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Salt-dependent structural changes of chromatin in isolated chicken liver nuclei as visualized by scanning electron microscopy.

MgCl2- and KCl-dependent structural changes of chromatin in isolated chicken liver nuclei were examined by scanning electron microscopy. In the absence of salts, the chromatin appeared as granular or knobby fibrillar structures, 15-30 nm in diameter. At 0.2 mM MgCl2, the structures condensed into 30-60 nm fibers. At 2 mM MgCl2, most chromatin was present as condensed chromatin masses. Above 50 mM MgCl2, the condensed chromatin masses were loosened. At KCl concentrations higher than 25 mM, the chromatin appeared as discrete 30-60 nm fibers. However, no condensed chromatin masses were formed at any of the KCl concentrations examined in this study. These results indicate that the MgCl2- or KCl-dependent transitions from a lower to a higher level of chromatin organization in the isolated nuclei are comparable to those reported in isolated chromatin fragments, and that KCl is ineffective in the formation of condensed chromatin.

Animals↗

The three-dimensional architecture of the myosalpinx in the cow as revealed by scanning electron microscopy.

The three-dimensional architecture of the myosalpinx in the cow has been investigated by means of scanning electron microscopy after removal of interstitial connective tissue with NaOH digestion. In the extramural portion of the tubo-uterine junction, in the isthmus and ampulla, the myosalpinx is made up of oblique bundles of variable orientation and length, which are loosely distributed in the tubo-uterine junction and densely packed in both isthmus and ampulla. These bundles intersect and merge into the surrounding musculature. Our observations demonstrate how myosalpinx consists mainly of bundles of muscular fibers independent one of another, which show a multiple spatial arrangement and form a complex network. Such a muscular architecture is likely more suitable for stirring rather than pushing the embryos and gametes through the tube.

Animals↗

Use of environmental scanning electron microscopy to evaluate dental stain removal.

The purpose of the study was to assess the usefulness of environmental scanning electron microscopy (ESEM) to evaluate stain removal from extracted teeth. The ESEM differs from conventional SEM in that no sample preparation is needed, eliminating artifactual changes. Furthermore, the same sample can be viewed on multiple occasions, allowing "before" and "after" pictures of the same tooth. As a model stain removal device, we tested the Sonicare sonic toothbrush, which has previously been shown to remove dental stain in vivo. Twelve freshly extracted teeth with extrinsic coffee, tea or tobacco stain were obtained for the study. Nine of these had heavy stain (stain covering more than one-third buccal or lingual surface) and were used without further modification. Three teeth were treated in vitro with chlorhexidine and a mixture of coffee and tea to enhance staining. All teeth were examined by ESEM at three times: prior to brushing, after 15-30 seconds of brushing, and after 60-80 seconds of brushing. Light microscopy and 35 mm photography was also done to correlate the ultrastructural changes with those visible at low magnification. Water, mouthwash and 30% slurry of toothpaste were used as fluid vehicles during brushing, but little difference in stain removal was noted among these three fluids. Approximately half the stain was removed within 15-30 seconds, and most visible stain was removed in 60-80 seconds of brushing. Pits and crevices of tooth enamel that were smaller than the bristle diameter, and thus would be inaccessible to abrasive cleaning by direct bristle contact, were generally found to be stain-free. These findings confirm previous reports of the stain removal effectiveness of the Sonicare, and demonstrate the usefulness of ESEM for stain removal studies.

Chlorhexidine↗

Scanning electron microscopy evaluation of the effect of etching agents on human enamel surface.

Acid etching promotes microporosities on enamel surface, which provide a better bonding surface to adhesive materials. The purpose of this study was to comparatively analyze the microstructure of enamel surface after etching with 37% phosphoric acid or with two self-etching primers, Non-rinse conditioner (NRC) and Clearfil SE Bond (CSEB) using scanning electron microscopy. Thirty sound premolars were divided into 3 groups with ten teeth each: Group 1: the buccal surface was etched with 37% phosphoric acid for 15 seconds; Group 2: the buccal surface was etched with NRC for 20 seconds; Group 3: the buccal surface was etched with CSEB for 20 seconds. Teeth from Group 1 were rinsed with water; teeth from all groups were air-dried for 15 seconds. After that, all specimens were processed for scanning electron microscopy and analyzed in a Jeol 6100 SEM. The results showed deeper etching when the enamel surface was etched with 37% phosphoric acid, followed by NRC and CSEB. It is concluded that 37% phosphoric acid is still the best agent for a most effective enamel etching.

Acid Etching, Dental↗

Scanning electron microscopy of hydroxylated polyvinyl acetal and conventional gauze strip nasal packing materials.

The application of nasal packing is one of the most commonly performed procedures in rhinology. Various materials have been used as nasal packing, including conventional gauze strips and hydroxylated polyvinyl acetal. Complications related to nasal packing may cause problems that lead to increased morbidity. Among those complications, infectious ones range from localized infection in the nasal cavity to toxic shock syndrome. The purpose of this study was to evaluate conventional gauze strips and hydroxylated polyvinyl acetal nasal packing materials by scanning electron microscopy, to reveal their surface characteristics that would promote or prevent the development of infectious complications. The two types of materials were examined before and after application into the nasal cavity. Scanning electron microscopy demonstrated that hydroxylated polyvinyl acetal material had a smooth surface, whereas conventional gauze strips had an irregular surface with fibers projecting, thus increasing the surface area for bacterial adherence and allowing possible evasion of bacteria from the elements of the immune system within this fibrillar structure.

Bandages↗

Zinc deficiency: specific scalp hair defects seen by scanning electron microscopy may provide a valuable new test.

Zinc deficiency in man results in multisystem disease. It may be acquired or hereditary; the latter can be fatal if left untreated. Premature babies are particularly susceptible to zinc deficiency. Unfortunately no simple, reliable test for zinc status exists at present. Short, newly-emerging scalp hair samples from 3 classical cases of zinc deficiency all showed the same characteristic abnormalities when examined by scanning electron microscopy, i.e. straight, blunt tips bearing scales, unusually thick cuticular scales with jagged free-edges, and very fine longitudinal corrugations in individual scales. These abnormal features occurring together appear to be specific for zinc deficiency; they also varied in severity with marked variations in zinc status during follow-up studies. Due to the relative rarity of classic cases of zinc deficiency, it is not possible for one centre of our catchment size to conduct a preplanned study. However, if the present findings can be confirmed elsewhere, it is concluded that scanning electron microscopy of appropriately-selected hairs may provide a valuable new test for the diagnosis of zinc deficiency and for monitoring the response to zinc therapy.

Child, Preschool↗

[Scanning electron microscopy of the lung in experimental collapse].

The author induced experimental lung collapse by means of ligating a bronch of a dog. The material was divided into six groups and was examined by a scanning electrone microscopy. Two phases in the development of the collapse were found. She indicated the advantages of the scanning electrone microscope in comparison with the light and the remaining electrone optical systems.

Animals↗

Starch granule surface imaging using low-voltage scanning electron microscopy and atomic force microscopy.

High resolution imaging of wheat and potato starch granule surfaces has been performed using low-voltage scanning electron microscopy and atomic force microscopy. The complimentary images of uncoated granules demonstrate that the two starch types possess substantially different surface topologies; potato starch has many protrusions (100-300 nm in diameter), above a flatter surface containing 20-50 nm structures, whilst wheat starch possesses far fewer protrusions and generally has a smoother surface composed of approximately 20 nm structures. The protrusions are believed to be carbohydrate in nature and thus represent the ends of amylopectin side-chain clusters at the granule surface.

Microscopy, Atomic Force↗

The role of scanning electron microscopy in the diagnosis of human laryngeal cancer.

Malignant and non-malignant human laryngeal tissue samples (31 cases) were examined first by light microscopy (LM) and then the specimens were reprocessed for scanning electron microscopy (SEM). The presence of LM and SEM signs of malignancy were compared with each other. In all the 22 histologically unambiguous cases the LM and SEM results were in accordance with each other. From the 9 histologically doubtful cases, 2 were found malignant and 3 benign by retrospective SEM. The case histories of these 5 patients have confirmed the SEM results. This preliminary study suggests the application of SEM in histologically doubtful laryngeal biopsies.

Carcinoma, Squamous Cell↗

Adhesive interactions between CD34(+)-derived dendritic cell precursors and dermal microvascular endothelial cells studied by scanning electron microscopy.

Dendritic cells are migratory cells. Before they extravasate from the circulation into the skin across capillary blood vessel walls, they have to interact with endothelial cells. Using a fluorimetric adhesion assay, we have recently shown that CD34(+)-derived dendritic cell precursors are able to bind to resting and stimulated dermal microvascular endothelial cells. In the present study, we attempted to visualize this process at an ultrastructural level. CD34(+) progenitor cells were purified from human cord blood samples by means of immunomagnetic beads, and dendritic cells were generated by culture in the presence of GM-CSF, TNF-alpha and hSCF for 5 days. Immature CD83(-) CD86(low) dendritic cells were added to human dermal microvascular endothelial cells grown to confluence on membrane chambers. After 2 h, unbound dendritic cell precursors were removed, and bound cells were prepared for routine scanning electron microscopy. We found that (1). dendritic cell precursors firmly adhere to microvascular endothelial cells, enveloping them with their surface processes; (2). dendritic cell precursors are extremely deformable as they squeeze through the dense network of microvascular endothelial cells; (3). microvascular endothelial cells form, in part, a multi-layered network rather than the typical cobblestone pattern as seen by phase-contrast microscopy. The morphology of dendritic cell precursors and of human dermal microvascular endothelial cells was examined here, for the first time, by scanning electron microscopy. These data further emphasize that CD34(+)-derived dendritic cells efficiently adhere to dermal microvascular endothelial cells.

Antigens, CD34↗

Atrial endocardial changes in mitral valve disease: a scanning electron microscopy study.

BACKGROUND: The precise contribution of left atrial appendage (LAA) endocardial damage and dysfunction to the process of thrombus formation in patients with mitral valve (MV) disease, especially in the presence of atrial fibrillation (AF), has not as yet been clearly described. This may be important because the LAA is the usual site for thrombus formation. METHODS: The purpose of this study was to describe endocardial surface changes, through the use of scanning electron microscopy, in the left and right atrial appendages of patients with MV disease and the differences, if any, between patients with mitral stenosis and mitral regurgitation as well as between those with AF and sinus rhythm. Our second objective was to relate endocardial changes to plasma levels of von Willebrand factor (vWf), an established marker for endothelial damage. LAA specimens were obtained immediately after commencement of cardiopulmonary bypass from 35 patients (18 men; mean age 65 years, range 20 to 85) during surgery for MV repair or replacement. Right atrial appendage (RAA) specimens were similarly obtained as controls for individual patients. The specimens were fixed in 2.5% glutaraldehyde solution overnight, stored in Sorensen's phosphate buffer, and examined by means of scanning electron microscopy. Two independent observers documented the most advanced lesion in each specimen as follows: (1) "minimal" changes, with minimal disruption of the endocardium; (2) "intermediate" changes or prethrombotic lesions; and (3) "advanced" changes, with endocardial disruption and thrombotic lesions. Plasma levels of vWf were also measured (enzyme-linked immunosorbent assay) in all patients, and results were compared with those of age- and sex-matched healthy control patients. RESULTS: Advanced changes were more frequently seen in the endocardium of the LAA when compared with the RAA (31% vs 6%), whereas minimal changes were more frequently seen in the RAA compared with the LAA (23% vs 6%) (P =.00167). Similarly, the LAA from patients with mitral stenosis had a higher proportion of "advanced" endocardial changes when compared with patients with mitral regurgitation (67% vs 24%; P =.0066). The LAA in patients with AF had more "advanced" changes (39% vs 27%), but this was not statistically significant. Plasma vWf levels were significantly higher in patients with MV disease compared with healthy control patients (132 +/- 33 IU/dL vs 99 +/- 37 IU/dL; P =.0004) and in patients with advanced LAA changes compared with earlier changes (149 +/- 34 IU/dL vs 121 +/- 31 IU/dL; P =.042). CONCLUSIONS: Endocardial damage occurs in the atrial appendages of patients with MV disease. Potentially thrombogenic changes are more commonly seen in the LAA compared with the RAA and in patients with mitral stenosis compared with mitral regurgitation. These anatomic appearances may contribute to the risk of intra-atrial thrombus formation in patients with mitral valve disease, especially if AF is present.

Adult↗

Analysis of microvascular trees by means of scanning electron microscopy of vascular casts and 3D-morphometry.

Arterial and capillary trees form by consecutive branching (mostly bifurcations) from a stem vessel, venous trees form by repeated merging of blood vessels. Diameters of stem (parent, mother) vessels and daughter vessels (branches), interbranching distances and branching angles between stem and daughter vessels lastly define the overall three-dimensional structure of the vascular network as well as the basic transport capacity of the system. Here we use scanning electron microscopy and 3D-morphometry to measure these variables from stereo paired images of vascular corrosion casts of the anterior cerebral artery and its main branches and from arteriolar bifurcations of the mesencephalic optic tectum in the actinopterygian fish, Acipenser ruthenus. We then calculate bifurcation indices, area ratios, asymmetry ratios and test for the optimality principles underlying the bifurcations studied. Our results show that arteriolar bifurcations in the optic tectum are in favor of the principles of minimum pumping power and minimum volume rather than the principles of minimum surface and minimum drag. We conclude that scanning electron microscopy of vascular corrosion casts in conjunction with 3D-morphometry is an excellent tool to thoroughly analyze vascular trees in healthy and diseased tissues and organs, as well as on an ontogenetic and phylogenetic scale.

Algorithms↗

Coalescence of endothelial cells in the traumatized cornea. III. Correlation between specular and scanning electron microscopy.

Sequential specular microscopy has shown that coalescence of endothelial cells occurs after thawing in cryopreserved rabbit corneas. Scanning electron microscopy of corneas studied in vitro by specular microscopy showed the same large, fused endothelial cells. As observed in the electron micrographs, these cells showed no evidence of the cell borders that had disappeared during the specular microscopic examination. These data support the reliability of our specular microscopic observations and provide additional evidence for the occurrence of fusion or coalescence of corneal endothelial cells.

Animals↗

Quantitative scanning electron microscopy for the evaluation of thrombosis in extracorporeal circuits.

Methods devised in the past to evaluate blood-material interactions in extracorporeal devices assess small segments of test materials and do not analyze the entire circuit. In order to develop an effective method for the control of thrombosis in extracorporeal circulation, it is important to perform a quantitative evaluation of random samples of all the circuit elements. In an effort to quantify thrombosis in experimental extracorporeal membrane oxygenation circuits, random samples of silicone rubber membrane and polyvinyl chloride tubing were examined using scanning electron microscopy (SEM) and quantitative morphometry. The results indicate no statistically significant differences in the amounts of various adhered blood components between the tubing and the membrane. Greater than 60% of the examined surface had no blood components present. Platelet adhesion on the surface was predominant. Evidence of fibrin, gross thrombi, red cells, and white cells in small quantities were noted. It is concluded that quantitative scanning electron microscopy (QSEM) not only provides accurate information about the morphologic changes of blood cells due to surface contact, but also quantifies the individual elements and therefore assesses their contributions to thrombosis. This quantitative description will allow more meaningful comparison of blood-compatible materials and surfaces.

Animals↗

Iron carbonyl, a tracer for phagocytosis in scanning electron microscopy.

Human peripheral blood mononuclear cells and polymorphonuclear (PMN) leukocytes, separated by Ficoll-Hypaque have been incubated with a preparation containing iron carbonyl, and studied by scanning electron microscopy (SEM) in the backscattered electron imaging (BEI) mode. Iron carbonyl particles were visible only on the cell surface when studied in the conventional secondary electron (SE) mode of the scanning electron microscope. However, ingested iron carbonyl particles were observed in high contrast when the backscattered electrons were selectively collected. The BEI mode of the scanning electron microscope appeared very useful in identifying phagocytic activity in mixed cell populations and in establishing direct correlations, at the level of each individual cell, between surface morphology and phagocytic activity.

Humans↗

Scanning electron microscopy of the wall of the third ventricle of the brain of Rana temporia. II. Electron microscopy of the ventricular surface of the pars ventralis of the tuber cinereum.

Scanning electrom microscopy of the ventricular surface of the pars ventralis of the tuber cinereum of the frog Rana temporaria shows that it can be divided in (1) a dorsolateral area, characterized by the presence of very numerous large, solid, bulbous protrusions (2) a medial area, where the large bulbous structures are completely absent and which is diffusely covered with very numerous cilia, (3) a transiotional area located between the two other regions. The large bulbs, shown by scanning electron microscopy, correspond with the end-bulbs of thick ventricular dendrites of nerve cells revealed by transmission electron microscopy. At least many of these intra-ventricular bulbs are dendritic endings of peptidergic neurosecretory neurons, which have been tentatively identified as adenohypophysiotropic neuro-hormone producing neurons (Derickx et al., 1972, 1973a, 1973b). The structural features of the dendritic endings plead in favour of a possible receptive role.

Animals↗

Uncombable hair (cheveux incoiffables, pili trianguli et canaliculi) syndrome: brief review and role of scanning electron microscopy in diagnosis.

Uncombable hair syndrome was first described some 3 decades ago as "cheveux incoiffables" and is also known as spun-glass hair and pili trianguli et canaliculi. Both inherited (autosomal dominant and recessive with variable levels of penetrance) and sporadic forms of uncombable hair syndrome have been described, both being characterized by scalp hair that is impossible to comb due to the haphazard arrangement of the hair bundles. A characteristic morphologic feature of hair in this syndrome is a triangular to reniform to heart shape on cross-sections, and a groove, canal or flattening along the entire length of the hair in at least 50% of hairs examined by scanning electron microscopy. Most individuals are affected early in childhood and the hair takes on a spun-glass appearance with the hair becoming dry, curly, glossy, lighter in color, and progressively uncombable. Only the scalp hair is affected. Several conditions are associated with uncombable hair, such as ectodermal dysplasia, retinal dysplasia/pigmentary dystrophy, juvenile cataract, digit abnormalities, tooth enamel anomalies, oligodontia, and phalangoepiphyseal dysplasia. Other syndromes with hair abnormalities may also mimic uncombable hair syndrome clinically and these include, Rapp-Hodgkin ectodermal dysplasia; loose anagen hair syndrome; ectodermal dysplasia, ectrodatyly, cleft lip/palate (EEC) syndrome; and familial tricho-odonto-onchyial ectodermal dysplasia with syndactyly. Unlike other conditions with an uncombable hair component, uncombable hair syndrome alone (cheveux incoiffables, pili trianguli et canaliculi) is not associated with physical, neurologic, or mental abnormalities. In most cases of uncombable hair syndrome, the hair is grossly abnormal in infancy and early childhood, but may have improved manageability later in life. Scanning electron microscopy of hair samples provides definitive evidence for diagnosis of clinically suspected uncombable hair syndrome and eliminates other hair abnormalities from the differential diagnosis.

Chemical Phenomena↗

[Fornicate structure of blind saccular lymph vessel segments in the conjunctiva--scanning electron microscopy studies].

BACKGROUND: The accepted fundamental element of the initial lymphatic system is a tubular structure (lymphatic capillary), which is present either in the form of a finger-shaped blind-ending protuberance, or in the nature of a plexiform composite arrangement. We were able to demonstrate experimentally that the finger-shaped protuberances, which are known as "initial segments" are in fact temporary filling states. As the filling process continues, these vascular elements become intermediate segments, via which other parts of the vascular network take up the dye. To date there have been no investigations of the internal structure of these apparently blind-ending vessel segments. In this study, lymphographically represented "terminal segments" should be cut away for the purpose of examining the fornix by scanning electron microscopy. Is the internal surface of the fornix regular and unbroken, or are there fissure-like structures which could explain the observed filling processes? MATERIAL AND METHODS: In conjunctiva of bovine eyes (n = 80), interstitial double-contrast lymphography (Berlinblue solution/air) under a slit-lamp microscope was used specifically to search for finger-shaped terminal segments. The conjunctivae had previously been prepared by fixing in polymeric resins. A proportion (n = 21) of the specimens were amenable to examination under the scanning electron microscope. RESULTS: Notwithstanding the observed great variation in the shape of the lymphographically obtained blind-ending structures-in the form of terminations shaped variously like fingers, balloons, domes, pistons or pyramids, terminations with two humps, and terminations shaped like spear heads-scanning electron microscopy revealed within the fornices many relatively uniformly shaped structures in the form of fissures, configured with lips and saw-tooth edges, rather like zip fasteners. These findings are suggestive of preformed connections to neighbouring segments. This appears to be another element, in addition to the familiar flap-like structures, for controlling the circulation of lymph. Does the "initial part" of the lymphatic system now have to be redefined?

Animals↗