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Sample records for two-photon scanning microscopy

  1. Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2

    International Nuclear Information System (INIS)

    McConnell, Gail; Riis, Erling

    2004-01-01

    We report on a novel and compact reliable laser source capable of short-wavelength two-photon laser scanning fluorescence microscopy based on soliton self-frequency shift effects in photonic crystal fibre. We demonstrate the function of the system by performing two-photon microscopy of smooth muscle cells and cardiac myocytes from the rat pulmonary vein and Chinese hamster ovary cells loaded with the fluorescent calcium indicator fura-2/AM

  2. Optimizing pulse compressibility in completely all-fibered Ytterbium chirped pulse amplifiers for in vivo two photon laser scanning microscopy.

    Science.gov (United States)

    Fernández, A; Grüner-Nielsen, L; Andreana, M; Stadler, M; Kirchberger, S; Sturtzel, C; Distel, M; Zhu, L; Kautek, W; Leitgeb, R; Baltuska, A; Jespersen, K; Verhoef, A

    2017-08-01

    A simple and completely all-fiber Yb chirped pulse amplifier that uses a dispersion matched fiber stretcher and a spliced-on hollow core photonic bandgap fiber compressor is applied in nonlinear optical microscopy. This stretching-compression approach improves compressibility and helps to maximize the fluorescence signal in two-photon laser scanning microscopy as compared with approaches that use standard single mode fibers as stretcher. We also show that in femtosecond all-fiber systems, compensation of higher order dispersion terms is relevant even for pulses with relatively narrow bandwidths for applications relying on nonlinear optical effects. The completely all-fiber system was applied to image green fluorescent beads, a stained lily-of-the-valley root and rat-tail tendon. We also demonstrated in vivo imaging in zebrafish larvae, where we simultaneously measure second harmonic and fluorescence from two-photon excited red-fluorescent protein. Since the pulses are compressed in a fiber, this source is especially suited for upgrading existing laser scanning (confocal) microscopes with multiphoton imaging capabilities in space restricted settings or for incorporation in endoscope-based microscopy.

  3. Extended two-photon microscopy in live samples with Bessel beams: steadier focus, faster volume scans, and simpler stereoscopic imaging.

    Science.gov (United States)

    Thériault, Gabrielle; Cottet, Martin; Castonguay, Annie; McCarthy, Nathalie; De Koninck, Yves

    2014-01-01

    Two-photon microscopy has revolutionized functional cellular imaging in tissue, but although the highly confined depth of field (DOF) of standard set-ups yields great optical sectioning, it also limits imaging speed in volume samples and ease of use. For this reason, we recently presented a simple and retrofittable modification to the two-photon laser-scanning microscope which extends the DOF through the use of an axicon (conical lens). Here we demonstrate three significant benefits of this technique using biological samples commonly employed in the field of neuroscience. First, we use a sample of neurons grown in culture and move it along the z-axis, showing that a more stable focus is achieved without compromise on transverse resolution. Second, we monitor 3D population dynamics in an acute slice of live mouse cortex, demonstrating that faster volumetric scans can be conducted. Third, we acquire a stereoscopic image of neurons and their dendrites in a fixed sample of mouse cortex, using only two scans instead of the complete stack and calculations required by standard systems. Taken together, these advantages, combined with the ease of integration into pre-existing systems, make the extended depth-of-field imaging based on Bessel beams a strong asset for the field of microscopy and life sciences in general.

  4. Microsphere imaging with confocal microscopy and two photon microscopy

    International Nuclear Information System (INIS)

    Chun, Hyung Su; An, Kyung Won; Lee, Jai Hyung

    2002-01-01

    We have acquired images of polystyrene and fused-silica microsphere by using conventional optical microscopy, confocal microscopy and two-photon microscopy, and performed comparative analysis of these images. Different from conventional optical microscopy, confocal and two-photon microscopy had good optical sectioning capability. In addition, confocal microscopy and two-photon microscopy had better lateral resolution than conventional optical microscopy. These results are attributed to confocality and nonlinearity of confocal microscopy and two photon microscopy, respectively.

  5. A high performance, cost-effective, open-source microscope for scanning two-photon microscopy that is modular and readily adaptable.

    Directory of Open Access Journals (Sweden)

    David G Rosenegger

    Full Text Available Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems.

  6. A High Performance, Cost-Effective, Open-Source Microscope for Scanning Two-Photon Microscopy that Is Modular and Readily Adaptable

    Science.gov (United States)

    Rosenegger, David G.; Tran, Cam Ha T.; LeDue, Jeffery; Zhou, Ning; Gordon, Grant R.

    2014-01-01

    Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems. PMID:25333934

  7. Evaluation of transdermal delivery of nanoemulsions in ex vivo porcine skin using two-photon microscopy and confocal laser-scanning microscopy

    Science.gov (United States)

    Choi, Sanghoon; Kim, Jin Woong; Lee, Yong Joong; Delmas, Thomas; Kim, Changhwan; Park, Soyeun; Lee, Ho

    2014-10-01

    This study experimentally evaluates the self-targeting ability of asiaticoside-loaded nanoemulsions compared with nontargeted nanoemulsions in ex vivo experiments with porcine skin samples. Homebuilt two-photon and confocal laser-scanning microscopes were employed to noninvasively examine the transdermal delivery of two distinct nanoemulsions. Prior to the application of nanoemulsions, we noninvasively observed the morphology of porcine skin using two-photon microscopy. We have successfully visualized the distributions of the targeted and nontargeted nanoemulsions absorbed into the porcine skin samples. Asiaticoside-loaded nanoemulsions showed an improved ex vivo transdermal delivery through the stratum corneum compared with nonloaded nanoemulsions. As a secondary measure, nanoemulsions-applied samples were sliced in the depth direction with a surgical knife in order to obtain the complete depth-direction distribution profile of Nile red fluorescence. XZ images demonstrated that asiaticoside-loaded nanoemulsion penetrated deeper into the skin compared with nontargeted nanoemulsions. The basal layer boundary is clearly visible in the case of the asiaticoside-loaded skin sample. These results reaffirm the feasibility of using self-targeting ligands to improve permeation through the skin barrier for cosmetics and topical drug applications.

  8. Two-photon excitation laser scanning microscopy of rabbit nasal septal cartilage following Nd:YAG-laser-mediated stress relaxation

    Science.gov (United States)

    Kim, Charlton C.; Wallace, Vincent P.; Coleno, Mariah L.; Dao, Xavier; Tromberg, Bruce J.; Wong, Brian J.

    2000-04-01

    Laser irradiation of hyaline cartilage result in stable shape changes due to temperature dependent stress relaxation. In this study, we determined the structural changes in chondrocytes within rabbit nasal septal cartilage tissue over a 12-day period using a two-photon laser scanning microscope (TPM) following Nd:YAG laser irradiation. During laser irradiation surface temperature, stress relaxation, and diffuse reflectance, were measured dynamically. Each specimen received one or two sequential laser exposures. The cartilage reached a peak surface temperature of about 61 degrees C during irradiation. Cartilage denatured in 50 percent EtOH was used as a positive control. TPM was performed to detect the fluorescence emission from the chondrocytes. Images of chondrocytes were obtained at depths up to 150 microns, immediately following laser exposure, and also following 12 days in culture. Few differences in the pattern or intensity of fluorescence was observed between controls and irradiated specimens imaged immediately following exposure, regardless of the number of laser pulses. However, following twelve days in tissue culture, the irradiated specimens increase, whereas the native tissue diminishes, in intensity and distribution of fluorescence in the cytoplasm. In contrast, the positive control shows only extracellular matrices and empty lacuna, feature consistent with cell membrane lysis.

  9. Correlative two-photon and serial block face scanning electron microscopy in neuronal tissue using 3D near-infrared branding maps.

    Science.gov (United States)

    Lees, Robert M; Peddie, Christopher J; Collinson, Lucy M; Ashby, Michael C; Verkade, Paul

    2017-01-01

    Linking cellular structure and function has always been a key goal of microscopy, but obtaining high resolution spatial and temporal information from the same specimen is a fundamental challenge. Two-photon (2P) microscopy allows imaging deep inside intact tissue, bringing great insight into the structural and functional dynamics of cells in their physiological environment. At the nanoscale, the complex ultrastructure of a cell's environment in tissue can be reconstructed in three dimensions (3D) using serial block face scanning electron microscopy (SBF-SEM). This provides a snapshot of high resolution structural information pertaining to the shape, organization, and localization of multiple subcellular structures at the same time. The pairing of these two imaging modalities in the same specimen provides key information to relate cellular dynamics to the ultrastructural environment. Until recently, approaches to relocate a region of interest (ROI) in tissue from 2P microscopy for SBF-SEM have been inefficient or unreliable. However, near-infrared branding (NIRB) overcomes this by using the laser from a multiphoton microscope to create fiducial markers for accurate correlation of 2P and electron microscopy (EM) imaging volumes. The process is quick and can be user defined for each sample. Here, to increase the efficiency of ROI relocation, multiple NIRB marks are used in 3D to target ultramicrotomy. A workflow is described and discussed to obtain a data set for 3D correlated light and electron microscopy, using three different preparations of brain tissue as examples. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Enhanced Emission from Single Isolated Gold Quantum Dots Investigated Using Two-Photon-Excited Fluorescence Near-Field Scanning Optical Microscopy.

    Science.gov (United States)

    Abeyasinghe, Neranga; Kumar, Santosh; Sun, Kai; Mansfield, John F; Jin, Rongchao; Goodson, Theodore

    2016-12-21

    New approaches in molecular nanoscopy are greatly desired for interrogation of biological, organic, and inorganic objects with sizes below the diffraction limit. Our current work investigates emergent monolayer-protected gold quantum dots (nanoclusters, NCs) composed of 25 Au atoms by utilizing two-photon-excited fluorescence (TPEF) near-field scanning optical microscopy (NSOM) at single NC concentrations. Here, we demonstrate an approach to synthesize and isolate single NCs on solid glass substrates. Subsequent investigation of the NCs using TPEF NSOM reveals that, even when they are separated by distances of several tens of nanometers, we can excite and interrogate single NCs individually. Interestingly, we observe an enhanced two-photon absorption (TPA) cross section for single Au 25 NCs that can be attributed to few-atom local field effects and to local field-induced microscopic cascading, indicating their potential for use in ultrasensitive sensing, disease diagnostics, cancer cell therapy, and molecular computers. Finally, we report room-temperature aperture-based TPEF NSOM imaging of these NCs for the first time at 30 nm point resolution, which is a ∼5-fold improvement compared to the previous best result for the same technique. This report unveils the unique combination of an unusually large TPA cross section and the high photostability of Au NCs to (non-destructively) investigate stable isolated single NCs using TPEF NSOM. This is the first reported optical study of monolayer-protected single quantum clusters, opening some very promising opportunities in spectroscopy of nanosized objects, bioimaging, ultrasensitive sensing, molecular computers, and high-density data storage.

  11. Two-Photon Fluorescence Microscopy Developed for Microgravity Fluid Physics

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2004-01-01

    Recent research efforts within the Microgravity Fluid Physics Branch of the NASA Glenn Research Center have necessitated the development of a microscope capable of high-resolution, three-dimensional imaging of intracellular structure and tissue morphology. Standard optical microscopy works well for thin samples, but it does not allow the imaging of thick samples because of severe degradation caused by out-of-focus object structure. Confocal microscopy, which is a laser-based scanning microscopy, provides improved three-dimensional imaging and true optical sectioning by excluding the out-of-focus light. However, in confocal microscopy, out-of-focus object structure is still illuminated by the incoming beam, which can lead to substantial photo-bleaching. In addition, confocal microscopy is plagued by limited penetration depth, signal loss due to the presence of a confocal pinhole, and the possibility of live-cell damage. Two-photon microscopy is a novel form of laser-based scanning microscopy that allows three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon microscopy, it utilizes the nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption because of the nonlinear (i.e., quadratic) electric field dependence, so an ultrafast pulsed laser source must typically be employed. On the other hand, this stringent energy density requirement effectively localizes fluorophore excitation to the focal volume. Consequently, two-photon microscopy provides optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction in photo-damage because of the longer excitation wavelength, a reduction in background fluorescence, and a 4 increase in penetration depth over confocal methods because of the reduction in Rayleigh scattering.

  12. Two-Photon Excitation Microscopy for the Study of Living Cells and Tissues

    Science.gov (United States)

    Benninger, Richard K.P.; Piston, David W.

    2013-01-01

    Two-photon excitation microscopy is an alternative to confocal microscopy that provides advantages for three-dimensional and deep tissue imaging. This unit will describe the basic physical principles behind two-photon excitation and discuss the advantages and limitations of its use in laser-scanning microscopy. The principal advantages of two-photon microscopy are reduced phototoxicity, increased imaging depth, and the ability to initiate highly localized photochemistry in thick samples. Practical considerations for the application of two-photon microscopy will then be discussed, including recent technological advances. This unit will conclude with some recent applications of two-photon microscopy that highlight the key advantages over confocal microscopy and the types of experiments which would benefit most from its application. PMID:23728746

  13. Wide-field two-photon microscopy with temporal focusing and HiLo background rejection

    Science.gov (United States)

    Yew, Elijah Y. S.; Choi, Heejin; Kim, Daekeun; So, Peter T. C.

    2011-03-01

    Scanningless depth-resolved microscopy is achieved through spatial-temporal focusing and has been demonstrated previously. The advantage of this method is that a large area may be imaged without scanning resulting in higher throughput of the imaging system. Because it is a widefield technique, the optical sectioning effect is considerably poorer than with conventional spatial focusing two-photon microscopy. Here we propose wide-field two-photon microscopy based on spatio-temporal focusing and employing background rejection based on the HiLo microscope principle. We demonstrate the effects of applying HiLo microscopy to widefield temporally focused two-photon microscopy.

  14. Fluorescence excitation analysis by two-photon confocal laser scanning microscopy: a new method to identify fluorescent nanoparticles on histological tissue sections

    Directory of Open Access Journals (Sweden)

    Kahn E

    2012-10-01

    Full Text Available Edmond Kahn,1 Nicolas Tissot,3 Perrine Frere,3 Aurélien Dauphin,3 Mohamed Boumhras,2,4 Claude-Marie Bachelet,3 Frédérique Frouin,1 Gérard Lizard21Institut National de la Santé et de la Recherche Médicale (INSERM U678/UMR-S UPMC, CHU Pitié-Salpêtrière, Paris, France; 2Equipe Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique EA7270, Faculté des Sciences Gabriel, Université de Bourgogne-INSERM Dijon, France; 3Plateforme d'Imagerie cellulaire, UPMC, Paris, France; 4Laboratory of Biochemistry and Neuroscience, Applied Toxicology Group, Faculty of Science and Technology, Settat, MoroccoAbstract: In the present study, we make use of the ability of two-photon confocal laser scanning microscopes (CLSMs equipped with tunable lasers to produce spectral excitation image sequences. Furthermore, unmixing, which is usually performed on emission image sequences, is performed on these excitation image sequences. We use factor analysis of medical image sequences (FAMIS, which produces factor images, to unmix spectral image sequences of stained structures in tissue sections to provide images of characterized stained cellular structures. This new approach is applied to histological tissue sections of mouse aorta containing labeled iron nanoparticles stained with Texas Red and counterstained with SYTO13, to obtain visual information about the accumulation of these nanoparticles in the arterial wall. The possible presence of Texas Red is determined using a two-photon CLSM associated with FAMIS via the excitation spectra. Texas Red and SYTO13 are thus differentiated, and corresponding factor images specify their possible presence and cellular localization. In conclusion, the designed protocol shows that sequences of images obtained by excitation in a two-photon CLSM enables characterization of Texas Red-stained nanoparticles and other markers. This methodology offers an alternative and complementary solution to the conventional use of emission

  15. Two-photon excitation laser scanning microscopy of porcine nasal septal cartilage following Nd:YAG laser-mediated stress relaxation

    Science.gov (United States)

    Kim, Charlton C.; Wallace, Vincent P.; Rasouli, Alexandre; Coleno, Mariah L.; Dao, Xavier; Tromberg, Bruce J.; Wong, Brian J.

    2000-05-01

    Laser irradiation of hyaline cartilage result in stable shape changes due to temperature dependent stress relaxation. In this study, we determined the structural changes in chondrocytes within porcine nasal septal cartilage tissue over a 4-day period using a two-photon laser scanning microscope (TPM) following Nd:YAG laser irradiation (lambda equals 1.32 micrometer) using parameters that result in mechanical stress relaxation (6.0 W, 5.4 mm spot diameter). TPM excitation (780 nm) result in induction of fluorescence from endogenous agents such as NADH, NADPH, and flavoproteins in the 400 - 500 nm spectral region. During laser irradiation diffuse reflectance (from a probe HeNe laser, (lambda) equals 632.8 nm), surface temperature, and stress relaxation were measured dynamically. Each specimen received one, two, or three sequential laser exposures (average irradiation times of 5, 6, and 8 seconds). The cartilage reached a peak surface temperature of about 70 degrees Celsius during irradiation. Cartilage denatured in 50% EtOH (20 minutes) was used as a positive control. TPM was performed using a mode-locked 780 nm Titanium:Sapphire (Ti:Al203) beam with a, 63X, 1.2 N.A. water immersion objective (working distance of 200 mm) to detect the fluorescence emission from the chondrocytes. Images of chondrocytes were obtained at depths up to 150 microns (lateral resolution equals 35 micrometer X 35 micrometer). Images were obtained immediately following laser exposure, and also after 4 days in culture. In both cases, the irradiated and non-irradiated specimens do not show any discernible difference in general shape or auto fluorescence. In contrast, positive controls (immersed in 50% ethanol), show markedly increased fluorescence relative to both the native and irradiated specimens, in the cytoplasmic region.

  16. Circular dichroism probed by two-photon fluorescence microscopy in enantiopure chiral polyfluorene thin films

    NARCIS (Netherlands)

    Savoini, M.; Wu, Xiaofei; Celebrano, M.; Ziegler, J.; Biagioni, P.; Meskers, S.C.J.; Duo, L.; Hecht, B.; Finazzi, M.

    2012-01-01

    Two-photon fluorescence scanning confocal microscopy sensitive to circular dichroism with a diffraction-limited resolution well below 500 nm is demonstrated. With this method, the spatial variation of the circular dichroism of thermally annealed chiral polyfluorene thin films has been imaged. We

  17. Integrated single- and two-photon light sheet microscopy using accelerating beams

    DEFF Research Database (Denmark)

    Piksarv, Peeter; Marti, Dominik; Le, Tuan

    2017-01-01

    We demonstrate the first light sheet microscope using propagation invariant, accelerating Airy beams that operates both in single- and two-photon modes. The use of the Airy beam permits us to develop an ultra compact, high resolution light sheet system without beam scanning. In two-photon mode......, an increase in the field of view over the use of a standard Gaussian beam by a factor of six is demonstrated. This implementation for light sheet microscopy opens up new possibilities across a wide range of biomedical applications, especially for the study of neuronal processes....

  18. Insights into esophagus tissue architecture using two-photon confocal microscopy

    Science.gov (United States)

    Liu, Nenrong; Wang, Yue; Feng, Shangyuan; Chen, Rong

    2013-08-01

    In this paper, microstructures of human esophageal mucosa were evaluated using the two-photon laser scanning confocal microscopy (TPLSCM), based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). The distribution of epithelial cells, muscle fibers of muscularis mucosae has been distinctly obtained. Furthermore, esophageal submucosa characteristics with cancer cells invading into were detected. The variation of collagen, elastin and cancer cells is very relevant to the pathology in esophagus, especially early esophageal cancer. Our experimental results indicate that the MPM technique has the much more advantages for label-free imaging, and has the potential application in vivo in the clinical diagnosis and monitoring of early esophageal cancer.

  19. Imaging Live Drosophila Brain with Two-Photon Fluorescence Microscopy

    Science.gov (United States)

    Ahmed, Syeed Ehsan

    Two-photon fluorescence microscopy is an imaging technique which delivers distinct benefits for in vivo cellular and molecular imaging. Cyclic adenosine monophosphate (cAMP), a second messenger molecule, is responsible for triggering many physiological changes in neural system. However, the mechanism by which this molecule regulates responses in neuron cells is not yet clearly understood. When cAMP binds to a target protein, it changes the structure of that protein. Therefore, studying this molecular structure change with fluorescence resonance energy transfer (FRET) imaging can shed light on the cAMP functioning mechanism. FRET is a non-radiative dipole-dipole coupling which is sensitive to small distance change in nanometer scale. In this study we have investigated the effect of dopamine in cAMP dynamics in vivo. In our study two-photon fluorescence microscope was used for imaging mushroom bodies inside live Drosophila melanogaster brain and we developed a method for studying the change in cyclic AMP level.

  20. In vivo reactive neural plasticity investigation by means of correlative two photon: electron microscopy

    Science.gov (United States)

    Allegra Mascaro, A. L.; Cesare, P.; Sacconi, L.; Grasselli, G.; Mandolesi, G.; Maco, B.; Knott, G.; Huang, L.; De Paola, V.; Strata, P.; Pavone, F. S.

    2013-02-01

    In the adult nervous system, different populations of neurons correspond to different regenerative behavior. Although previous works showed that olivocerebellar fibers are capable of axonal regeneration in a suitable environment as a response to injury1, we have hitherto no details about the real dynamics of fiber regeneration. We set up a model of singularly axotomized climbing fibers (CF) to investigate their reparative properties in the adult central nervous system (CNS) in vivo. Time lapse two-photon imaging has been combined to laser nanosurgery2, 3 to define a temporal pattern of the degenerative event and to follow the structural rearrangement after injury. To characterize the damage and to elucidate the possible formation of new synaptic contacts on the sprouted branches of the lesioned CF, we combined two-photon in vivo imaging with block face scanning electron microscopy (FIB-SEM). Here we describe the approach followed to characterize the reactive plasticity after injury.

  1. Two-photon Microscopy and Polarimetry for Assessment of Myocardial Tissue Organization

    Science.gov (United States)

    Archambault-Wallenburg, Marika

    Optical methods can provide useful tissue characterization tools. For this project, two-photon microscopy and polarized light examinations (polarimetry) were used to assess the organizational state of myocardium in healthy, infarcted, and stem-cell regenerated states. Two-photon microscopy visualizes collagen through second-harmonic generation and myocytes through two-photon excitation autofluorescence, providing information on the composition and structure/organization of the tissue. Polarimetry measurements yield a value of linear retardance that can serve as an indicator of tissue anisotropy, and with a dual-projection method, information about the anisotropy axis orientation can also be extracted. Two-photon microscopy results reveal that stem-cell treated tissue retains more myocytes and structure than infarcted myocardium, while polarimetry findings suggest that the injury caused by temporary ligation of a coronary artery is less severe and more diffuse that than caused by a permanent ligation. Both these methods show potential for tissue characterization.

  2. A new approach to dual-color two-photon microscopy with fluorescent proteins

    Directory of Open Access Journals (Sweden)

    Rebane Aleks

    2010-02-01

    Full Text Available Abstract Background Two-photon dual-color imaging of tissues and cells labeled with fluorescent proteins (FPs is challenging because most two-photon microscopes only provide one laser excitation wavelength at a time. At present, methods for two-photon dual-color imaging are limited due to the requirement of large differences in Stokes shifts between the FPs used and their low two-photon absorption (2PA efficiency. Results Here we present a new method of dual-color two-photon microscopy that uses the simultaneous excitation of the lowest-energy electronic transition of a blue fluorescent protein and a higher-energy electronic transition of a red fluorescent protein. Conclusion Our method does not require large differences in Stokes shifts and can be extended to a variety of FP pairs with larger 2PA efficiency and more optimal imaging properties.

  3. Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

    Science.gov (United States)

    Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

    2011-07-01

    Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

  4. Femtosecond laser subsurface scleral treatment in cadaver human sclera and evaluation using two-photon and confocal microscopy

    Science.gov (United States)

    Sun, Hui; Fan, Zhongwei; Yan, Ying; Lian, Fuqiang; Kurtz, Ron; Juhasz, Tibor

    2016-03-01

    Glaucoma is the second-leading cause of blindness worldwide and is often associated with elevated intraocular pressure (IOP). Partial-thickness drainage channels can be created with femtosecond laser in the translucent sclera for the potential treatment of glaucoma. We demonstrate the creation of partial-thickness subsurface drainage channels with the femtosecond laser in the cadaver human eyeballs and describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. A femtosecond laser operating at a wavelength of 1700 nm was scanned along a rectangular raster pattern to create the partial thickness subsurface drainage channels in the sclera of cadaver human eyes. Analysis of the dimensions and location of these channels is important in understanding their effects. We describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. High-resolution images, hundreds of microns deep in the sclera, were obtained to allow determination of the shape and dimension of such partial thickness subsurface scleral channels. Our studies suggest that the confocal and two-photon microscopy can be used to investigate femtosecond-laser created partial-thickness drainage channels in the sclera of cadaver human eyes.

  5. QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY

    Directory of Open Access Journals (Sweden)

    Merete Krog Raarup

    2011-05-01

    Full Text Available This paper discusses recent advances in confocal laser scanning microscopy (CLSM for imaging of 3D structure as well as quantitative characterization of biomolecular interactions and diffusion behaviour by means of one- and two-photon excitation. The use of CLSM for improved stereological length estimation in thick (up to 0.5 mm tissue is proposed. The techniques of FRET (Fluorescence Resonance Energy Transfer, FLIM (Fluorescence Lifetime Imaging Microscopy, FCS (Fluorescence Correlation Spectroscopy and FRAP (Fluorescence Recovery After Photobleaching are introduced and their applicability for quantitative imaging of biomolecular (co-localization and trafficking in live cells described. The advantage of two-photon versus one-photon excitation in relation to these techniques is discussed.

  6. Visualization of laser tattoo removal treatment effects in a mouse model by two-photon microscopy.

    Science.gov (United States)

    Jang, Won Hyuk; Yoon, Yeoreum; Kim, Wonjoong; Kwon, Soonjae; Lee, Seunghun; Song, Duke; Choi, Jong Woon; Kim, Ki Hean

    2017-08-01

    Laser tattoo removal is an effective method of eliminating tattoo particles in the skin. However, laser treatment cannot always remove the unwanted tattoo completely, and there are risks of either temporary or permanent side effects. Studies using preclinical animal models could provide detailed information on the effects of laser treatment in the skin, and might help to minimize side effects in clinical practices. In this study, two-photon microscopy (TPM) was used to visualize the laser treatment effects on tattoo particles in both phantom specimens and in vivo mouse models. Fluorescent tattoo ink was used for particle visualization by TPM, and nanosecond (ns) and picosecond (ps) lasers at 532 nm were used for treatment. In phantom specimens, TPM characterized the fragmentation of individual tattoo particles by tracking them before and after the laser treatment. These changes were confirmed by field emission scanning electron microscopy (FE-SEM). TPM was used to measure the treatment efficiency of the two lasers at different laser fluences. In the mouse model, TPM visualized clusters of tattoo particles in the skin and detected their fragmentation after the laser treatment. Longitudinal TPM imaging observed the migration of cells containing tattoo particles after the laser treatment. These results show that TPM may be useful for the assessment of laser tattoo removal treatment in preclinical studies.

  7. Comparison of confocal microscopy and two-photon microscopy in mouse cornea in vivo.

    Science.gov (United States)

    Lee, Jun Ho; Lee, Seunghun; Gho, Yong Song; Song, In Seok; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

    2015-03-01

    High-resolution imaging of the cornea is important for studying corneal diseases at cellular levels. Confocal microscopy (CM) has been widely used in the clinic, and two-photon microscopy (TPM) has recently been introduced in various pre-clinical studies. We compared the performance of CM and TPM in normal mouse corneas and neovascularized mouse corneas induced by suturing. Balb/C mice and C57BL/6 mice expressing green fluorescent protein (GFP) were used to compare modalities based on intrinsic contrast and extrinsic fluorescence contrast. CM based on reflection (CMR), CM based on fluorescence (CMF), and TPM based on intrinsic/extrinsic fluorescence and second harmonic generation (SHG) were compared by imaging the same sections of mouse corneas sequentially in vivo. In normal mouse corneas, CMR visualized corneal cell morphologies with some background noise, and CMF visualized GFP expressing corneal cells clearly. TPM visualized corneal cells and collagen in the stroma based on fluorescence and SHG, respectively. However, in neovascularized mouse corneas, CMR could not resolve cells deep inside the cornea due to high background noise from the effects of increased structural irregularity induced by suturing. CMF and TPM visualized cells and induced vasculature better than CMR because both collect signals from fluorescent cells only. Both CMF and TPM had signal decays with depth due to the structural irregularity, with CMF having faster signal decay than TPM. CMR, CMF, and TPM showed different degrees of image degradation in neovascularized mouse corneas. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Two-photon luminescence microscopy of field enhancement at gold nanoparticles

    DEFF Research Database (Denmark)

    Beermann, Jonas; Bozhevolnyi, Sergey I.

    2005-01-01

    Using a reflection scanning optical microscope detecting two-photon luminescence (TPL) we have imaged square gold bumps positioned in a periodic array either on a smooth gold film or directly on a glass substrate. The second-harmonic (SH) and TPL response from these structures show both...

  9. Applications of two-photon fluorescence microscopy in deep-tissue imaging

    Science.gov (United States)

    Dong, Chen-Yuan; Yu, Betty; Hsu, Lily L.; Kaplan, Peter D.; Blankschstein, D.; Langer, Robert; So, Peter T. C.

    2000-07-01

    Based on the non-linear excitation of fluorescence molecules, two-photon fluorescence microscopy has become a significant new tool for biological imaging. The point-like excitation characteristic of this technique enhances image quality by the virtual elimination of off-focal fluorescence. Furthermore, sample photodamage is greatly reduced because fluorescence excitation is limited to the focal region. For deep tissue imaging, two-photon microscopy has the additional benefit in the greatly improved imaging depth penetration. Since the near- infrared laser sources used in two-photon microscopy scatter less than their UV/glue-green counterparts, in-depth imaging of highly scattering specimen can be greatly improved. In this work, we will present data characterizing both the imaging characteristics (point-spread-functions) and tissue samples (skin) images using this novel technology. In particular, we will demonstrate how blind deconvolution can be used further improve two-photon image quality and how this technique can be used to study mechanisms of chemically-enhanced, transdermal drug delivery.

  10. Two-Photon Probes for Lysosomes and Mitochondria: Simultaneous Detection of Lysosomes and Mitochondria in Live Tissues by Dual-Color Two-Photon Microscopy Imaging.

    Science.gov (United States)

    Lim, Chang Su; Hong, Seung Taek; Ryu, Seong Shick; Kang, Dong Eun; Cho, Bong Rae

    2015-10-01

    Novel two-photon (TP) probes were developed for lysosomes (PLT-yellow) and mitochondria (BMT-blue and PMT-yellow). These probes emitted strong TP-excited fluorescence in cells at widely separated wavelength regions and displayed high organelle selectivity, good cell permeability, low cytotoxicity, and pH insensitivity. The BMT-blue and PLT-yellow probes could be utilized to detect lysosomes and mitochondria simultaneously in live tissues by using dual-color two-photon microscopy, with minimum interference from each other. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Phosphorescent probes for two-photon microscopy of oxygen (Conference Presentation)

    Science.gov (United States)

    Vinogradov, Sergei A.; Esipova, Tatiana V.

    2016-03-01

    The ability to quantify oxygen in vivo in 3D with high spatial and temporal resolution is much needed in many areas of biological research. Our laboratory has been developing the phosphorescence quenching technique for biological oximetry - an optical method that possesses intrinsic microscopic capability. In the past we have developed dendritically protected oxygen probes for quantitative imaging of oxygen in tissue. More recently we expanded our design on special two-photon enhanced phosphorescent probes. These molecules brought about first demonstrations of the two-photon phosphorescence lifetime microscopy (2PLM) of oxygen in vivo, providing new information for neouroscience and stem cell biology. However, current two-photon oxygen probes suffer from a number of limitations, such as sub-optimal brightness and high cost of synthesis, which dramatically reduce imaging performance and limit usability of the method. In this paper we discuss principles of 2PLM and address the interplay between the probe chemistry, photophysics and spatial and temporal imaging resolution. We then present a new approach to brightly phosphorescent chromophores with internally enhanced two-photon absorption cross-sections, which pave a way to a new generation of 2PLM probes.

  12. Selective two-photon collagen crosslinking in situ measured by Brillouin microscopy (Conference Presentation)

    Science.gov (United States)

    Kwok, Sheldon J. J.; Kuznetsov, Ivan A.; Kim, Moonseok; Choi, Myunghwan; Scarcelli, Giuliano; Yun, Seok-Hyun

    2017-02-01

    Two-photon polymerization and crosslinking are commonly used methods for microfabrication of three-dimensional structures with applications spanning from photonic microdevices, drug delivery systems, to cellular scaffolds. However, the use of two-photon processes for precise, internal modification of biological tissues has not yet been reported. One of the major challenges has been a lack of appropriate tools to monitor and characterize crosslinked regions nondestructively. Here, we demonstrate spatially selective two-photon collagen crosslinking (2P-CXL) in intact tissue for the first time. Using riboflavin photosensitizer and femtosecond laser irradiation, we crosslinked a small volume of tissue within animal corneas. Collagen fiber orientations and photobleaching were characterized by second harmonic generation and two-photon fluorescence imaging, respectively. Using confocal Brillouin microscopy, we measured local changes in longitudinal mechanical moduli and visualized the cross-linked pattern without perturbing surrounding non-irradiated regions. 2P-CXL-induced tissue stiffening was comparable to that achieved with conventional one-photon CXL. Our results demonstrate the ability to selectively stiffen biological tissue in situ at high spatial resolution, with broad implications in ophthalmology, laser surgery, and tissue engineering.

  13. 3D imaging of intrinsic crystalline defects in zinc oxide by spectrally resolved two-photon fluorescence microscopy

    Science.gov (United States)

    Al-Tabich, A.; Inami, W.; Kawata, Y.; Jablonski, R.; Worasawat, S.; Mimura, H.

    2017-05-01

    We present a method for three-dimensional intrinsic defect imaging in zinc oxide (ZnO) by spectrally resolved two-photon fluorescence microscopy, based on the previously presented method of observing a photoluminescence distribution in wide-gap semiconductor crystals [Noor et al., Appl. Phys. Lett. 92(16), 161106 (2008)]. A tightly focused light beam radiated by a titanium-sapphire laser is used to obtain a two-photon excitation of selected area of the ZnO sample. Photoluminescence intensity of a specific spectral range is then selected by optical band pass filters and measured by a photomultiplier tube. Reconstruction of the specimen image is done by scanning the volume of interest by a piezoelectric positioning stage and measuring the spectrally resolved photoluminescence intensity at each point. The method has been proved to be effective at locating intrinsic defects of the ZnO crystalline structure in the volume of the crystal. The method was compared with other defect imaging and 3D imaging techniques like scanning tunneling microscopy and confocal microscopy. In both cases, our method shows superior penetration abilities and, as the only method, allows location of the defects of the chosen type in 3D. In this paper, we present the results of oxygen vacancies and zinc antisites imaging in ZnO nanorods.

  14. Ultra-large field-of-view two-photon microscopy

    OpenAIRE

    Tsai, Philbert S.; Mateo, Celine; Field, Jeffrey J.; Schaffer, Chris B.; Anderson, Matthew E.; Kleinfeld, David

    2015-01-01

    We present a two-photon microscope that images the full extent of murine cortex with an objective-limited spatial resolution across an 8 mm by 10 mm field. The lateral resolution is approximately 1 µm and the maximum scan speed is 5 mm/ms. The scan pathway employs large diameter compound lenses to minimize aberrations and performs near theoretical limits. We demonstrate the special utility of the microscope by recording resting-state vasomotion across both hemispheres of the murine brain thro...

  15. Neural plasticity explored by correlative two-photon and electron/SPIM microscopy

    Science.gov (United States)

    Allegra Mascaro, A. L.; Silvestri, L.; Costantini, I.; Sacconi, L.; Maco, B.; Knott, G. W.; Pavone, F. S.

    2013-06-01

    Plasticity of the central nervous system is a complex process which involves the remodeling of neuronal processes and synaptic contacts. However, a single imaging technique can reveal only a small part of this complex machinery. To obtain a more complete view, complementary approaches should be combined. Two-photon fluorescence microscopy, combined with multi-photon laser nanosurgery, allow following the real-time dynamics of single neuronal processes in the cerebral cortex of living mice. The structural rearrangement elicited by this highly confined paradigm of injury can be imaged in vivo first, and then the same neuron could be retrieved ex-vivo and characterized in terms of ultrastructural features of the damaged neuronal branch by means of electron microscopy. Afterwards, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, based on the use of major blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from its apical portion, the whole pyramidal neuron can then be segmented and located in the correct cortical layer. With the correlative approach presented here, researchers will be able to place in a three-dimensional anatomic context the neurons whose dynamics have been observed with high detail in vivo.

  16. Intravital two-photon microscopy of immune cell dynamics in corneal lymphatic vessels.

    Directory of Open Access Journals (Sweden)

    Philipp Steven

    Full Text Available BACKGROUND: The role of lymphatic vessels in tissue and organ transplantation as well as in tumor growth and metastasis has drawn great attention in recent years. METHODOLOGY/PRINCIPAL FINDINGS: We now developed a novel method using non-invasive two-photon microscopy to simultaneously visualize and track specifically stained lymphatic vessels and autofluorescent adjacent tissues such as collagen fibrils, blood vessels and immune cells in the mouse model of corneal neovascularization in vivo. The mouse cornea serves as an ideal tissue for this technique due to its easy accessibility and its inducible and modifiable state of pathological hem- and lymphvascularization. Neovascularization was induced by suture placement in corneas of Balb/C mice. Two weeks after treatment, lymphatic vessels were stained intravital by intrastromal injection of a fluorescently labeled LYVE-1 antibody and the corneas were evaluated in vivo by two-photon microscopy (TPM. Intravital TPM was performed at 710 nm and 826 nm excitation wavelengths to detect immunofluorescence and tissue autofluorescence using a custom made animal holder. Corneas were then harvested, fixed and analyzed by histology. Time lapse imaging demonstrated the first in vivo evidence of immune cell migration into lymphatic vessels and luminal transport of individual cells. Cells immigrated within 1-5.5 min into the vessel lumen. Mean velocities of intrastromal corneal immune cells were around 9 µm/min and therefore comparable to those of T-cells and macrophages in other mucosal surfaces. CONCLUSIONS: To our knowledge we here demonstrate for the first time the intravital real-time transmigration of immune cells into lymphatic vessels. Overall this study demonstrates the valuable use of intravital autofluorescence two-photon microscopy in the model of suture-induced corneal vascularizations to study interactions of immune and subsequently tumor cells with lymphatic vessels under close as possible

  17. Simultaneous morphological and functional imaging of the honeybee's brain by two-photon microscopy

    International Nuclear Information System (INIS)

    Haase, A.

    2011-01-01

    Thanks to its rather simply structured but highly performing brain, the honeybee (Apis mellifera) is an important model for neurobiological studies. Therefore there is a great need for new functional imaging modalities adapted to this species. Herein we give a detailed report on the development and performance of a platform for in vivo functional and morphological imaging of the honeybee's brain, focusing on its primary olfactory centres, the antennal lobes (ALs). The experimental setup consists of a two-photon microscope combined with a synchronized odour stimulus generator. Our imaging platform allows to simultaneously obtain both morphological measurements of the ALs functional units, the glomeruli, and in vivo calcium recording of their neural activity. We were able to record the characteristic glomerular response maps to odour stimuli applied to the bee's antennae. Our approach offers several advantages over the commonly used conventional fluorescence microscopy. Two-photon microscopy provides substantial enhancement in both spatial and temporal resolutions, while minimizing photo damage. Calcium recordings show a more than fourfold improvement in the functional signal with respect to the techniques available up to now. Finally, the extended penetration depth, thanks to the infrared excitation, allows the functional imaging of profound glomeruli which have not been optically accessible up to now.

  18. Imaging immune response of skin mast cells in vivo with two-photon microscopy

    Science.gov (United States)

    Li, Chunqiang; Pastila, Riikka K.; Lin, Charles P.

    2012-02-01

    Intravital multiphoton microscopy has provided insightful information of the dynamic process of immune cells in vivo. However, the use of exogenous labeling agents limits its applications. There is no method to perform functional imaging of mast cells, a population of innate tissue-resident immune cells. Mast cells are widely recognized as the effector cells in allergy. Recently their roles as immunoregulatory cells in certain innate and adaptive immune responses are being actively investigated. Here we report in vivo mouse skin mast cells imaging with two-photon microscopy using endogenous tryptophan as the fluorophore. We studied the following processes. 1) Mast cells degranulation, the first step in the mast cell activation process in which the granules are released into peripheral tissue to trigger downstream reactions. 2) Mast cell reconstitution, a procedure commonly used to study mast cells functioning by comparing the data from wild type mice, mast cell-deficient mice, and mast-cell deficient mice reconstituted with bone marrow-derived mast cells (BMMCs). Imaging the BMMCs engraftment in tissue reveals the mast cells development and the efficiency of BMMCs reconstitution. We observed the reconstitution process for 6 weeks in the ear skin of mast cell-deficient Kit wsh/ w-sh mice by two-photon imaging. Our finding is the first instance of imaging mast cells in vivo with endogenous contrast.

  19. Brain refractive index measured in vivo with high-NA defocus-corrected full-field OCT and consequences for two-photon microscopy.

    Science.gov (United States)

    Binding, Jonas; Ben Arous, Juliette; Léger, Jean-François; Gigan, Sylvain; Boccara, Claude; Bourdieu, Laurent

    2011-03-14

    Two-photon laser scanning microscopy (2PLSM) is an important tool for in vivo tissue imaging with sub-cellular resolution, but the penetration depth of current systems is potentially limited by sample-induced optical aberrations. To quantify these, we measured the refractive index n' in the somatosensory cortex of 7 rats in vivo using defocus optimization in full-field optical coherence tomography (ff-OCT). We found n' to be independent of imaging depth or rat age. From these measurements, we calculated that two-photon imaging beyond 200 µm into the cortex is limited by spherical aberration, indicating that adaptive optics will improve imaging depth.

  20. Automated filtering of intrinsic movement artifacts during two-photon intravital microscopy.

    Directory of Open Access Journals (Sweden)

    Denis Soulet

    Full Text Available In vivo imaging using two-photon microscopy is an essential tool to explore the dynamic of physiological events deep within biological tissues for short or extended periods of time. The new capabilities offered by this technology (e.g. high tissue penetrance, low toxicity have opened a whole new era of investigations in modern biomedical research. However, the potential of using this promising technique in tissues of living animals is greatly limited by the intrinsic irregular movements that are caused by cardiac and respiratory cycles and muscular and vascular tone. Here, we show real-time imaging of the brain, spinal cord, sciatic nerve and myenteric plexus of living mice using a new automated program, named Intravital_Microscopy_Toolbox, that removes frames corrupted with motion artifacts from time-lapse videos. Our approach involves generating a dissimilarity score against precalculated reference frames in a specific reference channel, thus allowing the gating of distorted, out-of-focus or translated frames. Since the algorithm detects the uneven peaks of image distortion caused by irregular animal movements, the macro allows a fast and efficient filtering of the image sequence. In addition, extra features have been implemented in the macro, such as XY registration, channel subtraction, extended field of view with maximum intensity projection, noise reduction with average intensity projections, and automated timestamp and scale bar overlay. Thus, the Intravital_Microscopy_Toolbox macro for ImageJ provides convenient tools for biologists who are performing in vivo two-photon imaging in tissues prone to motion artifacts.

  1. Nanoshells for in vivo imaging using two-photon excitation microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Gao Liang; Nammalvar, Vengadesan [Department of Bioengineering, Rice University, Houston, TX 77005 (United States); Vadakkan, Tegy J, E-mail: lg3@rice.edu, E-mail: venkyn@rice.edu [Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030 (United States)

    2011-09-07

    Gold nanoshells have been intensively investigated and applied to various biomedical fields because of their flexible optical tunability and biological compatibility. They hold great potential to serve as luminescent contrast agents excitable with near-infrared (NIR) lasers. In this paper, we describe the development of nanoshells with a peak of plasmon resonance at 800 nm and their subsequent use for in vivo blood vessel imaging using two-photon excitation microscopy at an excitation wavelength of 750 nm. We were able to image single nanoshell particles in blood vessels and generate optical contrast for blood vessel structure using luminescent signals. These results confirm the feasibility of engineering nanoshells with controlled optical properties for single-particle-based in vivo imaging.

  2. Multimodal reconstruction of microvascular-flow distributions using combined two-photon microscopy and Doppler optical coherence tomography.

    Science.gov (United States)

    Gagnon, Louis; Sakadžić, Sava; Lesage, Fréderic; Mandeville, Emiri T; Fang, Qianqian; Yaseen, Mohammad A; Boas, David A

    2015-01-01

    Computing microvascular cerebral blood flow ([Formula: see text]) in real cortical angiograms is challenging. Here, we investigated whether the use of Doppler optical coherence tomography (DOCT) flow measurements in individual vessel segments can help in reconstructing [Formula: see text] across the entire vasculature of a truncated cortical angiogram. A [Formula: see text] computational framework integrating DOCT measurements is presented. Simulations performed on a synthetic angiogram showed that the addition of DOCT measurements, especially close to large inflowing or outflowing vessels, reduces the impact of pressure boundary conditions and estimated vessel resistances resulting in a more accurate reconstruction of [Formula: see text]. Our technique was then applied to reconstruct microvascular flow distributions in the mouse cortex down to [Formula: see text] by combining two-photon laser scanning microscopy angiography with DOCT.

  3. Stratum corneum lipid organization as observed by atomic force, confocal and two-photon excitation fluorescence microscopy

    DEFF Research Database (Denmark)

    Norlén, Lars; Plasencia Gil, Maria Inés; Bagatolli, Luis

    2008-01-01

    -related biophysical techniques (e.g. atomic force microscopy and confocal/two-photon excitation fluorescence microscopy), it was recently shown that reconstituted membranes composed of extracted decontaminated human stratum corneum lipids do not form a fluid phase, but exclusively a single-gel phase that segregates...

  4. Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy

    Science.gov (United States)

    Silvestri, Ludovico; Allegra Mascaro, Anna Letizia; Costantini, Irene; Sacconi, Leonardo; Pavone, Francesco S.

    2014-02-01

    One of the unique features of the brain is that its activity cannot be framed in a single spatio-temporal scale, but rather spans many orders of magnitude both in space and time. A single imaging technique can reveal only a small part of this complex machinery. To obtain a more comprehensive view of brain functionality, complementary approaches should be combined into a correlative framework. Here, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, taking advantage of blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living thy1-GFP-M mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from the apical portion, the whole pyramidal neuron can then be segmented. The correlative approach presented here allows contextualizing within a three-dimensional anatomic framework the neurons whose dynamics have been observed with high detail in vivo.

  5. Two-photon microscopy measurement of CMRO2 using periarteriolar PO2 gradients(Conference Presentation)

    Science.gov (United States)

    Sakadžić, Sava; Yaseen, Mohammad A.; Jaswal, Rajeshwer S.; Roussakis, Emmanuel; Dale, Anders M.; Buxton, Richard B.; Vinogradov, Sergei A.; Boas, David A.; Devor, Anna

    2017-02-01

    The cerebral metabolic rate of oxygen (CMRO2) is an essential parameter for evaluating brain function and pathophysiology. Measurements of CMRO2 with high spatio-temporal resolution are critically important for understanding how the brain copes with metabolic and blood perfusion changes associated with various clinical conditions, such as stroke, periinfarct depolarizations, and various microvasculopathies (e.g., Alzheimer's disease, chronic hypertension). CMRO2 measurements are also important for understanding the physiological underpinnings of functional Magnetic Resonance Imaging signals. However, the currently available approaches for quantifying CMRO2 rely on complex multimodal imaging and mathematical modeling. Here, we introduce a novel method that allows estimation of CMRO2 based on a single measurement modality - two-photon phosphorescence lifetime microscopy (2PLM) imaging of the partial pressure of oxygen (PO2) in cortical tissue. CMRO2 is estimated by fitting the changes of tissue PO2 around cortical penetrating arterioles with the Krogh cylinder model of oxygen diffusion. We measured the baseline CMRO2 in anesthetized rats, and modulated tissue PO2 levels by manipulating the depth of anesthesia. This method has a spatial resolution of approximately 200 μm and it may provide CMRO2 measurements in individual cortical layers or within confined cortical regions such as in ischemic penumbra and the foci of functional activation.

  6. Brain Tissue PO2 Measurement During Normoxia and Hypoxia Using Two-Photon Phosphorescence Lifetime Microscopy.

    Science.gov (United States)

    Xu, Kui; Boas, David A; Sakadžić, Sava; LaManna, Joseph C

    2017-01-01

    Key to the understanding of the principles of physiological and structural acclimatization to changes in the balance between energy supply (represented by substrate and oxygen delivery, and mitochondrial oxidative phosphorylation) and energy demand (initiated by neuronal activity) is to determine the controlling variables, how they are sensed and the mechanisms initiated to maintain the balance. The mammalian brain depends completely on continuous delivery of oxygen to maintain its function. We hypothesized that tissue oxygen is the primary sensed variable. In this study two-photon phosphorescence lifetime microscopy (2PLM) was used to determine and define the tissue oxygen tension field within the cerebral cortex of mice to a cortical depth of between 200-250 μm under normoxia and acute hypoxia (FiO 2  = 0.10). High-resolution images can provide quantitative distributions of oxygen and intercapillary oxygen gradients. The data are best appreciated by quantifying the distribution histogram that can then be used for analysis. For example, in the brain cortex of a mouse, at a depth of 200 μm, tissue oxygen tension was mapped and the distribution histogram was compared under normoxic and mild hypoxic conditions. This powerful method can provide for the first time a description of the delivery and availability of brain oxygen in vivo.

  7. Two-photon microscopy for non-invasive, quantitative monitoring of stem cell differentiation.

    Directory of Open Access Journals (Sweden)

    William L Rice

    Full Text Available BACKGROUND: The engineering of functional tissues is a complex multi-stage process, the success of which depends on the careful control of culture conditions and ultimately tissue maturation. To enable the efficient optimization of tissue development protocols, techniques suitable for monitoring the effects of added stimuli and induced tissue changes are needed. METHODOLOGY/PRINCIPAL FINDINGS: Here, we present the quantitative use of two-photon excited fluorescence (TPEF and second harmonic generation (SHG as a noninvasive means to monitor the differentiation of human mesenchymal stem cells (hMSCs using entirely endogenous sources of contrast. We demonstrate that the individual fluorescence contribution from the intrinsic cellular fluorophores NAD(PH, flavoproteins and lipofuscin can be extracted from TPEF images and monitored dynamically from the same cell population over time. Using the redox ratio, calculated from the contributions of NAD(PH and flavoproteins, we identify distinct patterns in the evolution of the metabolic activity of hMSCs maintained in either propagation, osteogenic or adipogenic differentiation media. The differentiation of these cells is mirrored by changes in cell morphology apparent in high resolution TPEF images and by the detection of collagen production via SHG imaging. Finally, we find dramatic increases in lipofuscin levels in hMSCs maintained at 20% oxygen vs. those in 5% oxygen, establishing the use of this chromophore as a potential biomarker for oxidative stress. CONCLUSIONS/SIGNIFICANCE: In this study we demonstrate that it is possible to monitor the metabolic activity, morphology, ECM production and oxidative stress of hMSCs in a non-invasive manner. This is accomplished using generally available multiphoton microscopy equipment and simple data analysis techniques, such that the method can widely adopted by laboratories with a diversity of comparable equipment. This method therefore represents a powerful tool

  8. Arduino Due based tool to facilitate in vivo two-photon excitation microscopy.

    Science.gov (United States)

    Artoni, Pietro; Landi, Silvia; Sato, Sebastian Sulis; Luin, Stefano; Ratto, Gian Michele

    2016-04-01

    Two-photon excitation spectroscopy is a powerful technique for the characterization of the optical properties of genetically encoded and synthetic fluorescent molecules. Excitation spectroscopy requires tuning the wavelength of the Ti:sapphire laser while carefully monitoring the delivered power. To assist laser tuning and the control of delivered power, we developed an Arduino Due based tool for the automatic acquisition of high quality spectra. This tool is portable, fast, affordable and precise. It allowed studying the impact of scattering and of blood absorption on two-photon excitation light. In this way, we determined the wavelength-dependent deformation of excitation spectra occurring in deep tissues in vivo.

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

    Science.gov (United States)

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

    2018-02-01

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

  10. Review: two-photon scanning systems for clinical high resolution in vivo tissue imaging

    Science.gov (United States)

    König, K.; Müller, J.; Höfer, M.; Müller, C.; Weinigel, M.; Bückle, R.; Elsner, P.; Kaatz, M.; Messerschmidt, B.

    2008-02-01

    The femtosecond laser multiphoton tomograph DermaInspect as well as high NA two-photon GRIN microendoscopes for in vivo tomography of human skin have been used to detect malignant melanoma as well as to study the diffusion and intradermal accumulation of topically applied cosmetical and pharmaceutical components. So far, more than 500 patients and volunteers in Europe, Australia, and Asia have been investigated with this unique tomograph. Near infrared 80 MHz picojoule femtosecond laser pulses were employed to excite endogenous fluorophores such as NAD(P)H, flavoproteins, melanin, and elastin as well as fluorescent components of a variety of ointments via a twophoton excitation process. In addition, collagen has been imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Individual intratissue cells, mitochondria, melanosomes, and the morphology of the nuclei as well as extracellular matrix elements could be clearly visualized due to molecular imaging and the calculation of fluorescence lifetime images. Nanoparticles and intratissue drugs have been detected non-invasively, in situ and over a period of up to 3 months. In addition, hydration effects and UV effects were studied by monitoring modifications of cellular morphology and autofluorescence. The system was used to observe the diffusion through the stratum corneum and the accumulation and release of functionalized nanoparticles along hair shafts and epidermal ridges. The DermaInspect been also employed to gain information on skin age and wound healing in patients with ulcers. Novel developments include a galvo/piezo-scan driven flexible articulated arm as

  11. Lipophilic phosphorescent gold(I) clusters as selective probes for visualization of lipid droplets by two-photon microscopy

    Czech Academy of Sciences Publication Activity Database

    Koshel, E. I.; Cheluskin, P. S.; Melnikov, A. S.; Serdobintsev, P. Y.; Stolbovaia, A. Y.; Saifitdinova, A. F.; Scheslavskiy, V. I.; Chernyavskiy, Oleksandr; Gaginskaya, E. R.; Koshevoy, I. O.; Tunik, S. P.

    2017-01-01

    Roč. 332, Jan 1 (2017), s. 122-130 ISSN 1010-6030 R&D Projects: GA MŠk(CZ) LM2015062 Institutional support: RVO:67985823 Keywords : polynuclear gold-alkynyl cluster * lipophilic probe * phosphorescence * adipocyte * two-photon microscopy * PLIM Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Medical laboratory technology (including laboratory samples analysis Impact factor: 2.625, year: 2016

  12. Simple approach to three-color two-photon microscopy by a fiber-optic wavelength convertor.

    Science.gov (United States)

    Li, Kuen-Che; Huang, Lynn L H; Liang, Jhih-Hao; Chan, Ming-Che

    2016-11-01

    A simple approach to multi-color two-photon microscopy of the red, green, and blue fluorescent indicators was reported based on an ultra-compact 1.03-μm femtosecond laser and a nonlinear fiber. Inside the nonlinear fiber, the 1.03-μm laser pulses were simultaneously blue-shifted to 0.6~0.8 μm and red-shifted to 1.2~1.4 μm region by the Cherenkov radiation and fiber Raman gain effects. The wavelength-shifted 0.6~0.8 μm and 1.2~1.4 μm radiations were co-propagated with the residual non-converted 1.03-μm pulses inside the same nonlinear fiber to form a fiber-output three-color femtosecond source. The application of the multi-wavelength sources on multi-color two-photon fluorescence microscopy were also demonstrated. Overall, due to simple system configuration, convenient wavelength conversion, easy wavelength tunability within the entire 0.7~1.35 μm bio-penetration window and less requirement for high power and bulky light sources, the simple approach to multi-color two-photon microscopy could be widely applicable as an easily implemented and excellent research tool for future biomedical and possibly even clinical applications.

  13. Ultrafast scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Botkin, D.A. [California Univ., Berkeley, CA (United States). Dept. of Physics]|[Lawrence Berkeley Lab., CA (United States)

    1995-09-01

    I have developed an ultrafast scanning tunneling microscope (USTM) based on uniting stroboscopic methods of ultrafast optics and scanned probe microscopy to obtain nanometer spatial resolution and sub-picosecond temporal resolution. USTM increases the achievable time resolution of a STM by more than 6 orders of magnitude; this should enable exploration of mesoscopic and nanometer size systems on time scales corresponding to the period or decay of fundamental excitations. USTM consists of a photoconductive switch with subpicosecond response time in series with the tip of a STM. An optical pulse from a modelocked laser activates the switch to create a gate for the tunneling current, while a second laser pulse on the sample initiates a dynamic process which affects the tunneling current. By sending a large sequence of identical pulse pairs and measuring the average tunnel current as a function of the relative time delay between the pulses in each pair, one can map the time evolution of the surface process. USTM was used to measure the broadband response of the STM`s atomic size tunnel barrier in frequencies from tens to hundreds of GHz. The USTM signal amplitude decays linearly with the tunnel junction conductance, so the spatial resolution of the time-resolved signal is comparable to that of a conventional STM. Geometrical capacitance of the junction does not appear to play an important role in the measurement, but a capacitive effect intimately related to tunneling contributes to the measured signals and may limit the ultimate resolution of the USTM.

  14. In vivo imaging of cerebral energy metabolism with two-photon fluorescence lifetime microscopy of NADH.

    Science.gov (United States)

    Yaseen, Mohammad A; Sakadžić, Sava; Wu, Weicheng; Becker, Wolfgang; Kasischke, Karl A; Boas, David A

    2013-02-01

    Minimally invasive, specific measurement of cellular energy metabolism is crucial for understanding cerebral pathophysiology. Here, we present high-resolution, in vivo observations of autofluorescence lifetime as a biomarker of cerebral energy metabolism in exposed rat cortices. We describe a customized two-photon imaging system with time correlated single photon counting detection and specialized software for modeling multiple-component fits of fluorescence decay and monitoring their transient behaviors. In vivo cerebral NADH fluorescence suggests the presence of four distinct components, which respond differently to brief periods of anoxia and likely indicate different enzymatic formulations. Individual components show potential as indicators of specific molecular pathways involved in oxidative metabolism.

  15. Label-free imaging immune cells and collagen in atherosclerosis with two-photon and second harmonic generation microscopy

    Directory of Open Access Journals (Sweden)

    Chunqiang Li

    2016-01-01

    Full Text Available Atherosclerosis has been recognized as a chronic inflammation disease, in which many types of cells participate in this process, including lymphocytes, macrophages, dendritic cells (DCs, mast cells, vascular smooth muscle cells (SMCs. Developments in imaging technology provide the capability to observe cellular and tissue components and their interactions. The knowledge of the functions of immune cells and their interactions with other cell and tissue components will facilitate our discovery of biomarkers in atherosclerosis and prediction of the risk factor of rupture-prone plaques. Nonlinear optical microscopy based on two-photon excited autofluorescence and second harmonic generation (SHG were developed to image mast cells, SMCs and collagen in plaque ex vivo using endogenous optical signals. Mast cells were imaged with two-photon tryptophan autofluorescence, SMCs were imaged with two-photon NADH autofluorescence, and collagen were imaged with SHG. This development paves the way for further study of mast cell degranulation, and the effects of mast cell derived mediators such as induced synthesis and activation of matrix metalloproteinases (MMPs which participate in the degradation of collagen.

  16. Dynamic characterization of hydrophobic and hydrophilic solutes in oleic-acid enhanced transdermal delivery using two-photon fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tseng, Te-Yu; Yang, Chiu-Sheng; Chen, Yang-Fang [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Tsai, Tsung-Hua [Department of Dermatology, Far Eastern Memorial Hospital, New Taipei City, Taiwan (China); Dong, Chen-Yuan, E-mail: cydong@phys.ntu.edu.tw [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Center for Quantum Science and Engineering, National Taiwan University, Taipei, Taiwan (China); Center for Optoelectronic Biomedicine, National Taiwan University, Taipei, Taiwan (China)

    2014-10-20

    In this letter, we propose an efficient methodology of investigating dynamic properties of sulforhodamine B and rhodamine B hexyl ester molecules transporting across ex-vivo human stratum corneum with and without oleic acid enhancement. Three-dimensional, time-lapse fluorescence images of the stratum corneum can be obtained using two-photon fluorescence microscopy. Furthermore, temporal quantifications of transport enhancements in diffusion parameters can be achieved with the use of Fick's second law. Dynamic characterization of solutes transporting across the stratum corneum is an effective method for understanding transient phenomena in transdermal delivery of probe molecules, leading to improved delivery strategies of molecular species for therapeutic purposes.

  17. Cell and brain tissue imaging of the flavonoid fisetin using label-free two-photon microscopy.

    Science.gov (United States)

    Krasieva, Tatiana B; Ehren, Jennifer; O'Sullivan, Thomas; Tromberg, Bruce J; Maher, Pamela

    2015-10-01

    Over the last few years, we have identified an orally active, novel neuroprotective and cognition-enhancing molecule, the flavonoid fisetin. Fisetin not only has direct antioxidant activity but it can also increase the intracellular levels of glutathione, the major intracellular antioxidant. Fisetin can also activate key neurotrophic factor signaling pathways. In addition, it has anti-inflammatory activity against microglia and astrocytes and inhibits the activity of lipoxygenases, thereby reducing the production of pro-inflammatory eicosanoids and their by-products. However, key questions about its targets and brain penetration remain. In this study, we used label-free two-photon microscopy of intrinsic fisetin fluorescence to examine the localization of fisetin in living nerve cells and the brains of living mice. In cells, fisetin but not structurally related flavonols with different numbers of hydroxyl groups, localized to the nucleoli suggesting that key targets of fisetin may reside in this organelle. In the mouse brain, following intraperitoneal injection and oral administration, fisetin rapidly distributed to the blood vessels of the brain followed by a slower dispersion into the brain parenchyma. Thus, these results provide further support for the effects of fisetin on brain function. In addition, they suggest that label-free two-photon microscopy may prove useful for studying the intracellular and tissue distribution of other intrinsically-fluorescent flavonoids. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Development and design of up-to-date laser scanning two-photon microscope using in neuroscience

    Science.gov (United States)

    Doronin, Maxim; Popov, Alexander

    2017-02-01

    Today one of the main areas of application of two-photon microscopy is biology. This is due to the fact that this technique allows to obtain 3D images of tissues due to laser focus change, that is possible due to substantially greater penetration depth on the main wavelength into biological tissues. Self-developed microscopy system provides possibility to service it and modify the structure of microscope depending on highly specialized experimental design and scientific goals. This article may be regarded as a quick reference to laboratory staff who are wishing to develop their own microscopy system for self-service and modernization of the system and in order to save the lab budget.

  19. Confocal scanning microscopy

    DEFF Research Database (Denmark)

    Bariani, Paolo

    This report is based on a metrological investigation on confocal microscopy technique carried out by Uffe Rolf Arlø Theilade and Paolo Bariani. The purpose of the experimental activity was twofold a metrological instrument characterization and application to assessment of rough PP injection moulded...... replicated topography. Confocal microscopy is seen to be a promising technique in metrology of microstructures. Some limitations with respect to surface metrology were found during the experiments. The experiments were carried out using a Zeiss LSM 5 Pascal microscope owned by the Danish Polymer Centre...

  20. Scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Cox, B. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada)

    1970-05-15

    The JSM-11 scanning electron microscope at CRNL has been used extensively for topographical studies of oxidized metals, fracture surfaces, entomological and biological specimens. A non-dispersive X-ray attachment permits the microanalysis of the surface features. Techniques for the production of electron channeling patterns have been developed. (author)

  1. Scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Binnig, G.; Rohrer, H.

    1983-01-01

    Based on vacuum tunneling, a novel type of microscope, the scanning tunneling microscope (STM) was developed. It has an unprecedented resolution in real space on an atomic scale. The authors review the important technical features, illustrate the power of the STM for surface topographies and discuss its potential in other areas of science and technology. (Auth.)

  2. Novel nanocarriers for topical drug delivery: investigating delivery efficiency and distribution in skin using two-photon microscopy

    Science.gov (United States)

    Kirejev, Vladimir; Guldbrand, Stina; Bauer, Brigitte; Smedh, Maria; Ericson, Marica B.

    2011-03-01

    The complex structure of skin represents an effective barrier against external environmental factors, as for example, different chemical and biochemical compounds, yeast, bacterial and viral infections. However, this impermeability prevents efficient transdermal drug delivery which limits the number of drugs that are able to penetrate the skin efficiently. Current trends in drug application through skin focus on the design and use of nanocarriers for transport of active compounds. The transport systems applied so far have several drawbacks, as they often have low payload, high toxicity, a limited variability of inclusion molecules, or long degradation times. The aim of these current studies is to investigate novel topical drug delivery systems, e.g. nanocarriers based on cyclic oligosaccharides - cyclodextrins (CD) or iron (III)-based metal-organic frameworks (MOF). Earlier studies on cell cultures imply that these drug nanocarriers show promising characteristics compared to other drug delivery systems. In our studies, we use two-photon microscopy to investigate the ability of the nanocarriers to deliver compounds through ex-vivo skin samples. Using near infrared light for excitation in the so called optical window of skin allows deep-tissue visualization of drug distribution and localization. In addition, it is possible to employ two-photon based fluorescence correlation spectroscopy for quantitative analysis of drug distribution and concentrations in different cell layers.

  3. Scanning probe microscopy

    International Nuclear Information System (INIS)

    Mainsbridge, B.

    1994-01-01

    In late 1959, Richard Feynman observed that manoeuvring atoms was something that could be done in principle but has not been done, 'because we are too big'. In 1982, the scanning tunnelling microscope (STM) was invented and is now a central tool for the construction of nanoscale devices in what was known as molecular engineering, and now, nanotechnology. The principles of the microscope are outlined and references are made to other scanning devices which have evolved from the original invention. The method of employment of the STM as a machine tool is described and references are made to current speculations on applications of the instrument in nanotechnology. A short bibliography on this topic is included. 27 refs., 7 figs

  4. Scanning probe microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Mainsbridge, B [Murdoch Univ., WA (Australia). School of Mathematical and Physical Sciences

    1994-12-31

    In late 1959, Richard Feynman observed that manoeuvring atoms was something that could be done in principle but has not been done, `because we are too big`. In 1982, the scanning tunnelling microscope (STM) was invented and is now a central tool for the construction of nanoscale devices in what was known as molecular engineering, and now, nanotechnology. The principles of the microscope are outlined and references are made to other scanning devices which have evolved from the original invention. The method of employment of the STM as a machine tool is described and references are made to current speculations on applications of the instrument in nanotechnology. A short bibliography on this topic is included. 27 refs., 7 figs.

  5. Non-scanning fiber-optic near-infrared beam led to two-photon optogenetic stimulation in-vivo.

    Directory of Open Access Journals (Sweden)

    Kamal R Dhakal

    Full Text Available Stimulation of specific neurons expressing opsins in a targeted region to manipulate brain function has proved to be a powerful tool in neuroscience. However, the use of visible light for optogenetic stimulation is invasive due to low penetration depth and tissue damage owing to larger absorption and scattering. Here, we report, for the first time, in-depth non-scanning fiber-optic two-photon optogenetic stimulation (FO-TPOS of neurons in-vivo in transgenic mouse models. In order to optimize the deep-brain stimulation strategy, we characterized two-photon activation efficacy at different near-infrared laser parameters. The significantly-enhanced in-depth stimulation efficiency of FO-TPOS as compared to conventional single-photon beam was demonstrated both by experiments and Monte Carlo simulation. The non-scanning FO-TPOS technology will lead to better understanding of the in-vivo neural circuitry because this technology permits more precise and less invasive anatomical delivery of stimulation.

  6. Scanning Tunneling Microscopy - image interpretation

    International Nuclear Information System (INIS)

    Maca, F.

    1998-01-01

    The basic ideas of image interpretation in Scanning Tunneling Microscopy are presented using simple quantum-mechanical models and supplied with examples of successful application. The importance is stressed of a correct interpretation of this brilliant experimental surface technique

  7. An orthotopic glioblastoma mouse model maintaining brain parenchymal physical constraints and suitable for intravital two-photon microscopy.

    Science.gov (United States)

    Ricard, Clément; Stanchi, Fabio; Rougon, Geneviève; Debarbieux, Franck

    2014-04-21

    Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date. Murine models of this pathology rely on the injection of a suspension of glioma cells into the brain parenchyma following incision of the dura-mater. Whereas the cells have to be injected superficially to be accessible to intravital two-photon microscopy, superficial injections fail to recapitulate the physiopathological conditions. Indeed, escaping through the injection tract most tumor cells reach the extra-dural space where they expand abnormally fast in absence of mechanical constraints from the parenchyma. Our improvements consist not only in focally implanting a glioma spheroid rather than injecting a suspension of glioma cells in the superficial layers of the cerebral cortex but also in clogging the injection site by a cross-linked dextran gel hemi-bead that is glued to the surrounding parenchyma and sealed to dura-mater with cyanoacrylate. Altogether these measures enforce the physiological expansion and infiltration of the tumor cells inside the brain parenchyma. Craniotomy was finally closed with a glass window cemented to the skull to allow chronic imaging over weeks in absence of scar tissue development. Taking advantage of fluorescent transgenic animals grafted with fluorescent tumor cells we have shown that the dynamics of interactions occurring between glioma cells, neurons (e.g. Thy1-CFP mice) and vasculature (highlighted by an intravenous injection of a fluorescent dye) can be visualized by intravital two-photon microscopy during the progression of the disease. The possibility to image a tumor at microscopic resolution in a minimally compromised cerebral environment represents an improvement of current GBM animal models which should benefit the field of neuro-oncology and drug testing.

  8. Two-photon-induced hot-electron transfer to a single molecule in a scanning tunneling microscope

    International Nuclear Information System (INIS)

    Wu, S. W.; Ho, W.

    2010-01-01

    The junction of a scanning tunneling microscope (STM) operating in the tunneling regime was irradiated with femtosecond laser pulses. A photoexcited hot electron in the STM tip resonantly tunnels into an excited state of a single molecule on the surface, converting it from the neutral to the anion. The electron-transfer rate depends quadratically on the incident laser power, suggesting a two-photon excitation process. This nonlinear optical process is further confirmed by the polarization measurement. Spatial dependence of the electron-transfer rate exhibits atomic-scale variations. A two-pulse correlation experiment reveals the ultrafast dynamic nature of photoinduced charging process in the STM junction. Results from these experiments are important for understanding photoinduced interfacial charge transfer in many nanoscale inorganic-organic structures.

  9. Volumetric label-free imaging and 3D reconstruction of mammalian cochlea based on two-photon excitation fluorescence microscopy

    International Nuclear Information System (INIS)

    Zhang, Xianzeng; Zhan, Zhenlin; Xie, Shusen; Geng, Yang; Ye, Qing

    2013-01-01

    The visualization of the delicate structure and spatial relationship of intracochlear sensory cells has relied on the laborious procedures of tissue excision, fixation, sectioning and staining for light and electron microscopy. Confocal microscopy is advantageous for its high resolution and deep penetration depth, yet disadvantageous due to the necessity of exogenous labeling. In this study, we present the volumetric imaging of rat cochlea without exogenous dyes using a near-infrared femtosecond laser as the excitation mechanism and endogenous two-photon excitation fluorescence (TPEF) as the contrast mechanism. We find that TPEF exhibits strong contrast, allowing cellular and even subcellular resolution imaging of the cochlea, differentiating cell types, visualizing delicate structures and the radial nerve fiber. Our results further demonstrate that 3D reconstruction rendered with z-stacks of optical sections enables better revealment of fine structures and spatial relationships, and easily performed morphometric analysis. The TPEF-based optical biopsy technique provides great potential for new and sensitive diagnostic tools for hearing loss or hearing disorders, especially when combined with fiber-based microendoscopy. (paper)

  10. Cardiac muscle organization revealed in 3-D by imaging whole-mount mouse hearts using two-photon fluorescence and confocal microscopy.

    Science.gov (United States)

    Sivaguru, Mayandi; Fried, Glenn; Sivaguru, Barghav S; Sivaguru, Vignesh A; Lu, Xiaochen; Choi, Kyung Hwa; Saif, M Taher A; Lin, Brian; Sadayappan, Sakthivel

    2015-11-01

    The ability to image the entire adult mouse heart at high resolution in 3-D would provide enormous advantages in the study of heart disease. However, a technique for imaging nuclear/cellular detail as well as the overall structure of the entire heart in 3-D with minimal effort is lacking. To solve this problem, we modified the benzyl alcohol:benzyl benzoate (BABB) clearing technique by labeling mouse hearts with periodic acid Schiff (PAS) stain. We then imaged the hearts with a combination of two-photon fluorescence microscopy and automated tile-scan imaging/stitching. Utilizing the differential spectral properties of PAS, we could identify muscle and nuclear compartments in the heart. We were also able to visualize the differences between a 3-month-old normal mouse heart and a mouse heart that had undergone heart failure due to the expression of cardiac myosin binding protein-C (cMyBP-C) gene mutation (t/t). Using 2-D and 3-D morphometric analysis, we found that the t/t heart had anomalous ventricular shape, volume, and wall thickness, as well as a disrupted sarcomere pattern. We further validated our approach using decellularized hearts that had been cultured with 3T3 fibroblasts, which were tracked using a nuclear label. We were able to detect the 3T3 cells inside the decellularized intact heart tissue, achieving nuclear/cellular resolution in 3-D. The combination of labeling, clearing, and two-photon microscopy together with tiling eliminates laborious and time-consuming physical sectioning, alignment, and 3-D reconstruction.

  11. High Resolution Scanning Ion Microscopy

    NARCIS (Netherlands)

    Castaldo, V.

    2011-01-01

    The structure of the thesis is the following. The first chapter is an introduction to scanning microscopy, where the path that led to the Focused Ion Beam (FIB) is described and the main differences between electrons and ion beams are highlighted. Chapter 2 is what is normally referred to (which I

  12. NEAR-IR TWO PHOTON MICROSCOPY IMAGING OF SILICA NANOPARTICLES FUNCTIONALIZED WITH ISOLATED SENSITIZED Yb(III) CENTERS

    Energy Technology Data Exchange (ETDEWEB)

    Lapadula, Giuseppe; Bourdolle, Adrien; Allouche, Florian; Conley, Matthew P.; Maron, Laurent; Lukens, Wayne W.; Guyot, Yannick; Andraud, Chantal; Brasselet, Sophie; Copé; ret, Christophe; Maury, Olivier; Andersen, Richard A.

    2013-01-12

    Bright nano objects emitting in the near infrared with a maximal cross section of 41.4 x 103 GM (Goppert Mayer), were prepared by implanting ca. 180 4,4 diethylaminostyryl 2,2 bipyridine (DEAS) Yb(III) complexes on the surface of 12 nm silica nanoparticles. The surface complexes ([DEAS Ln SiO2], Ln =Y,Yb) were characterized using IR, solid state NMR, UV Vis, EXAFS spectroscopies in combination with the preparation and characterization of similar molecular analogues by analytical techniques (IR, solution NMR, UV Vis, X ray crystallography) as well as DFT calculations. Starting from the partial dehydroxylation of the silica at 700 C on high vacuum having 0.8 OH.nm 2, the grafting of Ln(N(SiMe3)2)3 generate ≤SiO Ln(N(SiMe3)2)2, which upon thermal step and coordination of the DEAS chromophore yields (≤SiO)3Ln(DEAS). Surface and molecular analogues display similar properties, in terms of DEAS binding constants absorption maxima and luminescence properties (intense emission band assigned to a ligand centered CT fluorescence and life time) in the solid state, consistent with the molecular nature of the surface species. The densely functionalized nanoparticles can be dispersed via ultra-sonication in small ca. 15-20 nm aggregates (1 to 6 elementary particles) that were detected using two photon microscopy imaging at 720 nm excitation, making them promising nano objects for bio imaging.

  13. A close look at brain dynamics: cells and vessels seen by in vivo two-photon microscopy.

    Science.gov (United States)

    Fumagalli, Stefano; Ortolano, Fabrizio; De Simoni, Maria-Grazia

    2014-10-01

    The cerebral vasculature has a unique role in providing a constant supply of oxygen and nutrients to ensure normal brain functions. Blood vessels that feed the brain are far from being simply channels for passive transportation of fluids. They form complex structures made up of different cell types. These structures regulate blood supply, local concentrations of O2 and CO2, transport of small molecules, trafficking of plasma cells and fine cerebral functions in normal and diseased brains. Until few years ago, analysis of these functions has been typically based on post mortem techniques, whose interpretation is limited by the need for tissue processing at specific times. For a reliable and effective picture of the dynamic processes in the central nervous system, real-time information in vivo is required. There are now few in vivo systems, among which two-photon microscopy (2-PM) is a truly innovative tool for studying the brain. 2-PM has been used to dissect specific aspects of vascular and immune cell dynamics in the context of neurological diseases, providing exciting results that could not have been obtained with conventional methods. This review summarizes the latest findings on vascular and immune system action in the brain, with particular focus on the dynamic responses after ischemic brain injury. 2-PM has helped define the hierarchical architecture of the brain vasculature, the dynamic interaction between the vasculature and immune cells recruited to lesion sites, the effects of blood flow on neuronal and microglial activity and the ability of cells of the neurovascular unit to regulate blood flow. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Two-photon microscopy imaging of thy1GFP-M transgenic mice: a novel animal model to investigate brain dendritic cell subsets in vivo.

    Directory of Open Access Journals (Sweden)

    Claudia Laperchia

    Full Text Available Transgenic mice expressing fluorescent proteins in specific cell populations are widely used for in vivo brain studies with two-photon fluorescence (TPF microscopy. Mice of the thy1GFP-M line have been engineered for selective expression of green fluorescent protein (GFP in neuronal populations. Here, we report that TPF microscopy reveals, at the brain surface of these mice, also motile non-neuronal GFP+ cells. We have analyzed the behavior of these cells in vivo and characterized in brain sections their immunophenotype.With TPF imaging, motile GFP+ cells were found in the meninges, subarachnoid space and upper cortical layers. The striking feature of these cells was their ability to move across the brain parenchyma, exhibiting evident shape changes during their scanning-like motion. In brain sections, GFP+ cells were immunonegative to antigens recognizing motile cells such as migratory neuroblasts, neuronal and glial precursors, mast cells, and fibroblasts. GFP+ non-neuronal cells exhibited instead the characteristic features and immunophenotype (CD11c and major histocompatibility complex molecule class II immunopositivity of dendritic cells (DCs, and were immunonegative to the microglial marker Iba-1. GFP+ cells were also identified in lymph nodes and blood of thy1GFP-M mice, supporting their identity as DCs. Thus, TPF microscopy has here allowed the visualization for the first time of the motile behavior of brain DCs in situ. The results indicate that the thy1GFP-M mouse line provides a novel animal model for the study of subsets of these professional antigen-presenting cells in the brain. Information on brain DCs is still very limited and imaging in thy1GFP-M mice has a great potential for analyses of DC-neuron interaction in normal and pathological conditions.

  15. Labeling Human Mesenchymal Stem Cells with Gold Nanocages for in vitro and in vivo Tracking by Two-Photon Microscopy and Photoacoustic Microscopy

    Science.gov (United States)

    Zhang, Yu Shrike; Wang, Yu; Wang, Lidai; Wang, Yucai; Cai, Xin; Zhang, Chi; Wang, Lihong V.; Xia, Younan

    2013-01-01

    Stem cell tracking is a highly important subject. Current techniques based on nanoparticle-labeling, such as magnetic resonance imaging, fluorescence microscopy, and micro-computed tomography, are plagued by limitations including relatively low sensitivity or penetration depth, involvement of ionizing irradiation, and potential cytotoxicity of the nanoparticles. Here we introduce a new class of contrast agents based on gold nanocages (AuNCs) with hollow interiors and porous walls to label human mesenchymal stem cells (hMSCs) for both in vitro and in vivo tracking using two-photon microscopy and photoacoustic microscopy. As demonstrated by the viability assay, the AuNCs showed negligible cytotoxicity under a reasonable dose, and did not alter the differentiation potential of the hMSCs into desired lineages. We were able to image the cells labeled with AuNCs in vitro for at least 28 days in culture, as well as to track the cells that homed to the tumor region in nude mice in vivo. PMID:23946820

  16. A preliminary investigation on the interaction between sol-gel immobilized glucose oxidase and freely diffusing glucose by means of two-photon microscopy

    Science.gov (United States)

    Delfino, I.; Portaccio, M.; De Rosa, M.; Lepore, M.

    2013-02-01

    To study immobilized protein interactions with dissolved substrates is a very important topic both from a fundamental and technological standpoint. In the present report we illustrate the preliminary results obtained on sol-gel immobilized glucose oxidase (GOD) using a standard de-scanned two-photon microscope based on a modified confocal scanhead with internal detectors and a Ti:sapphire laser as a source. Data acquisition conditions were preliminary defined using functionalized beads of different dimensions. Various sol-gel supports were then investigated by monitoring endogeneous fluorescence due to the flavoadenine (FAD) molecules, present in GOD. Linear absorption and fluorescence spectroscopy along with Fourier Transform Infrared microscopy were employed for a full-optical characterization of the samples. The results show that GOD immobilization processes can be successfully monitored in some cases and also the interaction with glucose could be studied by this approach. This assessment holds potentials to better understand the characteristic of immobilized enzymes biocatalysis and to develop new biosensing schemes.

  17. The translated conceptual survey of physics / stablization of the focal plane in two photon excitation fluorescence microscopy

    Science.gov (United States)

    Wada, Asma

    As a reflection of my career to be an effective college physics teacher, my thesis is in two parts. The first is in education research, the focus of this part is to have a tool to evaluate pedagogies I have learned at the school and plan to apply in my classrooms back home. Consequently, this resulted in the development of the translated conceptual survey of physics ( TCSP). (TCSP) was designed by combining some questions from the Force Conceptual Inventory (FCI), and the Conceptual Survey of Electricity and Magnetism (CSEM) to assess student's understanding of basic concepts of Newtonian mechanics and electricity and magnetism in introductory physics. The idea of developing this questionnaire is to use it in classrooms back home as a part of a long term objective to implement what has been realized in the area of education research to improve the quality of teaching physics there. The survey was initially written in English, validated with interviews with native English speakers, translated into Arabic, and then validated via an interview with a native Arabic speaker. We then administered the survey to two different English-speaking intro physics courses and analyzed the results for consistency. The objective of the second part in my thesis is to expand my knowledge in an area of physics that I have interest in, and getting involved in a scientific research to develop skills I need as a teacher. My research is in optical physics, in particular, I am working on one of the challenges in implementing two photon excitation luorescence (TPEF) microscopy in imaging living systems. (TPEF) microscopy has been shown to be an invaluable tool for investigating biological structure and function in living organisms. The utility of (TPEF) imaging for this application arises from several important factors including it's ability to image deep within tissue, and to do so without harming the organism. Both of these advantages arise from the fact that (TPEF) imaging is done with

  18. Multifunctional scanning ion conductance microscopy

    Science.gov (United States)

    Page, Ashley; Unwin, Patrick R.

    2017-01-01

    Scanning ion conductance microscopy (SICM) is a nanopipette-based technique that has traditionally been used to image topography or to deliver species to an interface, particularly in a biological setting. This article highlights the recent blossoming of SICM into a technique with a much greater diversity of applications and capability that can be used either standalone, with advanced control (potential–time) functions, or in tandem with other methods. SICM can be used to elucidate functional information about interfaces, such as surface charge density or electrochemical activity (ion fluxes). Using a multi-barrel probe format, SICM-related techniques can be employed to deposit nanoscale three-dimensional structures and further functionality is realized when SICM is combined with scanning electrochemical microscopy (SECM), with simultaneous measurements from a single probe opening up considerable prospects for multifunctional imaging. SICM studies are greatly enhanced by finite-element method modelling for quantitative treatment of issues such as resolution, surface charge and (tip) geometry effects. SICM is particularly applicable to the study of living systems, notably single cells, although applications extend to materials characterization and to new methods of printing and nanofabrication. A more thorough understanding of the electrochemical principles and properties of SICM provides a foundation for significant applications of SICM in electrochemistry and interfacial science. PMID:28484332

  19. A spatio-temporally compensated acousto-optic scanner for two-photon microscopy providing large field of view.

    Science.gov (United States)

    Kremer, Y; Léger, J-F; Lapole, R; Honnorat, N; Candela, Y; Dieudonné, S; Bourdieu, L

    2008-07-07

    Acousto-optic deflectors (AOD) are promising ultrafast scanners for non-linear microscopy. Their use has been limited until now by their small scanning range and by the spatial and temporal dispersions of the laser beam going through the deflectors. We show that the use of AOD of large aperture (13mm) compared to standard deflectors allows accessing much larger field of view while minimizing spatio-temporal distortions. An acousto-optic modulator (AOM) placed at distance of the AOD is used to compensate spatial and temporal dispersions. Fine tuning of the AOM-AOD setup using a frequency-resolved optical gating (GRENOUILLE) allows elimination of pulse front tilt whereas spatial chirp is minimized thanks to the large aperture AOD.

  20. Optical nonlinearities of colloidal InP@ZnS core-shell quantum dots probed by Z-scan and two-photon excited emission

    International Nuclear Information System (INIS)

    Wawrzynczyk, Dominika; Szeremeta, Janusz; Samoc, Marek; Nyk, Marcin

    2015-01-01

    Spectrally resolved nonlinear optical properties of colloidal InP@ZnS core-shell quantum dots of various sizes were investigated with the Z-scan technique and two-photon fluorescence excitation method using a femtosecond laser system tunable in the range from 750 nm to 1600 nm. In principle, both techniques should provide comparable results and can be interchangeably used for determination of the nonlinear optical absorption parameters, finding maximal values of the cross sections and optimizing them. We have observed slight differences between the two-photon absorption cross sections measured by the two techniques and attributed them to the presence of non-radiative paths of absorption or relaxation. The most significant value of two-photon absorption cross section σ 2 for 4.3 nm size InP@ZnS quantum dot was equal to 2200 GM, while the two-photon excitation action cross section σ 2 Φ was found to be 682 GM at 880 nm. The properties of these cadmium-free colloidal quantum dots can be potentially useful for nonlinear bioimaging

  1. Scanning Tunneling Optical Resonance Microscopy

    Science.gov (United States)

    Bailey, Sheila; Wilt, Dave; Raffaelle, Ryne; Gennett, Tom; Tin, Padetha; Lau, Janice; Castro, Stephanie; Jenkins, Philip; Scheiman, Dave

    2003-01-01

    Scanning tunneling optical resonance microscopy (STORM) is a method, now undergoing development, for measuring optoelectronic properties of materials and devices on the nanoscale by means of a combination of (1) traditional scanning tunneling microscopy (STM) with (2) tunable laser spectroscopy. In STORM, an STM tip probing a semiconductor is illuminated with modulated light at a wavelength in the visible-to-near-infrared range and the resulting photoenhancement of the tunneling current is measured as a function of the illuminating wavelength. The photoenhancement of tunneling current occurs when the laser photon energy is sufficient to excite charge carriers into the conduction band of the semiconductor. Figure 1 schematically depicts a proposed STORM apparatus. The light for illuminating the semiconductor specimen at the STM would be generated by a ring laser that would be tunable across the wavelength range of interest. The laser beam would be chopped by an achromatic liquid-crystal modulator. A polarization-maintaining optical fiber would couple the light to the tip/sample junction of a commercial STM. An STM can be operated in one of two modes: constant height or constant current. A STORM apparatus would be operated in the constant-current mode, in which the height of the tip relative to the specimen would be varied in order to keep the tunneling current constant. In this mode, a feedback control circuit adjusts the voltage applied to a piezoelectric actuator in the STM that adjusts the height of the STM tip to keep the tunneling current constant. The exponential relationship between the tunneling current and tip-to-sample distance makes it relatively easy to implement this mode of operation. The choice of method by which the photoenhanced portion of the tunneling current would be measured depends on choice of the frequency at which the input illumination would be modulated (chopped). If the frequency of modulation were low enough (typically tunneling current

  2. Large scale serial two-photon microscopy to investigate local vascular changes in whole rodent brain models of Alzheimer's disease

    Science.gov (United States)

    Delafontaine-Martel, P.; Lefebvre, J.; Damseh, R.; Castonguay, A.; Tardif, P.; Lesage, F.

    2018-02-01

    In this study, an automated serial two-photon microscope was used to image a fluorescent gelatin filled rodent's brain in 3D. A method to compute vascular density using automatic segmentation was combined with coregistration techniques to build group-level vasculature metrics. By studying the medial prefrontal cortex and the hippocampal formation of 3 age groups (2, 4.5 and 8 months old), we compared vascular density for both WT and an Alzheimer model transgenic brain (APP/PS1). We observe a loss of vascular density caused by the ageing process and we propose further analysis to confirm our results.

  3. Vacuum scanning capillary photoemission microscopy.

    Science.gov (United States)

    Aseyev, S A; Cherkun, A P; Mironov, B N; Petrunin, V V; Chekalin, S V

    2017-08-01

    We demonstrate the use of a conical capillary in a scanning probe microscopy for surface analysis. The probe can measure photoemission from a substrate by transmitting photoelectrons along the capillary as a function of probe position. The technique is demonstrated on a model substrate consisting of a gold reflecting layer on a compact disc which has been illuminated by an unfocused laser beam with a wavelength 400nm, from a femtosecond laser with a beam size of 4mm. A quartz capillary with a 2-µm aperture has been used in the experiments. The period of gold microstructure, shown to be 1.6µ, was measured by the conical probe operating in shear force mode. In shear force regime, the dielectric capillary has been used as a "classical" SPM tip, which provided images reflecting the surface topology. In a photoelectron regime photoelectrons passed through hollow tip and entered a detector. The spatial distribution of the recorded photoelectrons consisted of periodic mountain-valley strips, resembling the surface profile of the sample. Submicron spatial resolution has been achieved. This approach paves the way to study pulsed photodesorption of large organic molecular ions with high spatial and element resolution using the combination of a hollow-tip scanner with time-of-flight technique. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Details of the Collagen and Elastin Architecture in the Human Limbal Conjunctiva, Tenon's Capsule and Sclera Revealed by Two-Photon Excited Fluorescence Microscopy.

    Science.gov (United States)

    Park, Choul Yong; Marando, Catherine M; Liao, Jason A; Lee, Jimmy K; Kwon, Jiwon; Chuck, Roy S

    2016-10-01

    To investigate the architecture and distribution of collagen and elastin in human limbal conjunctiva, Tenon's capsule, and sclera. The limbal conjunctiva, Tenon's capsule, and sclera of human donor corneal buttons were imaged with an inverted two-photon excited fluorescence microscope. No fixation process was necessary. The laser (Ti:sapphire) was tuned at 850 nm for two-photon excitation. Backscatter signals of second harmonic generation (SHG) and autofluorescence (AF) were collected through a 425/30-nm and a 525/45-nm emission filter, respectively. Multiple, consecutive, and overlapping (z-stack) images were acquired. Collagen signals were collected with SHG, whereas elastin signals were collected with AF. The size and density of collagen bundles varied widely depending on depth: increasing from conjunctiva to sclera. In superficial image planes, collagen bundles were image planes (episclera and superficial sclera), collagen bundles were thicker (near 100 μm in width) and densely packed. Comparatively, elastin fibers were thinner and sparse. The orientation of elastin fibers was independent of collagen fibers in superficial layers; but in deep sclera, elastin fibers wove through collagen interbundle gaps. At the limbus, both collagen and elastin fibers were relatively compact and were distributed perpendicular to the limbal annulus. Two-photon excited fluorescence microscopy has enabled us to understand in greater detail the collagen and elastin architecture of the human limbal conjunctiva, Tenon's capsule, and sclera.

  5. A fully-automated multiscale kernel graph cuts based particle localization scheme for temporal focusing two-photon microscopy

    Science.gov (United States)

    Huang, Xia; Li, Chunqiang; Xiao, Chuan; Sun, Wenqing; Qian, Wei

    2017-03-01

    The temporal focusing two-photon microscope (TFM) is developed to perform depth resolved wide field fluorescence imaging by capturing frames sequentially. However, due to strong nonignorable noises and diffraction rings surrounding particles, further researches are extremely formidable without a precise particle localization technique. In this paper, we developed a fully-automated scheme to locate particles positions with high noise tolerance. Our scheme includes the following procedures: noise reduction using a hybrid Kalman filter method, particle segmentation based on a multiscale kernel graph cuts global and local segmentation algorithm, and a kinematic estimation based particle tracking method. Both isolated and partial-overlapped particles can be accurately identified with removal of unrelated pixels. Based on our quantitative analysis, 96.22% isolated particles and 84.19% partial-overlapped particles were successfully detected.

  6. Two-Photon Fluorescence Microscopy for Determination of the Riboflavin Concentration in the Anterior Corneal Stroma When Using the Dresden Protocol.

    Science.gov (United States)

    Seiler, Theo G; Ehmke, Tobias; Fischinger, Isaak; Zapp, Daniel; Stachs, Oliver; Seiler, Theo; Heisterkamp, Alexander

    2015-10-01

    To determine the riboflavin concentration gradient in the anterior corneal stroma when using the Dresden protocol with different dextran solutions. Three different groups of porcine corneas, five each, were compared regarding the riboflavin concentration in the anterior stroma. Before all experiments, stable hydration conditions were established for the corresponding solution. All groups were treated with 0.1% riboflavin in different dextran solutions (15%, 16%, 20%). After imbibition, two-photon microscopy was used to determine fluorescence intensity. For signal attenuation and concentration determination corneas were saturated and measured a second time by two-photon microscopy. Additionally, the distribution was calculated mathematically and compared to the empiric results. Riboflavin concentration is decreasing with depth for all dextran solutions. A nearly constant concentration could be determined over the first 75 μm. Analysis of the fit functions leads to diffusion coefficients of D = 2.97 × 10-7 cm2/s for the 15% dextran solution, D = 2.34 × 10-7 cm2/s for the 16% dextran solution, and D = 1.28 × 10-7 cm2/s for the 20% dextran solution. The riboflavin gradients of the 20% dextran group were statistically significantly different from 15% dextran starting at a depth of 220 μm and deeper (P = 0.047). The 16% dextran group differed statistically at a depth of 250 μm and deeper (P = 0.047). These results show a significant difference to those published previously. With correct settings two-photon microscopy is a precise way to determine the concentration of riboflavin in cornea. The measured gradient is excellently fit by a Gaussian distribution, which comes out as a solution of Fick's second law.

  7. Two-photon time-lapse microscopy of BODIPY-cholesterol reveals anomalous sterol diffusion in chinese hamster ovary cells

    Directory of Open Access Journals (Sweden)

    Lund Frederik W

    2012-10-01

    Full Text Available Abstract Background Cholesterol is an important membrane component, but our knowledge about its transport in cells is sparse. Previous imaging studies using dehydroergosterol (DHE, an intrinsically fluorescent sterol from yeast, have established that vesicular and non-vesicular transport modes contribute to sterol trafficking from the plasma membrane. Significant photobleaching, however, limits the possibilities for in-depth analysis of sterol dynamics using DHE. Co-trafficking studies with DHE and the recently introduced fluorescent cholesterol analog BODIPY-cholesterol (BChol suggested that the latter probe has utility for prolonged live-cell imaging of sterol transport. Results We found that BChol is very photostable under two-photon (2P-excitation allowing the acquisition of several hundred frames without significant photobleaching. Therefore, long-term tracking and diffusion measurements are possible. Two-photon temporal image correlation spectroscopy (2P-TICS provided evidence for spatially heterogeneous diffusion constants of BChol varying over two orders of magnitude from the cell interior towards the plasma membrane, where D ~ 1.3 μm2/s. Number and brightness (N&B analysis together with stochastic simulations suggest that transient partitioning of BChol into convoluted membranes slows local sterol diffusion. We observed sterol endocytosis as well as fusion and fission of sterol-containing endocytic vesicles. The mobility of endocytic vesicles, as studied by particle tracking, is well described by a model for anomalous subdiffusion on short time scales with an anomalous exponent α ~ 0.63 and an anomalous diffusion constant of Dα = 1.95 x 10-3 μm2/sα. On a longer time scale (t > ~5 s, a transition to superdiffusion consistent with slow directed transport with an average velocity of v ~ 6 x 10-3 μm/s was observed. We present an analytical model that bridges the two regimes and fit this model to vesicle

  8. Scanning Capacitance Microscopy | Materials Science | NREL

    Science.gov (United States)

    obtained using scanning capacitance microscopy. Top Right: Image of p-type and n-type material, obtained 'fingers' of light-colored n-type material on a yellow and blue background representing p-type material material, obtained using scanning capacitance microscopy, in a sample semiconductor device; the image shows

  9. Measurement of polarization dependence of two-photon absorption coefficient in InP using extended Z-scan technique for thick materials

    Science.gov (United States)

    Oishi, Masaki; Shinozaki, Tomohisa; Hara, Hikaru; Yamamoto, Kazunuki; Matsusue, Toshio; Bando, Hiroyuki

    2018-03-01

    The two-photon absorption coefficient β in InP has been measured in the wavelength range of 1640 to 1800 nm by the Z-scan technique in relatively thick materials. The values of β have been evaluated from the fit to the equation including the spatial and temporal profiles of the focused Gaussian beam. The polarization dependence of β has also been measured. The dependence has been expressed very well by the expression of β with the imaginary part of the third-order nonlinear susceptibility tensor χ(3).

  10. 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.

  11. 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.

  12. Scanning vector Hall probe microscopy

    International Nuclear Information System (INIS)

    Cambel, V.; Gregusova, D.; Fedor, J.; Kudela, R.; Bending, S.J.

    2004-01-01

    We have developed a scanning vector Hall probe microscope for mapping magnetic field vector over magnetic samples. The microscope is based on a micromachined Hall sensor and the cryostat with scanning system. The vector Hall sensor active area is ∼5x5 μm 2 . It is realized by patterning three Hall probes on the tilted faces of GaAs pyramids. Data from these 'tilted' Hall probes are used to reconstruct the full magnetic field vector. The scanning area of the microscope is 5x5 mm 2 , space resolution 2.5 μm, field resolution ∼1 μT Hz -1/2 at temperatures 10-300 K

  13. Scanning probe microscopy competency development

    Energy Technology Data Exchange (ETDEWEB)

    Hawley, M.E.; Reagor, D.W.; Jia, Quan Xi [and others

    1998-12-31

    This is the final report of a three-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). The project collaborators developed an ultra-high vacuum scanning tunneling microscope (UHV-STM) capability, integrated it with existing scanning probe microscopes, and developed new, advanced air-based scanning force techniques (SPMs). Programmatic, basic, and industrially related laboratory research requires the existence of SPMs, as well as expertise capable of providing local nano-scale information. The UHV-STM capability, equipped with load-lock system and several surface science techniques, will allow introduction, examination, and reaction of surfaces prepared under well-controlled vacuum conditions, including the examination of morphology and local bonding associated with the initial stages of film growth under controlled growth conditions. The resulting capabilities will enable the authors to respond to a variety of problems requiring local characterization of conducting and nonconducting surfaces in liquids, air, and UHV.

  14. Introduction to scanning tunneling microscopy

    CERN Document Server

    Chen, C Julian

    2008-01-01

    The scanning tunneling and the atomic force microscope, both capable of imaging individual atoms, were crowned with the Physics Nobel Prize in 1986, and are the cornerstones of nanotechnology today. This is a thoroughly updated version of this 'bible' in the field.

  15. Uniform silica nanoparticles encapsulating two-photon absorbing fluorescent dye

    International Nuclear Information System (INIS)

    Wu Weibing; Liu Chang; Wang Mingliang; Huang Wei; Zhou Shengrui; Jiang Wei; Sun Yueming; Cui Yiping; Xu Chunxinag

    2009-01-01

    We have prepared uniform silica nanoparticles (NPs) doped with a two-photon absorbing zwitterionic hemicyanine dye by reverse microemulsion method. Obvious solvatochromism on the absorption spectra of dye-doped NPs indicates that solvents can partly penetrate into the silica matrix and then affect the ground and excited state of dye molecules. For dye-doped NP suspensions, both one-photon and two-photon excited fluorescence are much stronger and recorded at shorter wavelength compared to those of free dye solutions with comparative overall dye concentration. This behavior is possibly attributed to the restricted twisted intramolecular charge transfer (TICT), which reduces fluorescence quenching when dye molecules are trapped in the silica matrix. Images from two-photon laser scanning fluorescence microscopy demonstrate that the dye-doped silica NPs can be actively uptaken by Hela cells with low cytotoxicity. - Graphical abstract: Water-soluble silica NPs doped with a two-photon absorbing zwitterionic hemicyanine dye were prepared. They were found of enhanced one-photon and two-photon excited fluorescence compared to free dye solutions. Images from two-photon laser scanning fluorescence microscopy demonstrate that the dye-doped silica NPs can be actively uptaken by Hela cells.

  16. Measurement of cortical functional activation in awake mice using two-photon microscopy and a novel pO2-sensitive probe(Conference Presentation)

    Science.gov (United States)

    Sencan, Ikbal; Esipova, Tatiana V.; Kilic, Kivilcim; Li, Baoqiang; Desjardins, Michèle; Yaseen, Mohammad A.; Wang, Hui; Jaswal, Rajeshwer S.; Kura, Sreekanth; Fu, Buyin; Boas, David A.; Devor, Anna; Sakadžić, Sava; Vinogradov, Sergei A.

    2017-02-01

    We characterized cortical microvascular PO2 and blood flow changes in response to whisker stimulation in awake mice. The measurements were performed by combining two-photon microscopy imaging of the cortical oxygenation and optical coherence tomography imaging of the cerebral blood flow. In order to perform fast spatio-temporally resolved measurements of PO2, we used a newly-developed oxygen-sensitive probe PtG-2P, which has significantly higher brightness than the established two-photon-enhanced oxygen sensor PtP-C343. We characterized the performance of the new probe in vivo and mapped the amplitudes and shapes (e.g. initial dip, overshoot, and post stimulus undershoot) of the PO2 changes as a function of the vessel type (e.g., arterioles, capillaries, and venules) and a distance from the activation center. The measurements in the awake mice are not affected by the confounding factors of anesthesia on the animal physiology, including the level of cerebral metabolism and the amplitude and speed of neuronal and vascular responses. Our results will help to understand changes in oxygenation and blood flow on the cortical microvascular scale, will lead to improved understanding of the cerebral physiology, pathophysiology and will improve quantitative interpretation of fMRI signals.

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

    Science.gov (United States)

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

    2013-01-01

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

  18. Effect of detergents on the physico-chemical properties of skin stratum corneum: A two-photon excitation fluorescence microscopy study

    DEFF Research Database (Denmark)

    Bloksgaard, Maria; Brewer, Jonathan R.; Pashkovski, Eugene

    2014-01-01

    OBJECTIVE: Understanding the structural and dynamical features of skin is critical for advancing innovation in personal care and drug discovery. Synthetic detergent mixtures used in commercially available body wash products are thought to be less aggressive towards the skin barrier when compared...... to conventional detergents. The aim of this work is to comparatively characterize the effect of a mild synthetic cleanser mixture (SCM) and sodium dodecyl sulphate (SDS) on the hydration state of the intercellular lipid matrix and on proton activity of excised skin stratum corneum (SC). METHOD: Experiments were...... performed using two-photon excitation fluorescence microscopy. Fluorescent images of fluorescence reporters sensitive to proton activity and hydration of SC were obtained in excised skin and examined in presence and absence of SCM and SDS detergents. RESULTS: Hydration of the intercellular lipid matrix...

  19. Time-resolved scanning tunnelling microscopy

    NARCIS (Netherlands)

    van Houselt, Arie; Zandvliet, Henricus J.W.

    2010-01-01

    Scanning tunneling microscopy has revolutionized our ability to image, study, and manipulate solid surfaces on the size scale of atoms. One important limitation of the scanning tunneling microscope (STM) is, however, its poor time resolution. Recording a standard image with a STM typically takes

  20. Monochromated scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Rechberger, W.; Kothleitner, G.; Hofer, F.

    2006-01-01

    Full text: Electron energy-loss spectroscopy (EELS) has developed into an established technique for chemical and structural analysis of thin specimens in the (scanning) transmission electron microscope (S)TEM. The energy resolution in EELS is largely limited by the stability of the high voltage supply, by the resolution of the spectrometer and by the energy spread of the source. To overcome this limitation a Wien filter monochromator was recently introduced with commercially available STEMs, offering the advantage to better resolve EELS fine structures, which contain valuable bonding information. The method of atomic resolution Z-contrast imaging within an STEM, utilizing a high-angle annular dark-field (HAADF) detector can perfectly complement the excellent energy resolution, since EELS spectra can be collected simultaneously. In combination with a monochromator microscope not only high spatial resolution images can be recorded but also high energy resolution EELS spectra are attainable. In this work we investigated the STEM performance of a 200 kV monochromated Tecnai F20 with a high resolution Gatan Imaging Filter (HR-GIF). (author)

  1. Scanning electron microscopy of bone.

    Science.gov (United States)

    Boyde, Alan

    2012-01-01

    This chapter described methods for Scanning Electron Microscopical imaging of bone and bone cells. Backscattered electron (BSE) imaging is by far the most useful in the bone field, followed by secondary electrons (SE) and the energy dispersive X-ray (EDX) analytical modes. This chapter considers preparing and imaging samples of unembedded bone having 3D detail in a 3D surface, topography-free, polished or micromilled, resin-embedded block surfaces, and resin casts of space in bone matrix. The chapter considers methods for fixation, drying, looking at undersides of bone cells, and coating. Maceration with alkaline bacterial pronase, hypochlorite, hydrogen peroxide, and sodium or potassium hydroxide to remove cells and unmineralised matrix is described in detail. Attention is given especially to methods for 3D BSE SEM imaging of bone samples and recommendations for the types of resin embedding of bone for BSE imaging are given. Correlated confocal and SEM imaging of PMMA-embedded bone requires the use of glycerol to coverslip. Cathodoluminescence (CL) mode SEM imaging is an alternative for visualising fluorescent mineralising front labels such as calcein and tetracyclines. Making spatial casts from PMMA or other resin embedded samples is an important use of this material. Correlation with other imaging means, including microradiography and microtomography is important. Shipping wet bone samples between labs is best done in glycerol. Environmental SEM (ESEM, controlled vacuum mode) is valuable in eliminating -"charging" problems which are common with complex, cancellous bone samples.

  2. Scanning probe microscopy experiments in microgravity

    International Nuclear Information System (INIS)

    Drobek, Tanja; Reiter, Michael; Heckl, Wolfgang M.

    2004-01-01

    The scanning probe microscopy setups are small, lightweight and do not require vacuum or high voltage supply. In addition, samples can be investigated directly without further preparation. Therefore, these techniques are well-suited for applications in space, in particular, for operation on the International Space Station (ISS) or for high resolution microscopy on planetary missions. A feasibility study for a scanning tunneling microscopy setup was carried out on a parabolic flight campaign in November 2001 in order to test the technical setup for microgravity applications. With a pocket-size design microscope, a graphite surface was imaged under ambient conditions. Atomic resolution was achieved although the quality of the images was inferior in comparison to laboratory conditions. Improvements for future scanning probe microscopy experiments in microgravity are suggested

  3. Label-free imaging of brain and brain tumor specimens with combined two-photon excited fluorescence and second harmonic generation microscopy

    Science.gov (United States)

    Jiang, Liwei; Wang, Xingfu; Wu, Zanyi; Du, Huiping; Wang, Shu; Li, Lianhuang; Fang, Na; Lin, Peihua; Chen, Jianxin; Kang, Dezhi; Zhuo, Shuangmu

    2017-10-01

    Label-free imaging techniques are gaining acceptance within the medical imaging field, including brain imaging, because they have the potential to be applied to intraoperative in situ identifications of pathological conditions. In this paper, we describe the use of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) microscopy in combination for the label-free detection of brain and brain tumor specimens; gliomas. Two independently detecting channels were chosen to subsequently collect TPEF/SHG signals from the specimen to increase TPEF/SHG image contrasts. Our results indicate that the combined TPEF/SHG microscopic techniques can provide similar rat brain structural information and produce a similar resolution like conventional H&E staining in neuropathology; including meninges, cerebral cortex, white-matter structure corpus callosum, choroid plexus, hippocampus, striatum, and cerebellar cortex. It can simultaneously detect infiltrating human brain tumor cells, the extracellular matrix collagen fiber of connective stroma within brain vessels and collagen depostion in tumor microenvironments. The nuclear-to-cytoplasmic ratio and collagen content can be extracted as quantitative indicators for differentiating brain gliomas from healthy brain tissues. With the development of two-photon fiberscopes and microendoscope probes and their clinical applications, the combined TPEF and SHG microcopy may become an important multimodal, nonlinear optical imaging approach for real-time intraoperative histological diagnostics of residual brain tumors. These occur in various brain regions during ongoing surgeries through the method of simultaneously identifying tumor cells, and the change of tumor microenvironments, without the need for the removal biopsies and without the need for tissue labelling or fluorescent markers.

  4. Sample preparation method for scanning force microscopy

    CERN Document Server

    Jankov, I R; Szente, R N; Carreno, M N P; Swart, J W; Landers, R

    2001-01-01

    We present a method of sample preparation for studies of ion implantation on metal surfaces. The method, employing a mechanical mask, is specially adapted for samples analysed by Scanning Force Microscopy. It was successfully tested on polycrystalline copper substrates implanted with phosphorus ions at an acceleration voltage of 39 keV. The changes of the electrical properties of the surface were measured by Kelvin Probe Force Microscopy and the surface composition was analysed by Auger Electron Spectroscopy.

  5. Two-photon physics

    International Nuclear Information System (INIS)

    Bardeen, W.A.

    1981-10-01

    A new experimental frontier has recently been opened to the study of two photon processes. The first results of many aspects of these reactions are being presented at this conference. In contrast, the theoretical development of research ito two photon processes has a much longer history. This talk reviews the many different theoretical ideas which provide a detailed framework for our understanding of two photon processes

  6. Electrochemical gating in scanning electrochemical microscopy

    NARCIS (Netherlands)

    Ahonen, P.; Ruiz, V.; Kontturi, K.; Liljeroth, P.; Quinn, B.M.

    2008-01-01

    We demonstrate that scanning electrochemical microscopy (SECM) can be used to determine the conductivity of nanoparticle assemblies as a function of assembly potential. In contrast to conventional electron transport measurements, this method is unique in that electrical connection to the film is not

  7. Quantifying the microvascular origin of BOLD-fMRI from first principles with two-photon microscopy and an oxygen-sensitive nanoprobe.

    Science.gov (United States)

    Gagnon, Louis; Sakadžić, Sava; Lesage, Frédéric; Musacchia, Joseph J; Lefebvre, Joël; Fang, Qianqian; Yücel, Meryem A; Evans, Karleyton C; Mandeville, Emiri T; Cohen-Adad, Jülien; Polimeni, Jonathan R; Yaseen, Mohammad A; Lo, Eng H; Greve, Douglas N; Buxton, Richard B; Dale, Anders M; Devor, Anna; Boas, David A

    2015-02-25

    The blood oxygenation level-dependent (BOLD) contrast is widely used in functional magnetic resonance imaging (fMRI) studies aimed at investigating neuronal activity. However, the BOLD signal reflects changes in blood volume and oxygenation rather than neuronal activity per se. Therefore, understanding the transformation of microscopic vascular behavior into macroscopic BOLD signals is at the foundation of physiologically informed noninvasive neuroimaging. Here, we use oxygen-sensitive two-photon microscopy to measure the BOLD-relevant microvascular physiology occurring within a typical rodent fMRI voxel and predict the BOLD signal from first principles using those measurements. The predictive power of the approach is illustrated by quantifying variations in the BOLD signal induced by the morphological folding of the human cortex. This framework is then used to quantify the contribution of individual vascular compartments and other factors to the BOLD signal for different magnet strengths and pulse sequences. Copyright © 2015 the authors 0270-6474/15/353663-13$15.00/0.

  8. Three-dimensional cell organization leads to almost immediate HRE activity as demonstrated by molecular imaging of MG-63 spheroids using two-photon excitation microscopy.

    Science.gov (United States)

    Indovina, Paola; Collini, Maddalena; Chirico, Giuseppe; Santini, Maria Teresa

    2007-02-20

    Hypoxia through HRE (hypoxia-responsive element) activity in MG-63 human osteosarcoma cells grown in monolayer and as very small, three-dimensional tumor spheroids was investigated using molecular imaging techniques. MG-63 cells were stably transfected with a vector constructed with multiple copies of the HRE sequence of the human vascular endothelial growth factor (VEGF) gene and with the enhanced green fluorescent protein (EGFP) coding sequence. During hypoxia when HIF-1alpha (hypoxia-inducible factor-1alpha) is stabilized, the binding of HIF-1 to the HRE sequences of the vector allows the transcription of EGFP and the appearance of fluorescence. Transfected monolayer cells were characterized by flow cytometric analysis in response to various hypoxic conditions and HIF-1alpha expression in these cells was assessed by Western blotting. Two-photon excitation (TPE) microscopy was then used to examine both MG-63-transfected monolayer cells and spheroids at 2 and 5 days of growth in normoxic conditions. Monolayer cells reveal almost no fluorescence, whereas even very small spheroids (HRE activation. This activation of the HRE sequences, which control a wide variety of genes, suggests that monolayer cells and spheroids of the MG-63 cell line have different genes activated and thus diverse functional activities.

  9. New directions in scanning-tunneling microscopy

    International Nuclear Information System (INIS)

    Ferrell, T.L.; Warmack, R.J.; Reddick, R.C.

    1989-01-01

    The tunneling of electrons in scanning-tunneling microscopy (STM) has permitted imaging of the electronic distribution about individual atoms on surfaces. The need for use of conducting surfaces in STM limits its applicability, and new forms of scanning microscopy have emerged as a result of interest in poorly conducting samples. Atomic force microscopy has demonstrated that the force between a surface and a probe tip can be used to image selected materials. Now being developed are magnetic probe STM's and photon tunneling microscopes in which the probe is a sharpened optical fiber. Also of great interest presently is the measurement of differential conductance of surfaces using electron STM's. This method supplies spectral information and contrast enhancement in images. At present there remains much theoretical work to be carried out in order to better characterize related data on inelastic electron tunneling, and valuable insight may be gained from data being gathered on the local work function of materials. As matters stand today, the key problems lie in determining tip and contamination effects, preparation of samples, and understanding conductivity mechanisms in very thin materials on conducting substrates. Resolution of these problems and introduction of new forms of scanning microscopy may permit novel and important applications in biology as well as surface science

  10. Developments in Scanning Hall Probe Microscopy

    Science.gov (United States)

    Chouinard, Taras; Chu, Ricky; David, Nigel; Broun, David

    2009-05-01

    Low temperature scanning Hall probe microscopy is a sensitive means of imaging magnetic structures with high spatial resolution and magnetic flux sensitivity approaching that of a Superconducting Quantum Interference Device. We have developed a scanning Hall probe microscope with novel features, including highly reliable coarse positioning, in situ optimization of sensor-sample alignment and capacitive transducers for linear, long range positioning measurement. This has been motivated by the need to reposition accurately above fabricated nanostructures such as small superconducting rings. Details of the design and performance will be presented as well as recent progress towards time-resolved measurements with sub nanosecond resolution.

  11. Scanning thermal microscopy of thermoelectric nanostructures

    Czech Academy of Sciences Publication Activity Database

    Vaniš, Jan; Zelinka, Jiří; Zeipl, Radek; Jelínek, Miroslav; Kocourek, Tomáš; Remsa, Jan; Navrátil, Jiří

    2016-01-01

    Roč. 45, č. 3 (2016), s. 1734-1739 ISSN 0361-5235 R&D Projects: GA ČR(CZ) GA15-05864S; GA ČR(CZ) GA13-33056S Institutional support: RVO:68378271 ; RVO:61389013 Keywords : thermoelectric layer * scanning thermal microscopy * pulsed laser deposition * laser deposition * secondary ion mass spectrometry Subject RIV: BM - Solid Matter Physics ; Magnetism; CA - Inorganic Chemistry (UMCH-V) Impact factor: 1.579, year: 2016

  12. Scanning electron microscopy of superficial white onychomycosis*

    Science.gov (United States)

    de Almeida Jr., Hiram Larangeira; Boabaid, Roberta Oliveira; Timm, Vitor; Silva, Ricardo Marques e; de Castro, Luis Antonio Suita

    2015-01-01

    Superficial white onychomycosis is characterized by opaque, friable, whitish superficial spots on the nail plate. We examined an affected halux nail of a 20-year-old male patient with scanning electron microscopy. The mycological examination isolated Trichophyton mentagrophytes. Abundant hyphae with the formation of arthrospores were found on the nail's surface, forming small fungal colonies. These findings showed the great capacity for dissemination of this form of onychomycosis. PMID:26560225

  13. Scanning electron microscopy of primary bone tumors

    International Nuclear Information System (INIS)

    Pool, R.R.; Kerner, B.

    1975-01-01

    Critical-point-drying of tumor tissue fixed in a glutaraldehyde-paraformaldehyde solution and viewed by scanning electron microscopy (SEM) provides a 3-dimensional view of tumor cells and their matrices. This report describes the SEM appearance of three primary bone tumors: a canine osteosarcoma of the distal radius, a feline chondrosarcoma of the proximal tibia and a canine fibrosarcoma of the proximal humerus. The ultrastructural morphology is compared with the histologic appearance of each tumor

  14. Spin-polarized scanning electron microscopy

    International Nuclear Information System (INIS)

    Kohashi, Teruo

    2014-01-01

    Spin-Polarized Scanning Electron Microscopy (Spin SEM) is one way for observing magnetic domain structures taking advantage of the spin polarization of the secondary electrons emitted from a ferromagnetic sample. This principle brings us several excellent capabilities such as high-spatial resolution better than 10 nm, and analysis of magnetization direction in three dimensions. In this paper, the principle and the structure of the spin SEM is briefly introduced, and some examples of the spin SEM measurements are shown. (author)

  15. Scanning Ion Conductance Microscopy of Live Keratinocytes

    International Nuclear Information System (INIS)

    Hegde, V; Mason, A; Saliev, T; Smith, F J D; McLean, W H I; Campbell, P A

    2012-01-01

    Scanning ion conductance microscopy (SICM) is perhaps the least well known technique from the scanning probe microscopy (SPM) family of instruments. As with its more familiar counterpart, atomic force microscopy (AFM), the technique provides high-resolution topographic imaging, with the caveat that target structures must be immersed in a conducting solution so that a controllable ion current may be utilised as the basis for feedback. In operation, this non-contact characteristic of SICM makes it ideal for the study of delicate structures, such as live cells. Moreover, the intrinsic architecture of the instrument, incorporating as it does, a scanned micropipette, lends itself to combination approaches with complementary techniques such as patch-clamp electrophysiology: SICM therefore boasts the capability for both structural and functional imaging. For the present observations, an ICnano S system (Ionscope Ltd., Melbourn, UK) operating in 'hopping mode' was used, with the objective of assessing the instrument's utility for imaging live keratinocytes under physiological buffers. In scans employing cultured HaCaT cells (spontaneously immortalised, human keratinocytes), we compared the qualitative differences of live cells imaged with SICM and AFM, and also with their respective counterparts after chemical fixation in 4% paraformaldehyde. Characteristic surface microvilli were particularly prominent in live cell imaging by SICM. Moreover, time lapse SICM imaging on live cells revealed that changes in the pattern of microvilli could be tracked over time. By comparison, AFM imaging on live cells, even at very low contact forces (< nN), could not routinely image microvilli: rather, an apparently convolved image of the underlying cytoskeleton was instead prevalent. We note that the present incarnation of the commercial instrument falls some way behind the market leading SPMs in terms of technical prowess and scanning speed, however, the intrinsic non-obtrusive nature of

  16. Scanning Near-Field Optical Microscopy

    Directory of Open Access Journals (Sweden)

    Dušan Vobornik

    2008-02-01

    Full Text Available An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today’s science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution.

  17. Scanning near-field optical microscopy.

    Science.gov (United States)

    Vobornik, Dusan; Vobornik, Slavenka

    2008-02-01

    An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today's science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM) is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution.

  18. Scanning electron microscopy of coal macerals

    Energy Technology Data Exchange (ETDEWEB)

    Davis, M.R.; White, A.; Deegan, M.D.

    1986-02-01

    Individual macerals separated from some United Kingdom coals of Carboniferous age and bituminous rank were examined by scanning electron microscopy. In each case a specific morphology characteristic of the macerals studied could be recognized. Collinite (a member of the vitrinite maceral group) was recognizable in all samples by its angular shape and characteristic fracture patterns, the particles (30-200 ..mu..m) frequently showing striated or laminated surface. Sporinite particles had no well defined shape and were associated with more detrital material than were the other macerals studied. This detritus was shown by conventional light microscopy to be the maceral micrinite. Fusinite was remarkable in having a chunky needle form, with lengths of up to 200 ..mu..m. 8 references.

  19. Scanning electron microscopy of semiconductor materials

    International Nuclear Information System (INIS)

    Bresse, J.F.; Dupuy, M.

    1978-01-01

    The use of scanning electron microscopy in semiconductors opens up a large field of use. The operating modes lending themselves to the study of semiconductors are the induced current, cathodoluminescence and the use of the potential contrast which can also be applied very effectively to the study of the devices (planar in particular). However, a thorough knowledge of the mechanisms of the penetration of electrons, generation and recombination of generated carriers in a semiconductor is necessary in order to attain a better understanding of the operating modes peculiar to semiconductors [fr

  20. Investigation into scanning tunnelling luminescence microscopy

    International Nuclear Information System (INIS)

    Manson-Smith, S.K.

    2001-01-01

    This work reports on the development of a scanning tunnelling luminescence (STL) microscope and its application to the study of Ill-nitride semiconductor materials used in the production of light emitting devices. STL microscopy is a technique which uses the high resolution topographic imaging capabilities of the scanning tunnelling microscope (STM) to generate high resolution luminescence images. The STM tunnelling current acts as a highly localised source of electrons (or holes) which generates luminescence in certain materials. Light generated at the STM tunnelling junction is collected concurrently with the height variation of the tunnelling probe as it is scanned across a sample surface, producing simultaneous topographic and luminescence images. Due to the very localised excitation source, high resolution luminescence images can be obtained. Spectroscopic resolution can be obtained by using filters. Additionally, the variation of luminescence intensity with tunnel current and with bias voltage can provide information on recombination processes and material properties. The design and construction of a scanning tunnelling luminescence microscope is described in detail. Operating under ambient conditions, the microscope has several novel features, including a new type of miniature inertial slider-based approach motor, large solid-angle light collection optical arrangement and a tip-height regulation system which requires the minimum of operator input. (author)

  1. Automated evaluation of liver fibrosis in thioacetamide, carbon tetrachloride, and bile duct ligation rodent models using second-harmonic generation/two-photon excited fluorescence microscopy.

    Science.gov (United States)

    Liu, Feng; Chen, Long; Rao, Hui-Ying; Teng, Xiao; Ren, Ya-Yun; Lu, Yan-Qiang; Zhang, Wei; Wu, Nan; Liu, Fang-Fang; Wei, Lai

    2017-01-01

    Animal models provide a useful platform for developing and testing new drugs to treat liver fibrosis. Accordingly, we developed a novel automated system to evaluate liver fibrosis in rodent models. This system uses second-harmonic generation (SHG)/two-photon excited fluorescence (TPEF) microscopy to assess a total of four mouse and rat models, using chemical treatment with either thioacetamide (TAA) or carbon tetrachloride (CCl 4 ), and a surgical method, bile duct ligation (BDL). The results obtained by the new technique were compared with that using Ishak fibrosis scores and two currently used quantitative methods for determining liver fibrosis: the collagen proportionate area (CPA) and measurement of hydroxyproline (HYP) content. We show that 11 shared morphological parameters faithfully recapitulate Ishak fibrosis scores in the models, with high area under the receiver operating characteristic (ROC) curve (AUC) performance. The AUC values of 11 shared parameters were greater than that of the CPA (TAA: 0.758-0.922 vs 0.752-0.908; BDL: 0.874-0.989 vs 0.678-0.966) in the TAA mice and BDL rat models and similar to that of the CPA in the TAA rat and CCl 4 mouse models. Similarly, based on the trends in these parameters at different time points, 9, 10, 7, and 2 model-specific parameters were selected for the TAA rats, TAA mice, CCl 4 mice, and BDL rats, respectively. These parameters identified differences among the time points in the four models, with high AUC accuracy, and the corresponding AUC values of these parameters were greater compared with those of the CPA in the TAA rat and mouse models (rats: 0.769-0.894 vs 0.64-0.799; mice: 0.87-0.93 vs 0.739-0.836) and similar to those of the CPA in the CCl 4 mouse and BDL rat models. Similarly, the AUC values of 11 shared parameters and model-specific parameters were greater than those of HYP in the TAA rats, TAA mice, and CCl 4 mouse models and were similar to those of HYP in the BDL rat models. The automated

  2. Tale of two photons

    International Nuclear Information System (INIS)

    Anon.

    1994-01-01

    A very profitable spinoff from electron- positron collisions is two-photon physics. Rather than the electron and positron interacting directly via an exchanged photon, two virtual (transient) photons, one from each particle, get tangled up. With new electron-positron colliders appearing on the scene, a topical meeting on two-photon physics - 'From DAPHNE to LEP 200 and beyond' - held from 2-4 February in Paris, in the premises of the Ministry of Higher Education and Research, was particularly timely. Some 60 physicists, both experimentalists and theorists, participated, with some thirty speakers

  3. Video-rate resonant scanning multiphoton microscopy

    Science.gov (United States)

    Kirkpatrick, Nathaniel D.; Chung, Euiheon; Cook, Daniel C.; Han, Xiaoxing; Gruionu, Gabriel; Liao, Shan; Munn, Lance L.; Padera, Timothy P.; Fukumura, Dai; Jain, Rakesh K.

    2013-01-01

    The abnormal tumor microenvironment fuels tumor progression, metastasis, immune suppression, and treatment resistance. Over last several decades, developments in and applications of intravital microscopy have provided unprecedented insights into the dynamics of the tumor microenvironment. In particular, intravital multiphoton microscopy has revealed the abnormal structure and function of tumor-associated blood and lymphatic vessels, the role of aberrant tumor matrix in drug delivery, invasion and metastasis of tumor cells, the dynamics of immune cell trafficking to and within tumors, and gene expression in tumors. However, traditional multiphoton microscopy suffers from inherently slow imaging rates—only a few frames per second, thus unable to capture more rapid events such as blood flow, lymphatic flow, and cell movement within vessels. Here, we report the development and implementation of a video-rate multiphoton microscope (VR-MPLSM) based on resonant galvanometer mirror scanning that is capable of recording at 30 frames per second and acquiring intravital multispectral images. We show that the design of the system can be readily implemented and is adaptable to various experimental models. As examples, we demonstrate the utility of the system to directly measure flow within tumors, capture metastatic cancer cells moving within the brain vasculature and cells in lymphatic vessels, and image acute responses to changes in a vascular network. VR-MPLSM thus has the potential to further advance intravital imaging and provide new insight into the biology of the tumor microenvironment. PMID:24353926

  4. Scanning probe recognition microscopy investigation of tissue scaffold properties

    Science.gov (United States)

    Fan, Yuan; Chen, Qian; Ayres, Virginia M; Baczewski, Andrew D; Udpa, Lalita; Kumar, Shiva

    2007-01-01

    Scanning probe recognition microscopy is a new scanning probe microscopy technique which enables selective scanning along individual nanofibers within a tissue scaffold. Statistically significant data for multiple properties can be collected by repetitively fine-scanning an identical region of interest. The results of a scanning probe recognition microscopy investigation of the surface roughness and elasticity of a series of tissue scaffolds are presented. Deconvolution and statistical methods were developed and used for data accuracy along curved nanofiber surfaces. Nanofiber features were also independently analyzed using transmission electron microscopy, with results that supported the scanning probe recognition microscopy-based analysis. PMID:18203431

  5. Two Photon Distribution Amplitudes

    International Nuclear Information System (INIS)

    El Beiyad, M.; Pire, B.; Szymanowski, L.; Wallon, S.

    2008-01-01

    The factorization of the amplitude of the process γ*γ→γγ in the low energy and high photon virtuality region is demonstrated at the Born order and in the leading logarithmic approximation. The leading order two photon (generalized) distribution amplitudes exhibit a characteristic ln Q 2 behaviour and obey new inhomogeneous evolution equations

  6. Scanning electron microscopy and micro-analyses

    International Nuclear Information System (INIS)

    Brisset, F.; Repoux, L.; Ruste, J.; Grillon, F.; Robaut, F.

    2008-01-01

    Scanning electron microscopy (SEM) and the related micro-analyses are involved in extremely various domains, from the academic environments to the industrial ones. The overall theoretical bases, the main technical characteristics, and some complements of information about practical usage and maintenance are developed in this book. high-vacuum and controlled-vacuum electron microscopes are thoroughly presented, as well as the last generation of EDS (energy dispersive spectrometer) and WDS (wavelength dispersive spectrometer) micro-analysers. Beside these main topics, other analysis or observation techniques are approached, such as EBSD (electron backscattering diffraction), 3-D imaging, FIB (focussed ion beams), Monte-Carlo simulations, in-situ tests etc.. This book, in French language, is the only one which treats of this subject in such an exhaustive way. It represents the actualized and totally updated version of a previous edition of 1979. It gathers the lectures given in 2006 at the summer school of Saint Martin d'Heres (France). Content: 1 - electron-matter interactions; 2 - characteristic X-radiation, Bremsstrahlung; 3 - electron guns in SEM; 4 - elements of electronic optics; 5 - vacuum techniques; 6 - detectors used in SEM; 7 - image formation and optimization in SEM; 7a - SEM practical instructions for use; 8 - controlled pressure microscopy; 8a - applications; 9 - energy selection X-spectrometers (energy dispersive spectrometers - EDS); 9a - EDS analysis; 9b - X-EDS mapping; 10 - technological aspects of WDS; 11 - processing of EDS and WDS spectra; 12 - X-microanalysis quantifying methods; 12a - quantitative WDS microanalysis of very light elements; 13 - statistics: precision and detection limits in microanalysis; 14 - analysis of stratified samples; 15 - crystallography applied to EBSD; 16 - EBSD: history, principle and applications; 16a - EBSD analysis; 17 - Monte Carlo simulation; 18 - insulating samples in SEM and X-ray microanalysis; 18a - insulating

  7. Emission sources in scanning electron microscopy

    International Nuclear Information System (INIS)

    Malkusch, W.

    1990-01-01

    Since the beginning of the commercial scanning electron microscopy, there are two kinds of emission sources generally used for generation of the electron beam. The first group covers the cathodes heated directly and indirectly (tungsten hair-needle cathodes and lanthanum hexaboride single crystals, LaB 6 cathode). The other group is the field emission cathodes. The advantages of the thermal sources are their low vacuum requirement and their high beam current which is necessary for the application of microanalysis units. Disadvantages are the short life and the low resolution. Advantages of the field emission cathode unambiguously are the possibilities of the very high resolution, especially in the case of low acceleration voltages. Disadvantages are the necessary ultra-high vacuum and the low beam current. An alternative source is the thermally induced ZrO/W field emission cathode which works stably as compared to the cold field emission and does not need periodic flashing for emitter tip cleaning. (orig.) [de

  8. Elemental mapping in scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Allen, L J; D'Alfonso, A J; Lugg, N R; Findlay, S D; LeBeau, J M; Stemmer, S

    2010-01-01

    We discuss atomic resolution chemical mapping in scanning transmission electron microscopy (STEM) based on core-loss electron energy loss spectroscopy (EELS) and also on energy dispersive X-ray (EDX) imaging. Chemical mapping using EELS can yield counterintuitive results which, however, can be understood using first principles calculations. Experimental chemical maps based on EDX bear out the thesis that such maps are always likely to be directly interpretable. This can be explained in terms of the local nature of the effective optical potential for ionization under those imaging conditions. This is followed by an excursion into the complementary technique of elemental mapping using energy-filtered transmission electron microscopy (EFTEM) in a conventional transmission electron microscope. We will then consider the widely used technique of Z-contrast or high-angle annular dark field (HAADF) imaging, which is based on phonon excitation, where it has recently been shown that intensity variations can be placed on an absolute scale by normalizing the measured intensities to the incident beam. Results, showing excellent agreement between theory and experiment to within a few percent, are shown for Z-contrast imaging from a sample of PbWO 4 .

  9. A dark mode in scanning thermal microscopy

    Science.gov (United States)

    Ramiandrisoa, Liana; Allard, Alexandre; Joumani, Youssef; Hay, Bruno; Gomés, Séverine

    2017-12-01

    The need for high lateral spatial resolution in thermal science using Scanning Thermal Microscopy (SThM) has pushed researchers to look for more and more tiny probes. SThM probes have consequently become more and more sensitive to the size effects that occur within the probe, the sample, and their interaction. Reducing the tip furthermore induces very small heat flux exchanged between the probe and the sample. The measurement of this flux, which is exploited to characterize the sample thermal properties, requires then an accurate thermal management of the probe-sample system and to reduce any phenomenon parasitic to this system. Classical experimental methodologies must then be constantly questioned to hope for relevant and interpretable results. In this paper, we demonstrate and estimate the influence of the laser of the optical force detection system used in the common SThM setup that is based on atomic-force microscopy equipment on SThM measurements. We highlight the bias induced by the overheating due to the laser illumination on the measurements performed by thermoresistive probes (palladium probe from Kelvin Nanotechnology). To face this issue, we propose a new experimental procedure based on a metrological approach of the measurement: a SThM "dark mode." The comparison with the classical procedure using the laser shows that errors between 14% and 37% can be reached on the experimental data exploited to determine the heat flux transferred from the hot probe to the sample.

  10. Scanning probe microscopy in material science and biology

    International Nuclear Information System (INIS)

    Cricenti, A; Colonna, S; Girasole, M; Gori, P; Ronci, F; Longo, G; Dinarelli, S; Luce, M; Rinaldi, M; Ortenzi, M

    2011-01-01

    A review of the activity of scanning probe microscopy at our Institute is presented, going from instrumentation to software development of scanning tunnelling microscopy, atomic force microscopy and scanning near-field optical microscopy (SNOM). Some of the most important experiments in material science and biology performed by our group through the years with these SPM techniques will be presented. Finally, infrared applications by coupling a SNOM with a free electron laser will also be presented.

  11. Application of Confocal Laser Scanning Microscopy in Biology and Medicine

    OpenAIRE

    I. A. Volkov; N. V. Frigo; L. F. Znamenskaya; O. R. Katunina

    2014-01-01

    Fluorescence confocal laser scanning microscopy and reflectance confocal laser scanning microscopy are up-to-date highend study methods. Confocal microscopy is used in cell biology and medicine. By using confocal microscopy, it is possible to study bioplasts and localization of protein molecules and other compounds relative to cell or tissue structures, and to monitor dynamic cell processes. Confocal microscopes enable layer-by-layer scanning of test items to create demonstrable 3D models. As...

  12. Two-Photon Fluorescence Microscope for Microgravity Research

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2005-01-01

    A two-photon fluorescence microscope has been developed for the study of biophysical phenomena. Two-photon microscopy is a novel form of laser-based scanning microscopy that enables three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon optical microscopy, two-photon microscopy utilizes the simultaneous nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption, so an ultra-fast pulsed laser source is typically employed. On the other hand, the critical energy threshold for two-photon absorption leads to fluorophore excitation that is intrinsically localized to the focal volume. Consequently, two-photon microscopy enables optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction (relative to one-photon optical microscopy) in photon-induced damage because of the longer excitation wavelength. This reduction is especially advantageous for in vivo studies. Relative to confocal microscopy, there is also a reduction in background fluorescence, and, because of a reduction in Rayleigh scattering, there is a 4 increase of penetration depth. The prohibitive cost of a commercial two-photon fluorescence-microscope system, as well as a need for modularity, has led to the construction of a custom-built system (see Figure 1). This system includes a coherent mode-locked titanium: sapphire laser emitting 120-fs-duration pulses at a repetition rate of 80 MHz. The pulsed laser has an average output power of 800 mW and a wavelength tuning range of 700 to 980 nm, enabling the excitation of a variety of targeted fluorophores. The output from the laser is attenuated, spatially filtered, and then directed into a confocal scanning head that has been modified to provide for side entry of the laser beam. The laser output coupler has been replaced with a dichroic filter that reflects the

  13. Scanning tunnel microscopy of semiconductor nanostructures

    International Nuclear Information System (INIS)

    Eder, C.

    1997-09-01

    In this work a scanning tunneling microscope (STM) is utilized as a surface sensitive tool for local characterization of internal potential profiles of GaAs/AlGaAs heterostructures. The STM is operated at variable temperatures under ambient conditions, i.e. either in air or in the variable temperature insert of a cryostat. Distinct local differences between current-voltage curves taken on inverted heterostructures, which were patterned by wet chemically etching, are found. The spectroscopic differences can be ascribed to the internal potential profile in the subsurface regions of the sample. Current imaging tunneling spectroscopy (CITS) is applied to study quantum wire regions. It is found that the magnitude of the CITS-current is an indirect measure of edge depletion zones, which are much larger at 4.2 K. Direct measurements of relevant energy levels in quantum structures were obtained by ballistic electron emission microscopy (BEEM). It is shown that this 3-terminal technique is an excellent tool for transport characterization of minibands formed in semiconductor superlattices. Furthermore, low dimensional electron gases are shown to act as very efficient collector electrodes at low temperatures. For the first time, BEEM experiments were performed at 4.2 K. The enhanced thermal resolution at 4.2 K allows an analysis of the relevant scattering processes. It is found that the collector current is strongly influenced by diffusive scattering at the metal/semiconductor interface. (author)

  14. Scanning transmission ion microscopy of polycarbonate nanocapillaries

    International Nuclear Information System (INIS)

    Gal, G.A.B.; Rajta, I.; Szilasi, S.Z.; Juhasz, Z.; Biri, S.; Csik, A.; Sulik, B.; Cserhati, Cs.

    2011-01-01

    Complete text of publication follows. Nanochanneled materials are of a great interest due to their peculiar properties and high potential impact for the fabrication of nanostructures and nanodevices. Polycarbonate membranes are produced by heavy ion irradiation followed by chemical etching of the ion tracks. The irradiation parameters determine the porosity (areal density of the capillaries) and angular spread, while the channel diameters and shapes depend on the chemical process parameters. Such polycarbonate (and other materials) membranes are commercially available from a few manufacturers. The primary use of the filters involves packaging and filtering applications. Moreover, they are used for collecting atmospheric aerosols for environmental research. The nanocapillaries formed in membranes are particularly suitable for ion and electron guiding studies of a recently discovered, but not yet completely understood capillary guiding phenomenon. This interesting guiding effect is very promising for patterning by parallel writing with ions and/or electrons through masks. In order to get a better understanding of this phenomenon, we need a better characterization of the capillaries themselves. This study is addressing the angular distribution of the nanochannels in the polycarbonate filters by using a nuclear microprobe facility and the method of scanning transmission ion microscopy (STIM). The STIM experiments in this work have been performed at ATOMKI. The proton energy was 2 MeV, the beam intensity was about 1000 protons s -1 , the beam spot size was about 1 x 1 μm, the scan size was 100 x 100 μm and the beam divergence was smaller than 0.07 deg. A scanning electron microscope (SEM, Hitachi S4300 CFE) was used to measure the capillary diameters and the membrane porosity. The sample thickness was determined by a profilometer (AMBIOS XP-I). We have investigated two different pieces of Millipore Isopore TM samples. A typical SEM image showed several overlapping

  15. Scanning transmission electron microscopy imaging and analysis

    CERN Document Server

    Pennycook, Stephen J

    2011-01-01

    Provides the first comprehensive treatment of the physics and applications of this mainstream technique for imaging and analysis at the atomic level Presents applications of STEM in condensed matter physics, materials science, catalysis, and nanoscience Suitable for graduate students learning microscopy, researchers wishing to utilize STEM, as well as for specialists in other areas of microscopy Edited and written by leading researchers and practitioners

  16. Two dimensional numerical simulations of carrier dynamics during time-resolved photoluminescence decays in two-photon microscopy measurements in semiconductors

    International Nuclear Information System (INIS)

    Kanevce, Ana; Kuciauskas, Darius; Levi, Dean H.; Johnston, Steven W.; Allende Motz, Alyssa M.

    2015-01-01

    We use two-dimensional numerical simulations to analyze high spatial resolution time-resolved spectroscopy data. This analysis is applied to two-photon excitation time-resolved photoluminescence (2PE-TRPL) but is broadly applicable to all microscopic time-resolved techniques. By solving time-dependent drift-diffusion equations, we gain insight into carrier dynamics and transport characteristics. Accurate understanding of measurement results establishes the limits and potential of the measurement and enhances its value as a characterization method. Diffusion of carriers outside of the collection volume can have a significant impact on the measured decay but can also provide an estimate of carrier mobility as well as lifetime. In addition to material parameters, the experimental conditions, such as spot size and injection level, can impact the measurement results. Although small spot size provides better resolution, it also increases the impact of diffusion on the decay; if the spot size is much smaller than the diffusion length, it impacts the entire decay. By reproducing experimental 2PE-TRPL decays, the simulations determine the bulk carrier lifetime from the data. The analysis is applied to single-crystal and heteroepitaxial CdTe, material important for solar cells, but it is also applicable to other semiconductors where carrier diffusion from the excitation volume could affect experimental measurements

  17. Probing cytotoxicity of nanoparticles and organic compounds using scanning proton microscopy, scanning electron microscopy and fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tong Yongpeng [Institute of Nuclear Techniques, Shenzhen University, Nanhai Avenue 3688, Shenzhen 518060 (China)], E-mail: yongpengt@yahoo.com.cn; Li Changming [School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457 (Singapore); Liang Feng [Institute Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200025 (China); Chen Jianmin [Shenzhen Municipal Hospital for Chronic Disease Control and Prevention, Guangdong 518020 (China); Zhang Hong; Liu Guoqing; Sun Huibin [Institute of Nuclear Techniques, Shenzhen University, Nanhai Avenue 3688, Shenzhen 518060 (China); Luong, John H.T. [Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec, H4P 2R2 (Canada)

    2008-12-15

    Scanning proton microscopy, scanning electron microscopy (SEM) and fluorescence microscopy have been used to probe the cytotoxicity effect of benzo[a]pyrene (BaP), ethidium bromide (EB) and nanoparticles (ZnO, Al{sub 2}O{sub 3} and TiO{sub 2}) on a T lymphoblastic leukemia Jurkat cell line. The increased calcium ion (from CaCl{sub 2}) in the culture medium stimulated the accumulation of BaP and EB inside the cell, leading to cell death. ZnO, Al{sub 2}O{sub 3} and TiO{sub 2} nanoparticles, however, showed a protective effect against these two organic compounds. Such inorganic nanoparticles complexed with BaP or EB which became less toxic to the cell. Fe{sub 2}O{sub 3} nanoparticles as an insoluble particle model scavenged by macrophage were investigated in rats. They were scavenged out of the lung tissue about 48 h after infection. This result suggest that some insoluble inorganic nanoparticles of PM (particulate matters) showed protective effects on organic toxins induced acute toxic effects as they can be scavenged by macrophage cells. Whereas, some inorganic ions such as calcium ion in PM may help environmental organic toxins to penetrate cell membrane and induce higher toxic effect.

  18. Scanning photoemission microscopy with synchrotron radiation

    Science.gov (United States)

    Ade, Harald W.

    1992-08-01

    Progress in photoemission spectro-microscopy at various synchrotron radiation facilities is reviewed. Microprobe devices such as MAXIMUM at the SRC in Wisconsin, the X1-SPEM at the NSLS at BNL, as well as the ellipsoidal ring mirror microscope at DESY in Hamburg, recorded first images during the last few years. The present status of these devices which achieve their lateral resolution by focusing X-rays to a small spot is the primary focus of this paper, but work representing other approaches to spectro-microscopy is also discussed.

  19. Combined frequency modulated atomic force microscopy and scanning tunneling microscopy detection for multi-tip scanning probe microscopy applications

    International Nuclear Information System (INIS)

    Morawski, Ireneusz; Spiegelberg, Richard; Korte, Stefan; Voigtländer, Bert

    2015-01-01

    A method which allows scanning tunneling microscopy (STM) tip biasing independent of the sample bias during frequency modulated atomic force microscopy (AFM) operation is presented. The AFM sensor is supplied by an electronic circuit combining both a frequency shift signal and a tunneling current signal by means of an inductive coupling. This solution enables a control of the tip potential independent of the sample potential. Individual tip biasing is specifically important in order to implement multi-tip STM/AFM applications. An extensional quartz sensor (needle sensor) with a conductive tip is applied to record simultaneously topography and conductivity of the sample. The high resonance frequency of the needle sensor (1 MHz) allows scanning of a large area of the surface being investigated in a reasonably short time. A recipe for the amplitude calibration which is based only on the frequency shift signal and does not require the tip being in contact is presented. Additionally, we show spectral measurements of the mechanical vibration noise of the scanning system used in the investigations

  20. Combined frequency modulated atomic force microscopy and scanning tunneling microscopy detection for multi-tip scanning probe microscopy applications

    Energy Technology Data Exchange (ETDEWEB)

    Morawski, Ireneusz [Peter Grünberg Institut (PGI-3) and JARA-Fundamentals of Future Information Technology, Forschungszentrum Jülich, 52425 Jülich (Germany); Institute of Experimental Physics, University of Wrocław, pl. M. Borna 9, 50-204 Wrocław (Poland); Spiegelberg, Richard; Korte, Stefan; Voigtländer, Bert [Peter Grünberg Institut (PGI-3) and JARA-Fundamentals of Future Information Technology, Forschungszentrum Jülich, 52425 Jülich (Germany)

    2015-12-15

    A method which allows scanning tunneling microscopy (STM) tip biasing independent of the sample bias during frequency modulated atomic force microscopy (AFM) operation is presented. The AFM sensor is supplied by an electronic circuit combining both a frequency shift signal and a tunneling current signal by means of an inductive coupling. This solution enables a control of the tip potential independent of the sample potential. Individual tip biasing is specifically important in order to implement multi-tip STM/AFM applications. An extensional quartz sensor (needle sensor) with a conductive tip is applied to record simultaneously topography and conductivity of the sample. The high resonance frequency of the needle sensor (1 MHz) allows scanning of a large area of the surface being investigated in a reasonably short time. A recipe for the amplitude calibration which is based only on the frequency shift signal and does not require the tip being in contact is presented. Additionally, we show spectral measurements of the mechanical vibration noise of the scanning system used in the investigations.

  1. Utilizing two-photon fluorescence and second harmonic generation microscopy to study human bone marrow mesenchymal stem cell morphogenesis in chitosan scaffold

    Science.gov (United States)

    Su, Ping-Jung; Huang, Chi-Hsiu; Huang, Yi-You; Lee, Hsuan-Sue; Dong, Chen-Yuan

    2008-02-01

    A major goal of tissue engineering is to cultivate the cartilage in vitro. One approach is to implant the human bone marrow mesenchymal stem cells into the three dimensional biocompatible and biodegradable material. Through the action of the chondrogenic factor TGF-β3, the stem cells can be induced to secrete collagen. In this study, mesenchymal stem cells are implanted on the chitosan scaffold and TGF-β3 was added to produce the cartilage tissue and TP autofluorescence and SHG microscopy was used to image the process of chondrogenesis. With additional development, multiphoton microscopy can be developed into an effective tool for evaluating the quality of tissue engineering products.

  2. In vivo non-invasive monitoring of collagen remodelling by two-photon microscopy after micro-ablative fractional laser resurfacing.

    Science.gov (United States)

    Cicchi, Riccardo; Kapsokalyvas, Dimitrios; Troiano, Michela; Campolmi, Piero; Morini, Cristiano; Massi, Daniela; Cannarozzo, Giovanni; Lotti, Torello; Pavone, Francesco Saverio

    2014-11-01

    Non-linear optical microscopy is becoming popular as a non-invasive in vivo imaging modality in dermatology. In this study, combined TPF and SHG microscopy were used to monitor collagen remodelling in vivo after micro-ablative fractional laser resurfacing. Papillary dermis of living subjects, covering a wide age range, was imaged immediately before and forty days after treatment. A qualitative visual examination of acquired images demonstrated an age-dependent remodelling effect on collagen. Additional quantitative analysis of new collagen production was performed by means of two image analysis methods. A higher increase in SHG to TPF ratio, corresponding to a stronger treatment effectiveness, was found in older subjects, whereas the effect was found to be negligible in young, and minimal in middle age subjects. Analysis of collagen images also showed a dependence of the treatment effectiveness with age but with controversial results. While the diagnostic potential of in vivo multiphoton microscopy has already been demonstrated for skin cancer and other skin diseases, here we first successfully explore its potential use for a non-invasive follow-up of a laser-based treatment. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. A Comparative Scanning Electron Microscopy Evaluation of Smear ...

    African Journals Online (AJOL)

    2018-02-07

    Feb 7, 2018 ... scanning electron microscopy evaluation of smear layer removal with chitosan and .... this compound has considerably increased its concentration in rivers and .... of the images was done by three investigators who calibrated ...

  4. Surface physics studied by means of scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Besenbacher, F.; Laegsgaard, E.; Stensgaard, I.

    1993-01-01

    Scanning tunneling microscopy has been applied to study silicon crystal structures, oxygen on Cu (110), and real industrial catalyst surfaces. For the latter purpose an Atomic Force Microscope is being developed. (EG)

  5. Biological applications of near-field scanning optical microscopy

    NARCIS (Netherlands)

    Moers, M.H.P.; Moers, Marco H.P.; Ruiter, A.G.T.; Jalocha, A.; Jalocha, Alain; van Hulst, N.F.

    1995-01-01

    Near-field Scanning Optical Microscopy (NSOM) is a true optical microscopic technique allowing fluorescence, absorption, reflection and polarization contrast with the additional advantage of nanometer lateral resolution, unlimited by diffraction and operation at ambient conditions. NSOM based on

  6. Full information acquisition in scanning probe microscopy and spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Jesse, Stephen; Belianinov, Alex; Kalinin, Sergei V.; Somnath, Suhas

    2017-04-04

    Apparatus and methods are described for scanning probe microscopy and spectroscopy based on acquisition of full probe response. The full probe response contains valuable information about the probe-sample interaction that is lost in traditional scanning probe microscopy and spectroscopy methods. The full probe response is analyzed post data acquisition using fast Fourier transform and adaptive filtering, as well as multivariate analysis. The full response data is further compressed to retain only statistically significant components before being permanently stored.

  7. Fluorescence microscopy.

    Science.gov (United States)

    Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

    2014-10-01

    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

  8. Atmospheric scanning electron microscope for correlative microscopy.

    Science.gov (United States)

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

    2012-01-01

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

  9. Optomechatronics Design and Control for Confocal Laser Scanning Microscopy

    NARCIS (Netherlands)

    Yoo, H.W.

    2015-01-01

    Confocal laser scanning microscopy (CLSM) is considered as one of the major advancements in microscopy in the last century and is widely accepted as a 3D fluorescence imaging tool for biological studies. For the emerging biological questions CLSM requires fast imaging to detect rapid biological

  10. Scanning tunneling microscopy of hexagonal BN grown on graphite

    International Nuclear Information System (INIS)

    Fukumoto, H.; Hamada, T.; Endo, T.; Osaka, Y.

    1991-01-01

    The microscopic surface topography of thin BN x films grown on graphite by electron cyclotron resonance plasma chemical vapor deposition have been imaged with scanning tunneling microscopy in air. The scanning tunneling microscope has generated images of hexagonal BN with atomic resolution

  11. Forensic document analysis using scanning microscopy

    Science.gov (United States)

    Shaffer, Douglas K.

    2009-05-01

    The authentication and identification of the source of a printed document(s) can be important in forensic investigations involving a wide range of fraudulent materials, including counterfeit currency, travel and identity documents, business and personal checks, money orders, prescription labels, travelers checks, medical records, financial documents and threatening correspondence. The physical and chemical characterization of document materials - including paper, writing inks and printed media - is becoming increasingly relevant for law enforcement agencies, with the availability of a wide variety of sophisticated commercial printers and copiers which are capable of producing fraudulent documents of extremely high print quality, rendering these difficult to distinguish from genuine documents. This paper describes various applications and analytical methodologies using scanning electron miscoscopy/energy dispersive (x-ray) spectroscopy (SEM/EDS) and related technologies for the characterization of fraudulent documents, and illustrates how their morphological and chemical profiles can be compared to (1) authenticate and (2) link forensic documents with a common source(s) in their production history.

  12. Probing superconductors. Spectroscopic-imaging scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Hanaguri, Tetsuo

    2011-01-01

    Discovery of high-temperature superconductivity in a cuprate triggered developments of various spectroscopic tools which have been utilized to elucidate electronic states of this mysterious compound. Particularly, angle-resolved photoemission spectroscopy and scanning-tunneling microscopy/spectroscopy are improved considerably. It is now possible to map the superconducting gap in both momentum and real spaces using these two techniques. Here we review spectroscopic-imaging scanning tunneling microscopy which is able to explore momentum-space phase structure of the superconducting gap, as well as real-space structure. Applications of this technique to a cuprate and an iron-based superconductor are discussed. (author)

  13. Menadione metabolism to thiodione in hepatoblastoma by scanning electrochemical microscopy

    Science.gov (United States)

    Mauzeroll, Janine; Bard, Allen J.; Owhadian, Omeed; Monks, Terrence J.

    2004-01-01

    The cytotoxicity of menadione on hepatocytes was studied by using the substrate generation/tip collection mode of scanning electrochemical microscopy by exposing the cells to menadione and detecting the menadione-S-glutathione conjugate (thiodione) that is formed during the cellular detoxication process and is exported from the cell by an ATP-dependent pump. This efflux was electrochemically detected and allowed scanning electrochemical microscopy monitoring and imaging of single cells and groups of highly confluent live cells. Based on a constant flux model, ≈6 × 106 molecules of thiodione per cell per second are exported from monolayer cultures of Hep G2 cells. PMID:15601769

  14. Enhancement of two-photon photoluminescence and SERS for low-coverage gold films

    DEFF Research Database (Denmark)

    Novikov, Sergey M.; Beermann, Jonas; Frydendahl, Christian

    2016-01-01

    Electromagnetic field enhancement (FE) effects occurring in thin gold films 3-12-nm are investigated with two-photon photoluminescence (TPL) and Raman scanning optical microscopies. The samples are characterized using scanning electron microscopy images and linear optical spectroscopy. TPL images...... exhibit a strong increase in the level of TPL signals for films thicknesses 3-8-nm, near the percolation threshold. For some thicknesses, TPL measurements reveal super-cubic dependences on the incident power. We ascribe this feature to the occurrence of very strongly localized and enhanced electromagnetic...

  15. Polarization contrast in photon scanning tunnelling microscopy combined with atomic force microscopy

    NARCIS (Netherlands)

    Propstra, K.; Propstra, K.; van Hulst, N.F.

    1995-01-01

    Photon scanning tunnelling microscopy combined with atomic force microscopy allows simultaneous acquisition and direct comparison of optical and topographical images, both with a lateral resolution of about 30 nm, far beyond the optical diffraction limit. The probe consists of a modified

  16. System and method for compressive scanning electron microscopy

    Science.gov (United States)

    Reed, Bryan W

    2015-01-13

    A scanning transmission electron microscopy (STEM) system is disclosed. The system may make use of an electron beam scanning system configured to generate a plurality of electron beam scans over substantially an entire sample, with each scan varying in electron-illumination intensity over a course of the scan. A signal acquisition system may be used for obtaining at least one of an image, a diffraction pattern, or a spectrum from the scans, the image, diffraction pattern, or spectrum representing only information from at least one of a select subplurality or linear combination of all pixel locations comprising the image. A dataset may be produced from the information. A subsystem may be used for mathematically analyzing the dataset to predict actual information that would have been produced by each pixel location of the image.

  17. Surface properties and microporosity of polyhydroxybutyrate under scanning electron microscopy

    International Nuclear Information System (INIS)

    Raouf, A.A.; Samsudin, A.R.; Samian, R.; Akool, K.; Abdullah, N.

    2004-01-01

    This study was designed to investigate the surface properties especially surface porosity of polyhydroxybutyrate (PHB) using scanning electron microscopy. PHB granules were sprinkled on the double-sided sticky tape attached on a SEM aluminium stub and sputtered with gold (10nm thickness) in a Polaron SC515 Coater, following which the samples were placed into the SEM specimen chamber for viewing and recording. Scanning electron micrographs with different magnification of PHB surface revealed multiple pores with different sizes. (Author)

  18. Re-scan confocal microscopy (RCM) improves the resolution of confocal microscopy and increases the sensitivity

    NARCIS (Netherlands)

    de Luca, Giulia; Breedijk, Ronald; Hoebe, Ron; Stallinga, Sjoerd; Manders, Erik

    2017-01-01

    Re-scan confocal microscopy (RCM) is a new super-resolution technique based on a standard confocal microscope extended with a re-scan unit in the detection path that projects the emitted light onto a sensitive camera. In this paper the fundamental properties of RCM, lateral resolution, axial

  19. Re-scan confocal microscopy (RCM) improves the resolution of confocal microscopy and increases the sensitivity

    NARCIS (Netherlands)

    De Luca, G.; Breedijk, R.; Hoebe, R.; Stallinga, S.; Manders, E.

    Re-scan confocal microscopy (RCM) is a new super-resolution technique based on a standard confocal microscope extended with a re-scan unit in the detection path that projects the emitted light onto a sensitive camera. In this paper the fundamental properties of RCM, lateral resolution, axial

  20. Novel triphenylamine-cored two-photon absorbing dyes for labeling of biomolecules

    International Nuclear Information System (INIS)

    Xiao Haibo; Mei Chong; Wang Yaochuan; Li, Hui; Qian Shixiong; Yin Hongyao; Xu Zhisong

    2011-01-01

    Highlights: → Two novel triphenylamine-cored chromophores were synthesized. → These two dyes have sizable two-photon absorption cross-section at 800 nm. → They possess reasonable water solubility and are suitable as labels in aqueous biological environments. → These dyes have strong chelating ability. → They display a large set of reactivity for coupling to biomolecules. - Abstract: Two novel, V-shaped and Y-shaped dipicolinate derivatives branched from triphenylamine, {4-[(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl)vinyl]}-N-phenyl-N-{4- [(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl)vinylphenyl]}aniline (1) and {4-[(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl) vinyl]}-N,N-bis {4-[(E)-2-(2,6-dimethoxycarbonyl pyridin-4-yl)vinylphenyl]}aniline (2) were synthesized. These compounds were designed for large two-photon absorption and in particular for labeling of biomolecules. Their linear absorption, fluorescence properties and their two-photon absorption properties as well as two-photon fluorescence cell imaging were examined. When excited at 800 nm, the two-photon absorption cross-section values of chromophores 1 and 2 in THF were 208 GM, 376 GM, respectively. These two-photon absorbing dyes possess reasonable water solubility, strong chelating ability and display a large set of reactivity for coupling to biomolecules, which are apparently due to the two methoxycarbonyl groups in pyridine ring. This work suggests that chromophores 1 and 2 are promising labels potentially applicable for the tracking of biomolecules using two-photon scanning microscopy.

  1. The art of SPM : scanning probe microscopy in materials science

    NARCIS (Netherlands)

    Loos, J.

    2005-01-01

    In this Progress Report, outstanding scientific applications of scanning probe microscopy (SPM) in the field of materials science and the latest technique developments are introduced and discussed. Besides being able to image the organization of matter with sub-nanometer resolution, SPM, owing to

  2. Study of Hydrated Lime in Environmental Scanning Electron Microscopy

    Czech Academy of Sciences Publication Activity Database

    Tihlaříková, Eva; Neděla, Vilém; Rovnaníková, P.

    2013-01-01

    Roč. 19, S2 (2013), s. 1644-1645 ISSN 1431-9276 R&D Projects: GA ČR GAP102/10/1410; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : Hydrated Lime * Environmental Scanning Electron Microscopy Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.757, year: 2013

  3. Oxygen-free in situ scanning tunnelling microscopy

    DEFF Research Database (Denmark)

    Zhang, Jingdong; Ulstrup, Jens

    2007-01-01

    Scanning tunneling microscopy under full electrochemical potential control (in situ STM) has been used extensively as an efficient method to characterize microstructures at solid/liquid interfaces at the atomic and molecular levels. However, under ambient conditions oxygen may interfere in open i...

  4. Nanochannel alignment analysis by scanning transmission ion microscopy

    DEFF Research Database (Denmark)

    Rajta, I.; Gál, G.A.B.; Szilasi, S.Z.

    2010-01-01

    In this paper a study on the ion transmission ratio of a nanoporous alumina sample is presented. The sample was investigated by scanning transmission ion microscopy (STIM) with different beam sizes. The hexagonally close-packed AlO nanocapillary array, realized as a suspended membrane of 15 νm...

  5. Ultrafast terahertz scanning tunneling microscopy with atomic resolution

    DEFF Research Database (Denmark)

    Jelic, Vedran; Iwaszczuk, Krzysztof; Nguyen, Peter H.

    2016-01-01

    We demonstrate that ultrafast terahertz scanning tunneling microscopy (THz-STM) can probe single atoms on a silicon surface with simultaneous sub-nanometer and sub-picosecond spatio-temporal resolution. THz-STM is established as a new technique for exploring high-field non-equilibrium tunneling...

  6. Characterization of Polycaprolactone Films Biodeterioration by Scanning Electron Microscopy

    Czech Academy of Sciences Publication Activity Database

    Hrubanová, Kamila; Voberková, S.; Hermanová, S.; Krzyžánek, Vladislav

    2014-01-01

    Roč. 20, S3 (2014), s. 1950-1951 ISSN 1431-9276 R&D Projects: GA MŠk EE.2.3.20.0103; GA MŠk(CZ) LO1212 Institutional support: RVO:68081731 Keywords : polycaprolactone films * biodeterioration * scanning electron microscopy Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.877, year: 2014

  7. Surface chemical reactions probed with scanning force microscopy

    NARCIS (Netherlands)

    Werts, M.P L; van der Vegte, E.W.; Hadziioannou, G

    1997-01-01

    In this letter we report the study of surface chemical reactions with scanning force microscopy (SFM) with chemical specificity. Using chemically modified SFM probes, we can determine the local surface reaction conversion during a chemical surface modification. The adhesion forces between a

  8. Scanning electron microscopy-energy dispersive X-ray spectrometer ...

    African Journals Online (AJOL)

    The distribution of arsenic (As) and cadmium (Cd) in himematsutake was analyzed using scanning electron microscopy-energy dispersive X-ray spectrometer (SEM-EDX). The atomic percentage of the metals was confirmed by inductively coupled plasma-mass spectrometer (ICP-MS). Results show that the accumulation of ...

  9. Ultrafast photon counting applied to resonant scanning STED microscopy.

    Science.gov (United States)

    Wu, Xundong; Toro, Ligia; Stefani, Enrico; Wu, Yong

    2015-01-01

    To take full advantage of fast resonant scanning in super-resolution stimulated emission depletion (STED) microscopy, we have developed an ultrafast photon counting system based on a multigiga sample per second analogue-to-digital conversion chip that delivers an unprecedented 450 MHz pixel clock (2.2 ns pixel dwell time in each scan). The system achieves a large field of view (∼50 × 50 μm) with fast scanning that reduces photobleaching, and advances the time-gated continuous wave STED technology to the usage of resonant scanning with hardware-based time-gating. The assembled system provides superb signal-to-noise ratio and highly linear quantification of light that result in superior image quality. Also, the system design allows great flexibility in processing photon signals to further improve the dynamic range. In conclusion, we have constructed a frontier photon counting image acquisition system with ultrafast readout rate, excellent counting linearity, and with the capacity of realizing resonant-scanning continuous wave STED microscopy with online time-gated detection. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

  10. Scanning tunneling microscopy III theory of STM and related scanning probe methods

    CERN Document Server

    Güntherodt, Hans-Joachim

    1996-01-01

    Scanning Tunneling Microscopy III provides a unique introduction to the theoretical foundations of scanning tunneling microscopy and related scanning probe methods. The different theoretical concepts developed in the past are outlined, and the implications of the theoretical results for the interpretation of experimental data are discussed in detail. Therefore, this book serves as a most useful guide for experimentalists as well as for theoreticians working in the filed of local probe methods. In this second edition the text has been updated and new methods are discussed.

  11. Atomic force microscopy and confocal laser scanning microscopy on the cytoskeleton of permeabilised and embedded cells

    International Nuclear Information System (INIS)

    Meller, Karl; Theiss, Carsten

    2006-01-01

    We describe a technical method of cell permeabilisation and embedding to study the organisation and distribution of intracellular proteins with aid of atomic force microscopy and confocal laser scanning microscopy in identical areas. While confocal laser scanning microscopy is useful for the identification of certain proteins subsequent labelling with markers or antibodies, atomic force microscopy allows the observation of macromolecular structures in fixed and living cells. To demonstrate the field of application of this preparatory technique, cells were permeabilised, fixed, and the actin cytoskeleton was stained with phalloidin-rhodamine. Confocal laser scanning microscopy was used to show the organisation of these microfilaments, e.g. geodesic dome structures. Thereafter, cells were embedded in Durcupan water-soluble resin, followed by UV-polymerisation of resin at 4 o C. This procedure allowed intracellular visualisation of the cell nucleus or cytoskeletal elements by atomic force microscopy, for instance to analyse the globular organisation of actin filaments. Therefore, this method offers a great potential to combine both microscopy techniques in order to understand and interpret intracellular protein relations, for example, the biochemical and morphological interaction of the cytoskeleton

  12. Molecular photosensitisers for two-photon photodynamic therapy.

    Science.gov (United States)

    Bolze, F; Jenni, S; Sour, A; Heitz, V

    2017-11-30

    Two-photon excitation has attracted the attention of biologists, especially after the development of two-photon excited microscopy in the nineties. Since then, new applications have rapidly emerged such as the release of biologically active molecules and photodynamic therapy (PDT) using two-photon excitation. PDT, which requires a light-activated drug (photosensitiser), is a clinically approved and minimally invasive treatment for cancer and for non-malignant diseases. This feature article focuses on the engineering of molecular two-photon photosensitisers for PDT, which should bring important benefits to the treatment, increase the treatment penetration depth with near-infrared light excitation, improve the spatial selectivity and reduce the photodamage to healthy tissues. After an overview of the two-photon absorption phenomenon and the methods to evaluate two-photon induced phototoxicity on cell cultures, the different classes of photosensitisers described in the literature are discussed. The two-photon PDT performed with historical one-photon sensitisers are briefly presented, followed by specifically engineered cyclic tetrapyrrole photosensitisers, purely organic photosensitisers and transition metal complexes. Finally, targeted two-photon photosensitisers and theranostic agents that should enhance the selectivity and efficiency of the treatment are discussed.

  13. Volumetry of human taste buds using laser scanning microscopy.

    Science.gov (United States)

    Just, T; Srur, E; Stachs, O; Pau, H W

    2009-10-01

    In vivo laser scanning confocal microscopy is a relatively new, non-invasive method for assessment of oral cavity epithelia. The penetration depth of approximately 200-400 microm allows visualisation of fungiform papillae and their taste buds. This paper describes the technique of in vivo volumetry of human taste buds. Confocal laser scanning microscopy used a diode laser at 670 nm for illumination. Digital laser scanning confocal microscopy equipment consisted of the Heidelberg Retina Tomograph HRTII and the Rostock Cornea Module. Volume scans of fungiform papillae were used for three-dimensional reconstruction of the taste bud. This technique supplied information on taste bud structure and enabled measurement and calculation of taste bud volume. Volumetric data from a 23-year-old man over a nine-day period showed only a small deviation in values. After three to four weeks, phenomenological changes in taste bud structures were found (i.e. a significant increase in volume, followed by disappearance of the taste bud and appearance of a new taste bud). The data obtained indicate the potential application of this non-invasive imaging modality: to evaluate variation of taste bud volume in human fungiform papillae with ageing; to study the effects of chorda tympani nerve transection on taste bud volume; and to demonstrate recovery of taste buds in patients with a severed chorda tympani nerve who show recovery of gustatory sensibility after surgery.

  14. Image scanning microscopy using a SPAD detector array (Conference Presentation)

    Science.gov (United States)

    Castello, Marco; Tortarolo, Giorgio; Buttafava, Mauro; Tosi, Alberto; Sheppard, Colin J. R.; Diaspro, Alberto; Vicidomini, Giuseppe

    2017-02-01

    The use of an array of detectors can help overcoming the traditional limitation of confocal microscopy: the compromise between signal and theoretical resolution. Each element independently records a view of the sample and the final image can be reconstructed by pixel reassignment or by inverse filtering (e.g. deconvolution). In this work, we used a SPAD array of 25 detectors specifically designed for this goal and our scanning microscopy control system (Carma) to acquire the partial images and to perform online image processing. Further work will be devoted to optimize the image reconstruction step and to improve the fill-factor of the detector.

  15. Scanning conductance microscopy investigations on fixed human chromosomes

    DEFF Research Database (Denmark)

    Clausen, Casper Hyttel; Lange, Jacob Moresco; Jensen, Linda Boye

    2008-01-01

    Scanning conductance microscopy investigations were carried out in air on human chromosomes fixed on pre-fabricated SiO2 surfaces with a backgate. The point of the investigation was to estimate the dielectric constant of fixed human chromosomes in order to use it for microfluidic device...... optimization. The phase shift caused by the electrostatic forces, together with geometrical measurements of the atomic force microscopy (AFM) cantilever and the chromosomes were used to estimate a value,for the dielectric constant of different human chromosomes....

  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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Phase modulation mode of scanning ion conductance microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Li, Peng; Zhang, Changlin [State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Liu, Lianqing, E-mail: lqliu@sia.cn, E-mail: gli@engr.pitt.edu; Wang, Yuechao; Yang, Yang [State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016 (China); Li, Guangyong, E-mail: lqliu@sia.cn, E-mail: gli@engr.pitt.edu [Department of Electrical and Computer Engineering, University of Pittsburgh, Pittsburgh, Pennsylvania 15261 (United States)

    2014-08-04

    This Letter reports a phase modulation (PM) mode of scanning ion conductance microscopy. In this mode, an AC current is directly generated by an AC voltage between the electrodes. The portion of the AC current in phase with the AC voltage, which is the current through the resistance path, is modulated by the tip-sample distance. It can be used as the input of feedback control to drive the scanner in Z direction. The PM mode, taking the advantages of both DC mode and traditional AC mode, is less prone to electronic noise and DC drift but maintains high scanning speed. The effectiveness of the PM mode has been proven by experiments.

  18. Scanning microscopies of superconductors at very low temperatures

    International Nuclear Information System (INIS)

    Crespo, V.; Maldonado, A.; Galvis, J.A.; Kulkarni, P.; Guillamon, I.; Rodrigo, J.G.; Suderow, H.; Vieira, S.; Banerjee, S.; Rodiere, P.

    2012-01-01

    We discuss basics of Scanning Tunneling Microscopy and Spectroscopy (STM/S) of the superconducting state with normal and superconducting tips. We present a new method to measure the local variations in the Andreev reflection amplitude between a superconducting tip and the sample. This method is termed Scanning Andreev Reflection Spectroscopy (SAS). We also briefly discuss vortex imaging with STM/S under an applied current through the sample, and show the vortex lattice as a function of the angle between the magnetic field and sample’s surface.

  19. Scanning transmission ion microscopy on Fudan SPM facility

    International Nuclear Information System (INIS)

    Li Yongqiang; Shen Hao; Zheng Yi; Li Xinyi; Liu Bo; Satoh Takahiro

    2011-01-01

    In this paper, we report a novel measurement system based on the development of Fudan Scanning Proton Microscopy (SPM) facility. By using Si-PIN diode(Hamamatsu S1223-01) detector, scanning transmission ion microscopy (STIM) measurement system has been set up. It can provide density and structural images with high probing efficiency and non-destruction by utilizing the energy loss of high energy (MeV) and focused ions penetrating through a thin sample. STIM measurement is able to map the density distribution of organic elements which mostly compose biology materials, such information can not be detected by using conventional Be-windowed Si (Li) X-ray detector in Particle Induced X-ray Emission (PIXE) technique. The spatial resolution capability of STIM is higher than PIXE technique at same accelerator status. As a result of STIM measurement, Paramecium attached on the top of Kapton tube was measured by STIM. (authors)

  20. Local crystallography analysis for atomically resolved scanning tunneling microscopy images

    International Nuclear Information System (INIS)

    Lin, Wenzhi; Li, Qing; Belianinov, Alexei; Gai, Zheng; Baddorf, Arthur P; Pan, Minghu; Jesse, Stephen; Kalinin, Sergei V; Sales, Brian C; Sefat, Athena

    2013-01-01

    Scanning probe microscopy has emerged as a powerful and flexible tool for atomically resolved imaging of surface structures. However, due to the amount of information extracted, in many cases the interpretation of such data is limited to being qualitative and semi-quantitative in nature. At the same time, much can be learned from local atom parameters, such as distances and angles, that can be analyzed and interpreted as variations of local chemical bonding, or order parameter fields. Here, we demonstrate an iterative algorithm for indexing and determining atomic positions that allows the analysis of inhomogeneous surfaces. This approach is further illustrated by local crystallographic analysis of several real surfaces, including highly ordered pyrolytic graphite and an Fe-based superconductor FeTe 0.55 Se 0.45 . This study provides a new pathway to extract and quantify local properties for scanning probe microscopy images. (paper)

  1. Dysprosium disilicide nanostructures on silicon(001) studied by scanning tunneling microscopy and transmission electron microscopy

    International Nuclear Information System (INIS)

    Ye Gangfeng; Nogami, Jun; Crimp, Martin A.

    2006-01-01

    The microstructure of self-assembled dysprosium silicide nanostructures on silicon(001) has been studied by scanning tunneling microscopy and transmission electron microscopy. The studies focused on nanostructures that involve multiple atomic layers of the silicide. Cross-sectional high resolution transmission electron microscopy images and fast Fourier transform analysis showed that both hexagonal and orthorhombic/tetragonal silicide phases were present. Both the magnitude and the anisotropy of lattice mismatch between the silicide and the substrate play roles in the morphology and epitaxial growth of the nanostructures formed

  2. Scanning Electron Microscopy with Samples in an Electric Field

    Czech Academy of Sciences Publication Activity Database

    Frank, Luděk; Hovorka, Miloš; Mikmeková, Šárka; Mikmeková, Eliška; Müllerová, Ilona; Pokorná, Zuzana

    2012-01-01

    Roč. 5, č. 12 (2012), s. 2731-2756 ISSN 1996-1944 R&D Projects: GA ČR GAP108/11/2270; GA TA ČR TE01020118; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : scanning electron microscopy * slow electrons * low energy SEM * low energy STEM * cathode lens Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 2.247, year: 2012

  3. Scanning tunneling microscopy - STM: history, principle, construction, and related techniques

    International Nuclear Information System (INIS)

    Ostadal, I.; Sobotik, P.

    1998-01-01

    The method of scanning tunneling microscopy (STM) is discussed. The principle of STM and some other related methods is highlighted, and spectroscopy using STM is dealt with. A brief summary is given of problems to be solved in microscope design, and the influence of these on the resolution obtained is presented. A comparison of STM with other methods used at present in the physics of thin films and surfaces confirms its irreplaceability

  4. Detecting damage in steel with scanning SQUID microscopy

    International Nuclear Information System (INIS)

    Lee, Tae-Kyu; Clatterbuck, D.M.; Morris, J.W. Jr.; Shaw, T.J.; Lee, Seungkyun; Clarke, John

    2002-01-01

    A 'Holy Grail' of NDE research is a non-destructive method for measuring fatigue damage prior to crack initiation. High-Tc scanning SQUID microscopy may be a useful tool. Because of the exceptional magnetic sensitivity of this technique, fatigue damage can be detected well before microcrack initiation, and in the absence of other obvious microstructure or property changes. Given the spatial resolution of the technique, undamaged material can be located and used to set internal standards

  5. Detecting damage in steel with scanning SQUID microscopy

    International Nuclear Information System (INIS)

    Lee, Tae-Kyu; Clatterbuck, David; Morris Jr., J.W.; Shaw, T.J.; McDermott R.; Clarke, John

    2001-01-01

    A ''Holy Grail'' of NDE research is a non-destructive method for measuring fatigue damage prior to crack initiation. High-Tc scanning SQUID microscopy may be a useful tool. Because of the exceptional magnetic sensitivity of this technique, fatigue damage can be detected well before microcrack initiation, and in the absence of other obvious microstructure or property changes. Given the spatial resolution of the technique, undamaged material can be located and used to set internal standards

  6. Solid-State Quantum Computer Based on Scanning Tunneling Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Berman, G. P.; Brown, G. W.; Hawley, M. E.; Tsifrinovich, V. I.

    2001-08-27

    We propose a solid-state nuclear-spin quantum computer based on application of scanning tunneling microscopy (STM) and well-developed silicon technology. It requires the measurement of tunneling-current modulation caused by the Larmor precession of a single electron spin. Our envisioned STM quantum computer would operate at the high magnetic field ({approx}10 T) and at low temperature {approx}1 K .

  7. Solid-State Quantum Computer Based on Scanning Tunneling Microscopy

    International Nuclear Information System (INIS)

    Berman, G. P.; Brown, G. W.; Hawley, M. E.; Tsifrinovich, V. I.

    2001-01-01

    We propose a solid-state nuclear-spin quantum computer based on application of scanning tunneling microscopy (STM) and well-developed silicon technology. It requires the measurement of tunneling-current modulation caused by the Larmor precession of a single electron spin. Our envisioned STM quantum computer would operate at the high magnetic field (∼10 T) and at low temperature ∼1 K

  8. Second-harmonic scanning optical microscopy of poled silica waveguides

    DEFF Research Database (Denmark)

    Pedersen, Kjeld; Bozhevolnyi, Sergey I.; Arentoft, Jesper

    2000-01-01

    Second-harmonic scanning optical microscopy (SHSOM) is performed on electric-field poled silica-based waveguides. Two operation modes of SHSOM are considered. Oblique transmission reflection and normal reflection modes are used to image the spatial distribution of nonlinear susceptibilities...... and limitations of the two operation modes when used for SHSOM studies of poled silica-based waveguides are discussed. The influence of surface defects on the resulting second-harmonic images is also considered. ©2000 American Institute of Physics....

  9. Abrasion of 6 dentifrices measured by vertical scanning interference microscopy.

    Science.gov (United States)

    Pascaretti-Grizon, Florence; Mabilleau, Guillaume; Chappard, Daniel

    2013-01-01

    The abrasion of dentifrices is well recognized to eliminate the dental plaque. The aims of this study were to characterize the abrasive powders of 6 dentifrices (3 toothpastes and 3 toothpowders) and to measure the abrasion on a test surface by Vertical Scanning Interference microscopy (VSI). Bright field and polarization microscopy were used to identify the abrasive particles on the crude dentifrices and after prolonged washes. Scanning electron microscopy and microanalysis characterized the shape and nature of the particles. Standardized and polished blocks of poly(methylmethacrylate) were brushed with a commercial electric toothbrush with the dentifrices. VSI quantified the mean roughness (Ra) and illustrated in 3D the abraded areas. Toothpastes induced a limited abrasion. Toothpowders induced a significantly higher roughness linked to the size of the abrasive particles. One powder (Gencix® produced a high abrasion when used with a standard testing weight. However, the powder is based on pumice particles covered by a plant homogenate that readily dissolves in water. When used in the same volume, or after dispersion in water, Ra was markedly reduced. Light and electron microscopy characterize the abrasive particles and VSI is a new tool allowing the analysis of large surface of abraded materials.

  10. Analysis of leaf surfaces using scanning ion conductance microscopy.

    Science.gov (United States)

    Walker, Shaun C; Allen, Stephanie; Bell, Gordon; Roberts, Clive J

    2015-05-01

    Leaf surfaces are highly complex functional systems with well defined chemistry and structure dictating the barrier and transport properties of the leaf cuticle. It is a significant imaging challenge to analyse the very thin and often complex wax-like leaf cuticle morphology in their natural state. Scanning electron microscopy (SEM) and to a lesser extent Atomic force microscopy are techniques that have been used to study the leaf surface but their remains information that is difficult to obtain via these approaches. SEM is able to produce highly detailed and high-resolution images needed to study leaf structures at the submicron level. It typically operates in a vacuum or low pressure environment and as a consequence is generally unable to deal with the in situ analysis of dynamic surface events at submicron scales. Atomic force microscopy also possess the high-resolution imaging required and can follow dynamic events in ambient and liquid environments, but can over exaggerate small features and cannot image most leaf surfaces due to their inherent roughness at the micron scale. Scanning ion conductance microscopy (SICM), which operates in a liquid environment, provides a potential complementary analytical approach able to address these issues and which is yet to be explored for studying leaf surfaces. Here we illustrate the potential of SICM on various leaf surfaces and compare the data to SEM and atomic force microscopy images on the same samples. In achieving successful imaging we also show that SICM can be used to study the wetting of hydrophobic surfaces in situ. This has potentially wider implications than the study of leaves alone as surface wetting phenomena are important in a range of fundamental and applied studies. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  11. Head-facial hemangiomas studied with scanning electron microscopy.

    Science.gov (United States)

    Cavallotti, Carlo; Cavallotti, Chiara; Giovannetti, Filippo; Iannetti, Giorgio

    2009-11-01

    Hemangiomas of the head or face are a frequent vascular pathology, consisting in an embryonic dysplasia that involves the cranial-facial vascular network. Hemangiomas show clinical, morphological, developmental, and structural changes during their course. Morphological, structural, ultrastructural, and clinical characteristics of head-facial hemangiomas were studied in 28 patients admitted in our hospital. Nineteen of these patients underwent surgery for the removal of the hemangiomas, whereas 9 patients were not operated on. All the removed tissues were transferred in our laboratories for the morphological staining. Light microscopy, transmission electron microscopy, and scanning electron microscopy techniques were used for the observation of all microanatomical details. All patients were studied for a clinical diagnosis, and many were subjected to surgical therapy. The morphological results revealed numerous microanatomical characteristics of the hemangiomatous vessels. The observation by light microscopy shows the afferent and the efferent vessels for every microhemangioma. All the layers of the arterial wall are uneven. The lumen of the arteriole is entirely used by a blood clot. The observation by transmission electron microscopy shows that it was impossible to see the limits of the different layers (endothelium, medial layer, and adventitia) in the whole wall of the vessels. Moreover, both the muscular and elastic components are disarranged and replaced with connective tissue. The observation by scanning electron microscopy shows that the corrosion cast of the hemangioma offers 3 periods of filling: initially with partial filling of the arteriolar and of the whole cast, intermediate with the entire filling of the whole cast (including arteriole and venule), and a last period with a partial emptying of the arteriolar and whole cast while the venule remains totally injected with resin. Our morphological results can be useful to clinicians for a precise

  12. Potential Applications of Scanning Probe Microscopy in Forensic Science

    Energy Technology Data Exchange (ETDEWEB)

    Watson, G S [Nanoscale Science and Technology Centre, School of Science, Griffith University, Kessels Rd, Nathan, QLD, 4111 (Australia); Watson, J A [Nanoscale Science and Technology Centre, School of Science, Griffith University, Kessels Rd, Nathan, QLD, 4111 (Australia)

    2007-04-15

    The forensic community utilises a myriad of techniques to investigate a wide range of materials, from paint flakes to DNA. The various microscopic techniques have provided some of the greatest contributions, e.g., FT-IR (Fourier-transform infrared) microspectroscopy utilised in copy toner discrimination, multi-layer automobile paint fragment examination, etc, SEM-EDA (scanning electron microscopy with energy dispersive analysis) used to investigate glass fragments, fibers, and explosives, and SEM in microsampling for elemental analysis, just to name a few. This study demonstrates the ability of the Scanning Probe Microscope (SPM) to analyse human fingerprints on surfaces utilising a step-and-scan feature, enabling analysis of a larger field-of-view. We also extend a line crossings study by incorporating height analysis and surface roughness measurements. The study demonstrates the potential for SPM techniques to be utilised for forensic analysis which could complement the more traditional methodologies used in such investigations.

  13. Imaging ballistic carrier trajectories in graphene using scanning gate microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Morikawa, Sei; Masubuchi, Satoru [Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153-8505 (Japan); Dou, Ziwei; Wang, Shu-Wei; Smith, Charles G.; Connolly, Malcolm R., E-mail: mrc61@cam.ac.uk [Cavendish Laboratory, Department of Physics, University of Cambridge, Cambridge CB3 0HE (United Kingdom); Watanabe, Kenji; Taniguchi, Takashi [National Institute for Materials Science, 1-1 Namiki, Tsukuba 305-0044 (Japan); Machida, Tomoki, E-mail: tmachida@iis.u-tokyo.ac.jp [Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153-8505 (Japan); Institute for Nano Quantum Information Electronics, University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153-8505 (Japan)

    2015-12-14

    We use scanning gate microscopy to map out the trajectories of ballistic carriers in high-mobility graphene encapsulated by hexagonal boron nitride and subject to a weak magnetic field. We employ a magnetic focusing geometry to image carriers that emerge ballistically from an injector, follow a cyclotron path due to the Lorentz force from an applied magnetic field, and land on an adjacent collector probe. The local electric field generated by the scanning tip in the vicinity of the carriers deflects their trajectories, modifying the proportion of carriers focused into the collector. By measuring the voltage at the collector while scanning the tip, we are able to obtain images with arcs that are consistent with the expected cyclotron motion. We also demonstrate that the tip can be used to redirect misaligned carriers back to the collector.

  14. Potential Applications of Scanning Probe Microscopy in Forensic Science

    International Nuclear Information System (INIS)

    Watson, G S; Watson, J A

    2007-01-01

    The forensic community utilises a myriad of techniques to investigate a wide range of materials, from paint flakes to DNA. The various microscopic techniques have provided some of the greatest contributions, e.g., FT-IR (Fourier-transform infrared) microspectroscopy utilised in copy toner discrimination, multi-layer automobile paint fragment examination, etc, SEM-EDA (scanning electron microscopy with energy dispersive analysis) used to investigate glass fragments, fibers, and explosives, and SEM in microsampling for elemental analysis, just to name a few. This study demonstrates the ability of the Scanning Probe Microscope (SPM) to analyse human fingerprints on surfaces utilising a step-and-scan feature, enabling analysis of a larger field-of-view. We also extend a line crossings study by incorporating height analysis and surface roughness measurements. The study demonstrates the potential for SPM techniques to be utilised for forensic analysis which could complement the more traditional methodologies used in such investigations

  15. Simultaneous correlative scanning electron and high-NA fluorescence microscopy.

    Directory of Open Access Journals (Sweden)

    Nalan Liv

    Full Text Available Correlative light and electron microscopy (CLEM is a unique method for investigating biological structure-function relations. With CLEM protein distributions visualized in fluorescence can be mapped onto the cellular ultrastructure measured with electron microscopy. Widespread application of correlative microscopy is hampered by elaborate experimental procedures related foremost to retrieving regions of interest in both modalities and/or compromises in integrated approaches. We present a novel approach to correlative microscopy, in which a high numerical aperture epi-fluorescence microscope and a scanning electron microscope illuminate the same area of a sample at the same time. This removes the need for retrieval of regions of interest leading to a drastic reduction of inspection times and the possibility for quantitative investigations of large areas and datasets with correlative microscopy. We demonstrate Simultaneous CLEM (SCLEM analyzing cell-cell connections and membrane protrusions in whole uncoated colon adenocarcinoma cell line cells stained for actin and cortactin with AlexaFluor488. SCLEM imaging of coverglass-mounted tissue sections with both electron-dense and fluorescence staining is also shown.

  16. Correlative Analysis of Immunoreactivity in Confocal Laser-Scanning Microscopy and Scanning Electron Microscopy with Focused Ion Beam Milling

    Directory of Open Access Journals (Sweden)

    Takahiro eSonomura

    2013-02-01

    Full Text Available Three-dimensional reconstruction of ultrastructure of rat brain with minimal effort has recently been realized by scanning electron microscopy combined with focused ion beam milling (FIB-SEM. Because application of immunohistochemical staining to electron microscopy has a great advantage in that molecules of interest are specifically localized in ultrastructures, we here tried to apply immunocytochemistry to FIB-SEM and correlate immunoreactivity in confocal laser-scanning microcopy (CF-LSM with that in FIB-SEM. The dendrites of medium-sized spiny neurons in rat neostriatum were visualized with a recombinant viral vector, which labeled the infected neurons with membrane-targeted GFP in a Golgi stain-like fashion, and thalamostriatal afferent terminals were immunolabeled with Cy5 fluorescence for vesicular glutamate transporter 2 (VGluT2. After detecting the sites of terminals apposed to the dendrites in CF-LSM, GFP and VGluT2 immunoreactivities were further developed for electron microscopy by the immunogold/silver enhancement and immunoperoxidase/diaminobenzidine (DAB methods, respectively. In the contrast-inverted FIB-SEM images, silver precipitation and DAB deposits were observed as fine dark grains and diffuse dense profiles, respectively, indicating that these immunoreactivities were easily recognizable as in the images of transmission electron microscopy. In the sites of interest, some appositions were revealed to display synaptic specialization of asymmetric type. The present method is thus useful in the three-dimensional analysis of immunocytochemically differentiated synaptic connection in the central neural circuit.

  17. Gold nanocone probes for near-field scanning optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Zeeb, Bastian; Schaefer, Christian; Nill, Peter; Fleischer, Monika; Kern, Dieter P. [Institute of Applied Physics, University of Tuebingen, Auf der Morgenstelle 10, 72076 Tuebingen (Germany)

    2010-07-01

    Apertureless near-field scanning optical microscopy (ANSOM) provides the possibility to collect simultaneously high-resolution topographical and sub-diffraction limited optical information from a surface. When optically excited, the scanning probes act as optical antennae with a strong near-field enhancement near the tip apex. Spatial resolution and optical near-field enhancement depend strongly on the properties and geometry of the scanning probe - in particular on very sharp tip radii. Various possibilities for fabricating good antennae have been pursued. Most commonly, scanning probes consist of electrochemically etched gold wires which are sharp but not well-defined in geometry. We present two different approaches for ultra sharp and well-defined antennae based upon fabricating gold nanocones with a tip radius smaller than 10 nm which can be used in ANSOM. A transfer process is presented that can be used to attach single gold nanocones to non-metallic probes such as sharp glass fiber tips. Alternatively, new processes are presented to fabricate cones directly on pillars of different materials such as silicon or bismuth, which can be applied to cantilever tips for ANSOM scanning applications.

  18. Stereoscopic and photometric surface reconstruction in scanning electron microscopy

    International Nuclear Information System (INIS)

    Scherer, S.

    2000-01-01

    The scanning electron microscope (SEM) is one of the most important devices to examine microscopic structures as it offers images of a high contrast range with a large depth of focus. Nevertheless, three-dimensional measurements, as desired in fracture mechanics, have previously not been accomplished. This work presents a system for automatic, robust and dense surface reconstruction in scanning electron microscopy combining new approaches in shape from stereo and shape from photometric stereo. The basic theoretical assumption for a known adaptive window algorithm is shown not to hold in scanning electron microscopy. A constraint derived from this observation yields a new, simplified, hence faster calculation of the adaptive window. The correlation measure itself is obtained by a new ordinal measure coefficient. Shape from photometric stereo in the SEM is formulated by relating the image formation process with conventional photography. An iterative photometric ratio reconstruction is invented based on photometric ratios of backscatter electron images. The performance of the proposed system is evaluated using ground truth data obtained by three alternative shape recovery devices. Most experiments showed relative height accuracy within the tolerances of the alternative devices. (author)

  19. Scanning Ion Conductance Microscopy for Studying Biological Samples

    Directory of Open Access Journals (Sweden)

    Irmgard D. Dietzel

    2012-11-01

    Full Text Available Scanning ion conductance microscopy (SICM is a scanning probe technique that utilizes the increase in access resistance that occurs if an electrolyte filled glass micro-pipette is approached towards a poorly conducting surface. Since an increase in resistance can be monitored before the physical contact between scanning probe tip and sample, this technique is particularly useful to investigate the topography of delicate samples such as living cells. SICM has shown its potential in various applications such as high resolution and long-time imaging of living cells or the determination of local changes in cellular volume. Furthermore, SICM has been combined with various techniques such as fluorescence microscopy or patch clamping to reveal localized information about proteins or protein functions. This review details the various advantages and pitfalls of SICM and provides an overview of the recent developments and applications of SICM in biological imaging. Furthermore, we show that in principle, a combination of SICM and ion selective micro-electrodes enables one to monitor the local ion activity surrounding a living cell.

  20. Integrated Confocal and Scanning Probe Microscopy for Biomedical Research

    Directory of Open Access Journals (Sweden)

    B.J. Haupt

    2006-01-01

    Full Text Available Atomic force microscopy (AFM continues to be developed, not only in design, but also in application. The new focus of using AFM is changing from pure material to biomedical studies. More frequently, it is being used in combination with other optical imaging methods, such as confocal laser scanning microscopy (CLSM and fluorescent imaging, to provide a more comprehensive understanding of biological systems. To date, AFM has been used increasingly as a precise micromanipulator, probing and altering the mechanobiological characteristics of living cells and tissues, in order to examine specific, receptor-ligand interactions, material properties, and cell behavior. In this review, we discuss the development of this new hybrid AFM, current research, and potential applications in diagnosis and the detection of disease.

  1. High resolution helium ion scanning microscopy of the rat kidney.

    Directory of Open Access Journals (Sweden)

    William L Rice

    Full Text Available Helium ion scanning microscopy is a novel imaging technology with the potential to provide sub-nanometer resolution images of uncoated biological tissues. So far, however, it has been used mainly in materials science applications. Here, we took advantage of helium ion microscopy to explore the epithelium of the rat kidney with unsurpassed image quality and detail. In addition, we evaluated different tissue preparation methods for their ability to preserve tissue architecture. We found that high contrast, high resolution imaging of the renal tubule surface is possible with a relatively simple processing procedure that consists of transcardial perfusion with aldehyde fixatives, vibratome tissue sectioning, tissue dehydration with graded methanol solutions and careful critical point drying. Coupled with the helium ion system, fine details such as membrane texture and membranous nanoprojections on the glomerular podocytes were visualized, and pores within the filtration slit diaphragm could be seen in much greater detail than in previous scanning EM studies. In the collecting duct, the extensive and striking apical microplicae of the intercalated cells were imaged without the shrunken or distorted appearance that is typical with conventional sample processing and scanning electron microscopy. Membrane depressions visible on principal cells suggest possible endo- or exocytotic events, and central cilia on these cells were imaged with remarkable preservation and clarity. We also demonstrate the use of colloidal gold probes for highlighting specific cell-surface proteins and find that 15 nm gold labels are practical and easily distinguishable, indicating that external labels of various sizes can be used to detect multiple targets in the same tissue. We conclude that this technology represents a technical breakthrough in imaging the topographical ultrastructure of animal tissues. Its use in future studies should allow the study of fine cellular details

  2. Very low energy scanning electron microscopy in nanotechnology

    Czech Academy of Sciences Publication Activity Database

    Müllerová, Ilona; Hovorka, Miloš; Mika, Filip; Mikmeková, Eliška; Mikmeková, Šárka; Pokorná, Zuzana; Frank, Luděk

    2012-01-01

    Roč. 9, 8/9 (2012), s. 695-716 ISSN 1475-7435 R&D Projects: GA MŠk OE08012; GA MŠk ED0017/01/01; GA AV ČR IAA100650902 Institutional research plan: CEZ:AV0Z20650511 Keywords : scanning electron microscopy * very low energy electrons * cathode lens * grain contrast * strain contrast * imaging of participates * dopant contrast * very low energy STEM * graphene Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.087, year: 2012

  3. Analysis of archaeological materials through Scanning electron microscopy

    International Nuclear Information System (INIS)

    Camacho, A.; Tenorio C, D.; Elizalde, S.; Mandujano, C.; Cassiano, G.

    2005-01-01

    With the purpose to know the uses and the chemical composition of some cultural objects in the pre hispanic epoch this work presents several types of analysis for identifying them by means of the Scanning electron microscopy and its techniques as the Functional analysis of artifacts based on the 'tracks of use' analysis, also the X-ray spectroscopy and the X-ray dispersive energy (EDS) are mentioned, all of them allowing a major approach to the pre hispanic culture in Mexico. (Author)

  4. Fracture characteristics of uranium alloys by scanning electron microscopy

    International Nuclear Information System (INIS)

    Koger, J.W.; Bennett, R.K. Jr.

    1976-10-01

    The fracture characteristics of uranium alloys were determined by scanning electron microscopy. The fracture mode of stress-corrosion cracking (SCC) of uranium-7.5 weight percent niobium-2.5 weight percent zirconium (Mulberry) alloy, uranium--niobium alloys, and uranium--molybdenum alloys in aqueous chloride solutions is intergranular. The SCC fracture surface of the Mulberry alloy is characterized by very clean and smooth grain facets. The tensile-overload fracture surfaces of these alloys are characteristically ductile dimple. Hydrogen-embrittlement failures of the uranium alloys are brittle and the fracture mode is transgranular. Fracture surfaces of the uranium-0.75 weight percent titanium alloys are quasi cleavage

  5. Analysis of main artifacts in scanning probe microscopy (1)

    International Nuclear Information System (INIS)

    Alekperov, S.D.; Alekperov, S.D.

    2012-01-01

    The analysis of experiment carrying methodology in the scanning probe microscopy (SPM) region is carried out, the main parameters influencing on image quality are revealed. In order to reveal the artifact reason the main components of SPM signal which are divided on 5 groups : the useful signal; noises connected with external influences and temperature drift; distortions connected with piezoceramics and piezo-scanner non-ideality; probe geometry influence; apparatus noises are considered. The main methods of removal and minimization of the given artifacts are considered. The second and third groups of main components of SPM signal are considered in the articles first part

  6. Contrast distortion induced by modulation voltage in scanning capacitance microscopy

    Science.gov (United States)

    Chang, M. N.; Hu, C. W.; Chou, T. H.; Lee, Y. J.

    2012-08-01

    With a dark-mode scanning capacitance microscopy (SCM), we directly observed the influence of SCM modulation voltage (MV) on image contrasts. For electrical junctions, an extensive modulated area induced by MV may lead to noticeable changes in the SCM signal phase and intensity, resulting in a narrowed junction image and a broadened carrier concentration profile. This contrast distortion in SCM images may occur even if the peak-to-peak MV is down to 0.3 V. In addition, MV may shift the measured electrical junction depth. The balance of SCM signals components explain these MV-induced contrast distortions.

  7. Multifrequency scanning probe microscopy study of nanodiamond agglomerates

    Science.gov (United States)

    Aravind, Vasudeva; Lippold, Stephen; Li, Qian; Strelcov, Evgheny; Okatan, Baris; Legum, Benjamin; Kalinin, Sergei; Clarion University Team; Oak Ridge National Laboratory Team

    Due to their rich surface chemistry and excellent mechanical properties and non-toxic nature, nanodiamond particles have found applications such as biomedicine, tribology and lubrication, targeted drug delivery systems, tissue scaffolds and surgical implants. Although single nanodiamond particles have diameters about 4-5nm, they tend to form agglomerates. While these agglomerates can be useful for some purposes, many applications of nanodiamonds require single particle, disaggregated nanodiamonds. This work is oriented towards studying forces and interactions that contribute to agglomeration in nanodiamonds. In this work, using multifrequency scanning probe microscopy techniques, we show that agglomerate sizes can vary between 50-100nm in raw nanodiamonds. Extremeties of particles and Interfaces between agglomerates show dissipative forces with scanning probe microscope tip, indicating agglomerates could act as points of increased adhesion, thus reducing lubricating efficiency when nanodiamonds are used as lubricant additives. This research was conducted at the Center for Nanophase Materials Sciences, which is a DOE Office of Science User Facility.

  8. Scanning tunneling microscopy and spectroscopy studies of graphite edges

    International Nuclear Information System (INIS)

    Niimi, Y.; Matsui, T.; Kambara, H.; Tagami, K.; Tsukada, M.; Fukuyama, Hiroshi

    2005-01-01

    We studied experimentally and theoretically the electronic local density of states (LDOS) near single-step edges at the surface of exfoliated graphite. In scanning tunneling microscopy measurements, we observed the (3x3)R30 o and honeycomb superstructures extending over 3-4-bar nm both from the zigzag and armchair edges. Calculations based on a density-functional-derived non-orthogonal tight-binding model show that these superstructures can coexist if the two types of edges admix each other in real graphite step edges. Scanning tunneling spectroscopy measurements near the zigzag edge reveal a clear peak in the LDOS at an energy below the Fermi energy by 20-bar meV. No such a peak was observed near the armchair edge. We concluded that this peak corresponds to the 'edge state' theoretically predicted for graphene ribbons, since a similar prominent LDOS peak due to the edge state is obtained by the first principles calculations

  9. Scanning gate microscopy on graphene: charge inhomogeneity and extrinsic doping

    International Nuclear Information System (INIS)

    Jalilian, Romaneh; Tian Jifa; Chen, Yong P; Jauregui, Luis A; Lopez, Gabriel; Roecker, Caleb; Jovanovic, Igor; Yazdanpanah, Mehdi M; Cohn, Robert W

    2011-01-01

    We have performed scanning gate microscopy (SGM) on graphene field effect transistors (GFET) using a biased metallic nanowire coated with a dielectric layer as a contact mode tip and local top gate. Electrical transport through graphene at various back gate voltages is monitored as a function of tip voltage and tip position. Near the Dirac point, the response of graphene resistance to the tip voltage shows significant variation with tip position, and SGM imaging displays mesoscopic domains of electron-doped and hole-doped regions. Our measurements reveal substantial spatial fluctuation in the carrier density in graphene due to extrinsic local doping from sources such as metal contacts, graphene edges, structural defects and resist residues. Our scanning gate measurements also demonstrate graphene's excellent capability to sense the local electric field and charges.

  10. A computer program for scanning transmission ion microscopy simulation

    International Nuclear Information System (INIS)

    Wu, R.; Shen, H.; Mi, Y.; Sun, M.D.; Yang, M.J.

    2005-01-01

    With the installation of the Scanning Proton Microprobe system at Fudan University, we are in the process of developing a three-dimension reconstruction technique based on scanning transmission ion microscopy-computed tomography (STIM-CT). As the first step, a related computer program of STIM simulation has been established. This program is written in the Visual C++[reg], using the technique of OOP (Object Oriented Programming) and it is a standard multiple-document Windows[reg] program. It can be run with all MS Windows[reg] operating systems. The operating mode is the menu mode, using a multiple process technique. The stopping power theory is based on the Bethe-Bloch formula. In order to simplify the calculation, the improved cylindrical coordinate model was introduced in the program instead of a usual spherical or cylindrical coordinate model. The simulated results of a sample at several rotation angles are presented

  11. Quantitative phase imaging with scanning holographic microscopy: an experimental assesment

    Directory of Open Access Journals (Sweden)

    Tada Yoshitaka

    2006-11-01

    Full Text Available Abstract This paper demonstrates experimentally how quantitative phase information can be obtained in scanning holographic microscopy. Scanning holography can operate in both coherent and incoherent modes, simultaneously if desired, with different detector geometries. A spatially integrating detector provides an incoherent hologram of the object's intensity distribution (absorption and/or fluorescence, for example, while a point detector in a conjugate plane of the pupil provides a coherent hologram of the object's complex amplitude, from which a quantitative measure of its phase distribution can be extracted. The possibility of capturing simultaneously holograms of three-dimensional specimens, leading to three-dimensional reconstructions with absorption contrast, reflectance contrast, fluorescence contrast, as was previously demonstrated, and quantitative phase contrast, as shown here for the first time, opens up new avenues for multimodal imaging in biological studies.

  12. Examination of living fungal spores by scanning electron microscopy

    International Nuclear Information System (INIS)

    Read, N.D.; Lord, K.M.

    1991-01-01

    Ascospores of Sordaria macrospora germinated and produced hyphae exhibiting normal growth and differentiation after examination by scanning electron microscopy and following numerous, different preparative protocols. Seventy-nine to ninety-nine percent of the ascospores retained normal viability after being observed in the fully frozen-hydrated, partially freeze-dried, and vacuum-dried states at accelerating voltages of 5 and 40 keV. Hyphae did not survive these treatments. From these observations it is concluded that ascospores of S. macrospora can remain in a state of suspended animation while being observed in the scanning electron microscope. The ascospores also survived, but with reduced viability: 6 h in glutaraldehyde and formaldehyde, 6 h in OsO4, or 2 h in glutaraldehyde and formaldehyde followed by 2 h in OsO 4 . However, the ascospores did not germinate after dehydration in ethanol. (author)

  13. Nanometrology using a through-focus scanning optical microscopy method

    International Nuclear Information System (INIS)

    Attota, Ravikiran; Silver, Richard

    2011-01-01

    We present an initial review of a novel through-focus scanning optical microscopy (TSOM pronounced as 'tee-som') imaging method that produces nanometer-dimensional measurement sensitivity using a conventional bright-field optical microscope. In the TSOM method a target is scanned through the focus of an optical microscope, acquiring conventional optical images at different focal positions. The TSOM images are constructed using the through-focus optical images. A TSOM image is unique under given experimental conditions and is sensitive to changes in the dimensions of a target in a distinct way. We use this characteristic for nanoscale-dimensional metrology. This technique can be used to identify the dimension which is changing between two nanosized targets and to determine the dimensions using a library-matching method. This methodology has potential utility for a wide range of target geometries and application areas, including nanometrology, nanomanufacturing, defect analysis, inspection, process control and biotechnology

  14. Through-focus scanning optical microscopy (TSOM) with adaptive optics

    Science.gov (United States)

    Lee, Jun Ho; Park, Gyunam; Jeong, Junhee; Park, Chris

    2018-03-01

    Through-focus optical microscopy (TSOM) with nanometer-scale lateral and vertical sensitivity levels matching those of scanning electron microscopy has been demonstrated to be useful both for 3D inspections and metrology assessments. In 2014, funded by two private companies (Nextin/Samsung Electronics) and the Korea Evaluation Institute of Industrial Technology (KEIT), a research team from four universities in South Korea set out to investigate core technologies for developing in-line TSOM inspection and metrology tools, with the respective teams focusing on optics implementation, defect inspection, computer simulation and high-speed metrology matching. We initially confirmed the reported validity of the TSOM operation through a computer simulation, after which we implemented the TSOM operation by throughfocus scanning of existing UV (355nm) and IR (800nm) inspection tools. These tools have an identical sampling distance of 150 nm but have different resolving distances (310 and 810 nm, respectively). We initially experienced some improvement in the defect inspection sensitivity level over TSV (through-silicon via) samples with 6.6 μm diameters. However, during the experiment, we noted sensitivity and instability issues when attempting to acquire TSOM images. As TSOM 3D information is indirectly extracted by differentiating a target TSOM image from reference TSOM images, any instability or mismatch in imaging conditions can result in measurement errors. As a remedy to such a situation, we proposed the application of adaptive optics to the TSOM operation and developed a closed-loop system with a tip/tilt mirror and a Shack-Hartmann sensor on an optical bench. We were able to keep the plane position within in RMS 0.4 pixel by actively compensating for any position instability which arose during the TSOM scanning process along the optical axis. Currently, we are also developing another TSOM tool with a deformable mirror instead of a tip/tilt mirror, in which case we

  15. The relevance of electrostatics for scanning-gate microscopy

    International Nuclear Information System (INIS)

    Schnez, S; Guettinger, J; Stampfer, C; Ensslin, K; Ihn, T

    2011-01-01

    Scanning-probe techniques have been developed to extract local information from a given physical system. In particular, conductance maps obtained by means of scanning-gate microscopy (SGM), where a conducting tip of an atomic-force microscope is used as a local and movable gate, seem to present an intuitive picture of the underlying physical processes. Here, we argue that the interpretation of such images is complex and not very intuitive under certain circumstances: scanning a graphene quantum dot (QD) in the Coulomb-blockaded regime, we observe an apparent shift of features in scanning-gate images as a function of gate voltages, which cannot be a real shift of the physical system. Furthermore, we demonstrate the appearance of more than one set of Coulomb rings arising from the graphene QD. We attribute these effects to screening between the metallic tip and the gates. Our results are relevant for SGM on any kind of nanostructure, but are of particular importance for nanostructures that are not covered with a dielectric, e.g. graphene or carbon nanotube structures.

  16. From Graphite to Graphene via Scanning Tunneling Microscopy

    Science.gov (United States)

    Qi, Dejun

    The primary objective of this dissertation is to study both graphene on graphite and pristine freestanding grapheme using scanning tunneling microscopy (STM) and density functional theory (DFT) simulation technique. In the experiment part, good quality tungsten metalic tips for experiment were fabricated using our newly developed tip making setup. Then a series of measurements using a technique called electrostatic-manipulation scanning tunneling microscopy (EM-STM) of our own development were performed on a highly oriented pyrolytic graphite (HOPG) surface. The electrostatic interaction between the STM tip and the sample can be tuned to produce both reversible and irreversible large-scale movement of the graphite surface. Under this influence, atomic-resolution STM images reveal that a continuous electronic transition between two distinct patterns can be systematically controlled. DFT calculations reveal that this transition can be related to vertical displacements of the top layer of graphite relative to the bulk. Evidence for horizontal shifts in the top layer of graphite is also presented. Excellent agreement is found between experimental STM images and those simulated using DFT. In addition, the EM-STM technique was also used to controllably and reversibly pull freestanding graphene membranes up to 35 nm from their equilibrium height. Atomic-scale corrugation amplitudes 20 times larger than the STM electronic corrugation for graphene on a substrate were observed. The freestanding graphene membrane responds to a local attractive force created at the STM tip as a highly conductive yet flexible grounding plane with an elastic restoring force.

  17. Spin-polarized scanning tunneling microscopy: breakthroughs and highlights.

    Science.gov (United States)

    Bode, Matthias

    2012-01-01

    The principle of scanning tunneling microscopy, an imaging method with atomic resolution capability invented by Binnig and Rohrer in 1982, can be adapted for surface magnetism studies by using magnetic probe tips. The contrast mechanism of this so-called spin-polarized scanning tunneling microscopy, or SP-STM, relies on the tunneling magneto-resistance effect, i.e. the tip-sample distance as well as the differential conductance depend on the relative magnetic orientation of tip and sample. To illustrate the working principle and the unique capabilities of SP-STM, this compilation presents some key experiments which have been performed on various magnetic surfaces, such as the topological antiferromagnet Cr(001), a double-layer of Fe which exhibits a stripe- domain pattern with about 50 nm periodicity, and the Mn monolayer on W(110), where the combination of experiment and theory reveal an antiferromagnetic spin cycloid. Recent experimental results also demonstrate the suitability of SP-STM for studies of dynamic properties, such as the spin relaxation time of single magnetic nanostructures.

  18. Humidity effects on scanning polarization force microscopy imaging

    Energy Technology Data Exchange (ETDEWEB)

    Shen, Yue, E-mail: shenyue@isl.ac.cn [Key Laboratory of Comprehensive and Highly Efficient Utilization of Salt Lake Resources, Key Laboratory of Salt Lake Resources Chemistry of Qinghai Province, Qinghai Institute of Salt Lakes, Chinese Academy of Sciences, Xining, Qinghai 810008 (China); Key Laboratory of Interfacial Physics and Technology of Chinese Academy of Sciences, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China); Zhou, Yuan, E-mail: zhouy@isl.ac.cn [Key Laboratory of Comprehensive and Highly Efficient Utilization of Salt Lake Resources, Key Laboratory of Salt Lake Resources Chemistry of Qinghai Province, Qinghai Institute of Salt Lakes, Chinese Academy of Sciences, Xining, Qinghai 810008 (China); Sun, Yanxia; Zhang, Lijuan [Key Laboratory of Comprehensive and Highly Efficient Utilization of Salt Lake Resources, Key Laboratory of Salt Lake Resources Chemistry of Qinghai Province, Qinghai Institute of Salt Lakes, Chinese Academy of Sciences, Xining, Qinghai 810008 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Wang, Ying; Hu, Jun; Zhang, Yi [Key Laboratory of Interfacial Physics and Technology of Chinese Academy of Sciences, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China)

    2017-08-01

    Highlights: • The humidity dramatically affects the contrast of scanning polarization force microscopy (SPFM) imaging on mica surface. • This influence roots in the sensitive dielectric constant of mica surface to the humidity change. • A strategy of controllable and repeatable imaging the local dielectric properties of nanomaterials with SPFM is proposed. - Abstract: Scanning polarization force microscopy (SPFM) is a useful surface characterization technique to visually characterize and distinguish nanomaterial with different local dielectric properties at nanometer scale. In this paper, taking the individual one-atom-thick graphene oxide (GO) and reduced graphene oxide (rGO) sheets on mica as examples, we described the influences of environmental humidity on SPFM imaging. We found that the apparent heights (AHs) or contrast of SPFM imaging was influenced significantly by relative humidity (RH) at a response time of a few seconds. And this influence rooted in the sensitive dielectric constant of mica surface to the RH change. While dielectric properties of GO and rGO sheets were almost immune to the humidity change. In addition, we gave the method to determine the critical humidity at which the contrast conversion happened under different conditions. And this is important to the contrast control and repeatable imaging of SPFM through RH adjusting. These findings suggest a strategy of controllable and repeatable imaging the local dielectric properties of nanomaterials with SPFM, which is critically important for further distinguishment, manipulation, electronic applications, etc.

  19. Scanning ion microscopy with low energy lithium ions

    International Nuclear Information System (INIS)

    Twedt, Kevin A.; Chen, Lei; McClelland, Jabez J.

    2014-01-01

    Using an ion source based on photoionization of laser-cooled lithium atoms, we have developed a scanning ion microscope with probe sizes of a few tens of nanometers and beam energies from 500 eV to 5 keV. These beam energies are much lower than the typical operating energies of the helium ion microscope or gallium focused ion beam systems. We demonstrate how low energy can be advantageous in ion microscopy when detecting backscattered ions, due to a decreased interaction volume and the potential for surface sensitive composition analysis. As an example application that demonstrates these advantages, we non-destructively image the removal of a thin residual resist layer during plasma etching in a nano-imprint lithography process. - Highlights: • We use an ion source based on photoionization of laser-cooled lithium atoms. • The ion source makes possible a low energy (500 eV to 5 keV) scanning ion microscope. • Low energy is preferred for ion microscopy with backscattered ions. • We use the microscope to image a thin resist used in nano-imprint lithography

  20. Helium ion microscopy and energy selective scanning electron microscopy - two advanced microscopy techniques with complementary applications

    Science.gov (United States)

    Rodenburg, C.; Jepson, M. A. E.; Boden, Stuart A.; Bagnall, Darren M.

    2014-06-01

    Both scanning electron microscopes (SEM) and helium ion microscopes (HeIM) are based on the same principle of a charged particle beam scanning across the surface and generating secondary electrons (SEs) to form images. However, there is a pronounced difference in the energy spectra of the emitted secondary electrons emitted as result of electron or helium ion impact. We have previously presented evidence that this also translates to differences in the information depth through the analysis of dopant contrast in doped silicon structures in both SEM and HeIM. Here, it is now shown how secondary electron emission spectra (SES) and their relation to depth of origin of SE can be experimentally exploited through the use of energy filtering (EF) in low voltage SEM (LV-SEM) to access bulk information from surfaces covered by damage or contamination layers. From the current understanding of the SES in HeIM it is not expected that EF will be as effective in HeIM but an alternative that can be used for some materials to access bulk information is presented.

  1. Confocal laser scanning microscopy in study of bone calcification

    Science.gov (United States)

    Nishikawa, Tetsunari; Kokubu, Mayu; Kato, Hirohito; Imai, Koichi; Tanaka, Akio

    2012-12-01

    Bone regeneration in mandible and maxillae after extraction of teeth or tumor resection and the use of rough surface implants in bone induction must be investigated to elucidate the mechanism of calcification. The calcified tissues are subjected to chemical decalcification or physical grinding to observe their microscopic features with light microscopy and transmission electron microscopy where the microscopic tissue morphology is significantly altered. We investigated the usefulness of confocal laser scanning microscopy (CLSM) for this purpose. After staggering the time of administration of calcein and alizarin red to experimental rats and dogs, rat alveolar bone and dog femur grafted with coral as scaffold or dental implants were observed with CLSM. In rat alveolar bone, the calcification of newly-formed bone and net-like canaliculi was observed at the mesial bone from the roots progressed at the rate of 15 μm/day. In dog femur grafted with coral, newly-formed bones along the space of coral were observed in an orderly manner. In dog femur with dental implants, after 8 weeks, newly-formed bone proceeded along the rough surface of the implants. CLSM produced high-magnification images of newly-formed bone and thin sections were not needed.

  2. Characterization of human breast cancer by scanning acoustic microscopy

    Science.gov (United States)

    Chen, Di; Malyarenko, Eugene; Seviaryn, Fedar; Yuan, Ye; Sherman, Mark; Bandyopadhyay, Sudeshna; Gierach, Gretchen; Greenway, Christopher W.; Maeva, Elena; Strumban, Emil; Duric, Neb; Maev, Roman

    2013-03-01

    Objectives: The purpose of this study was to characterize human breast cancer tissues by the measurement of microacoustic properties. Methods: We investigated eight breast cancer patients using acoustic microscopy. For each patient, seven blocks of tumor tissue were collected from seven different positions around a tumor mass. Frozen sections (10 micrometer, μm) of human breast cancer tissues without staining and fixation were examined in a scanning acoustic microscope with focused transducers at 80 and 200 MHz. Hematoxylin and Eosin (H and E) stained sections from the same frozen breast cancer tissues were imaged by optical microscopy for comparison. Results: The results of acoustic imaging showed that acoustic attenuation and sound speed in cancer cell-rich tissue regions were significantly decreased compared with the surrounding tissue regions, where most components are normal cells/tissues, such as fibroblasts, connective tissue and lymphocytes. Our observation also showed that the ultrasonic properties were influenced by arrangements of cells and tissue patterns. Conclusions: Our data demonstrate that attenuation and sound speed imaging can provide biomechanical information of the tumor and normal tissues. The results also demonstrate the potential of acoustic microscopy as an auxiliary method for operative detection and localization of cancer affected regions.

  3. Scanning probe microscope simulator for the assessment of noise in scanning probe microscopy controllers

    International Nuclear Information System (INIS)

    Wutscher, T.; Niebauer, J.; Giessibl, F. J.

    2013-01-01

    We present an electronic circuit that allows to calibrate and troubleshoot scanning probe microscopy (SPM) controllers with respect to their noise performance. The control signal in an SPM is typically highly nonlinear—the tunneling current in scanning tunneling microscopy (STM) varies exponentially with distance. The exponential current-versus-voltage characteristics of diodes allow to model the current dependence in STM. Additional inputs allow to simulate the effects of external perturbations and the reactions of the control electronics. We characterized the noise performance of the feedback controller using the apparent topography roughness of recorded images. For a comparison of different STM controllers, an optimal gain parameter was determined by exploring settling times through a rectangular perturbation signal. We used the circuit to directly compare the performance of two types of SPM controllers used in our laboratory

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

    Science.gov (United States)

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

    2018-04-01

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

  5. Qualitative doping area characterization of SONOS transistor utilizing scanning capacitance microscopy (SCM) and scanning spread resistance microscopy (SSRM)

    International Nuclear Information System (INIS)

    Heo, Jinhee; Kim, Deoksu; Kim, Chung woo; Chung, Ilsub

    2005-01-01

    Continuous shrinkage in the memory devices demands further understanding about the doping concentration variations at shallow junction and channel region. Scanning capacitance microscopy (SCM) and scanning spread resistance microscopy (SSRM) can provide reliable information about the electrical and physical junction structure simultaneously. In this work, we attempt to visualize the doping concentration variations of split-gate structure silicon-oxide-nitride-oxide-silicon (SONOS) transistor with thin oxide-nitride-oxide (ONO; 4/7/11 nm). From SCM image, we could identify the source and drain region, which have different doping concentrations from that at channel region. In addition, a gate oxide layer and a depletion region were also identified. Similar results were obtained using SSRM. However, SSRM shows a better resolution, in particular, for highly doped region. For this experiment, the cross-sectional sample has been prepared using focused ion beam (FIB) and hand-polishing method. The results show that SCM and SSRM are very useful methods to analyze the doping profile near the junction as well as the channel

  6. Electrical characterization of doped semiconductor nanostructures with scanning microwave microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Fenner, Matthias A.; Tanbakuchi, Hassan [Agilent Technologies, Kronberg (Germany); Streit, Stephan; Baumgart, Christine; Helm, Manfred; Schmidt, Heidemarie [Institute of Ion Beam Physics and Materials Research, Forschungszentrum Dresden-Rossendorf e.V., Dresden (Germany)

    2010-07-01

    Highly sensitive scanning microwave microscopy (SMM) with a capacitance resolution in the aF range has been used to investigate the electrical properties of doped semiconductor nanostructures in the microwave frequency range from 1.5 GHz to 6 GHz at different dc offset biases. The microwave signal S11 reflected by the sample is related to the impedance of the sample. Superimposing an ac voltage in the kHz range one also gains information about the derivative of the S11 signal (dC/dV), which is dependent on the doping density in the semiconductor, circuit resistance, and reactance. We investigated a static random access memory (SRAM) cell and one cross-sectionally prepared Si epilayer structured sample. The derivative of S11 strongly depends on the dc offset bias. The Si epilayer sample reveals the strongest dependence on f{sub ac} and also on the biasing history during the SMM measurements.

  7. Electronic structure classifications using scanning tunneling microscopy conductance imaging

    International Nuclear Information System (INIS)

    Horn, K.M.; Swartzentruber, B.S.; Osbourn, G.C.; Bouchard, A.; Bartholomew, J.W.

    1998-01-01

    The electronic structure of atomic surfaces is imaged by applying multivariate image classification techniques to multibias conductance data measured using scanning tunneling microscopy. Image pixels are grouped into classes according to shared conductance characteristics. The image pixels, when color coded by class, produce an image that chemically distinguishes surface electronic features over the entire area of a multibias conductance image. Such open-quotes classedclose quotes images reveal surface features not always evident in a topograph. This article describes the experimental technique used to record multibias conductance images, how image pixels are grouped in a mathematical, classification space, how a computed grouping algorithm can be employed to group pixels with similar conductance characteristics in any number of dimensions, and finally how the quality of the resulting classed images can be evaluated using a computed, combinatorial analysis of the full dimensional space in which the classification is performed. copyright 1998 American Institute of Physics

  8. Super-resolved terahertz microscopy by knife-edge scan

    Science.gov (United States)

    Giliberti, V.; Flammini, M.; Ciano, C.; Pontecorvo, E.; Del Re, E.; Ortolani, M.

    2017-08-01

    We present a compact, all solid-state THz confocal microscope operating at 0.30 THz that achieves super-resolution by using the knife-edge scan approach. In the final reconstructed image, a lateral resolution of 60 μm ≍ λ/17 is demonstrated when the knife-edge is deep in the near-field of the sample surface. When the knife-edge is lifted up to λ/4 from the sample surface, a certain degree of super-resolution is maintained with a resolution of 0.4 mm, i.e. more than a factor 2 if compared to the diffraction-limited scheme. The present results open an interesting path towards super-resolved imaging with in-depth information that would be peculiar to THz microscopy systems.

  9. Scanning spreading resistance microscopy of shallow doping profiles in silicon

    International Nuclear Information System (INIS)

    Suchodolskis, A.; Hallen, A.; Gran, J.; Hansen, T.-E.; Karlsson, U.O.

    2006-01-01

    We demonstrate the application of scanning spreading resistance microscopy (SSRM) for characterization of shallow highly-conductive layers formed by boron implantation of lowly doped n-type silicon substrate followed by a post-implantation annealing. The electrically active dopant concentration versus depth was obtained from a cross-section of freshly cleaved samples where the Si-surface could be clearly distinguished by depositing a SiO 2 -layer before cleavage. To quantify free carrier concentration we calibrated our data against samples with implanted/annealed boron profiles established by secondary ion mass spectrometry (SIMS). A good fit of SSRM and SIMS data is possible for free carrier concentrations lower than 10 20 cm -3 , but for higher concentrations there is a discrepancy indicating an incomplete activation of the boron

  10. Local deposition of anisotropic nanoparticles using scanning electrochemical microscopy (SECM).

    Science.gov (United States)

    Fedorov, Roman G; Mandler, Daniel

    2013-02-28

    We demonstrate localized electrodeposition of anisotropic metal nanoobjects, namely Au nanorods (GNR), on indium tin oxide (ITO) using scanning electrochemical microscopy (SECM). A gold microelectrode was the source of the gold ions whereby double pulse chronoamperometry was employed to generate initially Au seeds which were further grown under controlled conditions. The distance between the microelectrode and the ITO surface as well as the different experimental parameters (electrodeposition regime, solution composition and temperature) were optimized to produce faceted gold seeds with the required characteristics (size and distribution). Colloidal chemical synthesis was successfully exploited for better understanding the role of the surfactant and different additives in breaking the crystallographic symmetry and anisotropic growth of GNR. Experiments performed in a conventional three-electrode cell revealed the most appropriate electrochemical conditions allowing high yield synthesis of nanorods with well-defined shape as well as nanocubes and bipyramids.

  11. Visualization of bacterial polysaccharides by scanning transmission electron microscopy.

    Science.gov (United States)

    Wolanski, B S; McAleer, W J; Hilleman, M R

    1983-04-01

    Highly purified capsular polysaccharides of Neisseria meningitidis groups A, B, and C have been visualized by high resolution Scanning Transmission Electron Microscopy (STEM). Spheroidal macromolecules approximately 200 A in diameter are characteristic of the Meningococcus A and C polysaccharides whereas filaments that are 400-600 A in length are found in Meningococcus B polysaccharide preparations. Filaments are occasionally found associated with the spheroidal Meningococcus A and C polysaccharides and it is proposed that these structures are composed of a long (1-4 microns) filament or filaments that are arranged in spheroidal molecules or micelles of high molecular weight. The Meningococcus B polysaccharide, by contrast, is a short flexuous filament or strand of relatively low molecular weight. A relationship between morphology and antigenicity is proposed.

  12. Optimization of permanganic etching of polyethylenes for scanning electron microscopy

    International Nuclear Information System (INIS)

    Naylor, K.L.; Phillips, P.J.

    1983-01-01

    The permanganic etching technique has been studied as a function of time, temperature, and concentration for a series of polyethylenes. Kinetic studies show that a film of reaction products builds up on the surface, impeding further etching, an effect which is greatest for the lowest-crystallinity polymers. SEM studies combined with EDS show that the film contains sulfur, potassium and some manganese. An artifact is produced by the etching process which is impossible to remove by washing procedures if certain limits of time, temperature, and concentration are exceeded. For lower-crystallinity polyethylenes multiple etching and washing steps were required for optimal resolution. Plastic deformation during specimen preparation, whether from scratches or freeze fracturing, enhances artifact formation. When appropriate procedures are used, virtually artifact-free surfaces can be produced allowing a combination of permanganic etching and scanning electron microscopy to give a rapid method for detailed morphological characterization of bulk specimens

  13. Ultramicrosensors based on transition metal hexacyanoferrates for scanning electrochemical microscopy

    Directory of Open Access Journals (Sweden)

    Maria A. Komkova

    2013-10-01

    Full Text Available We report here a way for improving the stability of ultramicroelectrodes (UME based on hexacyanoferrate-modified metals for the detection of hydrogen peroxide. The most stable sensors were obtained by electrochemical deposition of six layers of hexacyanoferrates (HCF, more specifically, an alternating pattern of three layers of Prussian Blue and three layers of Ni–HCF. The microelectrodes modified with mixed layers were continuously monitored in 1 mM hydrogen peroxide and proved to be stable for more than 5 h under these conditions. The mixed layer microelectrodes exhibited a stability which is five times as high as the stability of conventional Prussian Blue-modified UMEs. The sensitivity of the mixed layer sensor was 0.32 A·M−1·cm−2, and the detection limit was 10 µM. The mixed layer-based UMEs were used as sensors in scanning electrochemical microscopy (SECM experiments for imaging of hydrogen peroxide evolution.

  14. Molecules on vicinal Au surfaces studied by scanning tunnelling microscopy

    International Nuclear Information System (INIS)

    Kroeger, J; Neel, N; Jensen, H; Berndt, R; Rurali, R; Lorente, N

    2006-01-01

    Using low-temperature scanning tunnelling microscopy and spectroscopy we investigated the adsorption characteristics of 3,4,9,10-perylenetetracarboxylic-dianhydride and fullerenes on Au(788), Au(433), and Au(778). On Au(788) and Au(778), 3,4,9,10-perylenetetracarboxylic-dianhydride exhibits three coexisting superstructures, which do not reflect the periodicity of the hosting substrate. The adsorption on Au(433) leads to the formation of molecule chains along the step edges after annealing the sample. Fullerene molecules on Au(788) arrange in a mesh of islands, which extends over several hundreds of nanometres with an extraordinarily high periodicity. A combination of fullerene adsorption and annealing leads to facetting of Au(433) and the formation of extraordinarily long fullerene stripes

  15. Signal and noise modeling in confocal laser scanning fluorescence microscopy.

    Science.gov (United States)

    Herberich, Gerlind; Windoffer, Reinhard; Leube, Rudolf E; Aach, Til

    2012-01-01

    Fluorescence confocal laser scanning microscopy (CLSM) has revolutionized imaging of subcellular structures in biomedical research by enabling the acquisition of 3D time-series of fluorescently-tagged proteins in living cells, hence forming the basis for an automated quantification of their morphological and dynamic characteristics. Due to the inherently weak fluorescence, CLSM images exhibit a low SNR. We present a novel model for the transfer of signal and noise in CLSM that is both theoretically sound as well as corroborated by a rigorous analysis of the pixel intensity statistics via measurement of the 3D noise power spectra, signal-dependence and distribution. Our model provides a better fit to the data than previously proposed models. Further, it forms the basis for (i) the simulation of the CLSM imaging process indispensable for the quantitative evaluation of CLSM image analysis algorithms, (ii) the application of Poisson denoising algorithms and (iii) the reconstruction of the fluorescence signal.

  16. 2-photon laser scanning microscopy on native human cartilage

    Science.gov (United States)

    Martini, Joerg; Toensing, Katja; Dickob, Michael; Anselmetti, Dario

    2005-08-01

    Native hyaline cartilage from a human knee joint was directly investigated with laser scanning microscopy via 2-photon autofluorescence excitation with no additional staining or labelling protocols in a nondestructive and sterile manner. Using a femtosecond, near-infrared (NIR) Ti:Sa laser for 2-photon excitation and a dedicated NIR long distance objective, autofluorescence imaging and measurements of the extracellular matrix (ECM) tissue with incorporated chondrocytes were possible with a penetration depth of up to 460 μm inside the sample. Via spectral autofluorescence separation these experiments allowed the discrimination of chondrocytes from the ECM and therefore an estimate of chondrocytic cell density within the cartilage tissue to approximately 0.2-2•107cm3. Furthermore, a comparison of the relative autofluorescence signals between nonarthritic and arthritic cartilage tissue exhibited distinct differences in tissue morphology. As these morphological findings are in keeping with the macroscopic diagnosis, our measurement has the potential of being used in future diagnostic applications.

  17. Scanning electron microscopy of Strongylus spp. in zebra.

    Science.gov (United States)

    Els, H J; Malan, F S; Scialdo-Krecek, R C

    1983-12-01

    The external ultrastructure of the anterior and posterior extremities of the nematodes, Strongylus asini , Strongylus vulgaris, Strongylus equinus and Strongylus edentatus, was studied with scanning electron microscopy (SEM). Fresh specimens of S. asini were collected from the caecum, ventral colon and vena portae of Equus burchelli and Equus zebra hartmannae ; S. vulgaris from the caecum, colon and arteria ileocolica of E. burchelli ; S. equinus from the ventral colon of E. z. hartmannae and S. edentatus from the caecum and ventral colon of both zebras , during surveys of parasites in zebras in the Etosha Game Reserve, South West Africa/Namibia, and the Kruger National Park, Republic of South Africa. The worms were cleaned, fixed and mounted by standard methods and photographed in a JEOL JSM - 35C scanning electron microscope (SEM) operating at 12kV . The SEM showed the following differences: the tips of the external leaf-crowns varied and were fine and delicate in S. asini , coarse and broad in S. vulgaris and, in S. equinus and S. edentatus, closely adherent, separating into single elements for half their length. The excretory pores showed only slight variation, and the morphology of the copulatory bursae did not differ from those seen with light microscopy. The genital cones differed markedly: S. asini had a ventral triangular projection and laterally 2 finger-like projections: in S. vulgaris there were numerous bosses on the lateral and ventral aspects of the cone; in S. equinus 2 finger-like processes projected laterocaudally ; and in S. edentatus 2 pairs of papilla-like processes projected laterally on the ventral aspects, and a pair of rounded projections and a pair of hair-like structures adorned the dorsal aspects.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Probing Individual Ice Nucleation Events with Environmental Scanning Electron Microscopy

    Science.gov (United States)

    Wang, Bingbing; China, Swarup; Knopf, Daniel; Gilles, Mary; Laskin, Alexander

    2016-04-01

    Heterogeneous ice nucleation is one of the processes of critical relevance to a range of topics in the fundamental and the applied science and technologies. Heterogeneous ice nucleation initiated by particles proceeds where microscopic properties of particle surfaces essentially control nucleation mechanisms. Ice nucleation in the atmosphere on particles governs the formation of ice and mixed phase clouds, which in turn influence the Earth's radiative budget and climate. Heterogeneous ice nucleation is still insufficiently understood and poses significant challenges in predictive understanding of climate change. We present a novel microscopy platform allowing observation of individual ice nucleation events at temperature range of 193-273 K and relative humidity relevant for ice formation in the atmospheric clouds. The approach utilizes a home built novel ice nucleation cell interfaced with Environmental Scanning Electron Microscope (IN-ESEM system). The IN-ESEM system is applied for direct observation of individual ice formation events, determining ice nucleation mechanisms, freezing temperatures, and relative humidity onsets. Reported microanalysis of the ice nucleating particles (INP) include elemental composition detected by the energy dispersed analysis of X-rays (EDX), and advanced speciation of the organic content in particles using scanning transmission x-ray microscopy with near edge X-ray absorption fine structure spectroscopy (STXM/NEXAFS). The performance of the IN-ESEM system is validated through a set of experiments with kaolinite particles with known ice nucleation propensity. We demonstrate an application of the IN-ESEM system to identify and characterize individual INP within a complex mixture of ambient particles.

  19. Scanning electron microscopy of the neuropathology of murine cerebral malaria

    Directory of Open Access Journals (Sweden)

    Brenneis Christian

    2006-11-01

    Full Text Available Abstract Background The mechanisms leading to death and functional impairments due to cerebral malaria (CM are yet not fully understood. Most of the knowledge about the pathomechanisms of CM originates from studies in animal models. Though extensive histopathological studies of the murine brain during CM are existing, alterations have not been visualized by scanning electron microscopy (SEM so far. The present study investigates the neuropathological features of murine CM by applying SEM. Methods C57BL/6J mice were infected with Plasmodium berghei ANKA blood stages. When typical symptoms of CM developed perfused brains were processed for SEM or light microscopy, respectively. Results Ultrastructural hallmarks were disruption of vessel walls, parenchymal haemorrhage, leukocyte sequestration to the endothelium, and diapedesis of macrophages and lymphocytes into the Virchow-Robin space. Villous appearance of observed lymphocytes were indicative of activated state. Cerebral oedema was evidenced by enlargement of perivascular spaces. Conclusion The results of the present study corroborate the current understanding of CM pathophysiology, further support the prominent role of the local immune system in the neuropathology of CM and might expose new perspectives for further interventional studies.

  20. Quadratic electromechanical strain in silicon investigated by scanning probe microscopy

    Science.gov (United States)

    Yu, Junxi; Esfahani, Ehsan Nasr; Zhu, Qingfeng; Shan, Dongliang; Jia, Tingting; Xie, Shuhong; Li, Jiangyu

    2018-04-01

    Piezoresponse force microscopy (PFM) is a powerful tool widely used to characterize piezoelectricity and ferroelectricity at the nanoscale. However, it is necessary to distinguish microscopic mechanisms between piezoelectricity and non-piezoelectric contributions measured by PFM. In this work, we systematically investigate the first and second harmonic apparent piezoresponses of a silicon wafer in both vertical and lateral modes, and we show that it exhibits an apparent electromechanical response that is quadratic to the applied electric field, possibly arising from ionic electrochemical dipoles induced by the charged probe. As a result, the electromechanical response measured is dominated by the second harmonic response in the vertical mode, and its polarity can be switched by the DC voltage with the evolving coercive field and maximum amplitude, in sharp contrast to typical ferroelectric materials we used as control. The ionic activity in silicon is also confirmed by the scanning thermo-ionic microscopy measurement, and the work points toward a set of methods to distinguish true piezoelectricity from the apparent ones.

  1. Three-Dimensional scanning transmission electron microscopy of biological specimens

    KAUST Repository

    De Jonge, Niels

    2010-01-18

    A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2-3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original dataset. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy. However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved dataset. © 2010 Microscopy Society of America.

  2. Scanning probe microscopy with vertically oriented cantilevers made easy

    International Nuclear Information System (INIS)

    Valdrè, G; Moro, D; Ulian, G

    2012-01-01

    Non-contact imaging in scanning probe microscopy (SPM) is becoming of great importance in particular for imaging biological matter and in general soft materials. Transverse dynamic force microscopy (TDFM) is an SPM-based methodology that exploiting a cantilever oriented in a vertical configuration with respect to the sample surface may work with very low tip to sample interaction forces. The probe is oscillated parallel to the sample surface, usually by a piezoelectric element. However, this methodology often requires complex microscope setups and detection systems, so it is usually developed in specific laboratories as a prototype microscope. Here, we present a very simple device that easily enables a commercial SPM head to be oriented in such a way to have the cantilever long axis perpendicular to the sample surface. No modifications of the SPM hardware and software are required and commercial available cantilevers can be used as probes. Performance tests using polystyrene spheres, muscovite crystallographic steps and DNA single molecules were successful and all resulted in agreement with other TDFM and SPM observations demonstrating the reliability of the device. (paper)

  3. Amyloid Structure and Assembly: Insights from Scanning Transmission Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Goldsbury, C.; Wall, J.; Baxa, U.; Simon, M. N.; Steven, A. C.; Engel, A.; Aebi, U.; Muller, S. A.

    2011-01-01

    Amyloid fibrils are filamentous protein aggregates implicated in several common diseases such as Alzheimer's disease and type II diabetes. Similar structures are also the molecular principle of the infectious spongiform encephalopathies such as Creutzfeldt-Jakob disease in humans, scrapie in sheep, and of the so-called yeast prions, inherited non-chromosomal elements found in yeast and fungi. Scanning transmission electron microscopy (STEM) is often used to delineate the assembly mechanism and structural properties of amyloid aggregates. In this review we consider specifically contributions and limitations of STEM for the investigation of amyloid assembly pathways, fibril polymorphisms and structural models of amyloid fibrils. This type of microscopy provides the only method to directly measure the mass-per-length (MPL) of individual filaments. Made on both in vitro assembled and ex vivo samples, STEM mass measurements have illuminated the hierarchical relationships between amyloid fibrils and revealed that polymorphic fibrils and various globular oligomers can assemble simultaneously from a single polypeptide. The MPLs also impose strong constraints on possible packing schemes, assisting in molecular model building when combined with high-resolution methods like solid-state nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR).

  4. Scanning Surface Potential Microscopy of Spore Adhesion on Surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Ida [University of Tennessee, Knoxville (UTK); Chung, Eunhyea [Georgia Institute of Technology; Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Tsouris, Costas [ORNL

    2012-01-01

    The adhesion of spores of Bacillus anthracis - the cause of anthrax and a likely biological threat - to solid surfaces is an important consideration in cleanup after an accidental or deliberate release. However, because of safety concerns, directly studying B. anthracis spores with advanced instrumentation is problematic. As a first step, we are examining the electrostatic potential of Bacillus thuringiensis (Bt), which is a closely related species that is often used as a simulant to study B. anthracis. Scanning surface potential microscopy (SSPM), also known as Kelvin probe force microscopy (KPFM), was used to investigate the influence of relative humidity (RH) on the surface electrostatic potential of Bt that had adhered to silica, mica, or gold substrates. AFM/SSPM side-by-side images were obtained separately in air, at various values of RH, after an aqueous droplet with spores was applied on each surface and allowed to dry before measurements. In the SSPM images, a negative potential on the surface of the spores was observed compared with that of the substrates. The surface potential decreased as the humidity increased. Spores were unable to adhere to a surface with an extremely negative potential, such as mica.

  5. Scanning electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1985-01-01

    The aim of this book is to outline the physics of image formation, electron­ specimen interactions, imaging modes, the interpretation of micrographs and the use of quantitative modes "in scanning electron microscopy (SEM). lt forms a counterpart to Transmission Electron Microscopy (Vol. 36 of this Springer Series in Optical Sciences) . The book evolved from lectures delivered at the University of Münster and from a German text entitled Raster-Elektronenmikroskopie (Springer-Verlag), published in collaboration with my colleague Gerhard Pfefferkorn. In the introductory chapter, the principles of the SEM and of electron­ specimen interactions are described, the most important imaging modes and their associated contrast are summarized, and general aspects of eiemental analysis by x-ray and Auger electron emission are discussed. The electron gun and electron optics are discussed in Chap. 2 in order to show how an electron probe of small diameter can be formed, how the elec­ tron beam can be blanked at high fre...

  6. Confocal laser scanning microscopy in study of bone calcification

    Energy Technology Data Exchange (ETDEWEB)

    Nishikawa, Tetsunari, E-mail: tetsu-n@cc.osaka-dent.ac.jp [Department of Oral Pathology, Osaka Dental University, Osaka (Japan); Kokubu, Mayu; Kato, Hirohito [Department of Oral Pathology, Osaka Dental University, Osaka (Japan); Imai, Koichi [Department of Biomaterials, Osaka Dental University, Osaka (Japan); Tanaka, Akio [Department of Oral Pathology, Osaka Dental University, Osaka (Japan)

    2012-12-01

    Highlights: Black-Right-Pointing-Pointer High-magnification images with depth selection, and thin sections were observed using CLSM. Black-Right-Pointing-Pointer The direction and velocity of calcification of the bone was observed by administration of 2 fluorescent dyes. Black-Right-Pointing-Pointer In dog femora grafted with coral blocks, newly-formed bone was observed in the coral block space with a rough surface. Black-Right-Pointing-Pointer Twelve weeks after dental implant was grafted in dog femora, the space between screws was filled with newly-formed bones. - Abstract: Bone regeneration in mandible and maxillae after extraction of teeth or tumor resection and the use of rough surface implants in bone induction must be investigated to elucidate the mechanism of calcification. The calcified tissues are subjected to chemical decalcification or physical grinding to observe their microscopic features with light microscopy and transmission electron microscopy where the microscopic tissue morphology is significantly altered. We investigated the usefulness of confocal laser scanning microscopy (CLSM) for this purpose. After staggering the time of administration of calcein and alizarin red to experimental rats and dogs, rat alveolar bone and dog femur grafted with coral as scaffold or dental implants were observed with CLSM. In rat alveolar bone, the calcification of newly-formed bone and net-like canaliculi was observed at the mesial bone from the roots progressed at the rate of 15 {mu}m/day. In dog femur grafted with coral, newly-formed bones along the space of coral were observed in an orderly manner. In dog femur with dental implants, after 8 weeks, newly-formed bone proceeded along the rough surface of the implants. CLSM produced high-magnification images of newly-formed bone and thin sections were not needed.

  7. Confocal laser scanning microscopy in study of bone calcification

    International Nuclear Information System (INIS)

    Nishikawa, Tetsunari; Kokubu, Mayu; Kato, Hirohito; Imai, Koichi; Tanaka, Akio

    2012-01-01

    Highlights: ► High-magnification images with depth selection, and thin sections were observed using CLSM. ► The direction and velocity of calcification of the bone was observed by administration of 2 fluorescent dyes. ► In dog femora grafted with coral blocks, newly-formed bone was observed in the coral block space with a rough surface. ► Twelve weeks after dental implant was grafted in dog femora, the space between screws was filled with newly-formed bones. - Abstract: Bone regeneration in mandible and maxillae after extraction of teeth or tumor resection and the use of rough surface implants in bone induction must be investigated to elucidate the mechanism of calcification. The calcified tissues are subjected to chemical decalcification or physical grinding to observe their microscopic features with light microscopy and transmission electron microscopy where the microscopic tissue morphology is significantly altered. We investigated the usefulness of confocal laser scanning microscopy (CLSM) for this purpose. After staggering the time of administration of calcein and alizarin red to experimental rats and dogs, rat alveolar bone and dog femur grafted with coral as scaffold or dental implants were observed with CLSM. In rat alveolar bone, the calcification of newly-formed bone and net-like canaliculi was observed at the mesial bone from the roots progressed at the rate of 15 μm/day. In dog femur grafted with coral, newly-formed bones along the space of coral were observed in an orderly manner. In dog femur with dental implants, after 8 weeks, newly-formed bone proceeded along the rough surface of the implants. CLSM produced high-magnification images of newly-formed bone and thin sections were not needed.

  8. Investigation of Nematode Diversity using Scanning Electron Microscopy and Fluorescent Microscopy

    Science.gov (United States)

    Seacor, Taylor; Howell, Carina

    2013-03-01

    Nematode worms account for the vast majority of the animals in the biosphere. They are colossally important to global public health as parasites, and to agriculture both as pests and as beneficial inhabitants of healthy soil. Amphid neurons are the anterior chemosensory neurons in nematodes, mediating critical behaviors including chemotaxis and mating. We are examining the cellular morphology and external anatomy of amphid neurons, using fluorescence microscopy and scanning electron microscopy, respectively, of a wide range of soil nematodes isolated in the wild. We use both classical systematics (e.g. diagnostic keys) and molecular markers (e.g. ribosomal RNA) to classify these wild isolates. Our ultimate aim is to build a detailed anatomical database in order to dissect genetic pathways of neuronal development and function across phylogeny and ecology. Research supported by NSF grants 092304, 0806660, 1058829 and Lock Haven University FPDC grants

  9. Morphological classification of bioaerosols from composting using scanning electron microscopy

    International Nuclear Information System (INIS)

    Tamer Vestlund, A.; Al-Ashaab, R.; Tyrrel, S.F.; Longhurst, P.J.; Pollard, S.J.T.; Drew, G.H.

    2014-01-01

    Highlights: • Bioaerosols were captured using the filter method. • Bioaerosols were analysed using scanning electron microscope. • Bioaerosols were classified on the basis of morphology. • Single small cells were found more frequently than aggregates and larger cells. • Smaller cells may disperse further than heavier aggregate structures. - Abstract: This research classifies the physical morphology (form and structure) of bioaerosols emitted from open windrow composting. Aggregation state, shape and size of the particles captured are reported alongside the implications for bioaerosol dispersal after release. Bioaerosol sampling took place at a composting facility using personal air filter samplers. Samples were analysed using scanning electron microscopy. Particles were released mainly as small (<1 μm) single, spherical cells, followed by larger (>1 μm) single cells, with aggregates occurring in smaller proportions. Most aggregates consisted of clusters of 2–3 particles as opposed to chains, and were <10 μm in size. No cells were attached to soil debris or wood particles. These small single cells or small aggregates are more likely to disperse further downwind from source, and cell viability may be reduced due to increased exposure to environmental factors

  10. Scanning X-ray microscopy of superconductor/ferromagnet bilayers

    Energy Technology Data Exchange (ETDEWEB)

    Stahl, Claudia; Ruoss, Stephen; Weigand, Markus; Schuetz, Gisela [Max Planck Institute for Intelligent Systems, Stuttgart (Germany); Zahn, Patrick; Bayer, Jonas [Max Planck Institute for Intelligent Systems, Stuttgart (Germany); Research Institute for Innovative Surfaces, FINO, Aalen University (Germany); Albrecht, Joachim [Research Institute for Innovative Surfaces, FINO, Aalen University (Germany)

    2016-07-01

    The magnetic flux distribution arising from a high-T{sub c} superconductor is detected and visualized with high spatial resolution using scanning x-ray microscopy (SXM). Therefore, we introduce a sensor layer, namely, an amorphous, soft-magnetic CoFeB cover layer. The magnetic stray fields of the supercurrents lead to a local reorientation of the magnetic moments in the ferromagnet, which is visualized using the large x-ray magnetic circular dichroism (XMCD) effect of the Co and Fe L3-edge. We show that the XMCD contrast in the sensor layer corresponds to the in-plane magnetic flux distribution of the superconductor and can hence be used to image magnetic structures in superconductors with high spatial resolution. Using the total electron yield (TEY) mode the surface structure and the magnetic domains can be imaged simultaneously and can be correlated. The measurements are carried out at our scanning x-ray microscope MAXYMUS at Bessy II, Berlin with the new low temperature setup.

  11. 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.

  12. Morphological classification of bioaerosols from composting using scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tamer Vestlund, A. [Institute for Energy and Resource Technology, Environmental Science and Technology Department, School of Applied Sciences, Cranfield University, Building 40, Bedfordshire MK43 0AL (United Kingdom); FIRA International Ltd., Maxwell Road, Stevenage, Herts SG1 2EW (United Kingdom); Al-Ashaab, R.; Tyrrel, S.F.; Longhurst, P.J.; Pollard, S.J.T. [Institute for Energy and Resource Technology, Environmental Science and Technology Department, School of Applied Sciences, Cranfield University, Building 40, Bedfordshire MK43 0AL (United Kingdom); Drew, G.H., E-mail: g.h.drew@cranfield.ac.uk [Institute for Energy and Resource Technology, Environmental Science and Technology Department, School of Applied Sciences, Cranfield University, Building 40, Bedfordshire MK43 0AL (United Kingdom)

    2014-07-15

    Highlights: • Bioaerosols were captured using the filter method. • Bioaerosols were analysed using scanning electron microscope. • Bioaerosols were classified on the basis of morphology. • Single small cells were found more frequently than aggregates and larger cells. • Smaller cells may disperse further than heavier aggregate structures. - Abstract: This research classifies the physical morphology (form and structure) of bioaerosols emitted from open windrow composting. Aggregation state, shape and size of the particles captured are reported alongside the implications for bioaerosol dispersal after release. Bioaerosol sampling took place at a composting facility using personal air filter samplers. Samples were analysed using scanning electron microscopy. Particles were released mainly as small (<1 μm) single, spherical cells, followed by larger (>1 μm) single cells, with aggregates occurring in smaller proportions. Most aggregates consisted of clusters of 2–3 particles as opposed to chains, and were <10 μm in size. No cells were attached to soil debris or wood particles. These small single cells or small aggregates are more likely to disperse further downwind from source, and cell viability may be reduced due to increased exposure to environmental factors.

  13. Two-photon mapping of localized field enhancements in thin nanostrip antennas

    DEFF Research Database (Denmark)

    Beermann, I.; Novikov, S.M.; Søndergaard, Thomas

    2008-01-01

    scanning optical microscopy, in which two-photon-excited photoluminescence (TPL) excited with a strongly focused laser beam at the wavelength 745 nm is detected. We use TPL images to map the local field enhancements from individual nanostrips at a resolution of 0.35µm and compare results with theoretical......Resonant scattering and local field enhancements by 11-nm-thin gold nanostrip antennas due to constructive interference of counter propagating slow surface plasmon polaritons is investigated. We characterize nanostrips of widths between 50-530 nm using both reflection spectroscopy and nonlinear...

  14. The scanning probe microscopy study of thin polymer films

    International Nuclear Information System (INIS)

    Harron, H.R.

    1995-08-01

    Scanning Tunnelling Microscopy and Atomic Force Microscopy were used systematically to investigate the morphology, uniformity, coverage and structure of the thin films of several commercially important insulating polymers. Despite the poorly conducting nature of the polymer sample, detailed and convincing images of this class of materials were achieved by STM without the need to coat the samples with a conductive layer. The polymer regions of the sample were further investigated by the use of surface profiling with 'line scans'. The fluctuations of the amplitude therein enabled important film characteristics to be assessed. An environmental stage was designed for the STM to enable the effect of various vapour-sample interactions to be observed during the imaging process. Using the data from the environmental stage in addition to the surface profiling with line scans, an insight into the conduction mechanism and image interpretation was gained. Results suggest that the water content of the sample and its immediate surroundings is an important factor in achieving reliable STM images in air. The initial study culminated with the observation by STM alone of the plasticizer induced crystallization of uncoated PC thin films. The 'amorphous' PC films were observed before crystallization and small ordered regions in roughly the same proportion as that predicted by diffraction studies [Prietschk, 1959 and Schnell, 1964] were imaged. This has never been observed by a microscopy technique. Furthermore, images of the crystalline film contained elongated units that were attributed to the lamellae formations that form the basic building blocks of polymer spherulites. The study continued with the AFM imaging of the growth of crystalline entities in a PC film, without the need for harsh sample treatment or metal coating. A method of casting and crystallizing the films was developed such that the growth was predominantly in two dimensions and consequently ideal for observation by

  15. Electron microscopy of intermediate filaments: teaming up with atomic force and confocal laser scanning microscopy.

    Science.gov (United States)

    Kreplak, Laurent; Richter, Karsten; Aebi, Ueli; Herrmann, Harald

    2008-01-01

    Intermediate filaments (IFs) were originally discovered and defined by electron microscopy in myoblasts. In the following it was demonstrated and confirmed that they constitute, in addition to microtubules and microfilaments, a third independent, general filament system in the cytoplasm of most metazoan cells. In contrast to the other two systems, IFs are present in cells in two principally distinct cytoskeletal forms: (i) extended and free-running filament arrays in the cytoplasm that are integrated into the cytoskeleton by associated proteins of the plakin type; and (ii) a membrane- and chromatin-bound thin 'lamina' of a more or less regular network of interconnected filaments made from nuclear IF proteins, the lamins, which differ in several important structural aspects from cytoplasmic IF proteins. In man, more than 65 genes code for distinct IF proteins that are expressed during embryogenesis in various routes of differentiation in a tightly controlled manner. IF proteins exhibit rather limited sequence identity implying that the different types of IFs have distinct biochemical properties. Hence, to characterize the structural properties of the various IFs, in vitro assembly regimes have been developed in combination with different visualization methods such as transmission electron microscopy of fixed and negatively stained samples as well as methods that do not use staining such as scanning transmission electron microscopy (STEM) and cryoelectron microscopy as well as atomic force microscopy. Moreover, with the generation of both IF-type specific antibodies and chimeras of fluorescent proteins and IF proteins, it has become possible to investigate the subcellular organization of IFs by correlative fluorescence and electron microscopic methods. The combination of these powerful methods should help to further develop our understanding of nuclear architecture, in particular how nuclear subcompartments are organized and in which way lamins are involved.

  16. Preliminary Study of In Vivo Formed Dental Plaque Using Confocal Microscopy and Scanning Electron Microscopy

    Directory of Open Access Journals (Sweden)

    KA. Al-Salihi

    2009-12-01

    Full Text Available Objective: Confocal laser scanning microscopy (CLSM is relatively a new light microscopical imaging technique with a wide range of applications in biological sciences. The primary value of CLSM for the biologist is its ability to provide optical sections from athree-dimensional specimen. The present study was designed to assess the thickness and content of in vivo accumulated dental plaque using CLSM and scanning electron microscopy (SEM.Materials and Methods: Acroflat lower arch splints (acrylic appliance were worn by five participants for three days without any disturbance. The formed plaques were assessed using CLSM combined with vital fluorescence technique and SEM.Results: In this study accumulated dental plaque revealed varied plaque microflora vitality and thickness according to participant’s oral hygiene. The thickness of plaque smears ranged from 40.32 to 140.72 μm and 65.00 to 128.88 μm for live (vital and dead accumulated microorganisms, respectively. Meanwhile, the thickness of plaque on the appliance ranged from 101 μm to 653 μm. CLSM revealed both dead and vital bacteria on the surface of the dental plaque. In addition, SEM revealed layers of various bacterial aggregations in all dental plaques.Conclusion: This study offers a potent non-invasive tool to evaluate and assess the dental plaque biofilm, which is a very important factor in the development of dental caries.

  17. Scanning tunneling microscopy studies of glucose oxidase on gold surface

    International Nuclear Information System (INIS)

    Losic, D.; Shapter, J.G.; Gooding, J.J.

    2002-01-01

    Full text: Three immobilization methods have been used for scanning tunneling microscopy (STM) studies of glucose oxidase (GOD) on gold. They are based on a) physical adsorption from solution, b) microcontact printing and c) covalent bonding onto self-assembled monolayers (SAM) of 3-mercaptopropionic acid (MPA). The STM images are used to provide information about the organization of individual GOD molecules and more densely packed monolayers of GOD on electrode surfaces, thus providing information of the role of interfacial structure on biosensor performance. The use of atomically flat gold substrates enables easy distinction of deposited enzyme features from the flat gold substrate. Microcontact printing is found to be a more reliable method than adsorption from solution for preparing individual GOD molecules on the gold surface STM images of printed samples reveal two different shapes of native GOD molecules. One is a butterfly shape with dimensions of 10 ± 1 nm x 6 ± 1 nm, assigned to the lying position of molecule while the second is an approximately spherical shape with dimensions of 6.5 ± 1 nm x 5 ± 1nm assigned to a standing position. Isolated clusters of 5 to 6 GOD molecules are also observed. With monolayer coverage, GOD molecules exhibit a tendency to organize themselves into a two dimensional array with adequate sample stability to obtain high-resolution STM images. Within these two-dimensional arrays are clearly seen repeating clusters of five to six enzyme molecules in a unit STM imaging of GOD monolayers covalently immobilized onto SAM (MPA) are considerably more difficult than when the enzyme is adsorbed directly onto the metal. Cluster structures are observed both high and low coverage despite the fact that native GOD is a negatively charged molecule. Copyright (2002) Australian Society for Electron Microscopy Inc

  18. Scanning near-field infrared microscopy on semiconductor structures

    Energy Technology Data Exchange (ETDEWEB)

    Jacob, Rainer

    2011-01-15

    literature. While the structures of the first system were in the micrometer regime, the capability to probe buried nanostructures is demonstrated at a sample of indium arsenide quantum dots. Those dots are covered by a thick layer of gallium arsenide. For the first time ever, it is shown experimentally that transitions between electron states in single quantum dots can be investigated by near-field microscopy. By monitoring the near-field response of these quantum dots while scanning the wavelength of the incident light beam, it was possible to obtain characteristic near-field signatures of single dots. Near-field contrasts up to 30 % could be measured for resonant excitation of electrons in the conduction band of the indium arsenide dots. (orig.)

  19. Scanning near-field infrared microscopy on semiconductor structures

    International Nuclear Information System (INIS)

    Jacob, Rainer

    2011-01-01

    literature. While the structures of the first system were in the micrometer regime, the capability to probe buried nanostructures is demonstrated at a sample of indium arsenide quantum dots. Those dots are covered by a thick layer of gallium arsenide. For the first time ever, it is shown experimentally that transitions between electron states in single quantum dots can be investigated by near-field microscopy. By monitoring the near-field response of these quantum dots while scanning the wavelength of the incident light beam, it was possible to obtain characteristic near-field signatures of single dots. Near-field contrasts up to 30 % could be measured for resonant excitation of electrons in the conduction band of the indium arsenide dots. (orig.)

  20. Accurate virus quantitation using a Scanning Transmission Electron Microscopy (STEM) detector in a scanning electron microscope.

    Science.gov (United States)

    Blancett, Candace D; Fetterer, David P; Koistinen, Keith A; Morazzani, Elaine M; Monninger, Mitchell K; Piper, Ashley E; Kuehl, Kathleen A; Kearney, Brian J; Norris, Sarah L; Rossi, Cynthia A; Glass, Pamela J; Sun, Mei G

    2017-10-01

    A method for accurate quantitation of virus particles has long been sought, but a perfect method still eludes the scientific community. Electron Microscopy (EM) quantitation is a valuable technique because it provides direct morphology information and counts of all viral particles, whether or not they are infectious. In the past, EM negative stain quantitation methods have been cited as inaccurate, non-reproducible, and with detection limits that were too high to be useful. To improve accuracy and reproducibility, we have developed a method termed Scanning Transmission Electron Microscopy - Virus Quantitation (STEM-VQ), which simplifies sample preparation and uses a high throughput STEM detector in a Scanning Electron Microscope (SEM) coupled with commercially available software. In this paper, we demonstrate STEM-VQ with an alphavirus stock preparation to present the method's accuracy and reproducibility, including a comparison of STEM-VQ to viral plaque assay and the ViroCyt Virus Counter. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Holographic Two-Photon Induced Photopolymerization

    Data.gov (United States)

    Federal Laboratory Consortium — Holographic two-photon-induced photopolymerization (HTPIP) offers distinct advantages over conventional one-photon-induced photopolymerization and current techniques...

  2. Band excitation method applicable to scanning probe microscopy

    Science.gov (United States)

    Jesse, Stephen [Knoxville, TN; Kalinin, Sergei V [Knoxville, TN

    2010-08-17

    Methods and apparatus are described for scanning probe microscopy. A method includes generating a band excitation (BE) signal having finite and predefined amplitude and phase spectrum in at least a first predefined frequency band; exciting a probe using the band excitation signal; obtaining data by measuring a response of the probe in at least a second predefined frequency band; and extracting at least one relevant dynamic parameter of the response of the probe in a predefined range including analyzing the obtained data. The BE signal can be synthesized prior to imaging (static band excitation), or adjusted at each pixel or spectroscopy step to accommodate changes in sample properties (adaptive band excitation). An apparatus includes a band excitation signal generator; a probe coupled to the band excitation signal generator; a detector coupled to the probe; and a relevant dynamic parameter extractor component coupled to the detector, the relevant dynamic parameter extractor including a processor that performs a mathematical transform selected from the group consisting of an integral transform and a discrete transform.

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

    Energy Technology Data Exchange (ETDEWEB)

    Joseph, C.H., E-mail: hadlee.joseph@artov.imm.cnr.it [National Research Council, Institute for Microelectronics and Microsystems (CNR-IMM), Via del Fosso del Cavaliere 100, 00133 Rome (Italy); Department of Electronics Engineering, University of Rome “Tor Vergata”, Via del Politecnico 1, 00133 Rome (Italy); Sardi, G.M. [National Research Council, Institute for Microelectronics and Microsystems (CNR-IMM), Via del Fosso del Cavaliere 100, 00133 Rome (Italy); Tuca, S.S.; Gramse, G. [Johannes Kepler University, Institute for Biophysics, Gruberstrasse 40, A-4020 Linz (Austria); Lucibello, A.; Proietti, E. [National Research Council, Institute for Microelectronics and Microsystems (CNR-IMM), Via del Fosso del Cavaliere 100, 00133 Rome (Italy); Kienberger, F. [Keysight Technologies Austria GmbH, Keysight Laboratories, Gruberstrasse 40, A-4020 Linz (Austria); Marcelli, R. [National Research Council, Institute for Microelectronics and Microsystems (CNR-IMM), Via del Fosso del Cavaliere 100, 00133 Rome (Italy)

    2016-12-15

    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 S{sub 11} 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 S{sub 11} 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.

  4. Scanning electron microscopy and roughness study of dental composite degradation.

    Science.gov (United States)

    Soares, Luís Eduardo Silva; Cortez, Louise Ribeiro; Zarur, Raquel de Oliveira; Martin, Airton Abrahão

    2012-04-01

    Our aim was to test the hypothesis that the use of mouthwashes, consumption of soft drinks, as well as the type of light curing unit (LCU), would change the surface roughness (Ra) and morphology of a nanofilled composite resin (Z350® 3M ESPE). Samples (80) were divided into eight groups: Halogen LCU, group 1, saliva (control); group 2, Pepsi Twist®; group 3, Listerine®; group 4, Colgate Plax®; LED LCU, group 5, saliva; group 6, Pepsi Twist®; group 7, Listerine®; group 8, Colgate Plax®. Ra values were measured at baseline, and after 7 and 14 days. One specimen of each group was prepared for scanning electron microscopy analysis after 14 days. The data were subjected to multifactor analysis of variance at a 95% confidence followed by Tukey's honestly significant difference post-hoc test. All the treatments resulted in morphological changes in composite resin surface, and the most significant change was in Pepsi Twist® groups. The samples of G6 had the greatest increase in Ra. The immersion of nanofilled resin in mouthwashes with alcohol and soft drink increases the surface roughness. Polymerization by halogen LCU (reduced light intensity) associated with alcohol contained mouthwash resulted in significant roughness on the composite.

  5. Biological applications of near-field scanning optical microscopy

    Science.gov (United States)

    Moers, Marco H. P.; Ruiter, A. G. T.; Jalocha, Alain; van Hulst, Niko F.; Kalle, W. H. J.; Wiegant, J. C. A. G.; Raap, A. K.

    1995-09-01

    Near-field Scanning Optical Microscopy (NSOM) is a true optical microscopic technique allowing fluorescence, absorption, reflection and polarization contrast with the additional advantage of nanometer lateral resolution, unlimited by diffraction and operation at ambient conditions. NSOM based on metal coated adiabatically tapered fibers, combined with shear force feedback and operated in illumination mode, has proven to be the most powerful NSOM arrangement, because of its true localization of the optical interaction, its various optical contrast possibilities and its sensitivity down to the single molecular level. In this paper applications of `aperture' NSOM to Fluorescence In Situ Hybridization of human metaphase chromosomes are presented, where the localized fluorescence allows to identify specific DNA sequences. All images are accompanied by the simultaneously acquired force image, enabling direct comparison of the optical contrast with the sample topography on nanometer scale, far beyond the diffraction limit. Thus the unique combination of high resolution, specific optical contrast and ambient operation offers many new direction possibilities in biological studies.

  6. Non-thermal plasma mills bacteria: Scanning electron microscopy observations

    International Nuclear Information System (INIS)

    Lunov, O.; Churpita, O.; Zablotskii, V.; Jäger, A.; Dejneka, A.; Deyneka, I. G.; Meshkovskii, I. K.; Syková, E.; Kubinová, Š.

    2015-01-01

    Non-thermal plasmas hold great promise for a variety of biomedical applications. To ensure safe clinical application of plasma, a rigorous analysis of plasma-induced effects on cell functions is required. Yet mechanisms of bacteria deactivation by non-thermal plasma remain largely unknown. We therefore analyzed the influence of low-temperature atmospheric plasma on Gram-positive and Gram-negative bacteria. Using scanning electron microscopy, we demonstrate that both Gram-positive and Gram-negative bacteria strains in a minute were completely destroyed by helium plasma. In contrast, mesenchymal stem cells (MSCs) were not affected by the same treatment. Furthermore, histopathological analysis of hematoxylin and eosin–stained rat skin sections from plasma–treated animals did not reveal any abnormalities in comparison to control ones. We discuss possible physical mechanisms leading to the shred of bacteria under non-thermal plasma irradiation. Our findings disclose how helium plasma destroys bacteria and demonstrates the safe use of plasma treatment for MSCs and skin cells, highlighting the favorability of plasma applications for chronic wound therapy

  7. An overview on bioaerosols viewed by scanning electron microscopy

    International Nuclear Information System (INIS)

    Wittmaack, K.; Wehnes, H.; Heinzmann, U.; Agerer, R.

    2005-01-01

    Bioaerosols suspended in ambient air were collected with single-stage impactors at a semiurban site in southern Germany during late summer and early autumn. Sampling was mostly carried out at a nozzle velocity of 35 m/s, corresponding to a minimum aerodynamic diameter (cut-off diameter) of aerosol particles of 0.8 μm. The collected particles, sampled for short periods (∼15 min) to avoid pile-up, were characterized by scanning electron microscopy (SEM). The observed bioaerosols include brochosomes, fungal spores, hyphae, insect scales, hairs of plants and, less commonly, bacteria and epicuticular wax. Brochosomes, which serve as a highly water repellent body coating of leafhoppers, are hollow spheroids with diameters around 400 nm, resembling C 60 or footballs (soccer balls). They are usually airborne not as individuals but in the form of large clusters containing up to 10,000 individual species or even more. Various types of spores and scales were observed, but assignment turned out be difficult due to the large number of fungi and insects from which they may have originated. Pollens were observed only once. The absence these presumably elastic particles suggests that they are frequently lost, at the comparatively high velocities, due to bounce-off from the nonadhesive impaction surfaces

  8. Scanning probe microscopy of single Au ion implants in Si

    International Nuclear Information System (INIS)

    Vines, L.; Monakhov, E.; Maknys, K.; Svensson, B.G.; Jensen, J.; Hallen, A.; Kuznetsov, A. Yu.

    2006-01-01

    We have studied 5 MeV Au 2+ ion implantation with fluences between 7 x 10 7 and 2 x 10 8 cm -2 in Si by deep level transient spectroscopy (DLTS) and scanning capacitance microscopy (SCM). The DLTS measurements show formation of electrically active defects such as the two negative charge states of the divacancy (V 2 (=/-) and V 2 (-/0)) and the vacancy-oxygen (VO) center. It is observed that the intensity of the V 2 (=/-) peak is lower compared to that of V 2 (-/0) by a factor of 5. This has been attributed to a highly localized distribution of the defects along the ion tracks, which results in trapping of the carriers at V 2 (-/0) and incomplete occupancy of V 2 (=/-). The SCM measurements obtained in a plan view show a random pattern of regions with a reduced SCM signal for the samples implanted with fluence above 2 x 10 8 cm -2 . The reduced SCM signal is attributed to extra charges associated with acceptor states, such as V 2 (-/0), formed along the ion tracks in the bulk Si. Indeed, the electron emission rate from the V 2 (-/0) state is in the range of 10 kHz at room temperature, which is well below the probing frequency of the SCM measurements, resulting in 'freezing' of electrons at V 2 (-/0)

  9. Spectroscopic scanning tunneling microscopy insights into Fe-based superconductors

    International Nuclear Information System (INIS)

    Hoffman, Jennifer E

    2011-01-01

    In the first three years since the discovery of Fe-based high T c superconductors, scanning tunneling microscopy (STM) and spectroscopy have shed light on three important questions. First, STM has demonstrated the complexity of the pairing symmetry in Fe-based materials. Phase-sensitive quasiparticle interference (QPI) imaging and low temperature spectroscopy have shown that the pairing order parameter varies from nodal to nodeless s± within a single family, FeTe 1-x Se x . Second, STM has imaged C4 → C2 symmetry breaking in the electronic states of both parent and superconducting materials. As a local probe, STM is in a strong position to understand the interactions between these broken symmetry states and superconductivity. Finally, STM has been used to image the vortex state, giving insights into the technical problem of vortex pinning, and the fundamental problem of the competing states introduced when superconductivity is locally quenched by a magnetic field. Here we give a pedagogical introduction to STM and QPI imaging, discuss the specific challenges associated with extracting bulk properties from the study of surfaces, and report on progress made in understanding Fe-based superconductors using STM techniques.

  10. Scanning Auger microscopy for high lateral and depth elemental sensitivity

    Energy Technology Data Exchange (ETDEWEB)

    Martinez, E., E-mail: eugenie.martinez@cea.fr [CEA, LETI, MINATEC Campus, 17 rue des Martyrs, 38054 Grenoble Cedex 9 (France); Yadav, P. [CEA, LETI, MINATEC Campus, 17 rue des Martyrs, 38054 Grenoble Cedex 9 (France); Bouttemy, M. [Institut Lavoisier de Versailles, 45 av. des Etats-Unis, 78035 Versailles Cedex (France); Renault, O.; Borowik, Ł.; Bertin, F. [CEA, LETI, MINATEC Campus, 17 rue des Martyrs, 38054 Grenoble Cedex 9 (France); Etcheberry, A. [Institut Lavoisier de Versailles, 45 av. des Etats-Unis, 78035 Versailles Cedex (France); Chabli, A. [CEA, LETI, MINATEC Campus, 17 rue des Martyrs, 38054 Grenoble Cedex 9 (France)

    2013-12-15

    Highlights: •SAM performances and limitations are illustrated on real practical cases such as the analysis of nanowires and nanodots. •High spatial elemental resolution is shown with the analysis of reference semiconducting Al{sub 0.7}Ga{sub 0.3}As/GaAs multilayers. •High in-depth elemental resolution is also illustrated. Auger depth profiling with low energy ion beams allows revealing ultra-thin layers (∼1 nm). •Analysis of cross-sectional samples is another effective approach to obtain in-depth elemental information. -- Abstract: Scanning Auger microscopy is currently gaining interest for investigating nanostructures or thin multilayers stacks developed for nanotechnologies. New generation Auger nanoprobes combine high lateral (∼10 nm), energy (0.1%) and depth (∼2 nm) resolutions thus offering the possibility to analyze the elemental composition as well as the chemical state, at the nanometre scale. We report here on the performances and limitations on practical examples from nanotechnology research. The spatial elemental sensitivity is illustrated with the analysis of Al{sub 0.7}Ga{sub 0.3}As/GaAs heterostructures, Si nanowires and SiC nanodots. Regarding the elemental in-depth composition, two effective approaches are presented: low energy depth profiling to reveal ultra-thin layers (∼1 nm) and analysis of cross-sectional samples.

  11. Scanning tunneling microscopy and spectroscopy of twisted trilayer graphene

    Science.gov (United States)

    Zuo, Wei-Jie; Qiao, Jia-Bin; Ma, Dong-Lin; Yin, Long-Jing; Sun, Gan; Zhang, Jun-Yang; Guan, Li-Yang; He, Lin

    2018-01-01

    Twist, as a simple and unique degree of freedom, could lead to enormous novel quantum phenomena in bilayer graphene. A small rotation angle introduces low-energy van Hove singularities (VHSs) approaching the Fermi level, which result in unusual correlated states in the bilayer graphene. It is reasonable to expect that the twist could also affect the electronic properties of few-layer graphene dramatically. However, such an issue has remained experimentally elusive. Here, by using scanning tunneling microscopy/spectroscopy (STM/STS), we systematically studied a twisted trilayer graphene (TTG) with two different small twist angles between adjacent layers. Two sets of VHSs, originating from the two twist angles, were observed in the TTG, indicating that the TTG could be simply regarded as a combination of two different twisted bilayers of graphene. By using high-resolution STS, we observed a split of the VHSs and directly imaged the spatial symmetry breaking of electronic states around the VHSs. These results suggest that electron-electron interactions play an important role in affecting the electronic properties of graphene systems with low-energy VHSs.

  12. An overview on bioaerosols viewed by scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Wittmaack, K. [GSF-National Research Centre for Environment and Health, Institute of Radiation Protection, 85758 Neuherberg (Germany)]. E-mail: wittmaack@gsf.de; Wehnes, H. [GSF-National Research Centre for Environment and Health, Institute of Pathology, 85758 Neuherberg (Germany); Heinzmann, U. [GSF-National Research Centre for Environment and Health, Institute of Pathology, 85758 Neuherberg (Germany); Agerer, R. [Ludwig-Maximilians University Munich, Department Biology, Biodiversity Research: Mycology, Menzinger Stasse 67, 80638 Munich (Germany)

    2005-06-15

    Bioaerosols suspended in ambient air were collected with single-stage impactors at a semiurban site in southern Germany during late summer and early autumn. Sampling was mostly carried out at a nozzle velocity of 35 m/s, corresponding to a minimum aerodynamic diameter (cut-off diameter) of aerosol particles of 0.8 {mu}m. The collected particles, sampled for short periods ({approx}15 min) to avoid pile-up, were characterized by scanning electron microscopy (SEM). The observed bioaerosols include brochosomes, fungal spores, hyphae, insect scales, hairs of plants and, less commonly, bacteria and epicuticular wax. Brochosomes, which serve as a highly water repellent body coating of leafhoppers, are hollow spheroids with diameters around 400 nm, resembling C{sub 60} or footballs (soccer balls). They are usually airborne not as individuals but in the form of large clusters containing up to 10,000 individual species or even more. Various types of spores and scales were observed, but assignment turned out be difficult due to the large number of fungi and insects from which they may have originated. Pollens were observed only once. The absence these presumably elastic particles suggests that they are frequently lost, at the comparatively high velocities, due to bounce-off from the nonadhesive impaction surfaces.

  13. Specific surface area evaluation method by using scanning electron microscopy

    International Nuclear Information System (INIS)

    Petrescu, Camelia; Petrescu, Cristian; Axinte, Adrian

    2000-01-01

    Ceramics are among the most interesting materials for a large category of applications, including both industry and health. Among the characteristic of the ceramic materials, the specific surface area is often difficult to evaluate.The paper presents a method of evaluation for the specific surface area of two ceramic powders by means of scanning electron microscopy measurements and an original method of computing the specific surface area.Cumulative curves are used to calculate the specific surface area under assumption that the values of particles diameters follow a normal logarithmic distribution. For two powder types, X7R and NPO the results are the following: - for the density ρ (g/cm 2 ), 5.5 and 6.0, respectively; - for the average diameter D bar (μm), 0.51 and 0.53, respectively; - for σ, 1.465 and 1.385, respectively; - for specific surface area (m 2 /g), 1.248 and 1.330, respectively. The obtained results are in good agreement with the values measured by conventional methods. (authors)

  14. Reference nano-dimensional metrology by scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Dai, Gaoliang; Fluegge, Jens; Bosse, Harald; Heidelmann, Markus; Kübel, Christian; Prang, Robby

    2013-01-01

    Traceable and accurate reference dimensional metrology of nano-structures by scanning transmission electron microscopy (STEM) is introduced in the paper. Two methods, one based on the crystal lattice constant and the other based on the pitch of a feature pair, were applied to calibrate the TEM magnification. The threshold value, which was defined as the half-intensity of boundary materials, is suggested to extract the boundary position of features from the TEM image. Experimental investigations have demonstrated the high potential of the proposed methods. For instance, the standard deviation from ten repeated measurements of a line structure with a nominal 100 nm critical dimension (CD) reaches 1σ = 0.023 nm, about 0.02%. By intentionally introduced defocus and larger sample alignment errors, the investigation shows that these influences may reach 0.20 and 1.3 nm, respectively, indicating the importance of high-quality TEM measurements. Finally, a strategy for disseminating the destructive TEM results is introduced. Using this strategy, the CD of a reference material has been accurately determined. Its agreement over five independent TEM measurements is below 1.2 nm. (paper)

  15. Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS)

    Science.gov (United States)

    Song, Alexander; Charles, Adam S.; Koay, Sue Ann; Gauthier, Jeff L.; Thiberge, Stephan Y.; Pillow, Jonathan W.; Tank, David W.

    2017-01-01

    Two-photon laser scanning microscopy of calcium dynamics using fluorescent indicators is a widely used imaging method for large scale recording of neural activity in vivo. Here we introduce volumetric Two-photon Imaging of Neurons using Stereoscopy (vTwINS), a volumetric calcium imaging method that employs an elongated, V-shaped point spread function to image a 3D brain volume. Single neurons project to spatially displaced “image pairs” in the resulting 2D image, and the separation distance between images is proportional to depth in the volume. To demix the fluorescence time series of individual neurons, we introduce a novel orthogonal matching pursuit algorithm that also infers source locations within the 3D volume. We illustrate vTwINS by imaging neural population activity in mouse primary visual cortex and hippocampus. Our results demonstrate that vTwINS provides an effective method for volumetric two-photon calcium imaging that increases the number of neurons recorded while maintaining a high frame-rate. PMID:28319111

  16. Secondary electron spectroscopy and Auger microscopy at high spatial resolution. Application to scanning electron microscopy

    International Nuclear Information System (INIS)

    Le Gressus, Claude; Massignon, Daniel; Sopizet, Rene

    1979-01-01

    Secondary electron spectroscopy (SES), Auger electron spectroscopy (AES) and electron energy loss spectroscopy (ELS) are combined with ultra high vacuum scanning microscopy (SEM) for surface analysis at high spatial resolution. Reliability tests for the optical column for the vacuum and for the spectrometer are discussed. Furthermore the sensitivity threshold in AES which is compatible with a non destructive surface analysis at high spatial resolution is evaluated. This combination of all spectroscopies is used in the study of the beam damage correlated with the well known secondary electron image (SEI) darkening still observed in ultra high vacuum. The darkening is explained as a bulk decontamination of the sample rather than as a surface contamination from the residual vacuum gas [fr

  17. Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy.

    Science.gov (United States)

    Schulz, Olaf; Pieper, Christoph; Clever, Michaela; Pfaff, Janine; Ruhlandt, Aike; Kehlenbach, Ralph H; Wouters, Fred S; Großhans, Jörg; Bunt, Gertrude; Enderlein, Jörg

    2013-12-24

    We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions.

  18. Quantitative Scanning Transmission Electron Microscopy of Electronic and Nanostructured Materials

    Science.gov (United States)

    Yankovich, Andrew B.

    Electronic and nanostructured materials have been investigated using advanced scanning transmission electron microscopy (STEM) techniques. The first topic is the microstructure of Ga and Sb-doped ZnO. Ga-doped ZnO is a candidate transparent conducting oxide material. The microstructure of GZO thin films grown by MBE under different growth conditions and different substrates were examined using various electron microscopy (EM) techniques. The microstructure, prevalent defects, and polarity in these films strongly depend on the growth conditions and substrate. Sb-doped ZnO nanowires have been shown to be the first route to stable p-type ZnO. Using Z-contrast STEM, I have showed that an unusual microstructure of Sb-decorated head-to-head inversion domain boundaries and internal voids contain all the Sb in the nanowires and cause the p-type conduction. InGaN thin films and InGaN / GaN quantum wells (QW) for light emitting diodes are the second topic. Low-dose Z-contrast STEM, PACBED, and EDS on InGaN QW LED structures grown by MOCVD show no evidence for nanoscale composition variations, contradicting previous reports. In addition, a new extended defect in GaN and InGaN was discovered. The defect consists of a faceted pyramid-shaped void that produces a threading dislocation along the [0001] growth direction, and is likely caused by carbon contamination during growth. Non-rigid registration (NRR) and high-precision STEM of nanoparticles is the final topic. NRR is a new image processing technique that corrects distortions arising from the serial nature of STEM acquisition that previously limited the precision of locating atomic columns and counting the number of atoms in images. NRR was used to demonstrate sub-picometer precision in STEM images of single crystal Si and GaN, the best achieved in EM. NRR was used to measure the atomic surface structure of Pt nanoacatalysts and Au nanoparticles, which revealed new bond length variation phenomenon of surface atoms. In

  19. Quarkonium two-photon decays in QCD

    International Nuclear Information System (INIS)

    Dulyan, L.S.; Khodjamirian, A.Yu.; Magakian, A.D.

    1989-01-01

    The two-photon decay of tensor charmonium χ c2 → 2 γ is calculated with account of gluon condensate effects. The result is in good agreement with experiment. The two-photon width of pseudoscalar b-quarkonium η b → 2 γ is estimated. 19 refs.; 1 fig.; 1 tab

  20. Automated Quantitative Rare Earth Elements Mineralogy by Scanning Electron Microscopy

    Science.gov (United States)

    Sindern, Sven; Meyer, F. Michael

    2016-09-01

    Increasing industrial demand of rare earth elements (REEs) stems from the central role they play for advanced technologies and the accelerating move away from carbon-based fuels. However, REE production is often hampered by the chemical, mineralogical as well as textural complexity of the ores with a need for better understanding of their salient properties. This is not only essential for in-depth genetic interpretations but also for a robust assessment of ore quality and economic viability. The design of energy and cost-efficient processing of REE ores depends heavily on information about REE element deportment that can be made available employing automated quantitative process mineralogy. Quantitative mineralogy assigns numeric values to compositional and textural properties of mineral matter. Scanning electron microscopy (SEM) combined with a suitable software package for acquisition of backscatter electron and X-ray signals, phase assignment and image analysis is one of the most efficient tools for quantitative mineralogy. The four different SEM-based automated quantitative mineralogy systems, i.e. FEI QEMSCAN and MLA, Tescan TIMA and Zeiss Mineralogic Mining, which are commercially available, are briefly characterized. Using examples of quantitative REE mineralogy, this chapter illustrates capabilities and limitations of automated SEM-based systems. Chemical variability of REE minerals and analytical uncertainty can reduce performance of phase assignment. This is shown for the REE phases parisite and synchysite. In another example from a monazite REE deposit, the quantitative mineralogical parameters surface roughness and mineral association derived from image analysis are applied for automated discrimination of apatite formed in a breakdown reaction of monazite and apatite formed by metamorphism prior to monazite breakdown. SEM-based automated mineralogy fulfils all requirements for characterization of complex unconventional REE ores that will become

  1. Outwitting the series resistance in scanning spreading resistance microscopy

    International Nuclear Information System (INIS)

    Schulze, A.; Cao, R.; Eyben, P.; Hantschel, T.; Vandervorst, W.

    2016-01-01

    The performance of nanoelectronics devices critically depends on the distribution of active dopants inside these structures. For this reason, dopant profiling has been defined as one of the major metrology challenges by the international technology roadmap of semiconductors. Scanning spreading resistance microscopy (SSRM) has evolved as one of the most viable approaches over the last decade due to its excellent spatial resolution, sensitivity and quantification accuracy. However, in case of advanced device architectures like fins and nanowires a proper measurement of the spreading resistance is often hampered by the increasing impact of parasitic series resistances (e.g. bulk series resistance) arising from the confined nature of the aforementioned structures. In order to overcome this limitation we report in this paper the development and implementation of a novel SSRM mode (fast Fourier transform-SSRM: FFT-SSRM) which essentially decouples the spreading resistance from parasitic series resistance components. We show that this can be achieved by a force modulation (leading to a modulated spreading resistance signal) in combination with a lock-in deconvolution concept. In this paper we first introduce the principle of operation of the technique. We discuss in detail the underlying physical mechanisms as well as the technical implementation on a state-of-the-art atomic force microscope (AFM). We demonstrate the performance of FFT-SSRM and its ability to remove substantial series resistance components in practice. Eventually, the possibility of decoupling the spreading resistance from the intrinsic probe resistance will be demonstrated and discussed. - Highlights: • A novel electrical AFM mode for carrier profiling in confined volumes is presented. • Thereby the force and hence the contact area between AFM probe and sample is modulated. • Information on the spreading resistance is derived using a lock-in approach. • Bulk series resistance components are

  2. Folate receptor targeting silica nanoparticle probe for two-photon fluorescence bioimaging

    Science.gov (United States)

    Wang, Xuhua; Yao, Sheng; Ahn, Hyo-Yang; Zhang, Yuanwei; Bondar, Mykhailo V.; Torres, Joseph A.; Belfield, Kevin D.

    2010-01-01

    Narrow dispersity organically modified silica nanoparticles (SiNPs), diameter ~30 nm, entrapping a hydrophobic two-photon absorbing fluorenyl dye, were synthesized by hydrolysis of triethoxyvinylsilane and (3-aminopropyl)triethoxysilane in the nonpolar core of Aerosol-OT micelles. The surface of the SiNPs were functionalized with folic acid, to specifically deliver the probe to folate receptor (FR) over-expressing Hela cells, making these folate two-photon dye-doped SiNPs potential candidates as probes for two-photon fluorescence microscopy (2PFM) bioimaging. In vitro studies using FR over-expressing Hela cells and low FR expressing MG63 cells demonstrated specific cellular uptake of the functionalized nanoparticles. One-photon fluorescence microscopy (1PFM) imaging, 2PFM imaging, and two-photon fluorescence lifetime microscopy (2P-FLIM) imaging of Hela cells incubated with folate-modified two-photon dye-doped SiNPs were demonstrated. PMID:21258480

  3. Scanning tunneling microscopy III theory of STM and related scanning probe methods

    CERN Document Server

    Güntherodt, Hans-Joachim

    1993-01-01

    While the first two volumes on Scanning Tunneling Microscopy (STM) and its related scanning probe (SXM) methods have mainly concentrated on intro­ ducing the experimental techniques, as well as their various applications in different research fields, this third volume is exclusively devoted to the theory of STM and related SXM methods. As the experimental techniques including the reproducibility of the experimental results have advanced, more and more theorists have become attracted to focus on issues related to STM and SXM. The increasing effort in the development of theoretical concepts for STM/SXM has led to considerable improvements in understanding the contrast mechanism as well as the experimental conditions necessary to obtain reliable data. Therefore, this third volume on STM/SXM is not written by theorists for theorists, but rather for every scientist who is not satisfied by just obtaining real­ space images of surface structures by STM/SXM. After a brief introduction (Chap. 1), N. D. Lang first co...

  4. Enzymatic hydrolysis of Amaranth flour - differential scanning calorimetry and scanning electron microscopy studies

    Energy Technology Data Exchange (ETDEWEB)

    Barba de la Rosa, A.P.; Paredes-Lopez, O.; Carabez-Trejo, A.; Ordorica-Falomir, C. (Instituto Politecnico Nacional, Irapuato (Mexico). Centro de Investigacion y de Estudios Avanzados)

    1989-11-01

    High-protein amaranth flour (HPAF) and carbohydrate rich fraction (CRF) were produced from raw flour in a single-step process using a heat-stable alpha-amylase preparation. Protein content of flour increased from 15 to about 30 or 39% at liquefaction temperatures of 70 or 90{sup 0}C, respectively and 30 min hydrolysis time. CRF exhibited 14-22 DE. Enzymatic action at 70{sup 0}C increased endotherm temperature and gelatinization enthalpy of HPAF, in relation to gelatinized flour, as assessed by differential scanning calorimetry (DSC). Hydrolysis at 90{sup 0}C did not affect significantly (P > 0.05) DSC peak temperature. It is suggested that these changes in DSC performance might result from differences in amount and type of low-molecular weight carbohydrates and residual starch. Scanning electron microscopy (SEM) demonstrated that hydrolysis temperature changed substantially the structural appearance of flour particles. HPAF and CRF might find applications as dry milk extender and sweetener, respectively. (orig.).

  5. Mechanisms of biliary stent clogging: confocal laser scanning and scanning electron microscopy.

    Science.gov (United States)

    van Berkel, A M; van Marle, J; Groen, A K; Bruno, M J

    2005-08-01

    Endoscopic insertion of plastic biliary endoprostheses is a well-established treatment for obstructive jaundice. The major limitation of this technique is late stent occlusion. In order to compare events involved in biliary stent clogging and identify the distribution of bacteria in unblocked stents, confocal laser scanning (CLS) and scanning electron microscopy (SEM) were carried out on two different stent materials - polyethylene (PE) and hydrophilic polymer-coated polyurethane (HCPC). Ten consecutive patients with postoperative benign biliary strictures were included in the study. Two 10-Fr stents 9 cm in length, one made of PE and the other of HCPC, were inserted. The stents were electively exchanged after 3 months and examined using CLS and SEM. No differences were seen between the two types of stent. The inner stent surface was covered with a uniform amorphous layer. On top of this layer, a biofilm of living and dead bacteria was found, which in most cases was unstructured. The lumen was filled with free-floating colonies of bacteria and crystals, surrounded by mobile laminar structures of mucus. An open network of large dietary fibers was seen in all of the stents. The same clogging events occurred in both PE and HCPC stents. The most remarkable observation was the identification of networks of large dietary fibers, resulting from duodenal reflux, acting as a filter. The build-up of this intraluminal framework of dietary fibers appears to be a major factor contributing to the multifactorial process of stent clogging.

  6. Wide-band acousto-optic deflectors for large field of view two-photon microscope.

    Science.gov (United States)

    Jiang, Runhua; Zhou, Zhenqiao; Lv, Xiaohua; Zeng, Shaoqun

    2012-04-01

    Acousto-optic deflector (AOD) is an attractive scanner for two-photon microscopy because it can provide fast and versatile laser scanning and does not involve any mechanical movements. However, due to the small scan range of available AOD, the field of view (FOV) of the AOD-based microscope is typically smaller than that of the conventional galvanometer-based microscope. Here, we developed a novel wide-band AOD to enlarge the scan angle. Considering the maximum acceptable acoustic attenuation in the acousto-optic crystal, relatively lower operating frequencies and moderate aperture were adopted. The custom AOD was able to provide 60 MHz 3-dB bandwidth and 80% peak diffraction efficiency at 840 nm wavelength. Based on a pair of such AOD, a large FOV two-photon microscope was built with a FOV up to 418.5 μm (40× objective). The spatiotemporal dispersion was compensated simultaneously with a single custom-made prism. By means of dynamic power modulation, the variation of laser intensity within the FOV was reduced below 5%. The lateral and axial resolution of the system were 0.58-2.12 μm and 2.17-3.07 μm, respectively. Pollen grain images acquired by this system were presented to demonstrate the imaging capability at different positions across the entire FOV. © 2012 American Institute of Physics

  7. Surface confined metallosupramolecular architectures: formation and scanning tunneling microscopy characterization.

    Science.gov (United States)

    Li, Shan-Shan; Northrop, Brian H; Yuan, Qun-Hui; Wan, Li-Jun; Stang, Peter J

    2009-02-17

    Metallosupramolecular compounds have attracted a great deal of attention over the past two decades largely because of their unique, highly complex structural characteristics and their potential electronic, magnetic, optical, and catalytic properties. These molecules can be prepared with relative ease using coordination-driven self-assembly techniques. In particular, the use of electron-poor square-planar Pt(II) transition metals in conjunction with rigid, electron-rich pyridyl donors has enabled the spontaneous self-assembly of a rich library of 2D metallacyclic and 3D metallacage assemblies via the directional-bonding approach. With this progress in the preparation and characterization of metallosupramolecules, researchers have now turned their attention toward fully exploring and developing their materials properties. Assembling metallosupramolecular compounds on solid supports represents a vitally important step toward developing their materials properties. Surfaces provide a means of uniformly aligning and orienting these highly symmetric metallacycles and metallacages. This uniformity increases the level of coherence between molecules above that which can be achieved in the solution phase and provides a way to integrate adsorbed layers, or adlayers, into a solid-state materials setting. The dynamic nature of kinetically labile Pt(II)-N coordination bonds requires us to adjust deposition and imaging conditions to retain the assemblies' stability. Toward these aims, we have used scanning tunneling microscopy (STM) to image these adlayers and to understand the factors that govern surface self-assembly and the interactions that influence their structure and stability. This Account describes our efforts to deposit 2D rectangular and square metallacycles and 3D trigonal bipyramidal and chiral trigonal prism metallacages on highly oriented pyrolytic graphite (HOPG) and Au(111) substrates to give intact assemblies and ordered adlayers. We have investigated the effects

  8. Two-photon high-resolution measurement of partial pressure of oxygen in cerebral vasculature and tissue

    Science.gov (United States)

    Sakadžić, Sava; Roussakis, Emmanuel; Yaseen, Mohammad A.; Mandeville, Emiri T.; Srinivasan, Vivek J.; Arai, Ken; Ruvinskaya, Svetlana; Devor, Anna; Lo, Eng H.; Vinogradov, Sergei A.; Boas, David A.

    2010-01-01

    The ability to measure oxygen partial pressure (pO2) with high temporal and spatial resolution in three dimensions is crucial for understanding oxygen delivery and consumption in normal and diseased brain. Among existing pO2 measurement methods, phosphorescence quenching is optimally suited for the task. However, previous attempts to couple phosphorescence with two-photon laser scanning microscopy have faced substantial difficulties because of extremely low two-photon absorption cross-sections of conventional phosphorescent probes. Here, we report the first practical in vivo two-photon high-resolution pO2 measurements in small rodents’ cortical microvasculature and tissue, made possible by combining an optimized imaging system with a two-photon-enhanced phosphorescent nanoprobe. The method features a measurement depth of up to 250 µm, sub-second temporal resolution and requires low probe concentration. Most importantly, the properties of the probe allowed for the first direct high-resolution measurement of cortical extravascular (tissue) pO2, opening numerous possibilities for functional metabolic brain studies. PMID:20693997

  9. Two-photon high-resolution measurement of partial pressure of oxygen in cerebral vasculature and tissue.

    Science.gov (United States)

    Sakadzić, Sava; Roussakis, Emmanuel; Yaseen, Mohammad A; Mandeville, Emiri T; Srinivasan, Vivek J; Arai, Ken; Ruvinskaya, Svetlana; Devor, Anna; Lo, Eng H; Vinogradov, Sergei A; Boas, David A

    2010-09-01

    Measurements of oxygen partial pressure (pO(2)) with high temporal and spatial resolution in three dimensions is crucial for understanding oxygen delivery and consumption in normal and diseased brain. Among existing pO(2) measurement methods, phosphorescence quenching is optimally suited for the task. However, previous attempts to couple phosphorescence with two-photon laser scanning microscopy have faced substantial difficulties because of extremely low two-photon absorption cross-sections of conventional phosphorescent probes. Here we report to our knowledge the first practical in vivo two-photon high-resolution pO(2) measurements in small rodents' cortical microvasculature and tissue, made possible by combining an optimized imaging system with a two-photon-enhanced phosphorescent nanoprobe. The method features a measurement depth of up to 250 microm, sub-second temporal resolution and requires low probe concentration. The properties of the probe allowed for direct high-resolution measurement of cortical extravascular (tissue) pO(2), opening many possibilities for functional metabolic brain studies.

  10. Time gated fluorescence lifetime imaging and micro-volume spectroscopy using two-photon excitation

    NARCIS (Netherlands)

    Sytsma, J.; Vroom, J.M.; de Grauw, C.J.; Gerritsen, H.C.

    A scanning microscope utilizing two-photon excitation in combination with fluorescence lifetime contrast is presented. The microscope makes use of a tunable femtosecond titanium:sapphire laser enabling the two-photon excitation of a broad range of fluorescent molecules, including UV probes.

  11. Nanoscale electrical property studies of individual GeSi quantum rings by conductive scanning probe microscopy.

    Science.gov (United States)

    Lv, Yi; Cui, Jian; Jiang, Zuimin M; Yang, Xinju

    2012-11-29

    The nanoscale electrical properties of individual self-assembled GeSi quantum rings (QRs) were studied by scanning probe microscopy-based techniques. The surface potential distributions of individual GeSi QRs are obtained by scanning Kelvin microscopy (SKM). Ring-shaped work function distributions are observed, presenting that the QRs' rim has a larger work function than the QRs' central hole. By combining the SKM results with those obtained by conductive atomic force microscopy and scanning capacitance microscopy, the correlations between the surface potential, conductance, and carrier density distributions are revealed, and a possible interpretation for the QRs' conductance distributions is suggested.

  12. Study of Perylenetetracarboxylic Acid Dimethylimide Films by Cyclic Thermal Desorption and Scanning Probe Microscopy

    Science.gov (United States)

    Pochtennyi, A. E.; Lappo, A. N.; Il'yushonok, I. P.

    2018-02-01

    Some results of studying the direct-current (DC) conductivity of perylenetetracarboxylic acid dimethylimide films by cyclic oxygen thermal desorption are presented. The microscopic parameters of hopping electron transport over localized impurity and intrinsic states were determined. The bandgap width and the sign of major current carriers were determined by scanning probe microscopy methods (atomic force microscopy, scanning probe spectroscopy, and photoassisted Kelvin probe force microscopy). The possibility of the application of photoassisted scanning tunneling microscopy for the nanoscale phase analysis of photoconductive films is discussed.

  13. Direct observation of atoms on surfaces by scanning tunnelling microscopy

    International Nuclear Information System (INIS)

    Baldeschwieler, J.D.

    1989-01-01

    The scanning tunnelling microscope is a non-destructive means of achieving atomic level resolution of crystal surfaces in real space to elucidate surface structures, electronic properties and chemical composition. Scanning tunnelling microscope is a powerful, real space surface structure probe complementary to other techniques such as x-ray diffraction. 21 refs., 8 figs

  14. 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...

  15. Scanning probe microscopy techniques for mechanical characterization at nanoscale

    International Nuclear Information System (INIS)

    Passeri, D.; Anastasiadis, P.; Tamburri, E.; Gugkielmotti, V.; Rossi, M.

    2013-01-01

    Three atomic force microscopy (AFM)-based techniques are reviewed that allow one to conduct accurate measurements of mechanical properties of either stiff or compliant materials at a nanometer scale. Atomic force acoustic microscopy, AFM-based depth sensing indentation, and torsional harmonic AFM are briefly described. Examples and results of quantitative characterization of stiff (an ultrathin SeSn film), soft polymeric (polyaniline fibers doped with detonation nanodiamond) and biological (collagen fibers) materials are reported.

  16. Atomic force and scanning near-field optical microscopy study of carbocyanine dye J-aggregates

    Czech Academy of Sciences Publication Activity Database

    Prokhorov, V.V.; Petrova, M.G.; Kovaleva, Natalia; Demikhov, E.I.

    2014-01-01

    Roč. 10, č. 5 (2014), s. 700-704 ISSN 1573-4137 Institutional support: RVO:68378271 Keywords : carbocyanine dye * elementary fibri * high-resolution atomic force microscopy * J-aggregate * probe microscopy * scanning near-field optical microscopy Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.096, year: 2014

  17. Charm production in two-photon collisions

    International Nuclear Information System (INIS)

    Linde, F.L.

    1988-01-01

    The analysis focuses on the production of charmonium resonances η c , χ 0 and χ 2 in two-photon interactions. The measurement of the inclusive production of charged D* mesons is described. 97 refs.; 54 figs.; 15 tabs

  18. Exclusive hadron production in two photon reactions

    International Nuclear Information System (INIS)

    Poppe, M.

    1986-02-01

    This paper summarises experimental results on exclusive hadron production in two photon collisions at electron positron storage rings and attempts some interpretation. Experimental know how is described and new suggestions are made for future analyses. New model calculations on resonance form factors and pair production amplitudes are presented. The two photon vertex is decomposed such that experiments can be parameterised with the minimal number of free parameters. Selection rules for off shell photon collisions are given in addition to Yang's theorems. (orig.)

  19. Stimulated emission depletion following two photon excitation

    OpenAIRE

    Marsh, R. J.; Armoogum, D. A.; Bain, A. J.

    2002-01-01

    The technique of stimulated emission depletion of fluorescence (STED) from a two photon excited molecular population is demonstrated in the S, excited state of fluorescein in ethylene glycol and methanol. Two photon excitation (pump) is achieved using the partial output of a regeneratively amplified Ti:Sapphire laser in conjunction with an optical parametric amplifier whose tuneable output provides a synchronous depletion (dump) pulse. Time resolved fluorescence intensity and anisotropy measu...

  20. Observation of Magnetic Induction Distribution by Scanning Interference Electron Microscopy

    Science.gov (United States)

    Takahashi, Yoshio; Yajima, Yusuke; Ichikawa, Masakazu; Kuroda, Katsuhiro

    1994-09-01

    A scanning interference electron microscope (SIEM) capable of observing magnetic induction distribution with high sensitivity and spatial resolution has been developed. The SIEM uses a pair of fine coherent scanning probes and detects their relative phase change by magnetic induction, giving raster images of microscopic magnetic distributions. Its performance has been demonstrated by observing magnetic induction distributed near the edge of a recorded magnetic storage medium. Obtained images are compared with corresponding images taken in the scanning Lorentz electron microscope mode using the same microscope, and the differences between them are discussed.

  1. A compact two photon light sheet microscope for applications in neuroscience

    DEFF Research Database (Denmark)

    Piksarv, Peeter; Marti, Dominik; Le, Tuan

    2016-01-01

    We present a compact setup for two photon light sheet microscopy. By using pulsed Airy beam illumination we demonstrate eight-fold increase of the FOV compared to Gaussian light sheet with the same axial resolution....

  2. Water-Soluble Triarylborane Chromophores for One- and Two-Photon Excited Fluorescence Imaging of Mitochondria in Cells.

    Science.gov (United States)

    Griesbeck, Stefanie; Zhang, Zuolun; Gutmann, Marcus; Lühmann, Tessa; Edkins, Robert M; Clermont, Guillaume; Lazar, Adina N; Haehnel, Martin; Edkins, Katharina; Eichhorn, Antonius; Blanchard-Desce, Mireille; Meinel, Lorenz; Marder, Todd B

    2016-10-04

    Three water-soluble tetracationic quadrupolar chromophores comprising two three-coordinate boron π-acceptor groups bridged by thiophene-containing moieties were synthesised for biological imaging applications. Compound 3 containing the bulkier 5-(3,5-Me2 C6 H2 )-2,2'-(C4 H2 S)2 -5'-(3,5-Me2 C6 H2 ) bridge is stable over a long period of time, exhibits a high fluorescence quantum yield and strong one- and two-photon absorption (TPA), and has a TPA cross section of 268 GM at 800 nm in water. Confocal laser scanning fluorescence microscopy studies in live cells indicated localisation of the chromophore at the mitochondria; moreover, cytotoxicity measurements proved biocompatibility. Thus, chromophore 3 has excellent potential for one- and two-photon-excited fluorescence imaging of mitochondrial function in cells. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. A series of fluorene-based two-photon absorbing molecules: synthesis, linear and nonlinear characterization, and bioimaging

    Science.gov (United States)

    Andrade, Carolina D.; Yanez, Ciceron O.; Rodriguez, Luis; Belfield, Kevin D.

    2010-01-01

    The synthesis, structural, and photophysical characterization of a series of new fluorescent donor–acceptor and acceptor-acceptor molecules, based on the fluorenyl ring system, with two-photon absorbing properties is described. These new compounds exhibited large Stokes shifts, high fluorescent quantum yields, and, significantly, high two-photon absorption cross sections, making them well suited for two-photon fluorescence microscopy (2PFM) imaging. Confocal and two-photon fluorescence microscopy imaging of COS-7 and HCT 116 cells incubated with probe I showed endosomal selectivity, demonstrating the potential of this class of fluorescent probes in multiphoton fluorescence microscopy. PMID:20481596

  4. Three-Dimensional scanning transmission electron microscopy of biological specimens

    KAUST Repository

    De Jonge, Niels; Sougrat, Rachid; Northan, Brian M.; Pennycook, Stephen J.

    2010-01-01

    A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM

  5. Commendable surface physics by means of scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Besenbacher, F.; Laegsgaard, E.; Stensgard, I.

    1995-01-01

    The scanning tunneling microscope, developed at the Aarhus University (Denmark) allows taking several STM images per second, as opposite to other similar microscopes, where the typical scanning time is 0,5-1 min. This new system enables collecting of important information concerning dynamic processes on the surfaces. The Aarhus microscope is very stable, hence atomic resolution is achievable even on close-packed metallic surfaces, while it is difficult to achieve by means of the conventional STM. (EG)

  6. Real-time high dynamic range laser scanning microscopy

    Science.gov (United States)

    Vinegoni, C.; Leon Swisher, C.; Fumene Feruglio, P.; Giedt, R. J.; Rousso, D. L.; Stapleton, S.; Weissleder, R.

    2016-04-01

    In conventional confocal/multiphoton fluorescence microscopy, images are typically acquired under ideal settings and after extensive optimization of parameters for a given structure or feature, often resulting in information loss from other image attributes. To overcome the problem of selective data display, we developed a new method that extends the imaging dynamic range in optical microscopy and improves the signal-to-noise ratio. Here we demonstrate how real-time and sequential high dynamic range microscopy facilitates automated three-dimensional neural segmentation. We address reconstruction and segmentation performance on samples with different size, anatomy and complexity. Finally, in vivo real-time high dynamic range imaging is also demonstrated, making the technique particularly relevant for longitudinal imaging in the presence of physiological motion and/or for quantification of in vivo fast tracer kinetics during functional imaging.

  7. Confocal scanning microscopy with multiple optical probes for high speed measurements and better imaging

    Science.gov (United States)

    Chun, Wanhee; Lee, SeungWoo; Gweon, Dae-Gab

    2008-02-01

    Confocal scanning microscopy (CSM) needs a scanning mechanism because only one point information of specimen can be obtained. Therefore the speed of the confocal scanning microscopy is limited by the speed of the scanning tool. To overcome this limitation from scanning tool we propose another scanning mechanism. We make three optical probes in the specimen under confocal condition of each point. Three optical probes are moved by beam scanning mechanism with shared resonant scanning mirror (RM) and galvanometer driven mirror (GM). As each optical probe scan allocated region of the specimen, information from three points is obtained simultaneously and image acquisition time is reduced. Therefore confocal scanning microscopy with multiple optical probes is expected to have three times faster speed of the image acquisition than conventional one. And as another use, multiple optical probes to which different light wavelength is applied can scan whole same region respectively. It helps to obtain better contrast image in case of specimens having different optical characteristics for specific light wavelength. In conclusion confocal scanning microscopy with multiple optical probes is useful technique for views of image acquisition speed and image quality.

  8. Scanning force microscopy and fluorescence microscopy of microcontact printed antibodies and antibody fragments.

    Science.gov (United States)

    LaGraff, John R; Chu-LaGraff, Quynh

    2006-05-09

    Unlabeled primary immunoglobulin G (IgG) antibodies and its F(ab')2 and Fc fragments were attached to oxygen-plasma-cleaned glass substrates using either microcontact printing (MCP) or physical adsorption during bath application from dilute solutions. Fluorescently labeled secondary IgGs were then bound to surface-immobilized IgG, and the relative surface coverage was determined by measuring the fluorescence intensity. Results indicated that the surface coverage of IgG increased with increasing protein solution concentration for both MCP and bath-applied IgG and that a greater concentration of IgG was transferred to a glass substrate using MCP than during physisorption during bath applications. Scanning force microscopy (SFM) showed that patterned MCP IgG monolayers were 5 nm in height, indicating that IgG molecules lie flat on the substrate. After incubation with a secondary IgG, the overall line thickness increased to around 15 nm, indicating that the secondary IgG was in a more vertical orientation with respect to the substrate. The surface roughness of these MCP patterned IgG bilayers as measured by SFM was observed to increase with increasing surface coverage. Physisorption of IgG to both unmodified patterned polydimethylsiloxane (PDMS) stamps and plasma-cleaned glass substrates was modeled by Langmuir adsorption kinetics yielding IgG binding constants of K(MCP) = 1.7(2) x 10(7) M(-1) and K(bath) = 7.8(7) x 10(5) M(-1), respectively. MCP experiments involving primary F(ab')2 and Fc fragments incubated in fluorescently labeled fragment-specific secondary IgGs were carried out to test for the function and orientation of IgG. Finally, possible origins of MCP stamping defects such as pits, pull outs, droplets, and reverse protein transfer are discussed.

  9. Ultrastructure of Proechinophthirus zumpti (Anoplura, Echinophthiriidae by scanning electron microscopy

    Directory of Open Access Journals (Sweden)

    Dolores del Carmen Castro

    2002-09-01

    Full Text Available The ultrastructure of Proechinophthirus zumpti Werneck, 1955, mainly the external chorionic features of the egg, is described through electronic microscopy techniques. This species was first cited in Argentina, infesting Arctocephalus australis (Zimmermann, 1873. The morphological adaptations of adults and nymphs are described in both species of Proechinophthirus parasitic on Otariidae: P. fluctus (Ferris, 1916 and P. zumpti.

  10. 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...... in transmission mode. In both cases the SH signal peaks at a pump wavelength of similar to 885 nm in correspondence to the maximum in the photoluminescence spectrum of the QD sample. SH near-field optical images exhibit spatial signal variations on a subwavelength scale that depend on the pump wavelength. We...

  11. Electrically Induced Two-Photon Transparency in Semiconductor Quantum Wells

    International Nuclear Information System (INIS)

    Hayat, Alex; Nevet, Amir; Orenstein, Meir

    2009-01-01

    We demonstrate experimentally two-photon transparency, achieved by current injection into a semiconductor quantum-well structure which exhibits two-photon emission. The two-photon induced luminescence is progressively reduced by the injected current, reaching the point of two-photon transparency - a necessary condition for semiconductor two-photon gain and lasing. These results agree with our calculations.

  12. Scanning Probe Microscopy at 650 °C in Air

    DEFF Research Database (Denmark)

    Hansen, Karin Vels; Jacobsen, Torben; Nørgaard, Anne-Mette

    2009-01-01

    The controlled atmosphere high temperature scanning probe microscope was designed to study the electrical properties of surfaces at elevated temperatures by using the probe as an electrode. The capability of a simultaneous acquisition of topographical and electrical data for the same surface area...

  13. Scanning Emitter Lifetime Imaging Microscopy for Spontaneous Emission Control

    DEFF Research Database (Denmark)

    Frimmer, Martin; Chen, Yuntian; Koenderink, A. Femius

    2011-01-01

    We report an experimental technique to map and exploit the local density of optical states of arbitrary planar nanophotonic structures. The method relies on positioning a spontaneous emitter attached to a scanning probe deterministically and reversibly with respect to its photonic environment while...

  14. Thermovoltages in vacuum tunneling investigated by scanning tunneling microscopy

    OpenAIRE

    Hoffmann, D. H.; Rettenberger, Armin; Grand, Jean Yves; Läuger, K.; Leiderer, Paul; Dransfeld, Klaus; Möller, Rolf

    1995-01-01

    By heating the tunneling tip of a scanning tunneling microscope the thermoelectric properties of a variable vacuum barrier have been investigated. The lateral variation of the observed thermovoltage will be discussed for polycrystalline gold, stepped surfaces of silver, as well as for copper islands on silver.

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

    DEFF Research Database (Denmark)

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

    1999-01-01

    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....

  16. Electromigration of single metal atoms observed by scanning tunneling microscopy

    NARCIS (Netherlands)

    Braun, K.-F.; Soe, W.H.; Flipse, C.F.J.

    2007-01-01

    The authors show in this letter that single metal atoms on a Ni(111) surface can be pushed by electromigration forces from a scanning tunneling microscope tip. This repulsive interaction is obsd. over a length scale of 6 nm. While for voltages above -300 mV the atoms are pulled by the microscope

  17. Photoelectrical properties of semiconductor tips in scanning tunneling microscopy

    NARCIS (Netherlands)

    Prins, M.W.J.; Jansen, R.; Groeneveld, R.H.M.; Gelder, Van A.P.; Kempen, van H.

    1996-01-01

    We describe a model as well as experiments on the electrical properties of a photoexcited tunnel junction between a metal and a semiconductor material, as is established in a scanning tunneling microscope. The model treats the case in which carrier transport is mediated by capture and relaxation in

  18. A Comparative Scanning Electron Microscopy Evaluation of Smear ...

    African Journals Online (AJOL)

    2018-02-07

    Feb 7, 2018 ... The aim of the present study was to compare the efficacy of chitosan and MTAD for the smear layer removal from the root canal through a scanning electron microscope (SEM). Thirty teeth were randomly divided into three groups according to the final irrigants: 0.2% chitosan, MTAD, saline (control group).

  19. Transmission/Scanning Transmission Electron Microscopy | Materials Science

    Science.gov (United States)

    crystallographic structure of a material. Amplitude-contrast images yield information about the chemistry and microstructure of a material and its defects. Phase-contrast imaging or high-resolution (HR) TEM imaging gives information about the microstructure of a material and its defects at an atomic resolution. With scanning

  20. A Comparative Scanning Electron Microscopy Evaluation of Smear ...

    African Journals Online (AJOL)

    The aim of the present study was to compare the efficacy of chitosan and MTAD for the smear layer removal from the root canal through a scanning electron microscope (SEM). Thirty teeth were randomly divided into three groups according to the final irrigants: 0.2% chitosan, MTAD, saline (control group). After the ...

  1. Photothermal modulation of the gap distance in scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Amer, N.M.; Skumanich, A.; Ripple, D.

    1986-01-01

    We have employed the photothermal effect to modulate the gap distance in a tunneling microscope. In this approach, optical heating induces the expansion and buckling of laser-illuminated sample surface. The surface displacement can be modulated over a wide frequency range, and its height (typically <1 A-circle) can be varied by changing the illumination intensity and modulation frequency. This novel method provides an alternative means for performing tunneling spectroscopy and microscopy, and for determining work functions of materials

  2. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    Science.gov (United States)

    Jesse, S.; Chi, M.; Belianinov, A.; Beekman, C.; Kalinin, S. V.; Borisevich, A. Y.; Lupini, A. R.

    2016-05-01

    Electron microscopy is undergoing a transition; from the model of producing only a few micrographs, through the current state where many images and spectra can be digitally recorded, to a new mode where very large volumes of data (movies, ptychographic and multi-dimensional series) can be rapidly obtained. Here, we discuss the application of so-called “big-data” methods to high dimensional microscopy data, using unsupervised multivariate statistical techniques, in order to explore salient image features in a specific example of BiFeO3 domains. Remarkably, k-means clustering reveals domain differentiation despite the fact that the algorithm is purely statistical in nature and does not require any prior information regarding the material, any coexisting phases, or any differentiating structures. While this is a somewhat trivial case, this example signifies the extraction of useful physical and structural information without any prior bias regarding the sample or the instrumental modality. Further interpretation of these types of results may still require human intervention. However, the open nature of this algorithm and its wide availability, enable broad collaborations and exploratory work necessary to enable efficient data analysis in electron microscopy.

  3. Effect of morphology and solvent on two-photon absorption of nano zinc oxide

    Energy Technology Data Exchange (ETDEWEB)

    Kavitha, M.K. [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India); Haripadmam, P.C.; Gopinath, Pramod; Krishnan, Bindu [Department of Physics, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India); John, Honey, E-mail: honey@iist.ac.in [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India)

    2013-05-15

    Highlights: ► ZnO nanospheres and triangular structures synthesis by novel precipitation technique. ► The effect of precursor concentration on the size and shape of nano ZnO. ► Open aperture Z-scan measurements of the ZnO nanoparticle dispersions. ► Nanospheres exhibit higher two photon absorption coefficient than triangular nanostructures. ► Nanospheres dispersed in water exhibit higher two photon absorption coefficient than its dispersion in 2-propanol. - Abstract: In this paper, we report the effect of morphology and solvent on the two-photon absorption of nano zinc oxide. Zinc oxide nanoparticles in two different morphologies like nanospheres and triangular nanostructures are synthesized by novel precipitation technique and their two-photon absorption coefficient is measured using open aperture Z-scan technique. Experimental results show that the zinc oxide nanospheres exhibit higher two-photon absorption coefficient than the zinc oxide triangular nanostructures. The zinc oxide nanospheres dispersed in water exhibit higher two-photon absorption coefficient than that of its dispersion in 2-propanol. The zinc oxide nanospheres dispersed in water shows a decrease in two-photon absorption coefficient with an increase in on-axis irradiance. The result confirms the dependence of shape and solvent on the two-photon absorption of nano zinc oxide.

  4. Atomic force microscopy and scanning electron microscopy analysis of daily disposable limbal ring contact lenses.

    Science.gov (United States)

    Lorenz, Kathrine Osborn; Kakkassery, Joseph; Boree, Danielle; Pinto, David

    2014-09-01

    Limbal ring (also known as 'circle') contact lenses are becoming increasingly popular, especially in Asian markets because of their eye-enhancing effects. The pigment particles that give the eye-enhancing effects of these lenses can be found on the front or back surface of the contact lens or 'enclosed' within the lens matrix. The purpose of this research was to evaluate the pigment location and surface roughness of seven types of 'circle' contact lenses. Scanning electron microscopic (SEM) analysis was performed using a variable pressure Hitachi S3400N instrument to discern the placement of lens pigments. Atomic force microscopy (Dimension Icon AFM from Bruker Nano) was used to determine the surface roughness of the pigmented regions of the contact lenses. Atomic force microscopic analysis was performed in fluid phase under contact mode using a Sharp Nitride Lever probe (SNL-10) with a spring constant of 0.06 N/m. Root mean square (RMS) roughness values were analysed using a generalised linear mixed model with a log-normal distribution. Least square means and their corresponding 95% confidence intervals were estimated for each brand, location and pigment combination. SEM cross-sectional images at 500× and 2,000× magnification showed pigment on the surface of six of the seven lens types tested. The mean depth of pigment for 1-DAY ACUVUE DEFINE (1DAD) lenses was 8.1 μm below the surface of the lens, while the remaining lens types tested had pigment particles on the front or back surface. Results of the atomic force microscopic analysis indicated that 1DAD lenses had significantly lower root mean square roughness values in the pigmented area of the lens than the other lens types tested. SEM and AFM analysis revealed pigment on the surface of the lens for all types tested with the exception of 1DAD. Further research is required to determine if the difference in pigment location influences on-eye performance. © 2014 The Authors. Clinical and Experimental

  5. Simulation study of secondary electron images in scanning ion microscopy

    CERN Document Server

    Ohya, K

    2003-01-01

    The target atomic number, Z sub 2 , dependence of secondary electron yield is simulated by applying a Monte Carlo code for 17 species of metals bombarded by Ga ions and electrons in order to study the contrast difference between scanning ion microscopes (SIM) and scanning electron microscopes (SEM). In addition to the remarkable reversal of the Z sub 2 dependence between the Ga ion and electron bombardment, a fine structure, which is correlated to the density of the conduction band electrons in the metal, is calculated for both. The brightness changes of the secondary electron images in SIM and SEM are simulated using Au and Al surfaces adjacent to each other. The results indicate that the image contrast in SIM is much more sensitive to the material species and is clearer than that for SEM. The origin of the difference between SIM and SEM comes from the difference in the lateral distribution of secondary electrons excited within the escape depth.

  6. Scanning tunneling microscopy: A powerful tool for surface analysis

    International Nuclear Information System (INIS)

    Walle, G.F.A. van de; Nelissen, B.J.; Soethout, L.L.; Kempen, H. van

    1987-01-01

    The invention of the Scanning Tunneling Microscope (STM) has opened a new area of surface analysis. A description of the principle of operation is given in this paper. Also the technical problems encountered and their solution are described. Two examples demonstrating the possibilities of the STM are presented: topographic and spectroscopic measurements on a stepped Ni (111) surface and photoconductive measurements on GaAs. (orig.)

  7. Two-photon superradiance in extended medium

    International Nuclear Information System (INIS)

    Branzan, V.; Enache, N.

    1993-01-01

    The possibility of collectivization of an ensemble of atoms of an extended system (the distance between atoms is larger or equal to the wave-length of a spontaneous emitted radiation) during two-photon spontaneous decay is theoretically investigated. It is demonstrated that such systems of inverted atoms should emit phase-correlated pairs of photons. The time-space correlation among atoms is realized due to the two-photon exchanging through the electromagnetic field's vacuum. An increase of the spontaneous decay rate of the two-atom inverted ensemble is demonstrated. The dependence of two-photon superradiance on the sample geometry is investigated. A non-equilibrium method of the elimination of the atoms level Fermi-operators is proposed. (Author)

  8. Molecular recognition of DNA-protein complexes: A straightforward method combining scanning force and fluorescence microscopy

    NARCIS (Netherlands)

    H. Sanchez (Humberto); R. Kanaar (Roland); C. Wyman (Claire)

    2010-01-01

    textabstractCombining scanning force and fluorescent microscopy allows simultaneous identification of labeled biomolecules and analysis of their nanometer level architectural arrangement. Fluorescent polystyrene nano-spheres were used as reliable objects for alignment of optical and topographic

  9. Quantitative detection of gold nanoparticles on individual, unstained cancer cells by Scanning Electron Microscopy

    NARCIS (Netherlands)

    Hartsuiker, Liesbeth; van Es, Peter; Petersen, Wilhelmina; van Leeuwen, Ton; Terstappen, Leonardus Wendelinus Mathias Marie; Otto, Cornelis

    2011-01-01

    Gold nanoparticles are rapidly emerging for use in biomedical applications. Characterization of the interaction and delivery of nanoparticles to cells through microscopy is important. Scanning electron microscopes have the intrinsic resolution to visualize gold nanoparticles on cells. A novel sample

  10. Quantitative detection of gold nanoparticles on individual, unstained cancer cells by scanning electron microscopy

    NARCIS (Netherlands)

    Hartsuiker, L.; van Es, P.; Petersen, W.; van Leeuwen, T. G.; Terstappen, L. W. M. M.; Otto, C.

    2011-01-01

    Gold nanoparticles are rapidly emerging for use in biomedical applications. Characterization of the interaction and delivery of nanoparticles to cells through microscopy is important. Scanning electron microscopes have the intrinsic resolution to visualize gold nanoparticles on cells. A novel sample

  11. Visualization of carbon nanotubes dispersion in composite by using confocal laser scanning microscopy

    Czech Academy of Sciences Publication Activity Database

    Ilčíková, M.; Danko, M.; Doroshenko, M.; Best, A.; Mrlík, M.; Csomorová, K.; Šlouf, Miroslav; Chorvát Jr., D.; Koynov, K.; Mosnáček, J.

    2016-01-01

    Roč. 79, June (2016), s. 187-197 ISSN 0014-3057 Institutional support: RVO:61389013 Keywords : confocal laser scanning microscopy * composites * carbon nanotubes dispersion Subject RIV: CD - Macromolecular Chemistry Impact factor: 3.531, year: 2016

  12. Formation of hot spots in a superconductor observed by low-temperature scanning electron microscopy

    International Nuclear Information System (INIS)

    Eichele, R.; Seifert, H.; Huebener, R.P.

    1981-01-01

    Low-temperature scanning electron microscopy can be used for the direct observation of hot spots in a superconductor. Experiments performed at 2.10 K with tim films demonstrating the method are reported

  13. Squeezing via two-photon transitions

    Science.gov (United States)

    Savage, C. M.; Walls, D. F.

    1986-05-01

    The squeezing spectrum for a cavity field mode interacting with an ensemble of three-level 'Lambda-configuration' atoms by an effective two-photon transition is calculated. The advantage of the three-level Lambda system as a squeezing medium, that is, optical nonlinearity without atomic saturation, has recently been pointed out by Reid, Walls, and Dalton. Perfect squeezing is predicted at the turning points for dispersive optical bistability and good squeezing for a range of other cases. Three-level ladder atoms interacting by an effective two-photon transition are also shown to give perfect squeezing in the dispersive limit.

  14. Two-photon excitation of argon

    International Nuclear Information System (INIS)

    Pindzola, P.S.; Payne, M.C.

    1982-01-01

    The authors calculate two photon excitation parameters for various excited states of argon assuming the absorption of near resonance broad-bandwidth laser radiation. Results are given for the case of two photons absorbed for the same laser beam as well as the case of absorbing photons of different frequency from each of two laser beams. The authors use multiconfiguration Hartree-Fock wave functions to evaluate the second-order sums over matrix elements. Various experimental laser schemes are suggested for the efficient excitation and subsequent ionization of argon

  15. A review of two photon physics

    International Nuclear Information System (INIS)

    Cooper, S.

    1982-08-01

    This talk is intended as an introduction for those not yet expert in two-photon physics, especially those e + e - one-photon physicists who still think of two-photon events as background. I concentrate on the physics questions involved, especially emphasizing the areas where I feel progress can be made in the near future, and of necessity leaving most experimental details to be found in the references. After a quick survey of the field and a few words about kinematics, I discuss in detail two major fields: the photon structure function and resonance production. (orig.)

  16. Vortex imaging in superconducting films by scanning Hall probe microscopy

    International Nuclear Information System (INIS)

    Oral, A.; Bending, S.J.; Humphreys, R.G.

    1996-01-01

    The authors have used a low noise Scanning Hall Probe Microscope (SHPM) to study vortex structures in superconducting films. The microscope has high magnetic field (∼2.9 x 10 -8 T/√Hz at 77K) and spatial resolution, ∼0.85 μm. Magnetic field profiles of single vortices in High T c YBa 2 Cu 3 O 7-δ thin films have been successfully measured and the microscopic penetration depth of the superconductor has been extracted as a function of temperature. Flux penetration into the superconductor has been imaged in real time (∼8s/frame)

  17. Scanning microwave microscopy applied to semiconducting GaAs structures

    Science.gov (United States)

    Buchter, Arne; Hoffmann, Johannes; Delvallée, Alexandra; Brinciotti, Enrico; Hapiuk, Dimitri; Licitra, Christophe; Louarn, Kevin; Arnoult, Alexandre; Almuneau, Guilhem; Piquemal, François; Zeier, Markus; Kienberger, Ferry

    2018-02-01

    A calibration algorithm based on one-port vector network analyzer (VNA) calibration for scanning microwave microscopes (SMMs) is presented and used to extract quantitative carrier densities from a semiconducting n-doped GaAs multilayer sample. This robust and versatile algorithm is instrument and frequency independent, as we demonstrate by analyzing experimental data from two different, cantilever- and tuning fork-based, microscope setups operating in a wide frequency range up to 27.5 GHz. To benchmark the SMM results, comparison with secondary ion mass spectrometry is undertaken. Furthermore, we show SMM data on a GaAs p-n junction distinguishing p- and n-doped layers.

  18. Apparent Barrier Height in Scanning Tunneling Microscopy Revisited

    DEFF Research Database (Denmark)

    Olesen, L.; Brandbyge, Mads; Sørensen, Mads Reinholdt

    1996-01-01

    The apparent barrier height phi(ap), that is, the rate of change of the logarithm of the conductance with tip-sample separation in a scanning tunneling microscope (STM), has been measured for Ni, Pt, and Au single crystal surfaces. The results show that phi(ap) is constant until point contact...... is reached rather than decreasing at small tunneling gap distances, as previously reported. The findings for phi(ap) can be accounted for theoretically by including the relaxations of the tip-surface junction in an STM due to the strong adhesive forces at close proximity. These relaxation effects are shown...

  19. Characterization of catalysts by scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Targos, W.M.; Bradley, S.A.

    1989-01-01

    The dedicated scanning transmission electron microscope (STEM) is an integral tool for characterizing catalysts because of its unique ability to image and analyze nanosized volumes. This information is valuable in optimizing catalyst formulations and determining causes for reduced catalyst performance. For many commercial catalysts direct correlations between structural features of metal crystallites and catalytic performance are not attainable. When these instances occur, determination of elemental distribution may be the only information available. In this paper the authors discuss some of the techniques employed and limitations associated with characterizing commercial catalysts

  20. Observation of diamond turned OFHC copper using Scanning Tunneling Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Grigg, D.A.; Russell, P.E.; Dow, T.A.

    1988-12-01

    Diamond turned OFHC copper samples have been observed within the past few months using the Scanning Tunneling Microscope. Initial results have shown evidence of artifacts which may be used to better understand the diamond turning process. The STM`s high resolution capability and three dimensional data representation allows observation and study of surface features unobtainable with conventional profilometry systems. Also, the STM offers a better quantitative means by which to analyze surface structures than the SEM. This paper discusses findings on several diamond turned OFHC copper samples having different cutting conditions. Each sample has been cross referenced using STM and SEM.

  1. [Scanning electron microscopy of heat-damaged bone tissue].

    Science.gov (United States)

    Harsanyl, L

    1977-02-01

    Parts of diaphyses of bones were exposed to high temperature of 200-1300 degrees C. Damage to the bone tissue caused by the heat was investigated. The scanning electron microscopic picture seems to be characteristic of the temperature applied. When the bones heated to the high temperature of 700 degrees C characteristic changes appear on the periostal surface, higher temperatura on the other hand causes damage to the compact bone tissue and can be observed on the fracture-surface. Author stresses the importance of this technique in the legal medicine and anthropology.

  2. Scanning Hall probe microscopy of a diluted magnetic semiconductor

    International Nuclear Information System (INIS)

    Kweon, Seongsoo; Samarth, Nitin; Lozanne, Alex de

    2009-01-01

    We have measured the micromagnetic properties of a diluted magnetic semiconductor as a function of temperature and applied field with a scanning Hall probe microscope built in our laboratory. The design philosophy for this microscope and some details are described. The samples analyzed in this work are Ga 0.94 Mn 0.06 As films grown by molecular beam epitaxy. We find that the magnetic domains are 2-4 μm wide and fairly stable with temperature. Magnetic clusters are observed above T C , which we ascribe to MnAs defects too small and sparse to be detected by a superconducting quantum interference device magnetometer.

  3. Scanning Hall probe microscopy of a diluted magnetic semiconductor

    Science.gov (United States)

    Kweon, Seongsoo; Samarth, Nitin; de Lozanne, Alex

    2009-05-01

    We have measured the micromagnetic properties of a diluted magnetic semiconductor as a function of temperature and applied field with a scanning Hall probe microscope built in our laboratory. The design philosophy for this microscope and some details are described. The samples analyzed in this work are Ga0.94Mn0.06As films grown by molecular beam epitaxy. We find that the magnetic domains are 2-4 μm wide and fairly stable with temperature. Magnetic clusters are observed above TC, which we ascribe to MnAs defects too small and sparse to be detected by a superconducting quantum interference device magnetometer.

  4. Near-field optical microscopy with a scanning tunneling microscope

    International Nuclear Information System (INIS)

    Barbara, A.; Lopez-Rios, T.; Quemerais, P.

    2005-01-01

    A homemade apertureless near-field optical microscope using a scanning tunneling microscope (STM) is described. The experimental set-up simultaneously provides optical and topographic images of the sample. Technical details and features of the set-up are presented, together with results demonstrating the sub-wavelength resolution achieved as well as its sensitivity to dielectric contrasts. We show that the use of a STM permits to precisely control very small distances between the tip and the sample which is a great advantage to excite localized optical resonances between the tip and the surface

  5. Precision controlled atomic resolution scanning transmission electron microscopy using spiral scan pathways

    Science.gov (United States)

    Sang, Xiahan; Lupini, Andrew R.; Ding, Jilai; Kalinin, Sergei V.; Jesse, Stephen; Unocic, Raymond R.

    2017-03-01

    Atomic-resolution imaging in an aberration-corrected scanning transmission electron microscope (STEM) can enable direct correlation between atomic structure and materials functionality. The fast and precise control of the STEM probe is, however, challenging because the true beam location deviates from the assigned location depending on the properties of the deflectors. To reduce these deviations, i.e. image distortions, we use spiral scanning paths, allowing precise control of a sub-Å sized electron probe within an aberration-corrected STEM. Although spiral scanning avoids the sudden changes in the beam location (fly-back distortion) present in conventional raster scans, it is not distortion-free. “Archimedean” spirals, with a constant angular frequency within each scan, are used to determine the characteristic response at different frequencies. We then show that such characteristic functions can be used to correct image distortions present in more complicated constant linear velocity spirals, where the frequency varies within each scan. Through the combined application of constant linear velocity scanning and beam path corrections, spiral scan images are shown to exhibit less scan distortion than conventional raster scan images. The methodology presented here will be useful for in situ STEM imaging at higher temporal resolution and for imaging beam sensitive materials.

  6. Dual-detection confocal fluorescence microscopy: fluorescence axial imaging without axial scanning.

    Science.gov (United States)

    Lee, Dong-Ryoung; Kim, Young-Duk; Gweon, Dae-Gab; Yoo, Hongki

    2013-07-29

    We propose a new method for high-speed, three-dimensional (3-D) fluorescence imaging, which we refer to as dual-detection confocal fluorescence microscopy (DDCFM). In contrast to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically over a sample volume to reconstruct a 3-D image, DDCFM can obtain the depth of a fluorescent emitter without depth scanning. DDCFM comprises two photodetectors, each with a pinhole of different size, in the confocal detection system. Axial information on fluorescent emitters can be measured by the axial response curve through the ratio of intensity signals. DDCFM can rapidly acquire a 3-D fluorescent image from a single two-dimensional scan with less phototoxicity and photobleaching than confocal fluorescence microscopy because no mechanical depth scans are needed. We demonstrated the feasibility of the proposed method by phantom studies.

  7. Graphene quantum dots probed by scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Morgenstern, Markus; Freitag, Nils; Nent, Alexander; Nemes-Incze, Peter; Liebmann, Marcus [II. Institute of Physics B and JARA-FIT, RWTH Aachen University, Aachen (Germany)

    2017-11-15

    Scanning tunneling spectroscopy results probing the electronic properties of graphene quantum dots are reviewed. After a short summary of the study of squared wave functions of graphene quantum dots on metal substrates, we firstly present data where the Landau level gaps caused by a perpendicular magnetic field are used to electrostatically confine electrons in monolayer graphene, which are probed by the Coulomb staircase revealing the consecutive charging of a quantum dot. It turns out that these quantum dots exhibit much more regular charging sequences than lithographically confined ones. Namely, the consistent grouping of charging peaks into quadruplets, both, in the electron and hole branch, portrays a regular orbital splitting of about 10meV. At low hole occupation numbers, the charging peaks are, partly, additionally grouped into doublets. The spatially varying energy separation of the doublets indicates a modulation of the valley splitting by the underlying BN substrate. We outline that this property might be used to eventually tune the valley splitting coherently. Afterwards, we describe graphene quantum dots with multiple contacts produced without lithographic resist, namely by local anodic oxidation. Such quantum dots target the goal to probe magnetotransport properties during the imaging of the corresponding wave functions by scanning tunneling spectroscopy. (copyright 2017 by WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  8. Automated rapid particle investigation using scanning electron microscopy

    Science.gov (United States)

    Wilkins, Jerod Laurence

    The chemical composition of fly ash particles has been known to vary significantly depending on a number of factors. Current bulk methods of investigation including X-Ray Fluorescence and X-Ray Diffraction are thought to be inadequate in determining the performance of fly ash in concrete. It is the goal of this research to develop a method of Automated Rapid Particle Investigation that will not look at fly ash as a bulk material but as individual particles. By examining each particle individually scientists and engineers will have the ability to study the variation in chemical composition by comparing the chemistry present in each particle. The method of investigation developed by this research provides a practical technique that will allow the automated chemical analysis of hundreds, or even thousands, of fly ash particles in a matter of minutes upon completion of sample preparation and automated scanning electron microscope (ASEM) scanning. This research does not examine the significance of the chemical compounds discovered; rather, only the investigation methodology is discussed. Further research will be done to examine the importance of the chemistry discovered with this automated rapid particle investigation technique.

  9. Analysis of Al2O3 Nanostructure Using Scanning Microscopy

    Directory of Open Access Journals (Sweden)

    Marek Kubica

    2018-01-01

    Full Text Available It has been reported that the size and shape of the pores depend on the structure of the base metal, the type of electrolyte, and the conditions of the anodizing process. The paper presents thin Al2O3 oxide layer formed under hard anodizing conditions on a plate made of EN AW-5251 aluminum alloy. The oxidation of the ceramic layer was carried out for 40–80 minutes in a three-component SAS electrolyte (aqueous solution of acids: sulphuric 33 ml/l, adipic 67 g/l, and oxalic 30 g/l at a temperature of 293–313 K, and the current density was 200–400 A/m2. Presented images were taken by a scanning microscope. A computer analysis of the binary images of layers showed different shapes of pores. The structure of ceramic Al2O3 layers is one of the main factors determining mechanical properties. The resistance to wear of specimen-oxide coating layer depends on porosity, morphology, and roughness of the ceramic layer surface. A 3D oxide coating model, based on the computer analysis of images from a scanning electron microscope (Philips XL 30 ESEM/EDAX, was proposed.

  10. Analysis of Al2O3 Nanostructure Using Scanning Microscopy

    Science.gov (United States)

    Kubica, Marek; Bara, Marek

    2018-01-01

    It has been reported that the size and shape of the pores depend on the structure of the base metal, the type of electrolyte, and the conditions of the anodizing process. The paper presents thin Al2O3 oxide layer formed under hard anodizing conditions on a plate made of EN AW-5251 aluminum alloy. The oxidation of the ceramic layer was carried out for 40–80 minutes in a three-component SAS electrolyte (aqueous solution of acids: sulphuric 33 ml/l, adipic 67 g/l, and oxalic 30 g/l) at a temperature of 293–313 K, and the current density was 200–400 A/m2. Presented images were taken by a scanning microscope. A computer analysis of the binary images of layers showed different shapes of pores. The structure of ceramic Al2O3 layers is one of the main factors determining mechanical properties. The resistance to wear of specimen-oxide coating layer depends on porosity, morphology, and roughness of the ceramic layer surface. A 3D oxide coating model, based on the computer analysis of images from a scanning electron microscope (Philips XL 30 ESEM/EDAX), was proposed. PMID:29861823

  11. Near-field scanning optical microscopy based nanostructuring of glass

    International Nuclear Information System (INIS)

    Chimmalgi, A; Hwang, D J; Grigoropoulos, C P

    2007-01-01

    Nanofabrication, at lateral resolutions beyond the capability of conventional optical lithography techniques, is demonstrated here. Femtosecond laser was used in conjunction with Near-field Scanning Optical Microscopes (NSOMs) to nanostructure thin metal films. Also, the possibility of using these nanostructured metal films as masks to effectively transfer the pattern to the underlying substrate by wet etching process is shown. Two different optical nearfiled processing schemes were studied for near-field nanostructuring. In the first scheme, local field enhancement in the near-field of a scanning probe microscope (SPM) probe tip irradiated with femtosecond laser pulses was utilized (apertureless NSOM mode) and as a second approach, femtosecond laser beam was spatially confined by cantilevered NSOM fiber tip (apertured NOSM mode). The minimized heat- and shock-affected areas introduced during ultrafast laser based machining process, allows processing of even high conductivity thin metal films with minimized formation of any interfacial compounds between the metal films and the underlying substrate. Potential applications of this method may be in the fields of nanolithography, nanofluidics, nanoscale chemical and gas sensors, high-density data storage, nano-opto-electronics, as well as biotechnology related applications

  12. Special raster scanning for reduction of charging effects in scanning electron microscopy.

    Science.gov (United States)

    Suzuki, Kazuhiko; Oho, Eisaku

    2014-01-01

    A special raster scanning (SRS) method for reduction of charging effects is developed for the field of SEM. Both a conventional fast scan (horizontal direction) and an unusual scan (vertical direction) are adopted for acquiring raw data consisting of many sub-images. These data are converted to a proper SEM image using digital image processing techniques. About sharpness of the image and reduction of charging effects, the SRS is compared with the conventional fast scan (with frame-averaging) and the conventional slow scan. Experimental results show the effectiveness of SRS images. By a successful combination of the proposed scanning method and low accelerating voltage (LV)-SEMs, it is expected that higher-quality SEM images can be more easily acquired by the considerable reduction of charging effects, while maintaining the resolution. © 2013 Wiley Periodicals, Inc.

  13. Local thermal conductivity of polycrystalline AlN ceramics measured by scanning thermal microscopy and complementary scanning electron microscopy techniques

    International Nuclear Information System (INIS)

    Zhang Yue-Fei; Wang Li; Wei Bin; Ji Yuan; Han Xiao-Dong; Zhang Ze; Heiderhoff, R.; Geinzer, A. K.; Balk, L. J.

    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 3ω method. A thermal conductivity of 308 W/m·K within grains 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. (condensed matter: structural, mechanical, and thermal properties)

  14. A nanoscale gigahertz source realized with Josephson scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Jäck, Berthold, E-mail: b.jaeck@fkf.mpg.de; Eltschka, Matthias; Assig, Maximilian; Etzkorn, Markus; Ast, Christian R. [Max-Planck-Institut für Festkörperforschung, 70569 Stuttgart (Germany); Hardock, Andreas [Institut für Theoretische Elektrotechnik, Technische Universität Hamburg-Harburg, 21079 Hamburg (Germany); Kern, Klaus [Max-Planck-Institut für Festkörperforschung, 70569 Stuttgart (Germany); Institut de Physique de la Matière Condensée, Ecole Polytechnique Fédérale de Lausanne, 1015 Lausanne (Switzerland)

    2015-01-05

    Using the AC Josephson effect in the superconductor-vacuum-superconductor tunnel junction of a scanning tunneling microscope (STM), we demonstrate the generation of GHz radiation. With the macroscopic STM tip acting as a λ/4-monopole antenna, we first show that the atomic scale Josephson junction in the STM is sensitive to its frequency-dependent environmental impedance in the GHz regime. Further, enhancing Cooper pair tunneling via excitations of the tip eigenmodes, we are able to generate high-frequency radiation. We find that for vanadium junctions, the enhanced photon emission can be tuned from about 25 GHz to 200 GHz and that large photon flux in excess of 10{sup 20 }cm{sup −2} s{sup −1} is reached in the tunnel junction. These findings demonstrate that the atomic scale Josephson junction in an STM can be employed as a full spectroscopic tool for GHz frequencies on the atomic scale.

  15. Analysis of artificial opals by scanning near field optical microscopy

    Science.gov (United States)

    Barrio, J.; Lozano, G.; Lamela, J.; Lifante, G.; Dorado, L. A.; Depine, R. A.; Jaque, F.; Míguez, H.

    2011-04-01

    Herein we present a detailed analysis of the optical response of artificial opal films realized employing a near-field scanning optical microscope in collection and transmission modes. Near-field patterns measured at the rear surface when a plane wave impinges on the front face are presented with the finding that optical intensity maps present a clear correlation with the periodic arrangement of the outer surface. Calculations based on the vector Korringa-Kohn-Rostoker method reproduce the different profiles experimentally observed as well as the response to the polarization of the incident field. These observations constitute the first experimental confirmation of the collective lattice resonances that give rise to the optical response of these three dimensional periodic structures in the high-energy range.

  16. Conditioning of mealybug (Hemiptera: Pseudococcidae) by Scanning Electron Microscopy

    International Nuclear Information System (INIS)

    Palma-Jimenez, Melissa; Blanco-Meneses, Monica

    2015-01-01

    The cleaning and correct observation of the mealybug specimens was determined by the conditioning methodology. The research was done in the Laboratorio del Centro de Investigacion en Estructuras Microscopicas (CIEMIC) of the Universidad de Costa Rica during the year 2012. A gradual improvement for the observation of the ultrastructures through the Scanning Electron Microscope was evidenced by the implementation of four types of methodologies. Each process was described in detail. The incorporation of 10% xylene (in some cases have been viable using ethanol at 95-100% ) was allowed to remove the wax from the body of the insect, to avoid this the collapse and to observe specific ultrastructures of the individual, they were the best results. The methodology used has reduced the time and costs in future taxonomic research of mealybug. (author) [es

  17. Optimization of the imaging response of scanning microwave microscopy measurements

    Energy Technology Data Exchange (ETDEWEB)

    Sardi, G. M.; Lucibello, A.; Proietti, E.; Marcelli, R., E-mail: romolo.marcelli@imm.cnr.it [National Research Council, Institute for Microelectronics and Microsystems, Via del Fosso del Cavaliere 100, 00133 Rome (Italy); Kasper, M.; Gramse, G. [Biophysics Institute, Johannes Kepler University, Gruberstrasse 40, 4020 Linz (Austria); Kienberger, F. [Keysight Technologies Austria GmbH, Gruberstrasse 40, 4020 Linz (Austria)

    2015-07-20

    In this work, we present the analytical modeling and preliminary experimental results for the choice of the optimal frequencies when performing amplitude and phase measurements with a scanning microwave microscope. In particular, the analysis is related to the reflection mode operation of the instrument, i.e., the acquisition of the complex reflection coefficient data, usually referred as S{sub 11}. The studied configuration is composed of an atomic force microscope with a microwave matched nanometric cantilever probe tip, connected by a λ/2 coaxial cable resonator to a vector network analyzer. The set-up is provided by Keysight Technologies. As a peculiar result, the optimal frequencies, where the maximum sensitivity is achieved, are different for the amplitude and for the phase signals. The analysis is focused on measurements of dielectric samples, like semiconductor devices, textile pieces, and biological specimens.

  18. Scanning electron microscopy of cells from periapical lesions.

    Science.gov (United States)

    Farber, P A

    1975-09-01

    Examination of lymphocytes from peripheral blood with the scanning electron microscope (SEM) has shown differences between B cells and T cells on the basis of their surface architecture. This study was initiated to determine whether the cellular components of periapical lesions could be identified with the use of similar criteria. Cells were dispersed from lesions by aspiration of fragments of tissue through syringe needles of decreasing diameters. The liberated cells were filtered on silver-coated Flotronic membranes and examined under the SEM. Lymphocytes, macrophages, epithelial cells, and mast cells were observed in granulomas and cysts. Most of the lymphocytes had smooth surfaces similar to that of T cells; others had villous projections similar to that of B cells. Epithelial nests were seen in the cyst linings while the cyst fluid was rich in lymphocytes. These findings suggest that SEM examination of periapical lesions can be a useful adjunct in studying cellular composition and possible immunological reactions in these tissues.

  19. Evaluation of the bleached human enamel by Scanning Electron Microscopy

    DEFF Research Database (Denmark)

    Miranda, Carolina Baptista; Pagani, Clovis; Benetti, Ana Raquel

    2005-01-01

    Since bleaching has become a popular procedure, the effect of peroxides on dental hard tissues is of great interest in research. Purpose: The aim of this in vitro study was to perform a qualitative analysis of the human enamel after the application of in-office bleaching agents, using Scanning......: 2h); G3- four 2-hour exposures to 35% carbamide peroxide (total exposure: 8h); G4- two applications of 35% hydrogen peroxide, which was light-activated with halogen lamp at 700mW/cm² during 7min and remained in contact with enamel for 20min (total exposure: 40min). All bleaching treatments adopted...... analysis performing gold sputter coating under vacuum and were examined using 15kV at 500x and 2000x magnification. Results: Morphological alterations on the enamel surface were similarly detected after bleaching with either 35% carbamide peroxide or 35% hydrogen peroxide. Surface porosities were...

  20. Aberration-corrected scanning transmission electron microscopy of semiconductors

    International Nuclear Information System (INIS)

    Krivanek, O L; Dellby, N; Murfitt, M F

    2011-01-01

    The scanning transmission electron microscope (STEM) has been able to image individual heavy atoms in a light matrix for some time. It is now able to do much more: it can resolve individual atoms as light as boron in monolayer materials; image atomic columns as light as hydrogen, identify the chemical type of individual isolated atoms from the intensity of their annular dark field (ADF) image and by electron energy loss spectroscopy (EELS); and map elemental composition at atomic resolution by EELS and energy-dispersive X-ray spectroscopy (EDXS). It can even map electronic states, also by EELS, at atomic resolution. The instrumentation developments that have made this level of performance possible are reviewed, and examples of applications to semiconductors and oxides are shown.

  1. Dopant profiling based on scanning electron and helium ion microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Chee, Augustus K.W., E-mail: kwac2@cam.ac.uk [Centre for Advanced Photonics and Electronics, Electrical Engineering Division, Department of Engineering, University of Cambridge, 9 JJ Thomson Avenue, Cambridge CB3 0FA (United Kingdom); Boden, Stuart A. [University of Southampton, Electronics and Computer Science, Highfield, Southampton SO17 1BJ (United Kingdom)

    2016-02-15

    In this paper, we evaluate and compare doping contrast generated inside the scanning electron microscope (SEM) and scanning helium ion microscope (SHIM). Specialised energy-filtering techniques are often required to produce strong doping contrast to map donor distributions using the secondary electron (SE) signal in the SEM. However, strong doping contrast can be obtained from n-type regions in the SHIM, even without energy-filtering. This SHIM technique is more sensitive than the SEM to donor density changes above its sensitivity threshold, i.e. of the order of 10{sup 16} or 10{sup 17} donors cm{sup −3} respectively on specimens with or without a p–n junction; its sensitivity limit is well above 2×10{sup 17} acceptors cm{sup −3} on specimens with or without a p–n junction. Good correlation is found between the widths and slopes of experimentally measured doping contrast profiles of thin p-layers and the calculated widths and slopes of the potential energy distributions across these layers, at a depth of 1 to 3 nm and 5 to 10 nm below the surface in the SHIM and the SEM respectively. This is consistent with the mean escape depth of SEs in silicon being about 1.8 nm and 7 nm in the SHIM and SEM respectively, and we conclude that short escape depth, low energy SE signals are most suitable for donor profiling. - Highlights: • Strong doping contrast from n-type regions in the SHIM without energy-filtering. • Sensitivity limits are established of the SHIM and SEM techniques. • We discuss the impact of SHIM imaging conditions on quantitative dopant profiling. • Doping contrast stems from different surface layer thicknesses in the SHIM and SEM.

  2. Review of Two-Photon Interactions

    International Nuclear Information System (INIS)

    Urner, David

    2004-01-01

    Presented are recent results of two-photon interactions. Topics include photon structure functions, inclusive hadron production, differential cross sections derived from tagged γγ fusion events and results in exclusive hadron production, particularly the observations of the η'c

  3. Two-photon transitions to exciton polaritons

    International Nuclear Information System (INIS)

    Hassan, A.R.

    1979-08-01

    A semiclassical theory for the creation of excitonic polariton states by two-photon absorption, via an intermediate exciton state, is given. A band model has been introduced which gives the dominant contribution to this process. A numerical calculation is found to be in good agreement with a recent observation in CuCl. (author)

  4. Pion-pair production by two photons

    International Nuclear Information System (INIS)

    Terazawa, Hidezumi.

    1994-07-01

    The cross section for pion-pair production by two photons is calculated approximately by using the low energy theorem previously derived from partially-conserved-axial-vector-current hypothesis and current algebra, and found to agree very well with the experimental data recently obtained by the Mark II, TPC/Two-Gamma and CLEO Collaborations. (author)

  5. Fluorescence in situ hybridization on human metaphase chromosomes detected by near-field scanning optical microscopy

    NARCIS (Netherlands)

    Moers, M.H.P.; Moers, M.H.P.; Kalle, W.H.J.; Kalle, W.H.J.; Ruiter, A.G.T.; Wiegant, J.C.A.G.; Raap, A.K.; Greve, Jan; de Grooth, B.G.; van Hulst, N.F.

    1996-01-01

    Fluorescence in situ hybridization o­n human metaphase chromosomes is detected by near-field scanning optical microscopy. This combination of cytochemical and scanning probe techniques enables the localization and identification of several fluorescently labelled genomic DNA fragments o­n a single

  6. 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.

  7. 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.

  8. Defect imaging and channeling studies using channeling scanning transmission ion microscopy

    NARCIS (Netherlands)

    King, PJC; Breese, MBH; Smulders, PJM; Wilshaw, PR; Grime, GW

    The technique of channeling scanning transmission ion microscopy (CSTIM) can be used to produce images of individual crystal defects (such as dislocations and stacking faults) using the scanned, focused ion beam from a nuclear microprobe. As well as offering a new method for studies of crystal

  9. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy. Rev. 1

    International Nuclear Information System (INIS)

    Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.; Schemer-Kohrn, Alan L.; Guzman, Anthony D.; Lavender, Curt A.

    2016-01-01

    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. Recent applications of scanning electron microscopy; Neueste Anwendungen der Rasterelektronenmikroskopie

    Energy Technology Data Exchange (ETDEWEB)

    Johansson, Sten; Moverare, Johan; Peng, Ru [Linkoeping Univ. (Sweden). Dept. of Management and Engineering

    2013-07-01

    A few examples were shown of how to use SEM to study phenomena that are not normally visible and possible to identify by introducing a known phenomenon called Electron Channeling. The channeling is best utilized in a FEG SEM not because of the in lens detection system but due to the fact that the highly coherent high electron density probe is creating a high contrast image with a resolution that is high enough to image crystal defects on a dislocation level. The fact that diffraction phenomena are involved in channeling is also of great importance for the contrast formation. The technique allows the user to choose to either just take a picture or decide if the image should be based on careful determination of the Bragg condition. The biggest advantage with channeling in addition the good contrast produced is the possibility to literally combine it with other techniques like EBSD. In fact, it is also possible to use thin foils to combine ECCI, EBSD, EDS and STEM in a modern FEG SEM. The development of a eucentric specimen stage of the same class as a TEM stage would allow even more advanced microscopy in SEM. (orig.)

  11. Atomic resolution scanning tunneling microscopy in a cryogen free dilution refrigerator at 15 mK

    International Nuclear Information System (INIS)

    Haan, A. M. J. den; Wijts, G. H. C. J.; Galli, F.; Oosterkamp, T. H.; Usenko, O.; Baarle, G. J. C. van; Zalm, D. J. van der

    2014-01-01

    Pulse tube refrigerators are becoming more common, because they are cost efficient and demand less handling than conventional (wet) refrigerators. However, a downside of a pulse tube system is the vibration level at the cold-head, which is in most designs several micrometers. We implemented vibration isolation techniques which significantly reduced vibration levels at the experiment. These optimizations were necessary for the vibration sensitive magnetic resonance force microscopy experiments at milli-kelvin temperatures for which the cryostat is intended. With these modifications we show atomic resolution scanning tunneling microscopy on graphite. This is promising for scanning probe microscopy applications at very low temperatures

  12. Magnetic imaging of unconventional superconductors by scanning SQUID microscopy

    International Nuclear Information System (INIS)

    Hykel, D.

    2011-01-01

    We present the development of a scanning SQUID/AFM microscope and measurements performed on different samples. The microscope can take topographic and magnetic images simultaneously. The magnetic resolution is of the order of 10 -4 Φ 0 √Hz and the spatial resolution of the SQUIDs used in this thesis goes up to 600 nm. The scanning range is 70 μm * 85 μm. The temperature range accessible is between 200 mK and 10 K at the time of writing. Measurements on a thin rhenium film (80 nm) give an estimate of the minimal pinning force of a vortex of about 3.9 * 10 -16 N. Furthermore, the penetration depth λ on this sample was determined as a function of temperature. For T → 0, λ →79 nm. We have for the first time shown local measurements of the domain structure of the superconducting ferromagnet UCoGe and determined the average domain size in the virgin state (10 μm). By magnetic imaging we were capable of determining the magnetic field difference above opposite domains along the c-axis to be 45 G and 16 G along the b-axis. Due to these magnetic field measurements we were able to give an upper limit for the domain wall width (∼ 1μm) and domain reconstruction depth (100 nm). This is supported by simple calculations leading to a domain wall width of several angstroms. Thus UCoGe can be considered an ideal Ising ferromagnet. Different possible domain structures for an Ising ferromagnet have been discussed. The complicated domain structure found in the zero field cooled virgin state corresponds to up domains embedded in larger down domains and vice versa. We have shown evidence for coexistence of superconductivity and ferromagnetism. The weak Meissner effect can be explained by a spontaneous vortex state, put forward by other groups. Numerical simulations suggest that the strong magnetic background signal and the limited spatial and magnetic resolution of the used SQUID made it difficult to resolve the expected spontaneous vortex state. The relaxation of the

  13. Building Practical Apertureless Scanning Near-Field Microscopy

    Science.gov (United States)

    Gungordu, M. Zeki

    The fundamental objective of this study is to establish a functional, practical apertureless type scanning near-field optical microscope, and to figure out the working mechanism behind it. Whereas a far-field microscope can measure the propagating field's components, this gives us little information about the features of the sample. The resolution is limited to about half of the wavelength of the illuminating light. On the other hand, the a-SNOM system enables achieving non-propagating components of the field, which provides more details about the sample's features. It is really difficult to measure because the amplitude of this field decays exponentially when the tip is moved away from the sample. The sharpness of the tip is the only limitation for resolution of the a-SNOM system. Consequently, the sharp tips are achieved by using electrochemical etching, and these tips are used to detect near-field signal. Separating the weak a-SNOM system signals from the undesired background signal, the higher demodulation background suppression is utilized by lock-in detection.

  14. Scanning transmission electron microscopy: Albert Crewe's vision and beyond

    International Nuclear Information System (INIS)

    Krivanek, Ondrej L.; Chisholm, Matthew F.; Murfitt, Matthew F.; Dellby, Niklas

    2012-01-01

    Some four decades were needed to catch up with the vision that Albert Crewe and his group had for the scanning transmission electron microscope (STEM) in the nineteen sixties and seventies: attaining 0.5 Å resolution, and identifying single atoms spectroscopically. With these goals now attained, STEM developments are turning toward new directions, such as rapid atomic resolution imaging and exploring atomic bonding and electronic properties of samples at atomic resolution. The accomplishments and the future challenges are reviewed and illustrated with practical examples. -- Highlights: ► TV-rate STEM imaging of heavy atoms is demonstrated. ► DNA sequencing by STEM dark field imaging should be possible at a rate of 10 6 bases/s. ► Individual silicon atom impurities in graphene are imaged atom-by-atom. ► Single atoms of nitrogen and boron incorporated in graphene are imaged spectroscopically. ► Bonding of individual atoms can be probed by analyzing the fine structures of their EEL spectra.

  15. Broadband two-photon absorption cross sections of benzothiazole derivatives and benzobisthiazolium salts

    Science.gov (United States)

    Noskovičova, Eva; Lorenc, Dušan; Magdolen, Peter; Sigmundová, Ivica; Zahradník, Pavol; Velič, Dušan

    2018-05-01

    Two-photon absorption (TPA) cross sections of conjugated donor-π-acceptor dipolar structures containing benzothiazole or benzobisthiazolium moieties are determined in a broad spectral range from 700 nm to 1000 nm using two-photon induced fluorescence technique. The TPA cross section values range from 150 GM to 4600 GM. The largest values are observed in near-infrared region. The dipolar derivative of benzothiazole has the largest TPA cross section of 4600 GM at wavelength of 890 nm. A combination of the large TPA in the near-infrared region and the high emission quantum yield makes these compounds excellent candidates for two-photon fluorescence microscopy.

  16. Enhancing dynamic scanning force microscopy in air: as close as possible

    International Nuclear Information System (INIS)

    Palacios-Lidon, E; Perez-Garcia, B; Colchero, J

    2009-01-01

    Frequency modulation dynamic scanning force microscopy has been implemented in ambient conditions using low oscillation amplitudes (<1 nm) to simultaneously record not only topographic but also additional channels of information, in particular contact potential images. The performance of this mode as compared to the conventional amplitude modulation mode is analyzed in detail using a biological molecule, turning yellow mosaic virus RNA, as the model sample. On the basis of scanning force microscopy imaging as well as spectroscopy experiments, we find that for such very small samples the frequency modulation mode is superior since it can be operated with smaller tip-sample interaction, smaller effective tip-sample distance and lower forces. Combined with Kelvin probe microscopy it results not only in considerably higher electrostatic resolution, but also in correct quantitative values for the contact potential as compared to traditional amplitude modulation scanning force microscopy.

  17. Generation of laser-induced periodic surface structures in indium-tin-oxide thin films and two-photon lithography of ma-N photoresist by sub-15 femtosecond laser microscopy for liquid crystal cell application

    Science.gov (United States)

    Klötzer, Madlen; Afshar, Maziar; Feili, Dara; Seidel, Helmut; König, Karsten; Straub, Martin

    2015-03-01

    Indium-tin-oxide (ITO) is a widely used electrode material for liquid crystal cell applications because of its transparency in the visible spectral range and its high electrical conductivity. Important examples of applications are displays and optical phase modulators. We report on subwavelength periodic structuring and precise laser cutting of 150 nm thick indium-tin-oxide films on glass substrates, which were deposited by magnetron reactive DC-sputtering from an indiumtin target in a low-pressure oxygen atmosphere. In order to obtain nanostructured electrodes laser-induced periodic surface structures with a period of approximately 100 nm were generated using tightly focused high-repetition rate sub-15 femtosecond pulsed Ti:sapphire laser light, which was scanned across the sample by galvanometric mirrors. Three-dimensional spacers were produced by multiphoton photopolymerization in ma-N 2410 negative-tone photoresist spin-coated on top of the ITO layers. The nanostructured electrodes were aligned in parallel to set up an electrically switchable nematic liquid crystal cell.

  18. Characterization of gold nanoparticle films: Rutherford backscattering spectroscopy, scanning electron microscopy with image analysis, and atomic force microscopy

    Directory of Open Access Journals (Sweden)

    Pia C. Lansåker

    2014-10-01

    Full Text Available Gold nanoparticle films are of interest in several branches of science and technology, and accurate sample characterization is needed but technically demanding. We prepared such films by DC magnetron sputtering and recorded their mass thickness by Rutherford backscattering spectroscopy. The geometric thickness dg—from the substrate to the tops of the nanoparticles—was obtained by scanning electron microscopy (SEM combined with image analysis as well as by atomic force microscopy (AFM. The various techniques yielded an internally consistent characterization of the films. In particular, very similar results for dg were obtained by SEM with image analysis and by AFM.

  19. Observation of self-assembled fluorescent beads by scanning near-field optical microscopy and atomic force microscopy

    International Nuclear Information System (INIS)

    Oh, Y.J.; Jo, W.; Kim, Min-Gon; Kyu Park, Hyun; Hyun Chung, Bong

    2006-01-01

    Optical response and topography of fluorescent latex beads both on flat self-assembled monolayer and on a micron-patterned surface with poly(dimethylsiloxane) are studied. Scanning near-field optical microscopy and atomic force microscopy were utilized together for detecting fluorescence and imaging topography of the patterned latex beads, respectively. As a result, the micro-patterned latex beads where a specific chemical binding occurred show a strong signal, whereas no signals are observed in the case of nonspecific binding. With fluorescein isothiocyanate (FITC), it is convenient to measure fluorescence signal from the patterned beads allowing us to monitor the small balls of fluorescent latex

  20. Cryo-Scanning Electron Microscopy (SEM) and Scanning Transmission Electron Microscopy (STEM)-in-SEM for Bio- and Organo-Mineral Interface Characterization in the Environment.

    Science.gov (United States)

    Wille, Guillaume; Hellal, Jennifer; Ollivier, Patrick; Richard, Annie; Burel, Agnes; Jolly, Louis; Crampon, Marc; Michel, Caroline

    2017-12-01

    Understanding biofilm interactions with surrounding substratum and pollutants/particles can benefit from the application of existing microscopy tools. Using the example of biofilm interactions with zero-valent iron nanoparticles (nZVI), this study aims to apply various approaches in biofilm preparation and labeling for fluorescent or electron microscopy and energy dispersive X-ray spectrometry (EDS) microanalysis for accurate observations. According to the targeted microscopy method, biofilms were sampled as flocs or attached biofilm, submitted to labeling using 4',6-diamidino-2-phenylindol, lectins PNA and ConA coupled to fluorescent dye or gold nanoparticles, and prepared for observation (fixation, cross-section, freezing, ultramicrotomy). Fluorescent microscopy revealed that nZVI were embedded in the biofilm structure as aggregates but the resolution was insufficient to observe individual nZVI. Cryo-scanning electron microscopy (SEM) observations showed nZVI aggregates close to bacteria, but it was not possible to confirm direct interactions between nZVI and cell membranes. Scanning transmission electron microscopy in the SEM (STEM-in-SEM) showed that nZVI aggregates could enter the biofilm to a depth of 7-11 µm. Bacteria were surrounded by a ring of extracellular polymeric substances (EPS) preventing direct nZVI/membrane interactions. STEM/EDS mapping revealed a co-localization of nZVI aggregates with lectins suggesting a potential role of EPS in nZVI embedding. Thus, the combination of divergent microscopy approaches is a good approach to better understand and characterize biofilm/metal interactions.

  1. In vivo two-photon imaging of retina in rabbits and rats.

    Science.gov (United States)

    Jayabalan, Gopal Swamy; Wu, Yi-Kai; Bille, Josef F; Kim, Samuel; Mao, Xiao Wen; Gimbel, Howard V; Rauser, Michael E; Fan, Joseph T

    2018-01-01

    The purpose of this study was to evaluate the retina using near-infrared (NIR) two-photon scanning laser ophthalmoscopy. New Zealand white rabbits, albino rats, and brown Norway rats were used in this study. An autofluorescence image of the retina, including the retinal cells and its associated vasculatures was obtained by a real-time scan using the ophthalmoscope. Furthermore, the retinal vessels, nerve fiber layers and the non-pigmented retina were recorded with two-photon fluorescein angiography (FA); and the choroidal vasculatures were recorded using two-photon indocyanine green angiography (ICGA). Two-photon ICGA was achieved by exciting a second singlet state at ∼398 nm. Simultaneous two-photon FA and two-photon ICGA were performed to characterize the retinal and choroidal vessels with a single injection. The minimum laser power threshold required to elicit two-photon fluorescence was determined. The two-photon ophthalmoscope could serve as a promising tool to detect and monitor the disease progression in animal models. Moreover, these high-resolution images of retinal and choroidal vessels can be acquired in a real-time scan with a single light source, requiring no additional filters for FA or ICGA. The combination of FA and ICGA using the two-photon ophthalmoscope will help researchers to characterize the retinal diseases in animal models, and also to classify the types (classic, occult or mixed) of choroidal neovascularization (CNV) in macular degeneration. Furthermore, the prototype can be adapted to image the retina of rodents and rabbits. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Clinical applications of in vivo fluorescence confocal laser scanning microscopy

    Science.gov (United States)

    Oh, Chilhwan; Park, Sangyong; Kim, Junhyung; Ha, Seunghan; Park, Gyuman; Lee, Gunwoo; Lee, Onseok; Chun, Byungseon; Gweon, Daegab

    2008-02-01

    Living skin for basic and clinical research can be evaluated by Confocal Laser Scanning Microscope (CLSM) non-invasively. CLSM imaging system can achieve skin image its native state either "in vivo" or "fresh biopsy (ex vivo)" without fixation, sectioning and staining that is necessary for routine histology. This study examines the potential fluorescent CLSM with a various exogenous fluorescent contrast agent, to provide with more resolution images in skin. In addition, in vivo fluorescent CLSM researchers will be extended a range of potential clinical application. The prototype of our CLSM system has been developed by Prof. Gweon's group. The operating parameters are composed of some units, such as illuminated wavelength 488 nm, argon illumination power up to 20mW on the skin, objective lens, 0.9NA oil immersion, axial resolution 1.0μm, field of view 200μm x 100μm (lateral resolution , 0.3μm). In human volunteer, fluorescein sodium was administrated topically and intradermally. Animal studies were done in GFP transgenic mouse, IRC mouse and pig skin. For imaging of animal skin, fluorescein sodium, acridine orange, and curcumine were used for fluorescein contrast agent. We also used the GFP transgenic mouse for fluorescein CLSM imaging. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. Acridin Orange can be highlight nuclei in viable keratinocyte. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, hair and eccrine gland. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. In

  3. Can Two-Photon Interference be Considered the Interference of Two Photons?

    International Nuclear Information System (INIS)

    Pittman, T.B.; Strekalov, D.V.; Migdall, A.; Rubin, M.H.; Sergienko, A.V.; Shih, Y.H.

    1996-01-01

    We report on a open-quote open-quote postponed compensation close-quote close-quote experiment in which the observed two-photon entangled state interference cannot be pictured in terms of the overlap of the two individual photon wave packets of a parametric down-conversion pair on a beam splitter. In the sense of a quantum eraser, the distinguishability of the different two-photon Feynman amplitudes leading to a coincidence detection is removed by delaying the compensation until after the output of an unbalanced two-photon interferometer. copyright 1996 The American Physical Society

  4. Circular mode: a new scanning probe microscopy method for investigating surface properties at constant and continuous scanning velocities.

    Science.gov (United States)

    Nasrallah, Hussein; Mazeran, Pierre-Emmanuel; Noël, Olivier

    2011-11-01

    In this paper, we introduce a novel scanning probe microscopy mode, called the circular mode, which offers expanded capabilities for surface investigations especially for measuring physical properties that require high scanning velocities and/or continuous displacement with no rest periods. To achieve these specific conditions, we have implemented a circular horizontal displacement of the probe relative to the sample plane. Thus the relative probe displacement follows a circular path rather than the conventional back and forth linear one. The circular mode offers advantages such as high and constant scanning velocities, the possibility to be combined with other classical operating modes, and a simpler calibration method of the actuators generating the relative displacement. As application examples of this mode, we report its ability to (1) investigate the influence of scanning velocity on adhesion forces, (2) measure easily and instantly the friction coefficient, and (3) generate wear tracks very rapidly for tribological investigations. © 2011 American Institute of Physics

  5. Two-photon absorption of a supramolecular pseudoisocyanine J-aggregate assembly

    International Nuclear Information System (INIS)

    Belfield, Kevin D.; Bondar, Mykhailo V.; Hernandez, Florencio E.; Przhonska, Olga V.; Yao, Sheng

    2006-01-01

    Linear spectral properties, including excitation anisotropy, of pseudoisocyanine or 1,1'-diethyl-2,2'-cyanine iodide (PIC) J-aggregates in aqueous solutions with J-band position at 573 nm were investigated. Two-photon absorption of PIC J-aggregates and monomer molecules was studied using an open aperture Z-scan technique. A strong enhancement of the two-photon absorption cross-section of PIC in the supramolecular J-aggregate assembly was observed in aqueous solution. This enhancement is attributed to a strong coupling of the molecular transition dipoles. No two-photon absorption at the peak of the J-band was detected

  6. Scanning electron microscopy and transmission electron microscopy study of hot-deformed gamma-TiAl-based alloy microstructure.

    Science.gov (United States)

    Chrapoński, J; Rodak, K

    2006-09-01

    The aim of this work was to assess the changes in the microstructure of hot-deformed specimens made of alloys containing 46-50 at.% Al, 2 at.% Cr and 2 at.% Nb (and alloying additions such as carbon and boron) with the aid of scanning electron microscopy and transmission electron microscopy techniques. After homogenization and heat treatment performed in order to make diverse lamellae thickness, the specimens were compressed at 1000 degrees C. Transmission electron microscopy examinations of specimens after the compression test revealed the presence of heavily deformed areas with a high density of dislocation. Deformation twins were also observed. Dynamically recrystallized grains were revealed. For alloys no. 2 and no. 3, the recovery and recrystallization processes were more extensive than for alloy no. 1.

  7. Probing two-photon exchange with OLYMPUS

    International Nuclear Information System (INIS)

    Kohl, M.

    2014-01-01

    Two-photon exchange is believed to be responsible for the discrepancies in the proton electric to magnetic form factor ratio found with the Rosenbluth and polarization transfer methods. If this explanation is correct, one expects significant differences in the lepton-proton cross sections between positrons and electrons. The OLYMPUS experiment at DESY in Hamburg, Germany was designed to measure the ratio of unpolarized positron-proton and electron-proton elastic scattering cross sections over a wide kinematic range with high precision, in order to quantify the effect of two-photon exchange. The experiment used intense beams of electrons and positrons stored in the DORIS ring at 2.0 GeV interacting with an internal windowless hydrogen gas target. The current status of OLYMPUS will be discussed. (authors)

  8. Resonant Scanning with Large Field of View Reduces Photobleaching and Enhances Fluorescence Yield in STED Microscopy.

    Science.gov (United States)

    Wu, Yong; Wu, Xundong; Lu, Rong; Zhang, Jin; Toro, Ligia; Stefani, Enrico

    2015-10-01

    Photobleaching is a major limitation of superresolution Stimulated Depletion Emission (STED) microscopy. Fast scanning has long been considered an effective means to reduce photobleaching in fluorescence microscopy, but a careful quantitative study of this issue is missing. In this paper, we show that the photobleaching rate in STED microscopy can be slowed down and the fluorescence yield be enhanced by scanning with high speed, enabled by using large field of view in a custom-built resonant-scanning STED microscope. The effect of scanning speed on photobleaching and fluorescence yield is more remarkable at higher levels of depletion laser irradiance, and virtually disappears in conventional confocal microscopy. With ≥6 GW∙cm(-2) depletion irradiance, we were able to extend the fluorophore survival time of Atto 647N and Abberior STAR 635P by ~80% with 8-fold wider field of view. We confirm that STED Photobleaching is primarily caused by the depletion light acting upon the excited fluorophores. Experimental data agree with a theoretical model. Our results encourage further increasing the linear scanning speed for photobleaching reduction in STED microscopy.

  9. Symmetry adaptation in two-photon spectroscopy

    International Nuclear Information System (INIS)

    Kibler, M.

    1991-11-01

    Symmetry adaptation techniques are applied to the determination of the intensity of two-photon transitions for transition ions in finite symmetry environments. The case of intra-configurational transitions are discussed with some details and some results on inter-configurational transitions are briefly reported. In particular, for intra-configurational transitions, a model is described which takes into account the following ingredients: (symmetry, second- plus third-order mechanisms, S-, L- and J-mixings). (author) 20 refs

  10. Two-photon spin generation and detection

    Energy Technology Data Exchange (ETDEWEB)

    Miah, M Idrish, E-mail: m.miah@griffith.edu.a [Nanoscale Science and Technology Centre, Griffith University, Nathan, Brisbane, QLD 4111 (Australia)

    2009-02-21

    A time- and polarization-resolved two-photon pump-probe investigation is performed in lightly doped GaAs. We generate spin-polarized electrons in bulk GaAs at various temperatures using right-circularly polarized two-photon excitation and detect them by probing the spin-dependent transmission of the sample. The spin polarization (P) of conduction band electrons, as measured using probe pulses with the same (right) and opposite (left) circular polarization, is measured in dependences of pump-probe delay ({Delta}t), lattice temperature (T{sub L}), doping density (n) as well as of the excess photon energy {Delta}E{sub 2{omega}}= {h_bar}2{omega} - E{sub g}, where E{sub g} is the band gap energy. P is found to be decayed with {Delta}t and enhanced with the decrease in T{sub L} or the increase in n. It is also found that P decreases with the increase in {Delta}E{sub 2{omega}}and depolarizes rapidly for {Delta}E{sub 2{omega}}> {Delta}E{sub SO}, where {Delta}E{sub SO} is the spin-orbit splitting energy. The results demonstrate that due to a much longer absorption depth highly polarized spins can be generated optically by two-photon pumping of bulk semiconductors.

  11. Two-photon spin generation and detection

    International Nuclear Information System (INIS)

    Miah, M Idrish

    2009-01-01

    A time- and polarization-resolved two-photon pump-probe investigation is performed in lightly doped GaAs. We generate spin-polarized electrons in bulk GaAs at various temperatures using right-circularly polarized two-photon excitation and detect them by probing the spin-dependent transmission of the sample. The spin polarization (P) of conduction band electrons, as measured using probe pulses with the same (right) and opposite (left) circular polarization, is measured in dependences of pump-probe delay (Δt), lattice temperature (T L ), doping density (n) as well as of the excess photon energy ΔE 2ω = ℎ2ω - E g , where E g is the band gap energy. P is found to be decayed with Δt and enhanced with the decrease in T L or the increase in n. It is also found that P decreases with the increase in ΔE 2ω and depolarizes rapidly for ΔE 2ω > ΔE SO , where ΔE SO is the spin-orbit splitting energy. The results demonstrate that due to a much longer absorption depth highly polarized spins can be generated optically by two-photon pumping of bulk semiconductors.

  12. Combined Raman and continuous-wave-excited two-photon fluorescence cell imaging

    NARCIS (Netherlands)

    Uzunbajakava, N.; Otto, Cornelis

    2003-01-01

    We demonstrate a confocal optical microscope that combines cw two-photon-excited fluorescence microscopy with confocal Raman microscopy. With this microscope fast image acquisition with fluorescence imaging can be used to select areas of interest for subsequent chemical analysis with spontaneous

  13. Emulation and design of terahertz reflection-mode confocal scanning microscopy based on virtual pinhole

    Science.gov (United States)

    Yang, Yong-fa; Li, Qi

    2014-12-01

    In the practical application of terahertz reflection-mode confocal scanning microscopy, the size of detector pinhole is an important factor that determines the performance of spatial resolution characteristic of the microscopic system. However, the use of physical pinhole brings some inconvenience to the experiment and the adjustment error has a great influence on the experiment result. Through reasonably selecting the parameter of matrix detector virtual pinhole (VPH), it can efficiently approximate the physical pinhole. By using this approach, the difficulty of experimental calibration is reduced significantly. In this article, an imaging scheme of terahertz reflection-mode confocal scanning microscopy that is based on the matrix detector VPH is put forward. The influence of detector pinhole size on the axial resolution of confocal scanning microscopy is emulated and analyzed. Then, the parameter of VPH is emulated when the best axial imaging performance is reached.

  14. Scanning ion conductance microscopy for visualizing the three-dimensional surface topography of cells and tissues.

    Science.gov (United States)

    Nakajima, Masato; Mizutani, Yusuke; Iwata, Futoshi; Ushiki, Tatsuo

    2018-01-01

    Scanning ion conductance microscopy (SICM), which belongs to the family of scanning probe microscopy, regulates the tip-sample distance by monitoring the ion current through the use of an electrolyte-filled nanopipette as the probing tip. Thus, SICM enables "contact-free" imaging of cell surface topography in liquid conditions. In this paper, we applied hopping mode SICM for obtaining topographical images of convoluted tissue samples such as trachea and kidney in phosphate buffered saline. Some of the SICM images were compared with the images obtained by scanning electron microscopy (SEM) after drying the same samples. We showed that the imaging quality of hopping mode SICM was excellent enough for investigating the three-dimensional surface structure of the soft tissue samples. Thus, SICM is expected to be used for imaging a wide variety of cells and tissues - either fixed or alive- at high resolution under physiologically relevant liquid conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Hyaline articular cartilage dissected by papain: light and scanning electron microscopy and micromechanical studies.

    OpenAIRE

    O'Connor, P; Brereton, J D; Gardner, D L

    1984-01-01

    Papain was used to digest the hyaline femoral condylar cartilages of 30 adult Wistar rats. Matrix proteoglycan degradation was assessed by the light microscopy of paraffin sections stained with toluidine blue. The extent of surface structural change was estimated by scanning electron microscopy, and the structural integrity of the hyaline cartilage tested by the controlled impact of a sharp pin. The results demonstrated an early loss of cartilage metachromasia, increasing with time of papain ...

  16. Single-step electrochemical method for producing very sharp Au scanning tunneling microscopy tips

    International Nuclear Information System (INIS)

    Gingery, David; Buehlmann, Philippe

    2007-01-01

    A single-step electrochemical method for making sharp gold scanning tunneling microscopy tips is described. 3.0M NaCl in 1% perchloric acid is compared to several previously reported etchants. The addition of perchloric acid to sodium chloride solutions drastically shortens etching times and is shown by transmission electron microscopy to produce very sharp tips with a mean radius of curvature of 15 nm

  17. Scanning tunneling microscopy and atomic force microscopy: application to biology and technology.

    Science.gov (United States)

    Hansma, P K; Elings, V B; Marti, O; Bracker, C E

    1988-10-14

    The scanning tunneling microscope (STM) and the atomic force microscope (AFM) are scanning probe microscopes capable of resolving surface detail down to the atomic level. The potential of these microscopes for revealing subtle details of structure is illustrated by atomic resolution images including graphite, an organic conductor, an insulating layered compound, and individual adsorbed oxygen atoms on a semiconductor. Application of the STM for imaging biological materials directly has been hampered by the poor electron conductivity of most biological samples. The use of thin conductive metal coatings and replicas has made it possible to image some biological samples, as indicated by recently obtained images of a recA-DNA complex, a phospholipid bilayer, and an enzyme crystal. The potential of the AFM, which does not require a conductive sample, is shown with molecular resolution images of a nonconducting organic monolayer and an amino acid crystal that reveals individual methyl groups on the ends of the amino acids. Applications of these new microscopes to technology are demonstrated with images of an optical disk stamper, a diffraction grating, a thin-film magnetic recording head, and a diamond cutting tool. The STM has even been used to improve the quality of diffraction gratings and magnetic recording heads.

  18. Cement paste surface roughness analysis using coherence scanning interferometry and confocal microscopy

    International Nuclear Information System (INIS)

    Apedo, K.L.; Munzer, C.; He, H.; Montgomery, P.; Serres, N.; Fond, C.; Feugeas, F.

    2015-01-01

    Scanning electron microscopy and scanning probe microscopy have been used for several decades to better understand the microstructure of cementitious materials. Very limited work has been performed to date to study the roughness of cementitious materials by optical microscopy such as coherence scanning interferometry (CSI) and chromatic confocal sensing (CCS). The objective of this paper is to better understand how CSI can be used as a tool to analyze surface roughness and topography of cement pastes. Observations from a series of images acquired using this technique on both polished and unpolished samples are described. The results from CSI are compared with those from a STIL confocal microscopy technique (SCM). Comparison between both optical techniques demonstrates the ability of CSI to measure both polished and unpolished cement pastes. - Highlights: • Coherence scanning interferometry (CSI) was used to analyze cement paste surfaces. • The results from the CSI were compared with those from a confocal microscopy. • 3D roughness parameters were obtained using the window resizing method. • Polished and unpolished cement pastes were studied

  19. Cement paste surface roughness analysis using coherence scanning interferometry and confocal microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Apedo, K.L., E-mail: apedo@unistra.fr [ICube, Université de Strasbourg, CNRS, 2 rue Boussingault, 67000 Strasbourg (France); Munzer, C.; He, H. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France); Montgomery, P. [ICube, Université de Strasbourg, CNRS, 23 rue du Loess, 67037 Strasbourg (France); Serres, N. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France); Fond, C. [ICube, Université de Strasbourg, CNRS, 2 rue Boussingault, 67000 Strasbourg (France); Feugeas, F. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France)

    2015-02-15

    Scanning electron microscopy and scanning probe microscopy have been used for several decades to better understand the microstructure of cementitious materials. Very limited work has been performed to date to study the roughness of cementitious materials by optical microscopy such as coherence scanning interferometry (CSI) and chromatic confocal sensing (CCS). The objective of this paper is to better understand how CSI can be used as a tool to analyze surface roughness and topography of cement pastes. Observations from a series of images acquired using this technique on both polished and unpolished samples are described. The results from CSI are compared with those from a STIL confocal microscopy technique (SCM). Comparison between both optical techniques demonstrates the ability of CSI to measure both polished and unpolished cement pastes. - Highlights: • Coherence scanning interferometry (CSI) was used to analyze cement paste surfaces. • The results from the CSI were compared with those from a confocal microscopy. • 3D roughness parameters were obtained using the window resizing method. • Polished and unpolished cement pastes were studied.

  20. Two-photon-based photoactivation in live zebrafish embryos.

    Science.gov (United States)

    Russek-Blum, Niva; Nabel-Rosen, Helit; Levkowitz, Gil

    2010-12-24

    Photoactivation of target compounds in a living organism has proven a valuable approach to investigate various biological processes such as embryonic development, cellular signaling and adult physiology. In this respect, the use of multi-photon microscopy enables quantitative photoactivation of a given light responsive agent in deep tissues at a single cell resolution. As zebrafish embryos are optically transparent, their development can be monitored in vivo. These traits make the zebrafish a perfect model organism for controlling the activity of a variety of chemical agents and proteins by focused light. Here we describe the use of two-photon microscopy to induce the activation of chemically caged fluorescein, which in turn allows us to follow cell's destiny in live zebrafish embryos. We use embryos expressing a live genetic landmark (GFP) to locate and precisely target any cells of interest. This procedure can be similarly used for precise light induced activation of proteins, hormones, small molecules and other caged compounds.

  1. Thin films of metal oxides on metal single crystals: Structure and growth by scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Galloway, H.C.

    1995-12-01

    Detailed studies of the growth and structure of thin films of metal oxides grown on metal single crystal surfaces using Scanning Tunneling Microscopy (STM) are presented. The oxide overlayer systems studied are iron oxide and titanium oxide on the Pt(III) surface. The complexity of the metal oxides and large lattice mismatches often lead to surface structures with large unit cells. These are particularly suited to a local real space technique such as scanning tunneling microscopy. In particular, the symmetry that is directly observed with the STM elucidates the relationship of the oxide overlayers to the substrate as well as distinguishing, the structures of different oxides

  2. Scanning tunneling microscopy I general principles and applications to clean and adsorbate-covered surfaces

    CERN Document Server

    Wiesendanger, Roland

    1992-01-01

    Scanning Tunneling Microscopy I provides a unique introduction to a novel and fascinating technique that produces beautiful images of nature on an atomic scale. It is the first of three volumes that together offer a comprehensive treatment of scanning tunneling microscopy, its diverse applications, and its theoretical treatment. In this volume the reader will find a detailed description of the technique itself and of its applications to metals, semiconductors, layered materials, adsorbed molecules and superconductors. In addition to the many representative results reviewed, extensive references to original work will help to make accessible the vast body of knowledge already accumulated in this field.

  3. Investigation of non-collinear spin states with scanning tunneling microscopy.

    Science.gov (United States)

    Wulfhekel, W; Gao, C L

    2010-03-05

    Most ferromagnetic and antiferromagnetic substances show a simple collinear arrangement of the local spins. Under certain circumstances, however, the spin configuration is non-collinear. Scanning tunneling microscopy with its potential atomic resolution is an ideal tool for investigating these complex spin structures. Non-collinearity can be due to topological frustration of the exchange interaction, due to relativistic spin-orbit coupling or can be found in excited states. Examples for all three cases are given, illustrating the capabilities of spin-polarized scanning tunneling microscopy.

  4. Superresolution upgrade for confocal spinning disk systems using image scanning microscopy (Conference Presentation)

    Science.gov (United States)

    Isbaner, Sebastian; Hähnel, Dirk; Gregor, Ingo; Enderlein, Jörg

    2017-02-01

    Confocal Spinning Disk Systems are widely used for 3D cell imaging because they offer the advantage of optical sectioning at high framerates and are easy to use. However, as in confocal microscopy, the imaging resolution is diffraction limited, which can be theoretically improved by a factor of 2 using the principle of Image Scanning Microscopy (ISM) [1]. ISM with a Confocal Spinning Disk setup (CSDISM) has been shown to improve contrast as well as lateral resolution (FWHM) from 201 +/- 20 nm to 130 +/- 10 nm at 488 nm excitation. A minimum total acquisition time of one second per ISM image makes this method highly suitable for 3D live cell imaging [2]. Here, we present a multicolor implementation of CSDISM for the popular Micro-Manager Open Source Microscopy platform. Since changes in the optical path are not necessary, this will allow any researcher to easily upgrade their standard Confocal Spinning Disk system at remarkable low cost ( 5000 USD) with an ISM superresolution option. [1]. Müller, C.B. and Enderlein, J. Image Scanning Microscopy. Physical Review Letters 104, (2010). [2]. Schulz, O. et al. Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy. Proceedings of the National Academy of Sciences of the United States of America 110, 21000-5 (2013).

  5. Two-photon polarization microscopy reveals protein structure and function

    Czech Academy of Sciences Publication Activity Database

    Lazar, Josef; Bondar, Alexey; Timr, S.; Firestein, S. J.

    2011-01-01

    Roč. 8, č. 8 (2011), s. 684-U120 ISSN 1548-7091 Institutional research plan: CEZ:AV0Z60870520 Keywords : green fluorescent proteins * living cells * in-vivo * indicators * anisotropy * activacion * dissociation * orientation * calmodulin * membranes Subject RIV: CE - Biochemistry Impact factor: 19.276, year: 2011

  6. Mapping the antioxidant activity of apple peels with soft probe scanning electrochemical microscopy

    OpenAIRE

    Lin, Tzu-En; Lesch, Andreas; Li, Chi-Lin; Girault, Hubert

    2017-01-01

    We present a non-invasive electrochemical strategy for mapping the antioxidant (AO) activity of apple peels, which counterbalances oxidative stress caused by various external effectors. Soft carbon microelectrodes were used for soft probe scanning electrochemical microscopy (SECM) enabling the gentle and scratch-free in contact mode scanning of the rough and delicate apple peels in an electrolyte solution. The SECM feedback mode was applied using ferrocene methanol (FcMeOH) as redox mediator ...

  7. A spirobifluorene-based two-photon fluorescence probe for mercury ions and its applications in living cells

    Energy Technology Data Exchange (ETDEWEB)

    Xiao, Haibo, E-mail: xiaohb@shnu.edu.cn; Zhang, Yanzhen; Zhang, Wu; Li, Shaozhi; Tan, Jingjing; Han, Zhongying

    2017-05-01

    A novel spirobifluorene derivative SPF-TMS, which containing dithioacetal groups and triphenylamine units, was synthesized. The probing behaviors toward various metal ions were investigated via UV/Vis absorption spectra as well as one-photon fluorescence changes. The results indicated that SPF-TMS exhibits high sensitivity and selectivity for mercury ions. The detection limit was at least 8.6 × 10{sup −8}M, which is excellent comparing with other optical sensors for Hg{sup 2+}. When measured by two-photon excited fluorescence technique in THF at 800 nm, the two-photon cross-section of SPF-TMS is 272 GM. Especially, upon reaction with mercury species, SPF-TMS yielded another two-photon dye SPF-DA. Both SPF-TMS and SPF-DA emit strong two-photon induced fluorescence and can be applied in cell imaging by two-photon microscopy. - Highlights: • We report a spirobifluorene-based molecule as two-photon fluorescent probe with large two-photon cross-section. • The molecule has exclusive selectivity and sensitivity for mercury species. • The molecule has large two-photon emission changes before and after addition of Hg{sup 2+}. • Both the probe and the mercury ion-promoted reaction product can be applied in cell imaging by two-photon microscopy.

  8. Scanning fluorescent microscopy is an alternative for quantitative fluorescent cell analysis.

    Science.gov (United States)

    Varga, Viktor Sebestyén; Bocsi, József; Sipos, Ferenc; Csendes, Gábor; Tulassay, Zsolt; Molnár, Béla

    2004-07-01

    Fluorescent measurements on cells are performed today with FCM and laser scanning cytometry. The scientific community dealing with quantitative cell analysis would benefit from the development of a new digital multichannel and virtual microscopy based scanning fluorescent microscopy technology and from its evaluation on routine standardized fluorescent beads and clinical specimens. We applied a commercial motorized fluorescent microscope system. The scanning was done at 20 x (0.5 NA) magnification, on three channels (Rhodamine, FITC, Hoechst). The SFM (scanning fluorescent microscopy) software included the following features: scanning area, exposure time, and channel definition, autofocused scanning, densitometric and morphometric cellular feature determination, gating on scatterplots and frequency histograms, and preparation of galleries of the gated cells. For the calibration and standardization Immuno-Brite beads were used. With application of shading compensation, the CV of fluorescence of the beads decreased from 24.3% to 3.9%. Standard JPEG image compression until 1:150 resulted in no significant change. The change of focus influenced the CV significantly only after +/-5 microm error. SFM is a valuable method for the evaluation of fluorescently labeled cells. Copyright 2004 Wiley-Liss, Inc.

  9. Telocytes and putative stem cells in the lungs: electron microscopy, electron tomography and laser scanning microscopy.

    Science.gov (United States)

    Popescu, Laurentiu M; Gherghiceanu, Mihaela; Suciu, Laura C; Manole, Catalin G; Hinescu, Mihail E

    2011-09-01

    This study describes a novel type of interstitial (stromal) cell - telocytes (TCs) - in the human and mouse respiratory tree (terminal and respiratory bronchioles, as well as alveolar ducts). TCs have recently been described in pleura, epicardium, myocardium, endocardium, intestine, uterus, pancreas, mammary gland, etc. (see www.telocytes.com ). TCs are cells with specific prolongations called telopodes (Tp), frequently two to three per cell. Tp are very long prolongations (tens up to hundreds of μm) built of alternating thin segments known as podomers (≤ 200 nm, below the resolving power of light microscope) and dilated segments called podoms, which accommodate mitochondria, rough endoplasmic reticulum and caveolae. Tp ramify dichotomously, making a 3-dimensional network with complex homo- and heterocellular junctions. Confocal microscopy reveals that TCs are c-kit- and CD34-positive. Tp release shed vesicles or exosomes, sending macromolecular signals to neighboring cells and eventually modifying their transcriptional activity. At bronchoalveolar junctions, TCs have been observed in close association with putative stem cells (SCs) in the subepithelial stroma. SCs are recognized by their ultrastructure and Sca-1 positivity. Tp surround SCs, forming complex TC-SC niches (TC-SCNs). Electron tomography allows the identification of bridging nanostructures, which connect Tp with SCs. In conclusion, this study shows the presence of TCs in lungs and identifies a TC-SC tandem in subepithelial niches of the bronchiolar tree. In TC-SCNs, the synergy of TCs and SCs may be based on nanocontacts and shed vesicles.

  10. Occurrence of two-photon absorption saturation in Ag nanocolloids, prepared by chemical reduction method

    Energy Technology Data Exchange (ETDEWEB)

    Rahulan, K. Mani, E-mail: krahul.au@gmail.com [Department of Physics, Anna University, Chennai (India); Ganesan, S. [Department of Physics, Anna University, Chennai (India); Aruna, P., E-mail: aruna@annauniv.edu [Department of Physics, Anna University, Chennai (India)

    2012-09-01

    Highlights: Black-Right-Pointing-Pointer Ag nanocolloids were synthesized via chemical reduction method. Black-Right-Pointing-Pointer The molecules of PVP play an important role in growth and agglomeration of silver nanocolloids. Black-Right-Pointing-Pointer Saturation behaviour followed by two photon absorption was responsible for good optical limiting characteristics in these nanocolloids. Black-Right-Pointing-Pointer The nonlinear optical parameters calculated from the data showed that these materials could be used as efficient optical limiters. - Abstract: Silver nanocolloids stabilized with polyvinyl pyrrolidone (PVP) have been prepared from (AgNO{sub 3}) by a chemical reduction method, involving the intermediate preparation of (Ag{sub 2}O) colloidal dispersions in the presence of sodium dodecycle sulfate as a surfactant and formaldehyde as reducing agent. The molecules of PVP play an important role in growth and agglomeration of silver nanocolloids. The formation of Ag nanocolloids was studied from the UV-vis absorption characteristics. An energy dispersive X-ray (EDX) spectrum and X-ray diffraction peak of the nanoparticles showed the highly crystalline nature of silver structure. The particle size was found to be 40 nm as analyzed from Field emission scanning electron microscopy (FESEM). The nonlinear optical and optical limiting properties of these nanoparticle dispersions were studied by using the Z-scan technique at 532 nm. Experimental results show that the Ag nanocolloids possess strong optical limiting effect, originated from absorption saturation followed by two-photon mechanism. The data show that Ag nanocolloids have great potential for nonlinear optical devices.

  11. Radiative corrections to two photon physics

    International Nuclear Information System (INIS)

    Neervan, W.L. van; Vermaseren, J.A.M.

    1983-06-01

    The authors develop a method to calculate radiative corrections to two photon reactions of the type e + e - →e + e - X where X is an arbitrary final state. To illustrate this they take the example where X stands for a point-like pseudoscalar. It will be shown that the method is an improvement on the standard way in adding real and virtual photon contributions to the (differential) cross-sections. This makes it possible to compute differential and total cross-sections to a very high precision and with a minimum of computer time which is not possible by using more conventional methods. (Auth.)

  12. Inclusive two-photon reactions at TRISTAN

    International Nuclear Information System (INIS)

    Drees, M.

    1995-01-01

    After briefly reviewing past accomplishments of TRISTAN experiments in the field of inclusive two-photon reactions, I discuss open problems in the Monte Carlo simulation of such reactions. The main emphasis is on multiple scattering, i.e. events where at least two pairs of partons scatter within the same γγ collision to form at least four (mini)jets. The cross section for such events might just be observable at TRISTAN. While theoretical arguments for the existence of such events are strong, they have not yet been directly observed experimentally, thereby potentially opening a new opportunity for TRISTAN experiments. (author)

  13. Two-photon physics at LEP

    International Nuclear Information System (INIS)

    Ginzburg, I.F.

    1988-01-01

    The two-photon production of hadrons in e + e - collisions e + e - →e + e - h from which the γγ→h cross sections are extracted is discussed. The common features of these processes are: hadrons move, as a rule, along e + e - beam axis, their total transverse momentum K perpendicular or perpendicular to is small; the total hadron energy is usually less than √S/2. Physical problems of soft processes, exotics, hard processes, semihard processes are considered. New possibilities of LEP, the most interesting and real are presented

  14. Two-photon cooling of magnesium atoms

    DEFF Research Database (Denmark)

    Malossi, N.; Damkjær, S.; Hansen, P. L.

    2005-01-01

    A two-photon mechanism for cooling atoms below the Doppler temperature is analyzed. We consider the magnesium ladder system (3s2)S01¿(3s3p)P11 at 285.2nm followed by the (3s3p)P11¿(3s3d)D21 transition at 880.7nm . For the ladder system quantum coherence effects may become important. Combined with...... and experiment is excellent. In addition, by properly choosing the Rabi frequencies of the two optical transitions a velocity independent atomic dark state is observed....

  15. Superresolution size determination in fluorescence microscopy: A comparison between spatially modulated illumination and confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Spoeri, Udo; Failla, Antonio Virgilio; Cremer, Christoph

    2004-01-01

    Recently developed far field light optical methods are a powerful tool to analyze biological nanostructures and their dynamics, in particular including the interior of three-dimensionally conserved cells. In this article, the recently described method of spatially modulated illumination (SMI) microscopy has been further extended to the online determination of the extension of small, subwavelength sized, fluorescent objects (nanosizing). Using fluorescence excitation with 488 nm, the determination of fluorescent labeled object diameters down to 40 nm corresponding to about 1/12th of the wavelength used for one-photon excitation could be shown. The results of the SMI nanosizing procedure for a detailed, systematic variation of the object diameter are presented together with a fast algorithm for online size evaluation. In addition, we show a direct comparison of the diameter of 'colocalization volumes' between SMI nanosizing and conventional confocal laser scanning microscopy

  16. Two-photon imaging and analysis of neural network dynamics

    International Nuclear Information System (INIS)

    Luetcke, Henry; Helmchen, Fritjof

    2011-01-01

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  17. Two-photon imaging and analysis of neural network dynamics

    Science.gov (United States)

    Lütcke, Henry; Helmchen, Fritjof

    2011-08-01

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  18. Two-photon imaging and analysis of neural network dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Luetcke, Henry; Helmchen, Fritjof [Brain Research Institute, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich (Switzerland)

    2011-08-15

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  19. Impulse excitation scanning acoustic microscopy for local quantification of Rayleigh surface wave velocity using B-scan analysis

    Science.gov (United States)

    Cherry, M.; Dierken, J.; Boehnlein, T.; Pilchak, A.; Sathish, S.; Grandhi, R.

    2018-01-01

    A new technique for performing quantitative scanning acoustic microscopy imaging of Rayleigh surface wave (RSW) velocity was developed based on b-scan processing. In this technique, the focused acoustic beam is moved through many defocus distances over the sample and excited with an impulse excitation, and advanced algorithms based on frequency filtering and the Hilbert transform are used to post-process the b-scans to estimate the Rayleigh surface wave velocity. The new method was used to estimate the RSW velocity on an optically flat E6 glass sample, and the velocity was measured at ±2 m/s and the scanning time per point was on the order of 1.0 s, which are both improvement from the previous two-point defocus method. The new method was also applied to the analysis of two titanium samples, and the velocity was estimated with very low standard deviation in certain large grains on the sample. A new behavior was observed with the b-scan analysis technique where the amplitude of the surface wave decayed dramatically on certain crystallographic orientations. The new technique was also compared with previous results, and the new technique has been found to be much more reliable and to have higher contrast than previously possible with impulse excitation.

  20. Determination of line edge roughness in low-dose top-down scanning electron microscopy images

    NARCIS (Netherlands)

    Verduin, T.; Kruit, P.; Hagen, C.W.

    2014-01-01

    We investigated the off-line metrology for line edge roughness (LER) determination by using the discrete power spectral density (PSD). The study specifically addresses low-dose scanning electron microscopy (SEM) images in order to reduce the acquisition time and the risk of resist shrinkage. The

  1. Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Peng; Behan, Gavin; Kirkland, Angus I. [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Nellist, Peter D., E-mail: peter.nellist@materials.ox.ac.uk [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Cosgriff, Eireann C.; D' Alfonso, Adrian J.; Morgan, Andrew J.; Allen, Leslie J. [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Hashimoto, Ayako [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Takeguchi, Masaki [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Mitsuishi, Kazutaka [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Quantum Dot Research Center, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Shimojo, Masayuki [High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Advanced Science Research Laboratory, Saitama Institute of Technology, 1690 Fusaiji, Fukaya 369-0293 (Japan)

    2011-06-15

    Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored. -- Research Highlights: {yields} The confocal probe image in a scanning confocal electron microscopy image reveals information about the thickness and height of the crystalline layer. {yields} The form of the contrast in a three-dimensional bright-field scanning confocal electron microscopy image can be explained in terms of the confocal probe image. {yields} Despite the complicated form of the contrast in bright-field scanning confocal electron microscopy, we see that depth information is transferred on a 10 nm scale.

  2. In situ scanning tunnelling microscopy of redox molecules. Coherent electron transfer at large bias voltages

    DEFF Research Database (Denmark)

    Zhang, Jingdong; Kuznetsov, A.M.; Ulstrup, Jens

    2003-01-01

    Theories of in situ scanning tunnelling microscopy (STM) of molecules with redox levels near the substrate and tip Fermi levels point to 'spectroscopic' current-overpotential features. Prominent features require a narrow 'probing tip', i.e. a small bias voltage, eV(bias), compared...

  3. Adsorption of Cu phthalocyanine on Pt modified Ge(001): A scanning tunneling microscopy study

    NARCIS (Netherlands)

    Saedi, A.; Berkelaar, Robin P.; Kumar, Avijit; Poelsema, Bene; Zandvliet, Henricus J.W.

    2010-01-01

    The adsorption configurations of copper phthalocyanine (CuPc) molecules on platinum-modified Ge(001) have been studied using scanning tunneling microscopy. After deposition at room temperature and cooling down to 77 K the CuPc molecules are still dynamic. However, after annealing at 550±50 K, the

  4. A simple methodology for obtaining X-ray color images in scanning electron microscopy

    International Nuclear Information System (INIS)

    Veiga, M.M. da; Pietroluongo, L.R.V.

    1985-01-01

    A simple methodology for obtaining at least 3 elements X-ray images in only one photography is described. The fluorescent X-ray image is obtained from scanning electron microscopy with energy dispersion analysis system. The change of detector analytic channels, color cellophane foils and color films are used sequentially. (M.C.K.) [pt

  5. Use of scanning electron microscopy and microanalysis to determine chloride content of concrete and raw materials.

    Science.gov (United States)

    2013-02-01

    Standard sample sets of cement and mortar formulations with known levels of Cl as well as concrete samples subject to Cl diffusion were all prepared for and analyzed with scanning electron microscopy (SEM) and electron microprobe (EPMA). Using x-ray ...

  6. Scanning tunneling microscopy studies of thin foil x-ray mirrors

    DEFF Research Database (Denmark)

    Christensen, Finn Erland; Besenbacher, Flemming; Garnaes, Jorgen

    1990-01-01

    In this paper scanning tunneling microscopy (STM) measurements of x-ray mirrors are presented. The x-ray mirrors are 0.3 mm thick dip-lacquered aluminum foils coated with gold by evaporation, as well as state-of-the-art polished surfaces coated with gold, platinum, or iridium. The measurements...

  7. Preparation of Chemically Etched Tips for Ambient Instructional Scanning Tunneling Microscopy

    Science.gov (United States)

    Zaccardi, Margot J.; Winkelmann, Kurt; Olson, Joel A.

    2010-01-01

    A first-year laboratory experiment that utilizes concepts of electrochemical tip etching for scanning tunneling microscopy (STM) is described. This experiment can be used in conjunction with any STM experiment. Students electrochemically etch gold STM tips using a time-efficient method, which can then be used in an instructional grade STM that…

  8. Dynamics of decanethiol self-assembled monolayers on Au(111) studied by Scanning tunnelling microscopy

    NARCIS (Netherlands)

    Wu, Hairong; Sotthewes, Kai; Kumar, Avijit; Vancso, Gyula J.; Schön, Peter Manfred; Zandvliet, Henricus J.W.

    2013-01-01

    We investigated the dynamics of decanethiol self-assembled monolayers on Au(111) surfaces using time-resolved scanning tunneling microscopy at room temperature. The expected ordered phases (β, δ, χ*, and ) and a disordered phase (ε) were observed. Current–time traces with the feedback loop disabled

  9. Fine structure of the endolymphatic duct in the rat. A scanning and transmission electron microscopy study

    DEFF Research Database (Denmark)

    Qvortrup, K; Rostgaard, Jørgen; Bretlau, P

    1995-01-01

    To investigate the surface morphology of the endolymphatic duct epithelium, 8 rats were vascularly perfused with glutaraldehyde in a buffered and oxygenated blood substitute. Optimal preservation of the epithelium for scanning electron microscopy was attained by coating of the specimens with OsO4...

  10. Transient gels in colloid-polymer mixtures studied with fluorescence confocal scanning laser microscopy

    NARCIS (Netherlands)

    Verhaegh, N.A.M.; Asnaghi, D.; Lekkerkerker, H.N.W.

    1999-01-01

    We study the structure and the time evolution of transient gels formed in colloid-polymer mixtures, by means of uorescence Confocal Scanning Laser Microscopy (CSLM). This technique is used in conjunction with novel colloidal silica particles containing a uorescent core. The confocal micrographs

  11. Visualization of magnetic dipolar interaction based on scanning transmission X-ray microscopy

    International Nuclear Information System (INIS)

    Ohtori, Hiroyuki; Iwano, Kaoru; Takeichi, Yasuo; Ono, Kanta; Mitsumata, Chiharu; Yano, Masao; Kato, Akira; Miyamoto, Noritaka; Shoji, Tetsuya; Manabe, Akira

    2014-01-01

    Using scanning transmission X-ray microscopy (STXM), in this report we visualized the magnetic dipolar interactions in nanocrystalline Nd-Fe-B magnets and imaged their magnetization distributions at various applied fields. We calculated the magnetic dipolar interaction by analyzing the interaction between the magnetization at each point and those at the other points on the STXM image.

  12. Multiterminal semiconductor/ferromagnet probes for spin-filter scanning tunneling microscopy

    NARCIS (Netherlands)

    Vera Marun, I.J.; Jansen, R.

    2009-01-01

    We describe the fabrication of multiterminal semiconductor/ferromagnet probes for a new technique to study magnetic nanostructures: spin-filter scanning tunneling microscopy. We describe the principle of the technique, which is based on spin-polarized tunneling and subsequent analysis of the spin

  13. Spectroscopy of surface adsorbed molecules (scanning tunneling microscopy). Progress report, May 1, 1985-April 30, 1986

    International Nuclear Information System (INIS)

    Coleman, R.V.

    1986-01-01

    A review of the scanning tunneling microscopy program is given. This article contains a description of the design and fabrication of the microscope in addition to description of studies which use the microscope: studies of charge-density waves and studies of tunnel junctions doped with metals and semiconductors. 48 refs., 26 figs

  14. Musculature of Notholca acuminata (Rotifera : Ploima : Brachionidae) revealed by confocal scanning laser microscopy

    DEFF Research Database (Denmark)

    Sørensen, M.V.; Funch, P.; Hooge, M.

    2003-01-01

    The body-wall and visceral musculature of Notholca acuminata was visualized using phalloidin-linked fluorescent dye under confocal laser scanning microscopy. The body-wall musculature includes dorsal, lateral, and ventral pairs of longitudinally oriented body retractor muscles, two pairs of head...

  15. Covalently Immobilised Cytochrome C Imaged by In Situ Scanning Tunnelling Microscopy

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Olesen, Klaus G.; Danilov, Alexey I.

    1997-01-01

    In situ scanning tunnelling microscopy (STM) imaging of cytochrome c (cyt c) on polycrystalline Pt surfaces and on Au(lll) was achieved first by covalent immobilisation of 3-aminopropyltriethoxysilane (3-APTS) brought to react with oxide present on the Pt surfaces. Covalently bound 3-APTS forms...

  16. Second-Harmonic Generation Scanning Microscopy on Domains in Al Surfaces

    DEFF Research Database (Denmark)

    Pedersen, Kjeld; Bozhevolnyi, Sergey I.

    1999-01-01

    Scanning optical second-harmonic generation microscopy has been used to investigate domains in the surface of polycrystaline Al. Strong contrast among the crystalline grains is obtained due to variations in their crystallographic orientations and thus also nonlinear response. The origin of the co...

  17. Supramolecular chemistry at the liquid/solid interface probed by scanning tunnelling microscopy

    NARCIS (Netherlands)

    Feyter, S. De; Uji-i, H.; Mamdouh, W.; Miura, A.; Zhang, J.; Jonkheijm, P.; Schenning, A.P.H.J.; Meijer, E.W.; Chen, Z.; Wurthner, F.; Schuurmans, N.; Esch, J. van; Feringa, B.L.; Dulcey, A.E.; Percec, V.; Schryver, F.C. De

    2006-01-01

    The liquid/solid interface provides an ideal environment to investigate self-assembly phenomena, and scanning tunnelling microscopy (STM) is one of the preferred methodologies to probe the structure and the properties of physisorbed monolayers on the nanoscale. Physisorbed monolayers are of

  18. Calibration-free quantitative surface topography reconstruction in scanning electron microscopy

    NARCIS (Netherlands)

    Faber, E.T.; Martinez-Martinez, D.; Mansilla, C.; Ocelik, V.; De Hosson, J. Th. M.

    This work presents a new approach to obtain reliable surface topography reconstructions from 2D Scanning Electron Microscopy (SEM) images. In this method a set of images taken at different tilt angles are compared by means of digital image correlation (DlC). It is argued that the strength of the

  19. A robust method for processing scanning probe microscopy images and determining nanoobject position and dimensions

    NARCIS (Netherlands)

    Silly, F.

    2009-01-01

    P>Processing of scanning probe microscopy (SPM) images is essential to explore nanoscale phenomena. Image processing and pattern recognition techniques are developed to improve the accuracy and consistency of nanoobject and surface characterization. We present a robust and versatile method to

  20. Surface x-ray scattering and scanning tunneling microscopy studies at the Au(111) electrode

    International Nuclear Information System (INIS)

    Ocko, B.M.; Magnussen, O.M.; Wang, J.X.; Adzic, R.R.

    1993-01-01

    This chapter reviews Surface X-ray Scattering and Scanning Tunneling Microscopy results carried out at the Au(111) surface under electrochemical conditions. Results are presented for the reconstructed surface, and for bromide and thallium monolayers. These examples are used to illustrate the complementary nature of the techniques

  1. Simultaneous scanning tunneling microscopy and synchrotron X-ray measurements in a gas environment

    NARCIS (Netherlands)

    Mom, R.V.; Onderwaater, W.G.; Rost, M.J.; Jankowski, M.; Wenzel, S.; Jacobse, L.; Alkemade, P.F.A.; Vandalon, V.; van Spronsen, M.A.; van Weeren, M.; Crama, B.; van der Tuijn, P.; Felici, R.; Kessels, W.M.M.; Carlà, F.; Frenken, J.W.M.; Groot, I.M.N.

    2017-01-01

    A combined X-ray and scanning tunneling microscopy (STM) instrument is presented that enables the local detection of X-ray absorption on surfaces in a gas environment. To suppress the collection of ion currents generated in the gas phase, coaxially shielded STM tips were used. The conductive outer

  2. Cytochrome C Dynamics at Gold and Glassy Carbon Surfaces Monitored by in Situ Scanning Tunnel Microscopy

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Møller, Per; Pedersen, Marianne Vind

    1995-01-01

    We have investigated the absorption of cytochrome c on gold and glassy carbon substrates by in situ scanning tunnel microscopy under potentiostatic control of both substrate and tip. Low ionic strength and potential ranges where no Faradaic current flows were used. Cyt c aggregates into flat...

  3. Nano-tomography of porous geological materials using focused ion beam-scanning electron microscopy

    NARCIS (Netherlands)

    Liu, Yang; King, Helen E.; van Huis, Marijn A.; Drury, Martyn R.; Plümper, Oliver

    2016-01-01

    Tomographic analysis using focused ion beam-scanning electron microscopy (FIB-SEM) provides three-dimensional information about solid materials with a resolution of a few nanometres and thus bridges the gap between X-ray and transmission electron microscopic tomography techniques. This contribution

  4. In Situ Scanning Probe Microscopy and New Perspectives in Analytical Chemistry

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Zhang, Jingdong; Chi, Qijin

    1999-01-01

    The resolution of scanning probe microscopies is unpresedented but the techniques are fraught with limitations as analytical tools. These limitations and their relationship to the physical mechanisms of image contrast are first discussed. Some new options based on in situ STM, which hold prospect...

  5. Resonant production in two photon collisions

    International Nuclear Information System (INIS)

    Butler, F.

    1988-12-01

    Using 220 picobarn/sup /minus/1/ of data collected by the Mark II detector at PEP, we have measured the width of the /eta/' for quasi real photons. The width is separately measured in the three reactions e + e/sup /minus// → e + e/sup /minus///eta/ 0 π + π/sup /minus//; /eta/ 0 → γγ, e + e/sup /minus// → e + e/sup /minus///eta/ 0 π + π/sup /minus//; /eta/ 0 → π + π/sup /minus//(π 0 ,γ) and e + e/sup /minus// → e + e/sup /minus///rho/ 0 γ, giving a statistically weighted average two-photon width of Γ/sub /eta/'→γγ/ = 5. 21+-0.28 keV. As a by-product of the measurement of Γ/sub /eta/'→γγ/ using the decay mode /eta/' → /rho/γ, we also measured a two-photon decay width for the a 2 (1320) of Γ/sub a 2(1320)→γγ/ = 1.17+-0. 15+-0.25 keV. 47 refs., 50 figs., 8 tabs

  6. Evaluation of Yogurt Microstructure Using Confocal Laser Scanning Microscopy and Image Analysis

    DEFF Research Database (Denmark)

    Skytte, Jacob Lercke; Ghita, Ovidiu; Whelan, Paul F.

    2015-01-01

    The microstructure of protein networks in yogurts defines important physical properties of the yogurt and hereby partly its quality. Imaging this protein network using confocal scanning laser microscopy (CSLM) has shown good results, and CSLM has become a standard measuring technique for fermented...... to image texture description. Here, CSLM images from a yogurt fermentation study are investigated, where production factors including fat content, protein content, heat treatment, and incubation temperature are varied. The descriptors are evaluated through nearest neighbor classification, variance analysis...... scanning microscopy images can be used to provide information on the protein microstructure in yogurt products. For large numbers of microscopy images, subjective evaluation becomes a difficult or even impossible approach, if the images should be incorporated in any form of statistical analysis alongside...

  7. Growth of Pd-Filled Carbon Nanotubes on the Tip of Scanning Probe Microscopy

    Directory of Open Access Journals (Sweden)

    Tomokazu Sakamoto

    2009-01-01

    Full Text Available We have synthesized Pd-filled carbon nanotubes (CNTs oriented perpendicular to Si substrates using a microwave plasma-enhanced chemical vapor deposition (MPECVD for the application of scanning probe microscopy (SPM tip. Prior to the CVD growth, Al thin film (10 nm was coated on the substrate as a buffer layer followed by depositing a 5∼40 nm-thick Pd film as a catalyst. The diameter and areal density of CNTs grown depend largely on the initial Pd thickness. Scanning electron microscopy (SEM and transmission electron microscopy (TEM images clearly show that Pd is successfully encapsulated into the CNTs, probably leading to higher conductivity. Using optimum growth conditions, Pd-filled CNTs are successfully grown on the apex of the conventional SPM cantilever.

  8. Electrocatalytic activity mapping of model fuel cell catalyst films using scanning electrochemical microscopy

    International Nuclear Information System (INIS)

    Nicholson, P.G.; Zhou, S.; Hinds, G.; Wain, A.J.; Turnbull, A.

    2009-01-01

    Scanning electrochemical microscopy has been employed to spatially map the electrocatalytic activity of model proton exchange membrane fuel cell (PEMFC) catalyst films towards the hydrogen oxidation reaction (the PEMFC anode reaction). The catalyst films were composed of platinum-loaded carbon nanoparticles, similar to those typically used in PEMFCs. The electrochemical characterisation was correlated with a detailed physical characterisation using dynamic light scattering, transmission electron microscopy and field-emission scanning electron microscopy. The nanoparticles were found to be reasonably mono-dispersed, with a tendency to agglomerate into porous bead-type structures when spun-cast. The number of carbon nanoparticles with little or no platinum was surprisingly higher than would be expected based on the platinum-carbon mass ratio. Furthermore, the platinum-rich carbon particles tended to agglomerate and the clusters formed were non-uniformly distributed. This morphology was reflected in a high degree of heterogeneity in the film activity towards the hydrogen oxidation reaction.

  9. Three axis vector magnet set-up for cryogenic scanning probe microscopy

    International Nuclear Information System (INIS)

    Galvis, J. A.; Herrera, E.; Buendía, A.; Guillamón, I.; Vieira, S.; Suderow, H.; Azpeitia, J.; Luccas, R. F.; Munuera, C.; García-Hernandez, M.

    2015-01-01

    We describe a three axis vector magnet system for cryogenic scanning probe microscopy measurements. We discuss the magnet support system and the power supply, consisting of a compact three way 100 A current source. We obtain tilted magnetic fields in all directions with maximum value of 5T along z-axis and of 1.2T for XY-plane magnetic fields. We describe a scanning tunneling microscopy-spectroscopy (STM-STS) set-up, operating in a dilution refrigerator, which includes a new high voltage ultralow noise piezodrive electronics and discuss the noise level due to vibrations. STM images and STS maps show atomic resolution and the tilted vortex lattice at 150 mK in the superconductor β-Bi 2 Pd. We observe a strongly elongated hexagonal lattice, which corresponds to the projection of the tilted hexagonal vortex lattice on the surface. We also discuss Magnetic Force Microscopy images in a variable temperature insert

  10. Three axis vector magnet set-up for cryogenic scanning probe microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Galvis, J. A. [Laboratorio de Bajas Temperaturas, Departamento de Física de la Materia Condensada, Instituto de Ciencia de Materiales Nicolás Cabrera, Condensed Matter Physics Center (IFIMAC), Facultad de Ciencias Universidad Autónoma de Madrid, 28049 Madrid (Spain); Departamento de Ciencias Naturales Facultad de Ingeniería Universidad Central, Bogotá (Colombia); Herrera, E.; Buendía, A. [Laboratorio de Bajas Temperaturas, Departamento de Física de la Materia Condensada, Instituto de Ciencia de Materiales Nicolás Cabrera, Condensed Matter Physics Center (IFIMAC), Facultad de Ciencias Universidad Autónoma de Madrid, 28049 Madrid (Spain); Guillamón, I.; Vieira, S.; Suderow, H. [Laboratorio de Bajas Temperaturas, Departamento de Física de la Materia Condensada, Instituto de Ciencia de Materiales Nicolás Cabrera, Condensed Matter Physics Center (IFIMAC), Facultad de Ciencias Universidad Autónoma de Madrid, 28049 Madrid (Spain); Unidad Asociada de Bajas Temperaturas y Altos Campos Magnéticos, UAM, CSIC, Cantoblanco, E-28049 Madrid (Spain); Azpeitia, J.; Luccas, R. F.; Munuera, C.; García-Hernandez, M. [Unidad Asociada de Bajas Temperaturas y Altos Campos Magnéticos, UAM, CSIC, Cantoblanco, E-28049 Madrid (Spain); Instituto de Ciencia de Materiales de Madrid, Consejo Superior de Investigaciones Científicas (ICMM-CSIC), Sor Juana Inés de la Cruz 3, 28049 Madrid (Spain); and others

    2015-01-15

    We describe a three axis vector magnet system for cryogenic scanning probe microscopy measurements. We discuss the magnet support system and the power supply, consisting of a compact three way 100 A current source. We obtain tilted magnetic fields in all directions with maximum value of 5T along z-axis and of 1.2T for XY-plane magnetic fields. We describe a scanning tunneling microscopy-spectroscopy (STM-STS) set-up, operating in a dilution refrigerator, which includes a new high voltage ultralow noise piezodrive electronics and discuss the noise level due to vibrations. STM images and STS maps show atomic resolution and the tilted vortex lattice at 150 mK in the superconductor β-Bi{sub 2}Pd. We observe a strongly elongated hexagonal lattice, which corresponds to the projection of the tilted hexagonal vortex lattice on the surface. We also discuss Magnetic Force Microscopy images in a variable temperature insert.

  11. EDITORIAL: Three decades of scanning tunnelling microscopy that changed the course of surface science Three decades of scanning tunnelling microscopy that changed the course of surface science

    Science.gov (United States)

    Ramachandra Rao, M. S.; Margaritondo, Giorgio

    2011-11-01

    Three decades ago, with a tiny tip of platinum, the scientific world saw the real space imaging of single atoms with unprecedented spatial resolution. This signalled the birth of one of the most versatile surface probes, based on the physics of quantum mechanical tunnelling: the scanning tunnelling microscope (STM). Invented in 1981 by Gerd Binnig and Heinrich Rohrer of IBM, Zurich, it led to their award of the 1986 Nobel Prize. Atoms, once speculated to be abstract entities used by theoreticians for mere calculations, can be seen to exist for real with the nano-eye of an STM tip that also gives real-space images of molecules and adsorbed complexes on surfaces. From a very fundamental perspective, the STM changed the course of surface science and engineering. STM also emerged as a powerful tool to study various fundamental phenomena relevant to the properties of surfaces in technological applications such as tribology, medical implants, catalysis, sensors and biology—besides elucidating the importance of local bonding geometries and defects, non-periodic structures and the co-existence of nano-scale phases. Atom-level probing, once considered a dream, has seen the light with the evolution of STM. An important off-shoot of STM was the atomic force microscope (AFM) for surface mapping of insulating samples. Then followed the development of a flurry of techniques under the general name of scanning probe microscopy (SPM). These techniques (STM, AFM, MFM, PFM etc) designed for atomic-scale-resolution imaging and spectroscopy, have led to brand new developments in surface analysis. All of these novel methods enabled researchers in recent years to image and analyse complex surfaces on microscopic and nanoscopic scales. All of them utilize a small probe for sensing the surface. The invention of AFM by Gerd Binnig, Calvin Quate and Christopher Gerber opened up new opportunities for characterization of a variety of materials, and various industrial applications could be

  12. Recommendations for the design and the installation of large laser scanning microscopy systems

    Science.gov (United States)

    Helm, P. Johannes

    2012-03-01

    Laser Scanning Microscopy (LSM) has since the inventions of the Confocal Scanning Laser Microscope (CLSM) and the Multi Photon Laser Scanning Microscope (MPLSM) developed into an essential tool in contemporary life science and material science. The market provides an increasing number of turn-key and hands-off commercial LSM systems, un-problematic to purchase, set up and integrate even into minor research groups. However, the successful definition, financing, acquisition, installation and effective use of one or more large laser scanning microscopy systems, possibly of core facility character, often requires major efforts by senior staff members of large academic or industrial units. Here, a set of recommendations is presented, which are helpful during the process of establishing large systems for confocal or non-linear laser scanning microscopy as an effective operational resource in the scientific or industrial production process. Besides the description of technical difficulties and possible pitfalls, the article also illuminates some seemingly "less scientific" processes, i.e. the definition of specific laboratory demands, advertisement of the intention to purchase one or more large systems, evaluation of quotations, establishment of contracts and preparation of the local environment and laboratory infrastructure.

  13. 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.

  14. 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.

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

    International Nuclear Information System (INIS)

    Bolker, Asaf; Kalish, Rafi; Saguy, Cecile

    2014-01-01

    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. (paper)

  16. Imaging by Electrochemical Scanning Tunneling Microscopy and Deconvolution Resolving More Details of Surfaces Nanomorphology

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov

    observed in high-resolution images of metallic nanocrystallites may be effectively deconvoluted, as to resolve more details of the crystalline morphology (see figure). Images of surface-crystalline metals indicate that more than a single atomic layer is involved in mediating the tunneling current......Upon imaging, electrochemical scanning tunneling microscopy (ESTM), scanning electrochemical micro-scopy (SECM) and in situ STM resolve information on electronic structures and on surface topography. At very high resolution, imaging processing is required, as to obtain information that relates...... to crystallographic-surface structures. Within the wide range of new technologies, those images surface features, the electrochemical scanning tunneling microscope (ESTM) provides means of atomic resolution where the tip participates actively in the process of imaging. Two metallic surfaces influence ions trapped...

  17. New directions in point-contact spectroscopy based on scanning tunneling microscopy techniques (Review Article)

    International Nuclear Information System (INIS)

    Tartaglini, E.; Verhagen, T.G.A.; Galli, F.; Trouwborst, M.L.; Aarts, J.; Van-Ruitebbeek, J.M.; Muller, R.; Shiota, T.

    2013-01-01

    Igor Yanson showed 38 years ago for the first time a point-contact measurement where he probed the energy resolved spectroscopy of the electronic scattering inside the metal. Since this first measurement, the pointcontact spectroscopy (PCS) technique improved enormously. The application of the scanning probe microscopy (SPM) techniques in the late 1980s allowed achieving contacts with a diameter of a single atom. With the introduction of the mechanically controlled break junction technique, even spectroscopy on freely suspended chains of atoms could be performed. In this paper, we briefly review the current developments of PCS and show recent experiments in advanced scanning PCS based on SPM techniques. We describe some results obtained with both needle-anvil type of point contacts and scanning tunneling microscopy (STM). We also show our first attempt to lift up with a STM a chain of single gold atoms from a Au(110) surface.

  18. On the kinematics of the two-photon Cherenkov effect

    International Nuclear Information System (INIS)

    Afanas'ev, G.N.; Stepanovskij, Yu.P.

    2003-01-01

    We study the kinematics of the two-photon Cherenkov effect. In the general case, the emission angles of two photons satisfy certain inequalities and the corresponding radiation intensities are rather diffused. In special cases, when the above inequalities reduce to equalities, the emission angles of two photons are fixed and the corresponding radiation intensities should have sharp maxima at these angles. This makes easier the experimental study of the two-photon Cherenkov effect

  19. Quantitating morphological changes in biological samples during scanning electron microscopy sample preparation with correlative super-resolution microscopy.

    Science.gov (United States)

    Zhang, Ying; Huang, Tao; Jorgens, Danielle M; Nickerson, Andrew; Lin, Li-Jung; Pelz, Joshua; Gray, Joe W; López, Claudia S; Nan, Xiaolin

    2017-01-01

    Sample preparation is critical to biological electron microscopy (EM), and there have been continuous efforts on optimizing the procedures to best preserve structures of interest in the sample. However, a quantitative characterization of the morphological changes associated with each step in EM sample preparation is currently lacking. Using correlative EM and superresolution microscopy (SRM), we have examined the effects of different drying methods as well as osmium tetroxide (OsO4) post-fixation on cell morphology during scanning electron microscopy (SEM) sample preparation. Here, SRM images of the sample acquired under hydrated conditions were used as a baseline for evaluating morphological changes as the sample went through SEM sample processing. We found that both chemical drying and critical point drying lead to a mild cellular boundary retraction of ~60 nm. Post-fixation by OsO4 causes at least 40 nm additional boundary retraction. We also found that coating coverslips with adhesion molecules such as fibronectin prior to cell plating helps reduce cell distortion from OsO4 post-fixation. These quantitative measurements offer useful information for identifying causes of cell distortions in SEM sample preparation and improving current procedures.

  20. Detection of secondary phases in duplex stainless steel by magnetic force microscopy and scanning Kelvin probe force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ramírez-Salgado, J. [Instituto Mexicano del Petróleo, Dirección de Investigación y Posgrado, Eje Central Norte Lázaro Cárdenas, No. 152, 07730 D.F., México (Mexico); Domínguez-Aguilar, M.A., E-mail: madoming@imp.mx [Instituto Mexicano del Petróleo, Dirección de Investigación y Posgrado, Eje Central Norte Lázaro Cárdenas, No. 152, 07730 D.F., México (Mexico); Castro-Domínguez, B. [University of Tokyo, Department of Chemical System Engineering, Faculty of Engineering Bldg. 5, 7F 722, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113–8656 (Japan); Hernández-Hernández, P. [Instituto Mexicano del Petróleo, Dirección de Investigación y Posgrado, Eje Central Norte Lázaro Cárdenas, No. 152, 07730 D.F., México (Mexico); Newman, R.C. [University of Toronto, Department of Chemical Engineering and Applied Chemistry, 200 College Street, Toronto M5S 3E5 (Canada)

    2013-12-15

    The secondary phase transformations in a commercial super duplex stainless steel were investigated by micro-chemical analyses and high resolution scanning probe microscopy. Energy dispersive X-ray and electron probe detected ferrite and austenite as well as secondary phases in unetched aged duplex stainless steel type 25Cr-7Ni-3Mo. Volta potential indicated that nitride and sigma appeared more active than ferrite, while secondary austenite and austenite presented a nobler potential. Reversal order in nobility is thought to be attributable to the potential ranking provided by oxide nature diversity as a result of secondary phase surface compositions on steel. After eutectoid transformation, secondary austenite was detected by electron probe microanalysis, whereas atomic force microscopy distinguished this phase from former austenite by image contrast. Magnetic force microscopy revealed a “ghosted” effect on the latter microstructure probably derived from metal memory reminiscence of mechanical polishing at passivity and long range magnetic forces of ferrite phase. - Highlights: • Nobility detection of secondary phases by SKPFM in DSS particles is not a straightforward procedure. • As Volta potential and contrast are not always consistent SKPFM surface oxides is thought played an important role in detection. • AFM distinguished secondary austenite from former austenite by image contrast though SEM required EPMA.

  1. The Use Of Scanning Probe Microscopy To Investigate Crystal-Fluid Interfaces

    International Nuclear Information System (INIS)

    Orme, C A; Giocondi, J L

    2007-01-01

    Over the past decade there has been a natural drive to extend the investigation of dynamic surfaces in fluid environments to higher resolution characterization tools. Various aspects of solution crystal growth have been directly visualized for the first time. These include island nucleation and growth using transmission electron microscopy and scanning tunneling microscopy; elemental step motion using scanning probe microscopy; and the time evolution of interfacial atomic structure using various diffraction techniques. In this lecture we will discuss the use of one such in situ method, scanning probe microscopy, as a means of measuring surface dynamics during crystal growth and dissolution. We will cover both practical aspects of imaging such as environmental control, fluid flow, and electrochemical manipulation, as well as the types of physical measurements that can be made. Measurements such as step motion, critical lengths, nucleation density, and step fluctuations, will be put in context of the information they provide about mechanistic processes at surfaces using examples from metal and mineral crystal growth

  2. Two-photon interference : spatial aspects of two-photon entanglement, diffraction, and scattering

    NARCIS (Netherlands)

    Peeters, Wouter Herman

    2010-01-01

    This dissertation contains scientific research within the realm of quantum optics, which is a branch of physics. An experimental and theoretical study is made of two-photon interference phenomena in various optical systems. Spatially entangled photon pairs are produced via the nonlinear optical

  3. Infrared spectroscopy of molecular submonolayers on surfaces by infrared scanning tunneling microscopy: tetramantane on Au111.

    Science.gov (United States)

    Pechenezhskiy, Ivan V; Hong, Xiaoping; Nguyen, Giang D; Dahl, Jeremy E P; Carlson, Robert M K; Wang, Feng; Crommie, Michael F

    2013-09-20

    We have developed a new scanning-tunneling-microscopy-based spectroscopy technique to characterize infrared (IR) absorption of submonolayers of molecules on conducting crystals. The technique employs a scanning tunneling microscope as a precise detector to measure the expansion of a molecule-decorated crystal that is irradiated by IR light from a tunable laser source. Using this technique, we obtain the IR absorption spectra of [121]tetramantane and [123]tetramantane on Au(111). Significant differences between the IR spectra for these two isomers show the power of this new technique to differentiate chemical structures even when single-molecule-resolved scanning tunneling microscopy (STM) images look quite similar. Furthermore, the new technique was found to yield significantly better spectral resolution than STM-based inelastic electron tunneling spectroscopy, and to allow determination of optical absorption cross sections. Compared to IR spectroscopy of bulk tetramantane powders, infrared scanning tunneling microscopy (IRSTM) spectra reveal narrower and blueshifted vibrational peaks for an ordered tetramantane adlayer. Differences between bulk and surface tetramantane vibrational spectra are explained via molecule-molecule interactions.

  4. High-resolution photoluminescence electro-modulation microscopy by scanning lock-in

    Science.gov (United States)

    Koopman, W.; Muccini, M.; Toffanin, S.

    2018-04-01

    Morphological inhomogeneities and structural defects in organic semiconductors crucially determine the charge accumulation and lateral transport in organic thin-film transistors. Photoluminescence Electro-Modulation (PLEM) microscopy is a laser-scanning microscopy technique that relies on the modulation of the thin-film fluorescence in the presence of charge-carriers to image the spatial distribution of charges within the active organic semiconductor. Here, we present a lock-in scheme based on a scanning beam approach for increasing the PLEM microscopy resolution and contrast. The charge density in the device is modulated by a sinusoidal electrical signal, phase-locked to the scanning beam of the excitation laser. The lock-in detection scheme is achieved by acquiring a series of images with different phases between the beam scan and the electrical modulation. Application of high resolution PLEM to an organic transistor in accumulation mode demonstrates its potential to image local variations in the charge accumulation. A diffraction-limited precision of sub-300 nm and a signal to noise ratio of 21.4 dB could be achieved.

  5. X-ray optics for scanning fluorescence microscopy and other applications

    International Nuclear Information System (INIS)

    Ryon, R.W.; Warburton, W.K.

    1992-05-01

    Scanning x-ray fluorescence microscopy is analogous to scanning electron microscopy. Maps of chemical element distribution are produced by scanning with a very small x-ray beam. Goal is to perform such scanning microscopy with resolution in the range of <1 to 10 μm, using standard laboratory x-ray tubes. We are investigating mirror optics in the Kirkpatrick-Baez (K-B) configuration. K-B optics uses two curved mirrors mounted orthogonally along the optical axis. The first mirror provides vertical focus, the second mirror provides horizontal focus. We have used two types of mirrors: synthetic multilayers and crystals. Multilayer mirrors are used with lower energy radiation such as Cu Kα. At higher energies such as Ag Kα, silicon wafers are used in order to increase the incidence angles and thereby the photon collection efficiency. In order to increase the surface area of multilayers which reflects x-rays at the Bragg angle, we have designed mirrors with the spacing between layers graded along the optic axis in order to compensate for the changing angle of incidence. Likewise, to achieve a large reflecting surface with silicon, the wafers are placed on a specially designed lever arm which is bent into a log spiral by applying force at one end. In this way, the same diffracting angle is maintained over the entire surface of the wafer, providing a large solid angle for photon collection

  6. Two-photon excited UV fluorescence for protein crystal detection

    International Nuclear Information System (INIS)

    Madden, Jeremy T.; DeWalt, Emma L.; Simpson, Garth J.

    2011-01-01

    Complementary measurements using SONICC and TPE-UVF allow the sensitive and selective detection of protein crystals. Two-photon excited ultraviolet fluorescence (TPE-UVF) microscopy is explored for sensitive protein-crystal detection as a complement to second-order nonlinear optical imaging of chiral crystals (SONICC). Like conventional ultraviolet fluorescence (UVF), TPE-UVF generates image contrast based on the intrinsic fluorescence of aromatic residues, generally producing higher fluorescence emission within crystals than the mother liquor by nature of the higher local protein concentration. However, TPE-UVF has several advantages over conventional UVF, including (i) insensitivity to optical scattering, allowing imaging in turbid matrices, (ii) direct compatibility with conventional optical plates and windows by using visible light for excitation, (iii) elimination of potentially damaging out-of-plane UV excitation, (iv) improved signal to noise through background reduction from out-of-plane excitation and (v) relatively simple integration into instrumentation developed for SONICC

  7. Optical microscope illumination analysis using through-focus scanning optical microscopy.

    Science.gov (United States)

    Attota, Ravi Kiran; Park, Haesung

    2017-06-15

    Misalignment of the aperture diaphragm present in optical microscopes results in angular illumination asymmetry (ANILAS) at the sample plane. Here we show that through-focus propagation of ANILAS results in a lateral image shift with a focus position. This could lead to substantial errors in quantitative results for optical methods that use through-focus images such as three-dimensional nanoparticle tracking, confocal microscopy, and through-focus scanning optical microscopy (TSOM). A correlation exists between ANILAS and the slant in TSOM images. Hence, the slant in the TSOM image can be used to detect, analyze, and rectify the presence of ANILAS.

  8. Reversal of atomic contrast in scanning probe microscopy on (111) metal surfaces

    Czech Academy of Sciences Publication Activity Database

    Ondráček, Martin; González, C.; Jelínek, Pavel

    2012-01-01

    Roč. 24, 08 (2012), 084003/1-084003/7 ISSN 0953-8984 R&D Projects: GA ČR(CZ) GPP204/11/P578; GA ČR GAP204/10/0952; GA ČR GA202/09/0545; GA MŠk(CZ) ME10076 Grant - others:AVČR(CZ) M100100904 Institutional research plan: CEZ:AV0Z10100521 Keywords : atomic force microscopy * metallic surfaces * atomic contrast * scanning tunneling microscopy Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 2.355, year: 2012 http://iopscience.iop.org/0953-8984/24/8/084003

  9. Development of an add-on kit for scanning confocal microscopy (Conference Presentation)

    Science.gov (United States)

    Guo, Kaikai; Zheng, Guoan

    2017-03-01

    Scanning confocal microscopy is a standard choice for many fluorescence imaging applications in basic biomedical research. It is able to produce optically sectioned images and provide acquisition versatility to address many samples and application demands. However, scanning a focused point across the specimen limits the speed of image acquisition. As a result, scanning confocal microscope only works well with stationary samples. Researchers have performed parallel confocal scanning using digital-micromirror-device (DMD), which was used to project a scanning multi-point pattern across the sample. The DMD based parallel confocal systems increase the imaging speed while maintaining the optical sectioning ability. In this paper, we report the development of an add-on kit for high-speed and low-cost confocal microscopy. By adapting this add-on kit to an existing regular microscope, one can convert it into a confocal microscope without significant hardware modifications. Compared with current DMD-based implementations, the reported approach is able to recover multiple layers along the z axis simultaneously. It may find applications in wafer inspection and 3D metrology of semiconductor circuit. The dissemination of the proposed add-on kit under $1000 budget could also lead to new types of experimental designs for biological research labs, e.g., cytology analysis in cell culture experiments, genetic studies on multicellular organisms, pharmaceutical drug profiling, RNA interference studies, investigation of microbial communities in environmental systems, and etc.

  10. Imaging of buried phosphorus nanostructures in silicon using scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Oberbeck, Lars [Centre for Quantum Computation and Communication Technology, School of Physics, University of New South Wales, Sydney, New South Wales 2052 (Australia); TOTAL Marketing Services, New Energies, La Défense 10, 92069 Paris La Défense Cedex (France); Reusch, Thilo C. G.; Hallam, Toby; Simmons, Michelle Y., E-mail: n.curson@ucl.ac.uk, E-mail: michelle.simmons@unsw.edu.au [Centre for Quantum Computation and Communication Technology, School of Physics, University of New South Wales, Sydney, New South Wales 2052 (Australia); Schofield, Steven R. [Centre for Quantum Computation and Communication Technology, School of Physics, University of New South Wales, Sydney, New South Wales 2052 (Australia); London Centre for Nanotechnology, UCL, London WC1H 0AH (United Kingdom); Department of Physics and Astronomy, UCL, London WC1E 6BT (United Kingdom); Curson, Neil J., E-mail: n.curson@ucl.ac.uk, E-mail: michelle.simmons@unsw.edu.au [Centre for Quantum Computation and Communication Technology, School of Physics, University of New South Wales, Sydney, New South Wales 2052 (Australia); London Centre for Nanotechnology, UCL, London WC1H 0AH (United Kingdom); Department of Electronic and Electrical Engineering, UCL, London WC1E 7JE (United Kingdom)

    2014-06-23

    We demonstrate the locating and imaging of single phosphorus atoms and phosphorus dopant nanostructures, buried beneath the Si(001) surface using scanning tunneling microscopy. The buried dopant nanostructures have been fabricated in a bottom-up approach using scanning tunneling microscope lithography on Si(001). We find that current imaging tunneling spectroscopy is suited to locate and image buried nanostructures at room temperature and with residual surface roughness present. From these studies, we can place an upper limit on the lateral diffusion during encapsulation with low-temperature Si molecular beam epitaxy.

  11. Group velocity measurement using spectral interference in near-field scanning optical microscopy

    International Nuclear Information System (INIS)

    Mills, John D.; Chaipiboonwong, Tipsuda; Brocklesby, William S.; Charlton, Martin D. B.; Netti, Caterina; Zoorob, Majd E.; Baumberg, Jeremy J.

    2006-01-01

    Near-field scanning optical microscopy provides a tool for studying the behavior of optical fields inside waveguides. In this experiment the authors measure directly the variation of group velocity between different modes of a planar slab waveguide as the modes propagate along the guide. The measurement is made using the spectral interference between pulses propagating inside the waveguide with different group velocities, collected using a near-field scanning optical microscope at different points down the guide and spectrally resolved. The results are compared to models of group velocities in simple guides

  12. Imaging of buried phosphorus nanostructures in silicon using scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Oberbeck, Lars; Reusch, Thilo C. G.; Hallam, Toby; Simmons, Michelle Y.; Schofield, Steven R.; Curson, Neil J.

    2014-01-01

    We demonstrate the locating and imaging of single phosphorus atoms and phosphorus dopant nanostructures, buried beneath the Si(001) surface using scanning tunneling microscopy. The buried dopant nanostructures have been fabricated in a bottom-up approach using scanning tunneling microscope lithography on Si(001). We find that current imaging tunneling spectroscopy is suited to locate and image buried nanostructures at room temperature and with residual surface roughness present. From these studies, we can place an upper limit on the lateral diffusion during encapsulation with low-temperature Si molecular beam epitaxy.

  13. Development of a two photon microscope for tracking Drosophila larvae

    Science.gov (United States)

    Karagyozov, Doycho; Mihovilovic Skanata, Mirna; Gershow, Marc

    Current in vivo methods for measuring neural activity in Drosophila larva require immobilization of the animal. Although we can record neural signals while stimulating the sensory organs, we cannot read the behavioral output because we have prevented the animal from moving. Many research questions cannot be answered without observation of neural activity in behaving (freely-moving) animals. We incorporated a Tunable Acoustic Gradient (TAG) lens into a two-photon microscope to achieve a 70kHz axial scan rate, enabling volumetric imaging at tens of hertz. We then implemented a tracking algorithm based on a Kalman filter to maintain the neurons of interest in the field of view and in focus during the rapid three dimensional motion of a free larva. Preliminary results show successful tracking of a neuron moving at speeds reaching 500 μm/s. NIH Grant 1DP2EB022359 and NSF Grant PHY-1455015.

  14. Molecular engineering of two-photon fluorescent probes for bioimaging applications

    Science.gov (United States)

    Liu, Hong-Wen; Liu, Yongchao; Wang, Peng; Zhang, Xiao-Bing

    2017-03-01

    During the past two decades, two-photon microscopy (TPM), which utilizes two near-infrared photons as the excitation source, has emerged as a novel, attractive imaging tool for biological research. Compared with one-photon microscopy, TPM offers several advantages, such as lowering background fluorescence in living cells and tissues, reducing photodamage to biosamples, and a photobleaching phenomenon, offering better 3D spatial localization, and increasing penetration depth. Small-molecule-based two-photon fluorescent probes have been well developed for the detection and imaging of various analytes in biological systems. In this review, we will give a general introduction of molecular engineering of two-photon fluorescent probes based on different fluorescence response mechanisms for bioimaging applications during the past decade. Inspired by the desired advantages of small-molecule two-photon fluorescent probes in biological imaging applications, we expect that more attention will be devoted to the development of new two-photon fluorophores and applications of TPM in areas of bioanalysis and disease diagnosis.

  15. The Use of Intravital Two-Photon and Thick Section Confocal Imaging to Analyze B Lymphocyte Trafficking in Lymph Nodes and Spleen.

    Science.gov (United States)

    Park, Chung; Hwang, Il-Young; Kehrl, John H

    2018-01-01

    Intravital two-photon laser scanning microscopy (TP-LSM) has allowed the direct observation of immune cells in intact organs of living animals. In the B cell biology field TP-LSM has detailed the movement of B cells in high endothelial venules and during their transmigration into lymph organs; described the movement and positioning of B cells within lymphoid organs; outlined the mechanisms by which antigen is delivered to B cells; observed B cell interacting with T cells, other cell types, and even with pathogens; and delineated the egress of B cells from the lymph node (LN) parenchyma into the efferent lymphatics. As the quality of TP-LSM improves and as new fluorescent probes become available additional insights into B cell behavior and function await new investigations. Yet intravital TP-LSM has some disadvantages including a lower resolution than standard confocal microscopy, a narrow imaging window, and a shallow depth of imaging. We have found that supplementing intravital TP-LSM with conventional confocal microscopy using thick LN sections helps to overcome some of these shortcomings. Here, we describe procedures for visualizing the behavior and trafficking of fluorescently labeled, adoptively transferred antigen-activated B cells within the inguinal LN of live mice using two-photon microscopy. Also, we introduce procedures for fixed thick section imaging using standard confocal microscopy, which allows imaging of fluorescently labeled cells deep in the LN cortex and in the spleen with high resolution.

  16. Atomic-scale structure of dislocations revealed by scanning tunneling microscopy and molecular dynamics

    DEFF Research Database (Denmark)

    Christiansen, Jesper; Morgenstern, K.; Schiøtz, Jakob

    2002-01-01

    The intersection between dislocations and a Ag(111) surface has been studied using an interplay of scanning tunneling microscopy (STM) and molecular dynamics. Whereas the STM provides atomically resolved information about the surface structure and Burgers vectors of the dislocations, the simulati......The intersection between dislocations and a Ag(111) surface has been studied using an interplay of scanning tunneling microscopy (STM) and molecular dynamics. Whereas the STM provides atomically resolved information about the surface structure and Burgers vectors of the dislocations......, the simulations can be used to determine dislocation structure and orientation in the near-surface region. In a similar way, the subsurface structure of other extended defects can be studied. The simulations show dislocations to reorient the partials in the surface region leading to an increased splitting width...

  17. Memory properties and charge effect study in Si nanocrystals by scanning capacitance microscopy and spectroscopy

    Directory of Open Access Journals (Sweden)

    Bassani Franck

    2011-01-01

    Full Text Available Abstract In this letter, isolated Si nanocrystal has been formed by dewetting process with a thin silicon dioxide layer on top. Scanning capacitance microscopy and spectroscopy were used to study the memory properties and charge effect in the Si nanocrystal in ambient temperature. The retention time of trapped charges injected by different direct current (DC bias were evaluated and compared. By ramp process, strong hysteresis window was observed. The DC spectra curve shift direction and distance was observed differently for quantitative measurements. Holes or electrons can be separately injected into these Si-ncs and the capacitance changes caused by these trapped charges can be easily detected by scanning capacitance microscopy/spectroscopy at the nanometer scale. This study is very useful for nanocrystal charge trap memory application.

  18. Morphologic differences observed by scanning electron microscopy according to the reason for pseudophakic IOL explantation

    DEFF Research Database (Denmark)

    Fernandez-Buenaga, Roberto; Alio, Jorge L.; Ramirez, Jose M.

    2015-01-01

    Purpose To compare variations in surface morphology, as studied by scanning electron microscopy (SEM), of explanted intraocular lenses (IOLs) concerning the cause leading to the explantation surgery. Methods In this prospective multicenter study, explanted IOLs were analyzed by SEM and energy...... explanted due to dislocation demonstrated calcifications in 8 lenses (50%), salt precipitates in 6 cases (37.5%), and erythrocytes and fibrosis/fibroblasts in 2 cases (12.5%). In the refractive error cases, the SEM showed proteins in 5 cases (45.5%) and salt precipitates in 4 lenses (36.4%). In IOL...... opacification, the findings were calcifications in 2 of the 3 lenses (66.6%) and proteins in 2 lenses (66.6%). Conclusions A marked variation in surface changes was observed by SEM. Findings did not correlate with cause for explantation. Scanning electron microscopy is a useful tool that provides exclusive...

  19. Analysis of polymer grafted inside the porous hydrogel using confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    2007-04-01

    Full Text Available Graft polymerization of glycidyl methacrylate onto the pore surface of polyacrylamide macroporous gel was implemented in DMSO-aqueous solution using diperiodatocuprate(III complexes as an initiator. The grafting densities up to 410% were achieved. The graft polymerization was confirmed by gravimetrical methods and FTIR. The graft polymerization of polymer inside the pores of the macroporous gel resulted in increased flow resistance through the gel matrix. The distribution of grafted polymer on the gel pore surface material was studied by scanning electron microscopy (SEM and confocal laser scanning microscopy (CLSM. CLSM is an alternative method for studying morphology of gel surface with grafted polymer having the advantages over the SEM allowing to investigate the distribution of grafted polymer inside the hydrogel in a native hydrated state. The microscopic techniques demonstrated uneven distribution of the grafted polymer inside the gel pores as a result of initiating the graft polymerization by insoluble initiator deposited on the pore surface.

  20. Tetrairon(III) Single Molecule Magnet Studied by Scanning Tunneling Microscopy and Spectroscopy

    Science.gov (United States)

    Oh, Youngtek; Jeong, Hogyun; Lee, Minjun; Kwon, Jeonghoon; Yu, Jaejun; Mamun, Shariful Islam; Gupta, Gajendra; Kim, Jinkwon; Kuk, Young

    2011-03-01

    Tetrairon(III) single-molecule magnet (SMM) on a clean Au(111) has studied using scanning tunneling microscopy (STM) and spectroscopy (STS) to understand quantum mechanical tunneling of magnetization and hysteresis of pure molecular origin. Before the STM studies, elemental analysis, proton nuclear magnetic resonance (NMR) measurement and Energy Dispersive X- ray Spectroscopy (EDS) were carried out to check the robustness of the sample. The STM image of this molecule shows a hexagonal shape, with a phenyl ring at the center and surrounding six dipivaloylmethane ligands. Two peaks are observed at 0.5 eV, 1.5 eV in the STS results, agreeing well with the first principles calculations. Spin-polarized scanning tunneling microscopy (SPSTM) measurements have been performed with a magnetic tip to get the magnetization image of the SMM. We could observe the antiferromagnetic coupling and a centered- triangular topology with six alkoxo bridges inside the molecule while applying external magnetic fields.

  1. Use of scanning electron microscopy to confirm the identity of lice infesting communally grazed goat herds

    Directory of Open Access Journals (Sweden)

    P.J. Sebei

    2004-11-01

    Full Text Available Lice have been described on goats in commercial farming systems in South Africa but not from flocks on communal grazing. During a longitudinal survey on the causes of goat kid mortality, conducted in Jericho district, North West Province, lice were collected from communally grazed indigenous goats. These lice were prepared for and viewed by scanning electron microscopy, and micromorphological taxonomic details are described. Three species of lice were found in the study area and identified as Bovicola caprae, Bovicola limbatus and Linognathus africanus. Sucking and biting lice were found in ten of the 12 herds of goats examined. Lice were found on both mature goats and kids. Bovicola caprae and L. africanus were the most common biting and sucking lice respectively in all herds examined. Scanning electron microscopy revealed additional features which aided in the identification of the louse species. Photomicrographs were more accurate aids to identification than the line drawings in the literature and facilitated identification using dissecting microscope.

  2. Use of scanning electron microscopy to confirm the identity of lice infesting communally grazed goat herds.

    Science.gov (United States)

    Sebei, P J; McCrindle, C M E; Green, E D; Turner, M L

    2004-06-01

    Lice have been described on goats in commercial farming systems in South Africa but not from flocks on communal grazing. During a longitudinal survey on the causes of goat kid mortality, conducted in Jericho district, North West Province, lice were collected from communally grazed indigenous goats. These lice were prepared for and viewed by scanning electron microscopy, and micro-morphological taxonomic details are described. Three species of lice were found in the study area and identified as Bovicola caprae, Bovicola limbatus and Linognathus africanus. Sucking and biting lice were found in ten of the 12 herds of goats examined. Lice were found on both mature goats and kids. Bovicola caprae and L. africanus were the most common biting and sucking lice respectively in all herds examined. Scanning electron microscopy revealed additional features which aided in the identification of the louse species. Photomicrographs were more accurate aids to identification than the line drawings in the literature and facilitated identification using dissecting microscope.

  3. In vivo visualization of microneedle conduits in human skin using laser scanning microscopy

    International Nuclear Information System (INIS)

    Bal, S; Kruithof, A C; Bouwstra, J; Liebl, H; Tomerius, M; Lademann, J; Meinke, M

    2010-01-01

    Solid microneedles enhance the penetration of drugs into the viable skin but little is known about the geometry of the conduits in vivo. Therefore, laser scanning microscopy was used to visualize the conduits of a microneedle system with needles at a length of 300 μm in 6 healthy subjects over a period of time. The model drug, a fluorescent dye was applied before and after piercing. Laser scanning microscopy was evaluated as being an excellent method to monitor the geometry and closure of the conduits over time. The used microneedle system was evaluated as suitable to enhance the transport of model drugs into the viable epidermis without bleeding and a short closure time of the conduits at the skin surface

  4. In vivo visualization of microneedle conduits in human skin using laser scanning microscopy

    Science.gov (United States)

    Bal, S.; Kruithof, A. C.; Liebl, H.; Tomerius, M.; Bouwstra, J.; Lademann, J.; Meinke, M.

    2010-03-01

    Solid microneedles enhance the penetration of drugs into the viable skin but little is known about the geometry of the conduits in vivo. Therefore, laser scanning microscopy was used to visualize the conduits of a microneedle system with needles at a length of 300 μm in 6 healthy subjects over a period of time. The model drug, a fluorescent dye was applied before and after piercing. Laser scanning microscopy was evaluated as being an excellent method to monitor the geometry and closure of the conduits over time. The used microneedle system was evaluated as suitable to enhance the transport of model drugs into the viable epidermis without bleeding and a short closure time of the conduits at the skin surface.

  5. Scanning Electron Microscopy of Nonconductive Specimens at Critical Energies in a Cathode Lens System

    Czech Academy of Sciences Publication Activity Database

    Frank, Luděk; Zadražil, Martin; Müllerová, Ilona

    2001-01-01

    Roč. 23, č. 1 (2001), s. 36-50 ISSN 0161-0457 R&D Projects: GA ČR GA202/96/0961; GA ČR GA202/99/0008 Institutional research plan: CEZ:AV0Z2065902 Keywords : scanning electron microscopy * specimen charging * nonconductive specimens Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 0.389, year: 2001

  6. Advanced scanning transmission stereo electron microscopy of structural and functional engineering materials

    Czech Academy of Sciences Publication Activity Database

    Agudo Jácome, L.; Eggeler, G.; Dlouhý, Antonín

    2012-01-01

    Roč. 122, NOV (2012), s. 48-59 ISSN 0304-3991 R&D Projects: GA ČR GA202/09/2073 Institutional research plan: CEZ:AV0Z20410507 Keywords : stereoscopy * scanning transmission electron microscopy * single crystal Ni-base superalloys * Dislocation substructures * Foil thickness measurement Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 2.470, year: 2012

  7. Imaging of block copolymer vesicles in solvated state by wet scanning transmission electron microscopy

    Czech Academy of Sciences Publication Activity Database

    Šlouf, Miroslav; Lapčíková, Monika; Štěpánek, M.

    2011-01-01

    Roč. 47, č. 6 (2011), s. 1273-1278 ISSN 0014-3057 R&D Projects: GA ČR GAP208/10/0353; GA AV ČR KAN200520704; GA AV ČR IAA400500703 Institutional research plan: CEZ:AV0Z40500505 Keywords : environmental scanning electron microscopy * self-assembly * amphiphilic block copolymers Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.739, year: 2011

  8. Effect of Autoclave Cycles on Surface Characteristics of S-File Evaluated by Scanning Electron Microscopy

    OpenAIRE

    Razavian, Hamid; Iranmanesh, Pedram; Mojtahedi, Hamid; Nazeri, Rahman

    2015-01-01

    Introduction: Presence of surface defects in endodontic instruments can lead to unwanted complications such as instrument fracture and incomplete preparation of the canal. The current study was conducted to evaluate the effect of autoclave cycles on surface characteristics of S-File by scanning electron microscopy (SEM). Methods and Materials: In this experimental study, 17 brand new S-Files (#30) were used. The surface characteristics of the files were examined in four steps (without autocla...

  9. Scanning electrochemical microscopy determination of hydrogen flux at liquid|liquid interface with potentiometric probe

    OpenAIRE

    Jedraszko, Justyna; Nogala, Wojciech; Adamiak, Wojciech; Girault, Hubert H.; Opallo, Marcin

    2014-01-01

    Scanning electrochemical microscopy potentiometric determination of local hydrogen concentration and its flux next to the liquid|liquid interface was demonstrated. This method is based on the shift of open circuit potential of Pt-based reversible hydrogen electrode. The detection system was verified with a system generating hydrogen under galvanostatic conditions. Then, it was applied to aqueous|1,2-dichloroethane interface where hydrogen is produced with decamethylferrocene as electron donor.

  10. Methods for studying biofilm formation: flow cells and confocal laser scanning microscopy

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim; Sternberg, Claus

    2014-01-01

    In this chapter methods for growing and analyzing biofilms under hydrodynamic conditions in flow cells are described. Use of flow cells allows for direct microscopic investigation of biofilm formation. The flow in these chambers is essentially laminar, which means that the biofilms can be grown u......, inoculation of the flow cells, running of the system, confocal laser scanning microscopy and image analysis, and disassembly and cleaning of the system....

  11. Use of scanning electron microscopy to confirm the identity of lice infesting communally grazed goat herds

    OpenAIRE

    P.J. Sebei; C.M.E. McCrindle; E.D. Green; M.L. Turner

    2004-01-01

    Lice have been described on goats in commercial farming systems in South Africa but not from flocks on communal grazing. During a longitudinal survey on the causes of goat kid mortality, conducted in Jericho district, North West Province, lice were collected from communally grazed indigenous goats. These lice were prepared for and viewed by scanning electron microscopy, and micromorphological taxonomic details are described. Three species of lice were found in the study area and identif...

  12. RGB color coded images in scanning electron microscopy of biological surfaces

    Czech Academy of Sciences Publication Activity Database

    Kofroňová, Olga; Benada, Oldřich

    2017-01-01

    Roč. 61, č. 3 (2017), s. 349-352 ISSN 0001-723X R&D Projects: GA MŠk(CZ) LO1509; GA ČR(CZ) GA16-20229S Institutional support: RVO:61388971 Keywords : Biological surfaces * Color image s * Scanning electron microscopy Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 0.673, year: 2016

  13. In-air scanning transmission ion microscopy of cultured cancer cells

    International Nuclear Information System (INIS)

    Ortega, R.; Deves, G.; Moretto, Ph.

    2001-01-01

    Scanning transmission ion microscopy (STIM) imaging of living cultured cells has been carried out using a proton external-beam with the nuclear microprobe of Bordeaux-Gradignan. STIM could be performed in air atmosphere after passage of a focused proton beam through a 150 nm thick silicon nitride window. Energy loss STIM images were obtained with a spatial resolution in the micrometer range and enabled the identification of sub-cellular ultrastructures

  14. Single Cell Oxygen Mapping (SCOM) by Scanning Electrochemical Microscopy Uncovers Heterogeneous Intracellular Oxygen Consumption

    OpenAIRE

    Santos, Carla Santana; Kowaltowski, Alicia J.; Bertotti, Mauro

    2017-01-01

    We developed a highly sensitive oxygen consumption scanning microscopy system using platinized platinum disc microelectrodes. The system is capable of reliably detecting single-cell respiration, responding to classical regulators of mitochondrial oxygen consumption activity as expected. Comparisons with commercial multi-cell oxygen detection systems show that the system has comparable errors (if not smaller), with the advantage of being able to monitor inter and intra-cell heterogeneity in ox...

  15. SCANNING ELECTRON MICROSCOPY STUDY OF THE DORSAL SURFACE OF THE TONGUE IN Chaetophractus vellerosus (MAMMALIA, DASYPODIDAE)

    OpenAIRE

    Estecondo, Silvia; Codón, Stella Maris; Casanave, Emma Beatriz

    2001-01-01

    The characteristics of the dorsal surface of Chaetophractus vellerosus tongue were studied by scanning electron microscopy. Simple or branched filiform, fungiform and vallate papillae are described. Simple conical filiform papillae appear in the apex, lateral edges and posterior third, caudally to the circumvallated ones. The branched papillae are densely distributed all over the dorsal surface of the lingual body. Fungiform ones are scattered among the branched filiform papillae. In the post...

  16. Examination of the Combustion Morphology of Ziconium Carbide Using Scanning Electron Microscopy

    OpenAIRE

    Newbold, Brian R.

    1997-01-01

    Calculation of viscous particle damping of acoustic combustion instability in solid propellant motors requires an understanding of the combustion behavior of added particles and oxides. A simple hydrogen/oxygen flame was used to ignite carefully sieved zirconium carbide particles which were impacted on slides at different levels below the burner. Scanning electron microscopy revealed that zirconium carbide has a complex heterogeneous combustion morphology. Initially, particles are partly v...

  17. Solid-state electrochemistry on the nanometer and atomic scales: the scanning probe microscopy approach

    Science.gov (United States)

    Strelcov, Evgheni; Yang, Sang Mo; Jesse, Stephen; Balke, Nina; Vasudevan, Rama K.; Kalinin, Sergei V.

    2016-01-01

    Energy technologies of the 21st century require understanding and precise control over ion transport and electrochemistry at all length scales – from single atoms to macroscopic devices. This short review provides a summary of recent works dedicated to methods of advanced scanning probe microscopy for probing electrochemical transformations in solids at the meso-, nano- and atomic scales. Discussion presents advantages and limitations of several techniques and a wealth of examples highlighting peculiarities of nanoscale electrochemistry. PMID:27146961

  18. Phosphorus ligand imaging with two-photon fluorescence spectroscopy: towards rational catalyst immobilization

    NARCIS (Netherlands)

    Marras, F.; Kluwer, A.M.; Siekierzycka, J.R.; Vozza, A.; Brouwer, A.M.; Reek, J.N.H.

    2010-01-01

    Spotless catalysts: Ligand immobilization was studied by two-photon fluorescence microscopy with a fluorescent nixantphos ligand as probe (see picture). In the immobilization process ligand aggregates form in solution and are deposited on the support, where they appear as bright spots in

  19. Analytical procedure for experimental quantification of carrier concentration in semiconductor devices by using electric scanning probe microscopy

    International Nuclear Information System (INIS)

    Fujita, Takaya; Matsumura, Koji; Itoh, Hiroshi; Fujita, Daisuke

    2014-01-01

    Scanning capacitance microscopy (SCM) is based on a contact-mode variant of atomic force microscopy, which is used for imaging two-dimensional carrier (electrons and holes) distributions in semiconductor devices. We introduced a method of quantification of the carrier concentration by experimentally deduced calibration curves, which were prepared for semiconductor materials such as silicon and silicon carbide. The analytical procedure was circulated to research organizations in a round-robin test. The effectiveness of the method was confirmed for practical analysis and for what is expected for industrial pre-standardization from the viewpoint of comparability among users. It was also applied to other electric scanning probe microscopy techniques such as scanning spreading resistance microscopy and scanning nonlinear dielectric microscopy. Their depth profiles of carrier concentration were found to be in good agreement with those characterized by SCM. These results suggest that our proposed method will be compatible with future next-generation microscopy. (paper)

  20. Electron transparent graphene windows for environmental scanning electron microscopy in liquids and dense gases.

    Science.gov (United States)

    Stoll, Joshua D; Kolmakov, Andrei

    2012-12-21

    Due to its ultrahigh electron transmissivity in a wide electron energy range, molecular impermeability, high electrical conductivity and excellent mechanical stiffness, suspended graphene membranes appear to be a nearly ideal window material for in situ (in vivo) environmental electron microscopy of nano- and mesoscopic objects (including bio-medical samples) immersed in liquids and/or in dense gaseous media. In this paper, taking advantage of a small modification of the graphene transfer protocol onto metallic and SiN supporting orifices, reusable environmental cells with exchangeable graphene windows have been designed. Using colloidal gold nanoparticles (50 nm) dispersed in water as model objects for scanning electron microscopy in liquids as proof of concept, different conditions for imaging through the graphene membrane were tested. Limiting factors for electron microscopy in liquids, such as electron beam induced water radiolysis and damage of the graphene membrane at high electron doses, are discussed.