N4/Si02 optical channel waveguides being used for integrated optical-micromechanical sensors. PSTM utilizes an optical fiber tapered to a fine point which is piezoelectrically positioned to measure the decay of the evanescent field intensity associated with the waveguide propagating mode. Evanescent field decays are recorded for both ridge channel waveguides and planar waveguide regions. Values for the local effective refractive index are calculated from the data for both polarizations and compared to model calculations.

  1. Visualization of mouse neuronal ganglia infected by Herpes Simplex Virus 1 (HSV-1 using multimodal non-linear optical microscopy.

    Directory of Open Access Journals (Sweden)

    Pierre-Alexandre Rochette

    Full Text Available Herpes simplex virus 1 (HSV-1 is a neurotropic virus that causes skin lesions and goes on to enter a latent state in neurons of the trigeminal ganglia. Following stress, the virus may reactivate from latency leading to recurrent lesions. The in situ study of neuronal infections by HSV-1 is critical to understanding the mechanisms involved in the biology of this virus and how it causes disease; however, this normally requires fixation and sectioning of the target tissues followed by treatment with contrast agents to visualize key structures, which can lead to artifacts. To further our ability to study HSV-1 neuropathogenesis, we have generated a recombinant virus expressing a second generation red fluorescent protein (mCherry, which behaves like the parental virus in vivo. By optimizing the application of a multimodal non-linear optical microscopy platform, we have successfully visualized in unsectioned trigeminal ganglia of mice both infected cells by two-photon fluorescence microscopy, and myelinated axons of uninfected surrounding cells by coherent anti-Stokes Raman scattering (CARS microscopy. These results represent the first report of CARS microscopy being combined with 2-photon fluorescence microscopy to visualize virus-infected cells deep within unsectioned explanted tissue, and demonstrate the application of multimodal non-linear optical microscopy for high spatial resolution biological imaging of tissues without the use of stains or fixatives.

  2. Visualization of mouse neuronal ganglia infected by Herpes Simplex Virus 1 (HSV-1) using multimodal non-linear optical microscopy.

    Science.gov (United States)

    Rochette, Pierre-Alexandre; Laliberté, Mathieu; Bertrand-Grenier, Antony; Houle, Marie-Andrée; Blache, Marie-Claire; Légaré, François; Pearson, Angela

    2014-01-01

    Herpes simplex virus 1 (HSV-1) is a neurotropic virus that causes skin lesions and goes on to enter a latent state in neurons of the trigeminal ganglia. Following stress, the virus may reactivate from latency leading to recurrent lesions. The in situ study of neuronal infections by HSV-1 is critical to understanding the mechanisms involved in the biology of this virus and how it causes disease; however, this normally requires fixation and sectioning of the target tissues followed by treatment with contrast agents to visualize key structures, which can lead to artifacts. To further our ability to study HSV-1 neuropathogenesis, we have generated a recombinant virus expressing a second generation red fluorescent protein (mCherry), which behaves like the parental virus in vivo. By optimizing the application of a multimodal non-linear optical microscopy platform, we have successfully visualized in unsectioned trigeminal ganglia of mice both infected cells by two-photon fluorescence microscopy, and myelinated axons of uninfected surrounding cells by coherent anti-Stokes Raman scattering (CARS) microscopy. These results represent the first report of CARS microscopy being combined with 2-photon fluorescence microscopy to visualize virus-infected cells deep within unsectioned explanted tissue, and demonstrate the application of multimodal non-linear optical microscopy for high spatial resolution biological imaging of tissues without the use of stains or fixatives.

  3. Insights into the prominent effect of mahanimbine on Acanthamoeba castellanii: Cell profiling analysis based on microscopy techniques

    Science.gov (United States)

    Hashim, Fatimah; Amin, Nakisah Mat

    2017-02-01

    Mahanimbine (MH), has been shown to have antiamoeba properties. Therefore, the aim of this study was to assess the growth inhibitory mechanisms of MH on Acanthamoeba castellanii, a causative agents for Acanthamoeba keratitis. The IC50 value obtained for MH against A. castellanii was 1.18 µg/ml. Light and scanning electron microscopy observation showed that most cells were in cystic appearance. While transmission electron microscopy observation revealed changes at the ultrastructural level and fluorescence microscopy observation indicated the induction of apoptosis and autophagic activity in the amoeba cytoplasms. In conclusion, MH has very potent anti-amoebic properties on A. castellanii as is shown by cytotoxicity analyses based on microscopy techniques.

  4. Application and development of advanced Lorentz microscopy techniques for the study of magnetic nanostructures

    Science.gov (United States)

    Beacham, Robert J.

    This PhD project presents an investigation into the development of magnetic imaging methods in the TEM and their application in imaging narrow domain walls in multilayer magnetic structures. Lorentz microscopy techniques are limited in quantitative magnetic imaging as this generally requires using scanning imaging modes which limits the capability of imaging dynamic processes. The first imaging method developed in this study is a phase gradient technique with the aim of producing quantitative magnetic contrast proportional to the magnetic induction of the sample whilst maintaining a live imaging mode. This method uses a specifically engineered, semi-electron-transparent graded wedge aperture to controllably perturb intensity in the back focal plane. The results of this study found that this method could produce magnetic contrast proportional to the sample induction, however the required gradient of the wedge aperture made this contrast close to the noise level with large associated errors. In the second part of this study we investigated the development of a technique aimed at gaining sub-microsecond temporal resolution within TEMs based on streak imaging. We are using ramped pulsed magnetic fields, applied across nanowire samples to both induce magnetic behaviour and detect the electron beam across the detector with respect to time. We are coupling this with a novel pixelated detector on the TEM in the form of a Medipix/Timepix chip capable of microsecond exposure times without adding noise. Running this detector in integral mode and allowing for practical limitations such as experiment time and aperture stability, the resultant streak images were taken in Fresnel, Foucault and low angle diffraction imaging modes. We found that while this method is theoretically viable, the limiting factor was the contrast of the magnetic signal in the streak and therefore the total image counts. Domain walls (DWs) in synthetic antiferromagnetically (SAF) coupled films patterned

  5. Parallelized multi-graphics processing unit framework for high-speed Gabor-domain optical coherence microscopy.

    Science.gov (United States)

    Tankam, Patrice; Santhanam, Anand P; Lee, Kye-Sung; Won, Jungeun; Canavesi, Cristina; Rolland, Jannick P

    2014-07-01

    Gabor-domain optical coherence microscopy (GD-OCM) is a volumetric high-resolution technique capable of acquiring three-dimensional (3-D) skin images with histological resolution. Real-time image processing is needed to enable GD-OCM imaging in a clinical setting. We present a parallelized and scalable multi-graphics processing unit (GPU) computing framework for real-time GD-OCM image processing. A parallelized control mechanism was developed to individually assign computation tasks to each of the GPUs. For each GPU, the optimal number of amplitude-scans (A-scans) to be processed in parallel was selected to maximize GPU memory usage and core throughput. We investigated five computing architectures for computational speed-up in processing 1000×1000 A-scans. The proposed parallelized multi-GPU computing framework enables processing at a computational speed faster than the GD-OCM image acquisition, thereby facilitating high-speed GD-OCM imaging in a clinical setting. Using two parallelized GPUs, the image processing of a 1×1×0.6  mm3 skin sample was performed in about 13 s, and the performance was benchmarked at 6.5 s with four GPUs. This work thus demonstrates that 3-D GD-OCM data may be displayed in real-time to the examiner using parallelized GPU processing.

  6. Parallelized multi–graphics processing unit framework for high-speed Gabor-domain optical coherence microscopy

    Science.gov (United States)

    Tankam, Patrice; Santhanam, Anand P.; Lee, Kye-Sung; Won, Jungeun; Canavesi, Cristina; Rolland, Jannick P.

    2014-01-01

    Abstract. Gabor-domain optical coherence microscopy (GD-OCM) is a volumetric high-resolution technique capable of acquiring three-dimensional (3-D) skin images with histological resolution. Real-time image processing is needed to enable GD-OCM imaging in a clinical setting. We present a parallelized and scalable multi-graphics processing unit (GPU) computing framework for real-time GD-OCM image processing. A parallelized control mechanism was developed to individually assign computation tasks to each of the GPUs. For each GPU, the optimal number of amplitude-scans (A-scans) to be processed in parallel was selected to maximize GPU memory usage and core throughput. We investigated five computing architectures for computational speed-up in processing 1000×1000 A-scans. The proposed parallelized multi-GPU computing framework enables processing at a computational speed faster than the GD-OCM image acquisition, thereby facilitating high-speed GD-OCM imaging in a clinical setting. Using two parallelized GPUs, the image processing of a 1×1×0.6  mm3 skin sample was performed in about 13 s, and the performance was benchmarked at 6.5 s with four GPUs. This work thus demonstrates that 3-D GD-OCM data may be displayed in real-time to the examiner using parallelized GPU processing. PMID:24695868

  7. Optical mapping of conduction in early embryonic quail hearts with light-sheet microscopy (Conference Presentation)

    Science.gov (United States)

    Ma, Pei; Gu, Shi; Wang, Yves T.; Jenkins, Michael W.; Rollins, Andrew M.

    2016-03-01

    Optical mapping (OM) using fluorescent voltage-sensitive dyes (VSD) to measure membrane potential is currently the most effective method for electrophysiology studies in early embryonic hearts due to its noninvasiveness and large field-of-view. Conventional OM acquires bright-field images, collecting signals that are integrated in depth and projected onto a 2D plane, not capturing the 3D structure of the sample. Early embryonic hearts, especially at looping stages, have a complicated, tubular geometry. Therefore, conventional OM cannot provide a full picture of the electrical conduction circumferentially around the heart, and may result in incomplete and inaccurate measurements. Here, we demonstrate OM of Hamburger and Hamilton stage 14 embryonic quail hearts using a new commercially-available VSD, Fluovolt, and depth sectioning using a custom built light-sheet microscopy system. Axial and lateral resolution of the system is 14µm and 8µm respectively. For OM imaging, the field-of-view was set to 900µm×900µm to cover the entire heart. 2D over time OM image sets at multiple cross-sections through the looping-stage heart were recorded. The shapes of both atrial and ventricular action potentials acquired were consistent with previous reports using conventional VSD (di-4-ANNEPS). With Fluovolt, signal-to-noise ratio (SNR) is improved significantly by a factor of 2-10 (compared with di-4-ANNEPS) enabling light-sheet OM, which intrinsically has lower SNR due to smaller sampling volumes. Electrophysiologic parameters are rate dependent. Optical pacing was successfully integrated into the system to ensure heart rate consistency. This will also enable accurately gated reconstruction of full four dimensional conduction maps and 3D conduction velocity measurements.

  8. Optical and morphological characterization by atomic force microscopy of luminescent 2-styrylpyridine derivative compounds with Poly(N-vinylcarbazole) films

    Energy Technology Data Exchange (ETDEWEB)

    Perez-Gutierrez, E., E-mail: cuper_enrique@msn.com [Centro de Quimica, Instituto de Ciencias, Universidad Autonoma de Puebla, Complejo de Ciencias, ICUAP, Edif. 103-F, 22 Sur y San Claudio, C.P. 72570 Puebla, Puebla (Mexico); Percino, M.J.; Chapela, V.M. [Centro de Quimica, Instituto de Ciencias, Universidad Autonoma de Puebla, Complejo de Ciencias, ICUAP, Edif. 103-F, 22 Sur y San Claudio, C.P. 72570 Puebla, Puebla (Mexico); Maldonado, J.L. [Centro de Investigaciones en Optica A.C. (CIO), Lomas del Bosque 115, Col. Lomas del Campestre, C.P. 37150, Leon Guanajuato (Mexico)

    2011-07-01

    The present work addresses the optical and morphological properties of organic films based on low molecular weight dyes styrylpyridine derivatives 2-styrylpyridine (A), 4-chlorophenyl-2-vinylpyridine (B) and 4-fluorophenyl-2-vinylpyridine (C), embedded in a polymeric matrix poly(N-vinylcarbazole) (PVK). The films were prepared by a spin-coating technique from solutions with dye:PVK ratios of 0.25:1, 0.5:1 and 1:1. Solvents were chloroform and toluene. The molar absorption coefficient ({epsilon}) spectra for a dye:PVK mixture in solution were a combination of the absorptions of both components separately, but for the deposited films, the shape of the spectrum showed that the poly(N-vinylcarbazole) absorption dominated. However, when the same films were dissolved again in CHCl{sub 3}, their spectra showed an absorption shape similar to that of the solution mixture before the deposition. Solution viscosity measurements were carried out with an Ubbelohde glass capillary viscometer to corroborate the results that showed a better mixture of the dye with the host in chloroform. The morphology of the prepared films was analyzed by atomic force microscopy and exhibited a solvent effect, with a pinhole-free, smooth surface when toluene was used and a wavy surface with chloroform. The ratio dye:matrix was the principal parameter for obtaining optical quality films; for 0.25:1 and 0.5:1 ratios, the films were of good quality, but for 1:1, the dye was expelled from the PVK and a crystallization was present over the surface of the films. Film thickness was also measured and films deposited from toluene solutions gave an average thickness of 54 nm while films from chloroform solutions had an average thickness greater than 160 nm that increased depending on chromophore concentration.

  9. A real-time technique for optical alignment of telescopes with segmented optics

    Science.gov (United States)

    DiVittorio, Michael; Gathright, John

    2003-02-01

    While telescopes with segmented optics (currently Keck and HET and in the future GTC, CELT, GSMT, NGST, etc) present extra challenges in terms of optical alignment, they also present the opportunity for using an alignment technique not available to telescopes with monolithic optics. We present a technique for aligning telescope secondary mirrors utilizing the segmented nature of the primary. The data required is gathered in direct image mode and can be collected from science instrument detectors (as compared to a wavefront sensor). From this data aberrations (focus and coma) are calculated from which secondary piston and tip/tilt (or decenter) corrections are determined. In addition, tip/tilt corrections for each of the primary mirror segments can also be calculated. Furthermore, other aberrations are available to determine other alignment or support issues including differentiating secondary tip/tilt from decenter, focal surface tilt, and instrument aberrations. This technique has been used nightly on the Keck I and II telescopes over the last 8 years and has made a significant improvement in image quality.

  10. Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

    Directory of Open Access Journals (Sweden)

    D. A. Healy

    2014-02-01

    Full Text Available Primary biological aerosol particles (PBAP can contribute significantly to the coarse particle burden in many environments, may thus influence climate and precipitation systems as cloud nuclei, and can spread disease to humans, animals, and plants. Measurements of PBAP in natural environments taken at high time- and size- resolution are, however, sparse and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in south western Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of the waveband integrated bioaerosol sensor (WIBS-4 with the ultraviolet aerodynamic particle sizer (UV-APS and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behaviour, with increased fluorescent bioparticle concentrations at night when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each were correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.. The WIBS FL1 channel exhibited variable multi-modal distributions turning into a broad featureless single mode after averaging and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent

  11. Exploring the limits of optical microscopy: live cell and superresolution fluorescence microscopy of HIV-1 Transfer Between T lymphocytes Across the Virological Synapse

    Science.gov (United States)

    McNerney, Gregory Paul

    Human immunodeficiency virus 1 (HIV-1) is a human retrovirus that efficiently, albeit gradually, overruns the immune system. An already infected T lymphocyte can latch onto another T lymphocyte whereby creating a virological synapse (VS); this junction drives viral assembly and transfer to the target cell in batches in an efficient, protective manor. My Ph.D. doctoral thesis focused on studying this transmission mechanism using advanced optical imaging modalities and the fully infectious fluorescent clone HIV Gag-iGFP. T lymphocytes are non-adherent cells (˜10 um thick) and the viral transmission process is fairly dynamic, hence we employed a custom spinning disk confocal microscope that revealed many interesting characteristics of this cooperative event. This methodology has low throughput as cell contact and transfer is at random. Optical tweezers was then added to the microscope to directly initiate cell contact at will. To assess when viral maturation occurs post-transfer, an optical assay based off of Forster resonance energy transfer was developed to monitor maturation. Structured illumination microscopy was further used to image the process at higher resolution and it showed that viral particles are not entering existing degradative compartments. Non-HIV-1 applications of the optical technologies are also reviewed.

  12. New metrology techniques improve the production of silicon diffractive optics

    Science.gov (United States)

    Brooks, Cynthia B.; Gully-Santiago, Michael; Grigas, Michelle; Jaffe, Daniel T.

    2014-07-01

    Silicon immersion gratings and grisms offer significant advantages in compactness and performance over frontsurface gratings and over grisms made from lower-index materials. At the same time, the high refractive index of Si (3.4) leads to very stringent constraints on the allowable groove position errors, typically rms error of errors and their origins, we could then implement process controls for each step. The plasma uniformity was improved for the silicon nitride mask etch process and the phase contribution of the plasma etch step was measured. We then used grayscale lithography, a technique in which the photoresist is deliberately underexposed, to measure large-scale nonuniformities in the UV exposure system to an accuracy of 3-5%, allowing us to make corrections to the optical alignment. Additionally, we used a new multiple-exposure technique combined with laser interferometry to measure the relationship between UV exposure dose and line edge shift. From these data we predict the contribution of the etching and photolithographic steps to phase error of the grating surface. These measurements indicate that the errors introduced during the exposure step dominate the contributions of all the other processing steps. This paper presents the techniques used to quantify individual process contributions to phase errors and steps that were taken to improve overall phase uniformity.

  13. Structural, Optical and Electrical Properties of Thin Films Using Nebulizer Spray Pyrolysis Technique

    Directory of Open Access Journals (Sweden)

    R. Mariappan

    2013-01-01

    Full Text Available thin films have been deposited on glass substrates at substrate temperature 400°C through nebulizer spray pyrolysis technique. X-ray diffraction (XRD analysis shows that the films structure is changed from hexagonal to tetragonal. The high-resolution scanning electron microscopy (HRSEM studies reveal that the substrate is well covered with a number of grains indicating compact morphology with an average grain size 50–79 nm. Energy dispersive X-ray analysis (EDAX reveals the average ratio of the atomic percentage. Optical transmittance study shows the presence of direct transition. Band gap energy decreases from 3.33 to 2.87 eV with respect to the rise of Sn content. The electrical resistivity of the thin films was found to be 106 Ω-m.

  14. Imaging of normal and pathologic joint synovium using nonlinear optical microscopy as a potential diagnostic tool

    Science.gov (United States)

    Tiwari, Nivedan; Chabra, Sanjay; Mehdi, Sheherbano; Sweet, Paula; Krasieva, Tatiana B.; Pool, Roy; Andrews, Brian; Peavy, George M.

    2010-09-01

    An estimated 1.3 million people in the United States suffer from rheumatoid arthritis (RA). RA causes profound changes in the synovial membrane of joints, and without early diagnosis and intervention, progresses to permanent alterations in joint structure and function. The purpose of this study is to determine if nonlinear optical microscopy (NLOM) can utilize the natural intrinsic fluorescence properties of tissue to generate images that would allow visualization of the structural and cellular composition of fresh, unfixed normal and pathologic synovial tissue. NLOM is performed on rabbit knee joint synovial samples using 730- and 800-nm excitation wavelengths. Less than 30 mW of excitation power delivered with a 40×, 0.8-NA water immersion objective is sufficient for the visualization of synovial structures to a maximum depth of 70 μm without tissue damage. NLOM imaging of normal and pathologic synovial tissue reveals the cellular structure, synoviocytes, adipocytes, collagen, vascular structures, and differential characteristics of inflammatory infiltrates without requiring tissue processing or staining. Further study to evaluate the ability of NLOM to assess the characteristics of pathologic synovial tissue and its potential role for the management of disease is warranted.

  15. Multifocus optical-resolution photoacoustic microscopy using stimulated Raman scattering and chromatic aberration.

    Science.gov (United States)

    Hajireza, Parsin; Forbrich, Alexander; Zemp, Roger J

    2013-08-01

    In this Letter, multifocus optical-resolution photoacoustic microscopy is demonstrated using wavelength tuning and chromatic aberration for depth scanning. Discrete focal zones at several depth locations were created by refocusing light from a polarization-maintaining single-mode fiber pumped by a nanosecond fiber laser. The fiber and laser parameters were chosen to take advantage of stimulated Raman scattering (SRS) in the fiber to create a multiwavelength output that could then be bandpass filtered. The collimator lens and objective lens are chosen to take advantage of chromatic aberration in which each generated SRS wavelength peak focuses at a slightly different depth. The maximum amplitude of photoacoustic signals is mapped to form C-scan images. Additionally, all wavelength peaks fired simultaneously offers improved depth-of-field structural imaging at the cost of slight degradation of mainlobe-to-sidelobe ratios. Wavelength-tuned depth scanning over more than 440 μm is demonstrated, significantly greater than the ~100 μm depth of field predicted from our focused Gaussian beams. The improved depth of focus could be valuable for structural imaging of microvascular morphology without the need for mechanical scanning in the depth direction.

  16. Visualization of neuritic plaques in Alzheimer’s disease by polarization-sensitive optical coherence microscopy

    Science.gov (United States)

    Baumann, Bernhard; Woehrer, Adelheid; Ricken, Gerda; Augustin, Marco; Mitter, Christian; Pircher, Michael; Kovacs, Gabor G.; Hitzenberger, Christoph K.

    2017-03-01

    One major hallmark of Alzheimer’s disease (AD) and cerebral amyloid angiopathy (CAA) is the deposition of extracellular senile plaques and vessel wall deposits composed of amyloid-beta (Aβ). In AD, degeneration of neurons is preceded by the formation of Aβ plaques, which show different morphological forms. Most of them are birefringent owing to the parallel arrangement of amyloid fibrils. Here, we present polarization sensitive optical coherence microscopy (PS-OCM) for imaging mature neuritic Aβ plaques based on their birefringent properties. Formalin-fixed, post-mortem brain samples of advanced stage AD patients were investigated. In several cortical brain regions, neuritic Aβ plaques were successfully visualized in tomographic and three-dimensional (3D) images. Cortical grey matter appeared polarization preserving, whereas neuritic plaques caused increased phase retardation. Consistent with the results from PS-OCM imaging, the 3D structure of senile Aβ plaques was computationally modelled for different illumination settings and plaque sizes. Furthermore, the birefringent properties of cortical and meningeal vessel walls in CAA were investigated in selected samples. Significantly increased birefringence was found in smaller vessels. Overall, these results provide evidence that PS-OCM is able to assess amyloidosis based on intrinsic birefringent properties.

  17. OBSERVATIONS ON DEFORMATION BEHAVIOR OF HIGH PERFORMANCE FIBERS BY POLARIZING OPTICAL MICROSCOPY

    Institute of Scientific and Technical Information of China (English)

    Chang-fa Xiao; Yu-feng Zhang

    2000-01-01

    By means of polarizing optical microscopy (POM), deformation behavior of four kinds of fibers, i.e. ultra high molecular weight polyethylene (UHMW-PE) fiber, polyvinyl alcohol (PVA) fiber, polyethylene terephthalate (PET) fiber,and wholly aromatic (p-hydroxybenzoic acid/2-hydroxy-6-naphthoic acid) copolyester [P(HBA/HNA)]/PET (ACPET blend) fiber, in axial compression, axial impacting, and bending was observed. In compression, kink bands formed at an angle of 55~60° to the fiber axis in 10-times-drawn UHMW-PE fiber, 75~80° in 40-times-drawn sample, 80° in PVA fiber, and 90°in the ACPET blend fiber. In impacting and bending, band angles of UHMW-PE, PVA and PET fibers are nearly the same as those formed in compression, indicating that slip systems do not change. For any of the four kinds of fiber, band spacing exhibits great differences in compression, in impacting, and in bending, which may be attributed to the differences in the degrees of strain or stress concentration.

  18. Visualization of neuritic plaques in Alzheimer’s disease by polarization-sensitive optical coherence microscopy

    Science.gov (United States)

    Baumann, Bernhard; Woehrer, Adelheid; Ricken, Gerda; Augustin, Marco; Mitter, Christian; Pircher, Michael; Kovacs, Gabor G.; Hitzenberger, Christoph K.

    2017-01-01

    One major hallmark of Alzheimer’s disease (AD) and cerebral amyloid angiopathy (CAA) is the deposition of extracellular senile plaques and vessel wall deposits composed of amyloid-beta (Aβ). In AD, degeneration of neurons is preceded by the formation of Aβ plaques, which show different morphological forms. Most of them are birefringent owing to the parallel arrangement of amyloid fibrils. Here, we present polarization sensitive optical coherence microscopy (PS-OCM) for imaging mature neuritic Aβ plaques based on their birefringent properties. Formalin-fixed, post-mortem brain samples of advanced stage AD patients were investigated. In several cortical brain regions, neuritic Aβ plaques were successfully visualized in tomographic and three-dimensional (3D) images. Cortical grey matter appeared polarization preserving, whereas neuritic plaques caused increased phase retardation. Consistent with the results from PS-OCM imaging, the 3D structure of senile Aβ plaques was computationally modelled for different illumination settings and plaque sizes. Furthermore, the birefringent properties of cortical and meningeal vessel walls in CAA were investigated in selected samples. Significantly increased birefringence was found in smaller vessels. Overall, these results provide evidence that PS-OCM is able to assess amyloidosis based on intrinsic birefringent properties. PMID:28262719

  19. Highly efficient plasmonic tip design for plasmon nanofocusing in near-field optical microscopy

    Science.gov (United States)

    Umakoshi, Takayuki; Saito, Yuika; Verma, Prabhat

    2016-03-01

    Near-field scanning optical microscopy (NSOM) combined with plasmon nanofocusing is a powerful nano-analytical tool due to its attractive feature of efficient background suppression as well as light energy compression to the nanoscale. In plasmon nanofocusing-based NSOM, the metallic tip plays an important role in inducing plasmon nanofocusing. It is, however, very challenging to control plasmonic properties of tips for plasmon nanofocusing with existing tip fabrication methods, even though the plasmonic properties need to be adjusted to experimental environments such as the sample or excitation wavelength. In this study, we propose an efficient tip design and fabrication which enable one to actively control plasmonic properties for efficient plasmon nanofocusing. Because our method offers flexibility in the material and structure of tips, one can easily modify the plasmonic properties depending on the requirements. Importantly, through optimization of the plasmonic properties, we achieve almost 100% reproducibility in plasmon nanofocusing in our experiments. This new approach of tip fabrication makes plasmon nanofocusing-based NSOM practical and reliable, and opens doors for many scientists working in related fields.

  20. Magnetic stage with environmental control for optical microscopy and high-speed nano- and microrheology

    Science.gov (United States)

    Aprelev, Pavel; McKinney, Bonni; Walls, Chadwick; Kornev, Konstanin G.

    2017-07-01

    A novel design of a low-field magnetic stage for optical microscopy of droplets and films within a controlled environment is described. The stage consists of five magnetic coils with a 3D magnetic sensor in a feedback control loop, which allows one to manipulate magnetic nano- and microprobes with microtesla fields. A locally uniform time-dependent field within the focal plane of the microscope objective enables one to rotate the probes in a precisely set manner and observe their motion. The probe tracking protocol was developed to follow the probe rotation in real time and relate it with the viscosity of the host liquid. Using this magnetic stage, a method for measuring mPa s-level viscosity of nanoliter droplets and micron thick films in a 10-20 s timeframe is presented and validated. The viscosity of a rapidly changing liquid can be tracked by using only a few visible probes rotating simultaneously. Vapor pressure and temperature around the sample can be controlled to directly measure viscosity as a function of equilibrium vapor pressure; this addresses a significant challenge in characterization of volatile nanodroplets and thin films. Thin films of surfactant solutions undergoing phase transitions upon solvent evaporation were studied and their rheological properties were related to morphological changes in the material.