A new method based on image analysis for determining cyanobacterial biomass by CLSM in stratified benthic sediments

Cyanobacteria are the dominant primary producers in microbial mats, which are stratified benthic microbial ecosystems found in coastal environments. Some cyanobacteria form long filaments, which make difficult to apply classical methods to estimate their biomass because they establish strong interactions with detritic particles. In a previous study, we described a method for determining cyanobacterial biomass by means of confocal laser scanning microscopy (CLSM). However, the manual method used, based on summa projection images, was difficult to apply when analyzing a large number of samples. In this paper, we described a new automated method, based on stacks and applying the plugin voxel counter in the ImageJ analysis system, more adequate for obtaining biomass data quickly from a large number of CLSM images.     

Extraglandular and intraglandular vascularization of canine prostate

The literature on the vascularization of the canine prostate is reviewed and the clinical significance of prostate morphology is described. Scanning Electron Microscopy (SEM), combined with improved corrosion casting methods, reveal new morphological details that promise better diagnostics and treatment but also require expansion of clinical nomenclature. A proposal is made for including two previously unnamed veins in Nomina Anatomica Veterinaria (NAV). The canine prostate has two lobes with independent vascularization. Each lobe is supplied through the left and right a. prostatica, respectively. The a. prostatica sprouts three small vessels (cranial, middle, and caudal) towards the prostate gland. A. prostatica is a small-size artery whose wall structure is similar to the arteries of the muscular type. V. prostatica is a small-size valved vein. The canine prostate has capsular, parenchymal, and urethral vascular zones. The surface vessels of the capsule are predominantly veins and the diameter of arterial vessels is larger than that of the veins. The trabecular vessels are of two types: direct and branched. The prostate parenchyma is supplied by branches of the trabecular vessels. The periacinary capillaries are fenestrated and form a net in a circular pattern. The processes of the myoepithelial cells embrace both the acins and the periacinar capillaries. In the prostate ductal system. there are spermatozoa. The prostatic part of the urethra is supplied by an independent branch of a. prostatica. The prostatic urethral part is drained by v. prostatica, the vein of the urethral bulb and the ventral prostate veins. M. urethralis begins as early as the urethral prostatic part. The greater part of the white muscle fibers in m. urethralis suggest an enhanced anaerobic metabolism.     

Penetration pathways of fluorescent dyes in human hair fibres investigated by scanning near-field optical microscopy

Thin cross-sections of human hairs were investigated by scanning near-field optical microscopy (SNOM) and confocal laser scanning microscopy (CLSM) after penetration of a fluorescent dye. The same samples were measured with both techniques to compare the observed structures. The images obtained from the two methods show nearly identical structures representing pathways of the dye molecules in hairs. The SNOM images provide a higher resolution than the CLSM images. Therefore, SNOM is believed to be a suitable method for investigations at a resolution of 100 nm on penetration pathways of fluorescent dyes such as the cell membrane complex pathway in cross-sections of hairs.     

Floc architecture of bioflocculation sediment by ESEM and CLSM

Sediment flocculation is a critical component for the understanding of cohesive sediment dynamics. Traditionally, the referred study has largely been devoted to forming mechanism, influencing factors and physicochemical sediment conditions of all kinds of organic-flocculation and inorganic-flocculation. However, during the last decade, the bioflocculation of sediment by biological activity has been given increasing attention. But most studies have focused on the interrelations between biological and sedimentological variables. With the assistance of a newly developed field kit and correlative microscopy (which includes environmental scanning electron microscopy and confocal laser scanning microscopy), this article begins to bridge the resolution gap between sediment particles and biological activities as well as its metabolic products biofilm, in order to better understand the role of polymeric material biofilm in floc ultrastructure and outward floc behavior of bioflocculation sediment. Results have demonstrated that bioflocculation sediment was observed to be composed of complex networks of biofilm and appeared to be of complicated physical floc structures. The biofilm was found to embed particles and permeate the void space, representing the dominant physical bridging mechanism of the flocs and contributed to the extensive surface area, architecture characteristics, and mechanical properties of bioflocculation sediment.     

Adsorption properties of aqueous methylene blue on mesocellular foam silica: Equilibrium,kinetic, isotherm,and thermodynamic characterization

Abstract

Mesocellular foam silica was prepared by a hydrothermal synthesis protocol. The surface and pore structure of mesocellular foam silica were characterized by low temperature nitrogen adsorption isotherms, X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and Fourier transform infrared (FT-IR) spectroscopy. The methylene blue was adsorbed by the synthetic mesocellular foam silica; the optimized adsorption conditions were evaluated and the maximum adsorption capacity of methylene blue by mesocellular foam silica was determined to be 215.5?mg g^?1. The kinetics of the adsorption of methylene blue by mesocellular foam silica were in accordance with a quasi-second-order kinetic equation. The results were analyzed by Langmuir and Freundlich adsorption isotherm models. The adsorption of methylene blue on mesocellular foam silica was shown to follow the Freundlich adsorption isotherm model. The Gibbs free energy change during adsorption showed that this process was spontaneous. The enthalpy change in the process was –28.868?J·mol^?1 K^?1, indicating that the adsorption is exothermic. The negative value of entropy –49.296?J·mol^?1 K^-1 shows that the system disorder decreases due to adsorption.     

Palynological characteristics of some selected species of family Cucurbitaceae (Juss.) using light and scanning electron microscopy techniques

The pollen morphology of 11 species distributed in seven genera belonging to family Cucurbitaceae has been studied using light microscopy (LM) as well as scanning electron microscopy. Pollen morphology of this family is diverse and is almost europalynous. Nearly all types of pollen belonging to different size class and shape class with a variety of exine sculpturing are found in the family. Pollen grains are usually triporate to tricolporate, prolate, oblate‐spheroidal, spheroidal, to suboblate, radially symmetrical, and isopolar. The common exine pattern is reticulate of various forms ranging from finely reticulate to coarsely reticulate with some species having echinate type of exine ornamentation, for example, Cucurbita pepo and Praecitrullus fistulosus. Most of the studied species are somewhat triangular with respect to equatorial outlook and circular to elliptic in polar outlook. The palynological characteristics of the members of the family are useful in identification of taxa at generic level mostly. Based on the present study it is not recommended to wholly solely depend upon the pollen morphology for the identification of taxa especially at specific levels because very minor differences regarding pollen characters exist among the species of the same genera.     

Visualization and characterization of engineered nanoparticles in complex environmental and food matrices using atmospheric scanning electron microscopy

Imaging and characterization of engineered nanoparticles (ENPs) in water, soils, sediment and food matrices is very important for research into the risks of ENPs to consumers and the environment. However, these analyses pose a significant challenge as most existing techniques require some form of sample manipulation prior to imaging and characterization, which can result in changes in the ENPs in a sample and in the introduction of analytical artefacts. This study therefore explored the application of a newly designed instrument, the atmospheric scanning electron microscope (ASEM), which allows the direct characterization of ENPs in liquid matrices and which therefore overcomes some of the limitations associated with existing imaging methods. ASEM was used to characterize the size distribution of a range of ENPs in a selection of environmental and food matrices, including supernatant of natural sediment, test medium used in ecotoxicology studies, bovine serum albumin and tomato soup under atmospheric conditions. The obtained imaging results were compared to results obtained using conventional imaging by transmission electron microscope (TEM) and SEM as well as to size distribution data derived from nanoparticle tracking analysis (NTA). ASEM analysis was found to be a complementary technique to existing methods that is able to visualize ENPs in complex liquid matrices and to provide ENP size information without extensive sample preparation. ASEM images can detect ENPs in liquids down to 30 nm and to a level of 1 mg L^?1 (9×10⁸ particles mL^?1, 50 nm Au ENPs). The results indicate ASEM is a highly complementary method to existing approaches for analyzing ENPs in complex media and that its use will allow those studying to study ENP behavior in situ, something that is currently extremely challenging to do.     

Dental gallium alloy composites studied by SEM and TEM

Analytical scanning and transmission electron microscopy (SEM and TEM) studies of dental gallium alloys have been carried out. The Ga alloys were made by triturating a LU powder (Ag–Sn–Cu rich alloy powder) and a GF powder (Ag–Sn–Cu–Pd rich alloy powder) with a liquid Ga alloy containing Ga, In and Sn. The dental materials were found to be composites consisting of remaining, undissolved particles from the Ag-based alloy powders in a matrix of reaction products with the Ga alloy. SEM studies have been carried out to give an overview of the composites. The distribution of the elements was found by the X-ray mapping technique. The phases in the matrix and the remaining alloy particles have been identified by electron diffraction, high-resolution electron microscopy and energy-dispersive X-ray spectroscopy. The following phases were identified in the LU alloy: orthorhombic Ag₃Sn, cubic Ag₉In₄, tetragonal β-Sn and hexagonal Ag₂Ga. In addition to these well-known phases Ga-rich regions were observed consisting of an intergrowth of tetragonal CuGa₂ and one of the cubic γ-Cu₉Ga₄ phases. In addition to these phases cubic Ga₇Pd₃ was found in the GF alloy. The anomalous setting expansion of the GF alloy may be explained by the presence of Ga₇Pd₃.     

Palynological investigation of some selected species of family Fabaceae from Pakistan,using light and scanning electron microscopy techniques

Pollen morphology of 11 species of family Fabaceae that is, Trifolium alexandrinum, Trifolium resupinatum, Arachis hypogaea, Lathyrus aphaca, Medicago lupulina, Vicia sativa, Lathyrus odoratus, Pongamia pinnata, Melilotus indicus, Medicago polymorpha, Medicago sativa from Pakistan has been investigated by light and scanning electron microscopy. Pollen were generally tricolporate, radially symmetrical, isopolar, elliptic in equatorial view and triangular in polar view under LM. Results showed that pollens were per‐prolate (T. alexandrinum), prolate (T. resupinatum, V. sativa, L. odoratus, Melilotus indicus, M. polymorpha, M. sativa) and sub‐prolate (A. hypogaea, L. aphaca, M. lupulina, P. pinnata). The larger polar/equatorial (P/E) ratio was found in T. alexandrinum (2.26 μm) and the smallest was found in M. lupulina (1.21 μm). The exine of T. resupinatum was 3.00 μm in thickness while others posses smaller exine thickness. The larger pore diameter was found in P. pinnata (16.01 μm) while others have smaller. The length of colpi was larger in Arachis hypogaea (32.24) while others posses smaller. Eight types of surface ornamentation (Psilate, faintly rugulate). Perforate and rugulate to verrucate have been observed under SEM. The pollens were europalynous type. Pollen morphology proved to be useful for the specific delimitation and serve as a tool for the identification and classification of taxa at specific and generic levels and can also be used as a key for the taxonomic features. Diversity in exine sculpture is helpful indicative characters for the isolation of closely related species. Hence, it is clear that both qualitative and quantitative characters of pollen can be useful for differentiating between taxa at specific level.     

Comparative pollen and foliar micromorphological studies using light microscopy and scanning electron microscopy of some selected species of Lamiaceae from Alpine Zone of Deosai Plateau,Western Himalayas

The study was conducted to highlight a detailed account of morphology of pollen chosen species of Lamiaceae through scanning electron microscopy, and the anatomical characteristics of leaf epidermis of seven species using simple light microscopy. In results, Anisomeles indica and Otostegia aucheri belong to subfamily Lamioideae because it has tricolpate pollen while the rest eight species belong to subfamily Nepetoideae (hexacolpate pollen). The exine sculpturing of pollen of studied species was found to be reticulate. In the family Lamiaceae, four kinds of stomata were found anomocytic, anisocytic, diacytic, and actinocytic, respectively. The cell wall patterns of epidermal cells were irregular or polygonal with straight or undulate walls. It was noted that the variety of the epidermal trichomes seems of taxonomically important for the identification of species of Lamiaceae. Both nonglandular and glandular trichomes were analyzed. The nonglandular trichomes were characterized with long, thin, and pointed apical unicellular cells. The nonglandular trichomes were A‐shaped in Thymus linearis. In Perovskia abrotanoides, stellate glandular trichomes were observed whereas in A. indica and Mentha royleana both glandular and nonglandular trichomes were found. In A. indica, the nonglandular trichomes were sessile and peltate in M. royleana. For the first time in this study, pollen and foliar micromorphological features of selected species of this area are carried out. These taxonomic characters were found to be important in discrimination of species from each other. In future, the detailed study with comprehensive morphology coupled with other important characters is required for delimitation of taxa at various levels.     

Morphological organization of the dorsal protuberance of Linepithema humile (Mayr, 1868) ant's larvae (Hymenoptera, Formicidae)

The Argentine ant Linepithema humile is an important invasive species because of the levels of infestation that it can reach; however, there is little information about its presence, histological organization, and function of the dorsal protuberance, which is found exclusively in their larvae. The objective of this study was to describe it in L. humile through scanning electron microscopy and transmission electron microscopy, bringing information about this structure. The epidermis of these larvae have cuticles covering the whole body, and is formed by a sequence of overlapping lamellas where the inner ones were thicker and presented lower electron density, whereas the outer ones were thinner and highly electron dense. Pores or pore-like channels were not observed. A thick and acellular region composed of granular material was found under the cuticular layer. Out of this region, the flattened epidermic cells formed an epithelial layer. For the dorsal protuberance region, these cells become prismatic, and similarly to the cuticle, presents significant thickening. These cells presented extended microvilli, as well as a great amount of lamellar rough endoplasmic reticulum. Under this epithelium was observed a concentration of fat body cells, more numerous in the dorsal protuberance region. This study indicated that the dorsal protuberance present in the first segment of L. humile larvae has apparently no secretory function because no pores were found. This fact allowed to conclude that in L. humile larvae the dorsal protuberance would have the function to make it easier for the worker ants to carry them within the colony.     

Morphological and palynological assessment of taxonomically problematic genus Paspalum based on light and scanning electron microscopy

Present research work was carried out to clarify the variations among species of genus Paspalum morphologically and palynologically as this genus is taxonomically difficult due to having multiple similar morphologically overlapping characters which make it difficult to identify. Henceforth present research work was carried out to delimit taxa within the same genus by morphological and palynological tools through light microscopy (LM) and scanning electron microscopy (SEM). Both these tools are considered as the most useful taxonomic characters for taxonomically problematic genera. The results showed a lot of variations among morphological characters. In Paspalum dilatatum, the upper glume was ovate whereas in the other two species, the upper glume was elliptic. The upper glume apex found in P. dilatatum and Paspalum scrobiculatum was obtuse whereas in Paspalum distichum, upper glume apex was acute. Glume nerves showed variation in all three species. Paspalum distichum was 3 nerved, P. scrobiculatum was 5–7 nerved, and P. dilatatum was 5–9 nerved. All three species showed variation in lemma nerves. Paspalum scrobiculatum had 3 nerved lemma whereas in P. distichum 3–5 nerved and P. dilatatum 5–9 nerved lemma were present. In polar and equatorial view, pollen grains ranged from 25 (20–30) to 37.5 (30–45) μm. Paspalum distichum appeared to be the smallest in size whereas P. dilatatum was the largest. Exine thickness ranged from 0.75 (0.5–1) to 1.35 (1.2–1.5) μm. The higher value of pollen fertility was found in P. scrobiculatum as 87.69% and the lowest value was in P. distichum as 78.08%. Morphological keys were also given for correct identification.     

Analysis of efficiency of two-photon versus single-photon absorption for fluorescence generation in biological objects

By microinjecting rhodamine-conjugated porcine tubulin into pea epidermis we recently showed how cortical microtubules reorientate from transverse to longitudinal in living cells (Yuan et al ., 1994, Proc. Nat. Acad. Sci, USA 91, 6050–6053). In the present paper we compare this reorientation with the contrary longitudinal to transverse realignment induced by adding gibberellic acid to pre-injected cells on the microscope slide. Both kinds of reorientation are initiated by the appearance of 'discordant' microtubules which do not share the existing alignment but anticipate the new direction. These increase in number as the existing microtubules depolymerize, one alignment apparently replacing the other in a continuous process.
By rotating stacks of confocal sections by computer methods we have previously shown that microtubules at the outer tangential cell wall do not necessarily have the same orientation as microtubules at the adjoining anticlinal walls of the same cell (Yuan et al ., 1995, Plant J. 7, 17–23). This suggests that microtubule reorientations in these epidermal cells occur mainly (or, at least, first) at the outer wall, indicating that the array may not reorientate as a whole. Collectively, these data emphasize the discontinuous nature of the realignment process, the importance of new microtubule polymerization, and the special property of the outer epidermal surface as a sensitive domain.     

Observations on fretting damage transition to cracking; state of the art and preliminary observations

Interrupted fretting fatigue experiments were performed on 7075-T6 aluminum alloy and fretting damage characterized by confocal and scanning electron microscopy. Strain, induced by specimen fatigue, produces a small amplitude oscillatory motion between the fatigue specimen and the fretting pad. A fretting fatigue damage threshold exists in this material. Hundred percent fretting fatigue was defined as the average total cycles to fracture based on specimens 1–5 (both axial and normal forces were applied). Specimens had fretting damage induced at 100%, 80%, 60%, 40%, and 20% of the maximum fretting fatigue cycles to fracture. A positive correlation was not found between the depth of fretting damage and crack formation, but there appeared to be a stronger relationship between the fretting damaged surface areas, proximity of pits and crack nucleation sites.     

Towards quantification of doping in gallium arsenide nanostructures by low-energy scanning electron microscopy and conductive atomic force microscopy

We calculate a universal shift in work function of 59.4 meV per decade of dopant concentration change that applies to all doped semiconductors and from this use Monte Carlo simulations to simulate the resulting change in secondary electron yield for doped GaAs. We then compare experimental images of doped GaAs layers from scanning electron microscopy and conductive atomic force microscopy. Kelvin probe force microscopy allows to directly measure and map local work function changes, but values measured are often smaller, typically only around half, of what theory predicts for perfectly clean surfaces.     

Artefacts arising during preparation of hydrated paper pulp samples for low-temperature SEM

Beating, a pulp treatment widely used in the paper industry, causes disruption of cell wall layers and fibrillation. Previous studies of the effects of beating on fibre morphology have used conventional methods of specimen preparation, with all the attendant problems of shrinkage and distortion during dehydration. Low-temperature scanning electron microscopy (LTSEM) therefore seemed to offer an ideal method for examining fully hydrated wood pulp fibres. Cryofixation of pulp followed by sublimation of superficial ice, however, is shown to generate artefacts indistinguishable from structures present in the samples. Fibrillar and membranous structures were generated in LTSEM-prepared sugar solutions; their presence in pulp samples was therefore attributed to the dissolved carbohydrates inherent in pulp suspensions. Since artefact and fact are currently impossible to distinguish in LTSEM-prepared pulp samples, it seems that the technique should be applied to wet paper or pulp samples with considerable circumspection.