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Journal of Microscopy

P Xiu, Q Liu, X Zhou, Y Xu, C Kuang, X Liu
The refractive index (RI) of a sample as an endogenous contrast agent plays an important role in transparent live cell imaging. In tomographic phase microscopy (TPM), 3D quantitative RI maps can be reconstructed based on the measured projections of the RI in multiple directions. The resolution of the RI maps not only depends on the numerical aperture of the employed objective lens, but also is determined by the accuracy of the quantitative phase of the sample measured at multiple scanning illumination angles...
January 11, 2018: Journal of Microscopy
E Hasani, J Parravicini, L Tartara, A Tomaselli, D Tomassini
We propose an innovative experimental approach to estimate the two-photon absorption (TPA) spectrum of a fluorescent material. Our method develops the standard indirect fluorescence-based method for the TPA measurement by employing a line-shaped excitation beam, generating a line-shaped fluorescence emission. Such a configuration, which requires a relatively high amount of optical power, permits to have a greatly increased fluorescence signal, thus avoiding the photon counterdetection devices usually used in these measurements, and allowing to employ detectors such as charge-coupled device (CCD) cameras...
January 11, 2018: Journal of Microscopy
D Marques, A Miranda, A G Silva, P R T Munro, P A A DE Beule
Some implementations of interference microscopy imaging use digital holographic measurements of complex scattered fields to reconstruct three-dimensional refractive index maps of weakly scattering, semi-transparent objects, frequently encountered in biological investigations. Reconstruction occurs through application of the object scattering potential which assumes an isotropic refractive index throughout the object. Here, we demonstrate that this assumption can in some circumstances be invalid for biological imaging due to the presence of lipid-induced optical anisotropy...
January 11, 2018: Journal of Microscopy
L Jones, A Varambhia, H Sawada, P D Nellist
In the scanning transmission electron microscope, an accurate knowledge of detector collection angles is paramount in order to quantify signals on an absolute scale. Here we present an optical configuration designed for the accurate measurement of collection angles for both image-detectors and energy-loss spectrometers. By deflecting a parallel electron beam, carefully calibrated using a diffraction pattern from a known material, we can directly observe the projection-distortion in the post-specimen lenses of probe-corrected instruments, the 3-fold caustic when an image-corrector is fitted, and any misalignment of imaging detectors or spectrometer apertures...
January 8, 2018: Journal of Microscopy
Y Wang, C Wang, Z Zhang
Automated cell segmentation plays a key role in characterisations of cell behaviours for both biology research and clinical practices. Currently, the segmentation of clustered cells still remains as a challenge and is the main reason for false segmentation. In this study, the emphasis was put on the segmentation of clustered cells in negative phase contrast images. A new method was proposed to combine both light intensity and cell shape information through the construction of grey-weighted distance transform (GWDT) within preliminarily segmented areas...
December 27, 2017: Journal of Microscopy
D Lee, D-G Gweon, H Yoo
We propose a multipoint scanning dual-detection confocal microscopy (MS-DDCM) system for fast 3D volumetric measurements. Unlike conventional confocal microscopy, MS-DDCM can accomplish surface profiling without axial scanning. Also, to rapidly obtain 2D images, the MS-DDCM employs a multipoint scanning technique, with a digital micromirror device used to produce arrays of effective pinholes, which are then scanned. The MS-DDCM is composed of two CCDs: one collects the conjugate images and the other collects nonconjugate images...
December 18, 2017: Journal of Microscopy
Y Dauphin, G Luquet, M Salome, L Bellot-Gurlet, J P Cuif
Mollusc shells are complex organomineral structures, the arrangement and composition depending on the species. Most studies are dedicated to shells composed of an aragonite nacreous and a calcite prismatic layer, so the nacreous prismatic model based on Pinctada and Atrina-Pinna. Here, we studied the micro- and nanostructure, the mineralogy and composition of a nacroprismatic bivalve species: Unio pictorum. The prismatic layer of Unio is aragonite, and the inner structure of the prismatic units strongly differs from those of the calcitic layers...
December 14, 2017: Journal of Microscopy
P Liu, J-Y Zhu, B Tang, Z-C Hu
This study describes how three-dimensional (3D) human skin tissue is reconstructed, and provides digital anatomical data for the physiological structure of human skin tissue based on large-scale thin serial sections. Human skin samples embedded in paraffin were cut serially into thin sections and then stained with hematoxylin-eosin. Images of serial sections obtained from lighting microscopy were scanned and aligned by the scale-invariant feature transform algorithm. 3D reconstruction of the skin tissue was generated using Mimics software...
December 14, 2017: Journal of Microscopy
T J Deerinck, T M Shone, E A Bushong, R Ramachandra, S T Peltier, M H Ellisman
A longstanding limitation of imaging with serial block-face scanning electron microscopy is specimen surface charging. This charging is largely due to the difficulties in making biological specimens and the resins in which they are embedded sufficiently conductive. Local accumulation of charge on the specimen surface can result in poor image quality and distortions. Even minor charging can lead to misalignments between sequential images of the block-face due to image jitter. Typically, variable-pressure SEM is used to reduce specimen charging, but this results in a significant reduction to spatial resolution, signal-to-noise ratio and overall image quality...
December 1, 2017: Journal of Microscopy
M Petaccia, G Castellano
In electron probe microanalysis, secondary fluorescence can occur leading to an increase of the volume analysed, degrading the lateral resolution of this technique. An adequate knowledge of the interaction volumes from where the different signals of interest are detected is determinant to estimate the minimum size of the zone that can be characterized. In this work, the size of the signal source volume is surveyed for a wide set of samples at different beam energies. To this aim, the PENELOPE software package was chosen to run Monte Carlo simulations for several experimental situations in order to produce the various lateral radiation distributions of interest...
November 27, 2017: Journal of Microscopy
Y Yao, X He, C Liu, Jianqiang Zhu
A new coded splitting imaging technique is proposed to reconstruct the complex amplitude of a light field iteratively using a single-shot measurement. In this technique, a specially designed coded splitting plate is adopted to diffract the illuminating beam into multiple beams of different orders and code their wavefronts independently and differently. From the diffraction pattern array recorded on the detector plane, both the modulus and phase distributions of the illuminating beam can be reconstructed iteratively using known transmission functions of different orders of the coded splitting plate...
November 10, 2017: Journal of Microscopy
Keiichi Moriguchi
When polymorphonuclear leukocytes (PMNs) phagocytose opsonised zymosan particles (OPZ), free radicals and reactive oxygen species (ROS) are formed in the phagosomes. ROS production is mediated by NADPH oxidase (Nox), which transfers electrons in converting oxygen to superoxide (O2(-) ). Nox-generated O2(-) is rapidly converted to other ROS. Free radical-forming secretory vesicles containing the Nox redox center flavocytochrome b558, a membrane protein, and azurophil granules with packaged myeloperoxidase (MPO) have been described...
November 10, 2017: Journal of Microscopy
Brian Dorvel, Praveenkumar Boopalachandran, Ida Chen, Andrew Bowling, Kerry Williams, Steve King
Decking is one of the largest applications for the treated wood market. The most challenging property to obtain for treated wood is dimensional stability, which can be achieved, in part, by cell wall bulking, cell wall polymer crosslinking and removal of hygroscopic components in the cell wall. A commonly accepted key requirement is for the actives to infuse through the cell wall, which has a microporosity of ∼5-13 nm. Equally as challenging is being able to measure and quantify the cell wall penetration...
November 8, 2017: Journal of Microscopy
I Ipatova, P T Wady, S M Shubeita, C Barcellini, A Impagnatiello, E Jimenez-Melero
The formation of radiation-induced dislocation loops and voids in tantalum at 180(2), 345(3) and 590(5)°C was assessed by 3MeV proton irradiation experiments and subsequent damage characterisation using transmission electron microscopy. Voids formed at 345(3)°C and were arranged into a body centred cubic lattice at a damage level of 0.55 dpa. The low vacancy mobility at 180(2)°C impedes enough vacancy clustering and therefore the formation of voids visible by TEM. At 590(5)°C the Burgers vector of the interstitial-type dislocation loops is a<100>, instead of the a/2 <111> Burgers vector characteristic of the loops at 180(2) and 345(3)°C...
November 1, 2017: Journal of Microscopy
A A Sheader, A M Varambhia, R A Fleck, S J L Flatters, P D Nellist
The chemotherapeutics cisplatin and oxaliplatin are important tools in the fight against cancer. Both compounds are platinum complexes. Aberration-corrected scanning transmission electron microscopy using the annular dark-field imaging mode now routinely provides single-atom sensitivity with atomic number contrast. Here, this imaging mode is used to directly image the platinum within the two drugs in their dried form on an amorphous carbon support film. The oxaliplatin is found to have wetted the supporting amorphous carbon, forming disordered clusters suggesting that the platinum has remained within the complex...
November 1, 2017: Journal of Microscopy
D Westhoff, D P Finegan, P R Shearing, V Schmidt
We describe a segmentation algorithm that is able to identify defects (cracks, holes and breakages) in particle systems. This information is used to segment image data into individual particles, where each particle and its defects are identified accordingly. We apply the method to particle systems that appear in Li-ion battery electrodes. First, the algorithm is validated using simulated data from a stochastic 3D microstructure model, where we have full information about defects. This allows us to quantify the accuracy of the segmentation result...
October 26, 2017: Journal of Microscopy
J Kuva, J Sammaljärvi, J Parkkonen, M Siitari-Kauppi, M Lehtonen, T Turpeinen, J Timonen, M Voutilainen
We set out to study connected porosity of crystalline rock using X-ray microtomography and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDS) with caesium chloride as a contrast agent. Caesium is an important radionuclide regarding the final deposition of nuclear waste and also forms dense phases that can be readily distinguished by X-ray microtomography and SEM-EDS. Six samples from two sites, Olkiluoto (Finland) and Grimsel (Switzerland), where transport properties of crystalline rock are being studied in situ, were investigated using X-ray microtomography and SEM-EDS...
October 26, 2017: Journal of Microscopy
K Dembele, S Moldovan, Ch Hirlimann, J Harmel, K Soulantica, P Serp, B Chaudret, A-S Gay, S Maury, A Berliet, A Fecant, O Ersen
In situ transmission electron microscopy (TEM) of samples in a controlled gas environment allows for the real time study of the dynamical changes in nanomaterials at high temperatures and pressures up to the ambient pressure (10(5) Pa) with a spatial resolution close to the atomic scale. In the field of catalysis, the implementation and quantitative use of in situ procedures are fundamental for a better understanding of the behaviour of catalysts in their environments and operating conditions. By using a microelectromechanical systems (MEMS)-based atmospheric gas cell, we have studied the thermal stability and the reactivity of crystalline cobalt nanostructures with initial 'urchin-like' morphologies sustained by native surface ligands that result from their synthesis reaction...
October 24, 2017: Journal of Microscopy
B R Gordon, D E Martin, K R Bambery, C A Motti
The symbiotic relationship between corals and Symbiodinium spp. is the key to the success and survival of coral reef ecosystems the world over. Nutrient exchange and chemical communication between the two partners provides the foundation of this key relationship, yet we are far from a complete understanding of these processes. This is due, in part, to the difficulties associated with studying an intracellular symbiosis at the small spatial scales required to elucidate metabolic interactions between the two partners...
October 24, 2017: Journal of Microscopy
R O'Connell, Y Chen, H Zhang, Y Zhou, D Fox, P Maguire, J J Wang, C Rodenburg
Images of Ga(+) -implanted amorphous silicon layers in a 110 n-type silicon substrate have been collected by a range of detectors in a scanning electron microscope and a helium ion microscope. The effects of the implantation dose and imaging parameters (beam energy, dwell time, etc.) on the image contrast were investigated. We demonstrate a similar relationship for both the helium ion microscope Everhart-Thornley and scanning electron microscope Inlens detectors between the contrast of the images and the Ga(+) density and imaging parameters...
December 2017: Journal of Microscopy
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