The development of biological preparative techniques for light microscopy, 1839–1989

The development of techniques of preparing biological material for microscopical observation is summarized, with much greater emphasis being given to the introduction and establishment of the various techniques, largely in the nineteenth century, than to their perfection and extremely widespread use in the twentieth century. The development of the microtome as an instrument of the greatest importance is summarized and an assessment is made of the impact of preparative techniques on the development of the medical sciences.     

Nonlinear ultrasonic techniques for nondestructive assessment of micro damage in material: A review

The nondestructive assessment of the damage that occurs in components during service plays a key role for condition monitoring and residual life estimation of in-service components/structures. Ultrasound has been widely utilized for this; however most of these conventional methods using ultrasonic characteristics in the linear elastic region are only sensitive to gross defects but much less sensitive to micro-damage. Recently, the nonlinear ultrasonic technique, which uses nonlinear ultrasonic behavior such as higher-harmonic generation, subharmonic generation, nonlinear resonance, or mixed frequency response, has been studied as a positive method for overcoming this limitation. In this paper, overall progress in this technique is reviewed with the brief introduction of basic principle in the application of each nonlinear ultrasonic phenomenon.     

Microscopic techniques for characterization and authentication of oil-yielding seeds

Investigation of alternative energy sources is need of current time due to growing power crisis and associated environmental issues. Biodiesel is considered as sustainable power source and promising alternative to fossil fuels. Therefore, our current investigation aimed to identify micromorphological characters of 10 novel nonedible oil-yielding seeds through scanning electron microscopy. It was revealed from light microscopic study that there is variation in seed size from 3 to 15 mm in length and 2 to 11 mm in width. Likewise, a huge variation in color was observed such as light green, greenish yellow, blackish brown, and various shades of brown. Presence and absence of Hilum was observed, and compression of seeds varied from depressed, lateral, and dorsoventral. Seed's shape differs from ovate, clavate, triangular ovate, cuneiform, ovoid, and elliptical shape. Seed oil content fall in range of 18–58% (wt/wt). Free fatty acid content of the seeds varies from 0.3 to 3.1 mg KOH/g. Ultrastructure of seeds exhibited huge variation in shape, size, periclinal wall, anticlinal wall, and surface ornamentation. Nonedible seeds varied in wall structure from angular, wavy, dentate entire, irregular, puzzled, elongated, even, and polygonal. The periclinal wall arrangements show alteration from flat, looped, raised, depressed, lofty, even, pentagonal, polygonal, and undulate seed margins. Outcomes of this investigation recommended that scanning electron microscopy could act as a helpful tool in disclosing the hidden micromorphological characters among nonedible oil-yielding seeds and subsequently helping in correct, authentic seed identification and classification as potential feedstock for biodiesel.     

Any-degrees-of-freedom (anyDOF) registration for the characterization of freeform surfaces

This paper presents an any-degrees-of-freedom (anyDOF) registration method for the characterization of freeform surfaces. The method attempts to fill the research gap regarding traditional surface registration methods which are normally dedicated to solving the global optimization problem with all DOF but they lack flexibility. The proposed anyDOF method is capable of registering surfaces with any specified combination of DOF. This is particularly useful when some of the DOF are known to be unchanged according to the a priori knowledge. The anyDOF surface registration method is regarded as a typical optimization problem of finding the minimum distance from target surface to the reference surface, with constraints of the unwanted DOF. The problem is solved by the Levenberg-Marquardt method. Simulated experiments for a two-dimensional (2D) profile and a three-dimensional (3D) surface were undertaken, together with three measurement experiments including a fluid-jet polished surface, a bonnet polished surface and a diamond machined freeform surface. Experimental results show that the anyDOF registration method is highly flexible in the characterization of freeform surfaces.     

Novel techniques of preparing TEM samples for characterization of irradiation damage

Focus ion beam preparation of transmission electron microscopy (TEM) samples has become increasingly popular due to the relative ease of extraction of TEM foils from specific locations within a larger sample. However the sputtering damage induced by Ga ion bombardment in focus ion beam means that traditional electropolishing may be a preferable method. First, we describe a special electropolishing method for the preparation of irregular TEM samples from ex‐service nuclear reactor components, spring‐shaped spacers. This method has also been used to prepare samples from a nonirradiated component for a TEM in situ heavy ion irradiation study. Because the specimen size is small (0.7 × 0.7 × 3 mm), a sandwich installation is adopted to obtain high quality polishing. Second, we describe some modifications to a conventional TEM cross‐section sample preparation method that employs Ni electroplating. There are limitations to this method when preparing cross‐section samples from either (1) metals which are difficult to activate for electroplating, or (2) a heavy ion irradiated foil with a very shallow damage layer close to the surface, which may be affected by the electroplating process. As a consequence, a novel technique for preparing cross‐section samples was developed and is described.     

Structural characterization of bacterial lipopolysaccharides with mass spectrometry and on‐ and off‐line separation techniques

The focus of this review is the application of mass spectrometry to the structural characterization of bacterial lipopolysaccharides (LPSs), also referred to as “endotoxins,” because they elicit the strong immune response in infected organisms. Recently, a wide variety of MS‐based applications have been implemented to the structure elucidation of LPS. Methodological improvements, as well as on‐ and off‐line separation procedures, proved the versatility of mass spectrometry to study complex LPS mixtures. Special attention is given in the review to the tandem mass spectrometric methods and protocols for the analyses of lipid A, the endotoxic principle of LPS. We compare and evaluate the different ionization techniques (MALDI, ESI) in view of their use in intact R‐ and S‐type LPS and lipid A studies. Methods for sample preparation of LPS prior to mass spectrometric analysis are also described. The direct identification of intrinsic heterogeneities of most intact LPS and lipid A preparations is a particular challenge, for which separation techniques (e.g., TLC, slab‐PAGE, CE, GC, HPLC) combined with mass spectrometry are often necessary. A brief summary of these combined methodologies to profile LPS molecular species is provided. © 2012 Wiley Periodicals, Inc., Mass Spec Rev 32:90–117, 2013     

Experimental methods for local-scale characterization of hydro-morphodynamic dam breach processes. Breach detection, 3D reconstruction,flow kinematics and spatial surface velocimetry

A set of tests on failure by overtopping of earth dams was performed under hydraulic and geotechnical controlled conditions in a medium-scale facility located at the Portuguese National Laboratory for Civil Engineering. The flow over the breaching dam is three-dimensional and exhibits several length scales. Instrumentation must be able to capture this complexity and scale amplitude and must be non-intrusive. This work presents a summary of the instrumentation layout as well as its main components and measuring methods that were used and/or developed to address these issues. The emphasis is placed on post-processing analysis of digital imagery, for which there is detailed exposition of the adopted methods and an evaluation of its suitability for its intended purpose.     

极紫外多层膜基底表面粗糙度综合表征技术

研究用不同清洗方法对硅基底表面粗糙度的影响,首先使用原子力显微镜(AFM)直接测量硅片表面粗糙度;然后,利用直流磁控溅射方法,在相同制备工艺条件下镀制M o/S i多层膜,使用X射线衍射仪(XRD)对多层膜二级衍射峰进行摇摆曲线扫描;最后,利用同步辐射测量多层膜的反射率,间接表征基底的粗糙度。结果表明,超声清洗后镀制的多层膜反射率最高,结论与AFM,XRD等表征方法一致。     

Complementary techniques for the characterization of thin film Ti/Nb multilayers

An aberration corrector on the probe-forming lens of a scanning TEM (STEM) equipped with an electron energy-loss spectrometer (EELS) and X-ray energy-dispersive spectrometer (XEDS) has been employed to investigate the compositional variations as a function of length scale in nanoscale Ti/Nb metallic multilayers. The composition profiles of EELS and XEDS were compared with the profiles obtained from the complementary technique of 3D atom probe tomography. At large layer widths (h≥7 nm, where h is the layer width) of Ti and Nb, XEDS composition profiles of Ti/Nb metallic multilayers are in good agreement with the EELS results. However, at reduced layer widths (h≈2 nm), profiles of EELS and atom probe exhibited similar compositional variations, whereas XEDS results have shown a marked difference. This difference in the composition profiling of the layers has been addressed with reference to the effects of beam broadening and the origin of the signals collected in these techniques. The advantage of using EELS over XEDS for these nanoscaled multilayered materials is demonstrated.     

Detailed characterisation of sulphur and nitrogen components in a multifunctional,ashless lubricant additive by spectroscopic and chromatographic techniques

The work reported here used infrared (IR), multinuclear nuclear magnetic resonance (NMR), and chromatographic techniques to identify and characterise a commercial sulphur‐ and nitrogen‐containing ashless multifunctional additive. The proper assignment of signals in the ¹H/¹³C NMR and IR spectra of the sample has facilitated the identification of different types of sulphur‐ and nitrogen‐containing components. The methodology involves investigation by NMR and IR spectroscopies, thin‐layer chromatography (TLC) and high‐performance liquid chromatogaphy (HPLC) techniques to elucidate the types of components present. This requires the generation of NMR and IR spectral data for standard compounds of alkyl sulphides with different sulphur content and alkyl chain, and nitrogen and sulphur components such as thiadiazole, imidazoline, triazole, etc., and spectral comparison with the spectra of the unknown sample. Further, these components have been separated by silica/alumina column chromatographic and preparative TLC techniques and subsequently analysed for their exact chemical structure by spectroscopic techniques. The combined spectroscopic and chromatographic analyses have yielded the presence of four types of components: di‐t‐octyl polysulphide, di‐t‐octyl thiadiazole, a component containing amine ether functionality, and alkyl amine salts of mono/dialkyl phosphoric acid.     

New application of air-drying techniques for studying Ephemeroptera and Plecoptera eggs by scanning electron microscopy

Hexamethyldisilizane (HMDS) and tetramethylsilane are organic compounds that are volatile at ambient temperature and which can therefore be used for air-drying biological samples for SEM studies. The techniques using these compounds provide results that are comparable with those obtained by critical point drying, but which involve a very simple process that saves time and money. Both techniques were applied to SEM studies of Ephemeroptera and Plecoptera eggs in order to assess their suitability as alternative methods to critical point drying for these kinds of biological material. The results show no morphological differences between eggs HMDS air-dried and critical point-dried.     

A method for the characterization of surface-modified asbestos fibres by electron energy-loss spectroscopy and electron spectroscopic imaging

A method for the characterization of surface-treated asbestos fibres with electron microscopy is presented. Electron spectroscopic imaging (ESI) of organosilane-treated chrysotile asbestos fibres has been carried out. Initially, the region below the carbon edge was inspected in ESI mode for its effectiveness as a background correction. Elemental mapping was performed on standard untreated fibres to take into account non-characteristic signals from extrapolation errors and camera artefacts. The highest resulting pixel value that results from non-characteristic signals was used as a threshold for further background correction in the net images. Samples for electron energy-loss spectroscopy were prepared in two different ways, either by gluing on grids, or by using perforated carbon foils. The results show that the use of a conducting carbon film is necessary for the analysis of such electrically insulating asbestos fibres. Focusing of the electron beam on the individual fibres results in a thermal effect promoting the evaporation of the organosilane reaction products.     

Structural characterization of TiN/NbN multilayers: X-ray diffraction, energy-filtered TEM and Fresnel contrast techniques compared

Two TiN/NbN multilayers with wavelength 13.6 and 6.15 nm have been characterized by X-ray diffraction (XRD), Fresnel contrast analysis (FCA) and energy-filtered transmission electron microscopy (EFTEM). Good agreement between the composition profile obtained by FCA and EFTEM is obtained if the lower resolution of the EFTEM images is taken into account. The relative advantages and disadvantages of the techniques are discussed. Used together the two TEM techniques provide a quantitative characterization that is consistent with, and for some parameters provides more precise values than, that from XRD. The analysis shows that the multilayers have narrow interfaces (< 1 nm) and a composition amplitude close to 95% for the longer wavelength.     

An innovative,interdisciplinary, and multi‐technique study of gilding and painting techniques in the decoration of the main altarpiece of Miranda do Douro Cathedral (XVII‐XVIIIth centuries,Portugal)

The research results presented in this paper are part of a larger study on the materials and techniques used in polychrome altarpieces of gilded woodcarving decoration (“talha dourada”) in Portugal. The paper focuses on a narrative Portuguese Altarpiece from Miranda do Douro, considered one of the masterpieces of “talha dourada” among all the retables of the Iberian Peninsula in XVII^th and XVIII^th centuries. Although on the Portuguese territory, the altarpiece was made by artists from the Royal Spanish school of Valladolid, under a mannerist style. Thus the study opens a window on the artists' circulation between Spain and Portugal and influences of the Spanish schools in Baroque epoch on the Portuguese “talha”. During its history this altarpiece underwent several transformations and extensive conservation treatments in 1989. On this occasion more than 50 samples were collected and analyzed using an interdisciplinary multi‐technique methodology. 27 of these samples are chosen for this study in order to investigate the chromatic palette, the materials and techniques used in the polychromy of the retable. A novel protocol of investigation using different conventional and unconventional analytical techniques (OM + fluorescent staining tests on cross‐sections, Raman microscopy, XRD, XRF, X‐ray micro‐CT, SEM‐EDX, MALDI‐TOF‐MS and LC‐MS/MS) was established within an innovative research project ( http://sites.fct.unl.pt/gilt‐teller/ ) and applied on these samples. This protocol is necessary to confirm the results obtained in the 1989 campaign and to have further insight into the gilding and polychrome decoration materials and techniques and the additional information reported in the historical documents. The material and technical history of this important altarpiece will be thus re‐documented from a scientific perspective, meant to confirm and bring new information on the decorative technique used in the creation of this complex Portuguese monument. Microsc. Res. Tech. 76:733–743, 2013. © 2013 Wiley Periodicals, Inc.     

Morphological characterization of Hypnaceae (Bryopsida,Hypnales): Investigating four genera from Western Himalayas by using LM and SEM techniques

The Hypnaceae is one of the largest and diversified family among the pleurocarpous mosses which consists of nearly 60 genera and 1,000 species around the world. In Pakistan, it is represented by 15 species and 8 genera. The current research article provides the detail morphological investigation of four different species (genera) of the family Hypnaceae that is, Gollania clarescens, Hypnum revolutum, Homomallium andoi, and Taxiphyllum taxirameum in the Western Himalayas, Pakistan. The research article reports the first SEM study of the leaf surfaces of the studied taxa, along with comprehensive morphological characters of the four species. The main objective of the research project is to present the comparative light and scanning electron microscopic study to discuss the morphology in detail because previously the family is just reported in different bryophyte checklists of the Western Himalayas. Based on results, morphological characters, micromorphological observations, qualitative and quantitative attributes, taxonomic keys for the studied taxa, distribution, and habitat are described. Stereoscope was used to analyze different qualitative characters, and light microscope was used to observe and measure the laminal and alar cells of the leaf. The SEM study reveals many important surface features like cell shape and cell wall. The laminal cells were mostly linear and elongated with thin cell walls. The specimens used for the SEM were air dried, so the laminal cells were somewhat constricted and concaved. The current study project will help to make the contribution in the taxonomy and morphology of this family.     

Scope of in-process measurement, monitoring and control techniques in machining processes—Part 3: In-process techniques for cutting processes and machine tools

In-process techniques for cutting processes and machine tools are surveyed through representative examples. Detection of cutting force and chatter vibration are especially important in cutting processes. As for machine tools, four measuring items will be important which include the driving system, the bearing and rotating systems, the temperature control system, and the monitoring system. They are the primary factors to be taken into consideration for achieving high machining accuracy.