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Discrimination of morphological findings in dentine from osteogenesis imperfecta patients using combinations of polarized light microscopy, microradiography and scanning electron microscopy.

OBJECTIVES: The aim of this study was to investigate the morphological appearance of dentine in teeth from individuals with osteogenesis imperfecta type I, III and IV using different histological techniques, and to correlate morphological findings to different types of osteogenesis imperfecta. SAMPLE AND METHODS: Extracted or exfoliated primary and permanent teeth were collected from 15 patients with the osteogenesis imperfecta diagnoses I, III or IV, with or without the additional diagnosis dentinogenesis imperfecta. Ground and decalcified sections were prepared from the teeth. Histo-morphological studies of the dentine were performed utilizing light and polarized light microscopy, microradiography and scanning electron microscopy. RESULTS: Characteristic findings were irregular tubules, remnants of capillary inclusions and obliterated pulps. All types of osteogenesis imperfecta exhibited similar types of dentine aberrations, but patients with type III or IV had a higher frequency of aberrations when compared to type I. CONCLUSIONS: The combination of either polarized light microscopy or micro-radiography, together with scanning electron microscopy, gave the most amount of morphological information from dentine samples. In addition, aberrations in dentine structure were more clearly observable. Light microscopy was not critical for the analyses.

Adolescent↗

Topographic examination of sister chromatid differential staining by Nomarski interference microscopy and scanning electron microscopy.

BrdU-substituted Chinese hamster chromosomes were treated with a hog Na2HPO4 solution and stained with Giemsa to produce sister chromatid differential staining (SCD). The process of SCD was examined with the Nomarski differential interference microscope and the scanning electron microscope. After the Na2HPO4 treatment alone, unifilarly BrdU-substituted (TB) chromatids appeared somewhat more severely collapsed than the bifilarly substituted (BB) chromatids. Subsequent Giemsa staining, however, brought about pronounced piling up of the Giemsa dye on the TB-chromatids but not on the BB-ones, causing highly distinct differential Giemsa staining as well as a marked differentiation in surface topography between the sister chromatids. Removal of the Giemsa dye from the differentially Giemsa stained chromosomes resulted in a disappearance of such a pronounced topographic differentiation.

Animals↗

Examination of injected specimens by confocal laser scanning microscopy and scanning electron microscopy.

Although corrosion casts commonly examined by scanning electron microscopy (SEM) offer good insights into the general arrangement of the microvasculature of certain organs, no information can be obtained from such specimens on special morphological data or dynamic events. Thus, most investigators are obliged to make their interpretations of cast structures only on the basis of indirect criteria. A synopsis, which considers the most important data in that respect, is given for the blood and lymphatic microvessels. Moreover, a methodological approach is introduced, based on the application of fluorescent resin, which allows the examination of cast structures in uncorroded specimens together with the surrounding tissue by confocal laser scanning microscopy (CLSM). When two differently stained resins are injected at different sites, e.g., Mercox-fluorescent yellow into an artery and Mercox-rhodamine into the interstice, blood vessels and lymphatics can be well distinguished. In lymph nodes, the system of lymphoid spaces and that of intranodal sinuses can be separately represented with the same technique. Applications and advantages of the CLSM method, which is a useful tool for an extended interpretation of corrosion casts in SEM, are shown in cast specimens from tongue, skin, and liver in rats. In this study, both corroded and uncorroded specimens were considered. In addition, a new approach is demonstrated which combines casting technique and fluorescence microscopy with investigations carried out under experimental conditions. This allows the examination of microvascular casts together with labelled cells in liver and spleen in a state of phagocytosis.

Animals↗

Scanning electron microscopy and X-ray microanalysis of foreign bodies associated with Silastic implants in humans.

Polydimethylsiloxane (Silastic) capsules containing megestrol acetate have been implanted in subcutaneous tissue as a method of long-term contraception. Histological studies revealed a granulomatous foreign body reaction around these capsules with birefringent crystals in multinucleated giant cells. Although several authors had interpreted these crystals as steroids, this seemed unlikely since the tissue had been processed with organic solvents. Analysis of these crystals by polarization microscopy, scanning electron microscopy, and X-ray microanalysis demonstrated that the material was talc. The talc was probably introduced as a contaminant from gloves during the implantation. Further analysis showed that capsule fragmentation could not have produced the material since Silastic particles could not be detected.

Contraceptive Agents↗

Techniques for combining light microscopy and scanning electron microscopy: a survey of the literature.

A survey of methods combining light microscopy and scanning electron microscopy is presented. A simple correlation is made when two preparations from adjacent parts of one specimen are investigated in two different microscopes. A more sophisticated method is the consecutive investigation of one specimen with two microscopes. A major problem in this method is the relocation of the area of interest. Several authors have presented solutions for this problem. It is preferable when one preparation is investigated in only one instrument, combining the two microscopical (LM and SEM) techniques, thus making relocation redundant.

Microscopy↗

Three-dimensional morphology of cerebellar climbing fibers. A study by means of confocal laser scanning microscopy and scanning electron microscopy.

The intracortical pathway of cerebellar climbing fibers have been traced by means of scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) to study the degree of lateral collateralization of these fibers in the granular Purkinje cell and molecular layers. Samples of teleost fish were processed for conventional and freeze-fracture SEM. Samples of hamster cerebellum were examined by means of CLSM using FM4-64 as an intracellular stain. High resolution in lens SEM of primate cerebellar cortex was carried out using chromium coating. At scanning electron and confocal laser microscopy levels, the climbing fibers appeared at the white matter and granular layer as fine fibers with a typical arborescence or crossing-over branching pattern, whereas the mossy fibers exhibited a characteristic dichotomous bifurcation. At the granular layer, the parent climbing fibers and their tendrils collaterals appeared to be surrounding granule and Golgi cells. At the interface between granule and Purkinje cell layers, the climbing fibers were observed giving off three types of collateral processes: those remaining in the granular layer, others approaching the Purkinje cell bodies, and a third type ascending directly to the molecular layer. At this layer, retrograde collaterals were seen descending to the granular layer. By field emission high-resolution SEM of primate cerebellar cortex, the climbing fiber terminal collaterals were appreciated ending by means of round synaptic knobs upon the spines of secondary and tertiary Purkinje cell dendrites.

Animals↗

A new approach for studying semithin sections of human pathological material: intermicroscopic correlation between light microscopy and scanning electron microscopy.

In order to obtain useful and complete information on the study of pathological material, we observed by scanning electron microscopy (SEM) the same semithin sections observed by light microscopy (LM). For this purpose, the specimen must have, at the same time, chromatic and electron dense characteristics. We thus developed different specimen preparation methods, subjecting the semithin sections to specific polychromatic staining with high atomic number (Z) elements, to monochromatic staining followed by routine contrasting with uranyl acetate and lead citrate, and to specific cytochemical and immunocytochemical procedures. The specimens were examined in sequence by LM, by SEM equipped with secondary electron, backscattered electron, transmitted electron detectors and by scanning transmission electron microscopy (S(T)EM).

Actins↗

Arrays of hemispheric surface projections on Chlamydia psittaci and Chlamydia trachomatis observed by scanning electron microscopy.

Scanning microscopy of two strains of Chlamydia psittaci and four strains of Chlamydia trachomatis representative of the wide diversity in origin and behavior of members of the genus revealed patches of regular arrays of hemispheric projections on the surfaces of elementary bodies of all six strains. These distinctive and perhaps unique surface structure are probably present in all populations of chlamydiae.

Cell Membrane↗

[Surface ultrastructure of the human uterine exfoliated endometrial cells--comparative studies of the same cell by light microscopy and scanning electron microscopy (author's transl)].

The preparations for observations of the same cells by light microscopy (LM) and scanning electron microscopy (SEM) were examined and the exfoliated endometrial cells of the uterus obtained by the newly introduced brush and cannula was evaluated. 1) The degree of shrinkage of the area was not significant between the air drying method and critical point drying method. In the air drying group, adhesion, entanglement and disappearance of microvilli (mv) and the adhesion of cilia, and the collapse of cells were noted morphologically. 2) Papanicolaou stain had no effect on the degree of shrinkage of the area and the shape of cilia or mv. 3) When the samples were transferred from ethanol to critical point drying, without immersion in isoamyl acetate, artifacts appeared in small granulated form. 4) The menopausal phase was divided into two groups which showed a shift of the maturation Index (MI) to center and had a slender mv and the other group which showed a shift to the left and granular mv. The nether surface of the cells showed a wrinkled and irregular undulations and did not show cilia or mv. 5) In adenomatous hyperplasia, the ratio of ciliated cell to no ciliated cell was four to one. It was two to one in cystic glandular hyperplasia. 6) The dense and short mv were observed in well-differentiated adenocarcinoma and poorly-differentiated adenocarcinoma showed neither mv nor cilia. Morphologic changes of mv and cilia in carcinomatous change of uterine endometrium commenced with a decrease in cilia, shifted to the shortening and disappearance disappearance of mv.

Adenocarcinoma↗

The dendritic cells of the guinea pig popliteal lymph node: identification and classification of cells observed by scanning electron microscopy.

Scanning electrom microscopy of guinea pig popliteal lymph nodes revealed that 3 types of cells adhered to glass and possessed extensive cytoplasmic processes. Two of these types were considered to represent extremes of a macrophage population on the basis of possession of surface receptors for cytophilic Abs, complement and immunoglobulin, as well as their distribution, quantity and morphology. The third cell type was characterized by possession of large numbers of filamentous dendritic processes. On the basis of morphology, quantity, distribution and the absence of receptors, this cell was considered to represent a cell type clearly distinct from the other cells observed in the normal lymph node.

Animals↗

Confocal light microscopy and scanning electron microscopy of the human eye lens.

The potential of confocal light microscopy (CLM) for in vivo observation of pathology in the anterior pole of the eye lenses was evaluated by performing an in vitro study of human lenses comparing this type of microscopy with scanning electron microscopy (SEM). In vitro CLM showed high resolution images of the epithelium which would enable early detection of pathology and easily allows cell counting and estimating cell size. Superficial lens fibres are well visualised and low and high frequency bands as well as vacuolar elements were easily detected. SEM observations fully supported the CLM observations. This study shows that CLM has the potential to become a useful tool for detecting lens changes, after suitable adaptation for clinical use.

Adult↗

Specific surface area of snow samples determined by CH4 adsorption at 77 K and estimated by optical microscopy and scanning electron microscopy.

Snow is a divided medium that can adsorb atmospheric trace gases. Evaluating the impact of the snow cover on atmospheric chemistry therefore requires the knowledge of the specific surface area (SSA) of snow. This paper compares the results of three methods used to measure or estimate the SSA of four snow samples: CH4 adsorption at 77 K, optical microscopy (OM), and scanning electron microscopy (SEM, used only on two samples). Within error bars, CH4 adsorption and OM yield similar results on three of the four snow samples. Values for the 4th sample are within a factor of 2. For both samples where CH4 adsorption, OM, and SEM are used, all three methods yield similar results, but CH4 adsorption always has a better accuracy and a much better precision. Thus, despite its ease of use, estimates from OM images are often not accurate enough to monitor the evolution of snow SSA. The main sources of error in the OM method are the difficulty to determine snow crystal thicknesses and to take into account the topography of the snow crystal surface. The combination of CH4 adsorption and OM or SEM can provide useful information on the evolution of both the SSA and the shape of snow crystals. This will be useful to evaluate the respective contributions of adsorption/desorption and sublimation/condensation processes to the impact of the snow cover on atmospheric chemistry.

Adsorption↗

Retinopathy of Great Pyrenees dogs: fluorescein angiography, light microscopy and transmitting and scanning electron microscopy.

OBJECTIVE: Investigation of the pathogenesis of Great Pyrenees retinopathy. ANIMALS: One male and two female puppies of parents who were affected with Great Pyrenees retinopathy and one 4-year-old affected adult male Great Pyrenees dog. PROCEDURE: The puppies were examined daily from 7 weeks of age by indirect ophthalmoscopy and their fundi were photographed until the lesions were static. Fluorescein angiography was completed at 7 weeks of age, within 24 h of detection of ophthalmoscopic lesions, and then weekly. The eyes of a 4-year-old and two 20-week-old puppies were examined with light microscopy, and transmitting and scanning electron microscopy. RESULTS: Blocked choroidal fluorescence was detected at 7 weeks of age. The blocked fluorescence enlarged, when the characteristic serous retinal detachments developed at 11 weeks of age. The detachments enlarged in size and number until the puppies were approximately 20 weeks old. Fluorescein pooling confirmed the presence of transient retinal pigment epithelial detachments. Leakage of dye into serous retinal detachments was not detected in this study. Light microscopy and transmitting and scanning electron microscopy confirmed the presence of multifocal serous retinal detachments with focal retinal degeneration that extended to the inner nuclear layer in a 4-year-old dog. The retinal detachments were accompanied by hypertrophy, hyperplasia, increased pigmentation, and vacuolation of the retinal pigment epithelium. CONCLUSIONS AND CLINICAL RELEVANCE: Great Pyrenees retinopathy is manifested by multifocal serous retinal and retinal pigment epithelial detachments. These detachments are similar to those noted with central serous retinopathy of humans. The sudden development of multifocal retinal and retinal pigment epithelial detachments, and the serous nature of these detachments, supports a theory that they develop secondary to focal secretion and absorption defects in retinal pigment epithelial cells. Given the age of the puppies when the blocked choroidal fluorescence was noted and maturation of the dog retina at 8 weeks postpartum, this retinopathy is considered to be a retinal pigment epithelial dysplasia. This unique inherited retinopathy offers an opportunity to study retinal pigment epithelial secretion.

Animals↗