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Scanning electron microscopy of the floor of the fourth ventricle in rats subjected to graded impact injury to the sensorimotor cortex.

OBJECT: Respiratory dysfunction including apnea frequently follows head injury in humans. The purpose of this study was to identify any structural alterations in the region of brainstem respiratory nuclei that might account for immediate postinjury respiratory abnormalities in anesthetized experimental animals. METHODS: Using scanning electron microscopy, the authors examined the floor of the fourth ventricle in injured rats after a piston strike to the sensorimotor cortex that depressed the dura 1, 2, or 4 mm. The rats were killed within minutes of injury. Cortical impact depths measuring either 1 or 2 mm (eight rats) produced no respiratory abnormalities, and the structural integrity of the ependymal lining of the ventricular floor in these animals was not compromised. Thirteen rats were subjected to impact to a 4-mm depth and 10 of these exhibited immediate temporary or permanent apnea. The medullae of nine of these rats were studied using scanning electron microscopy, and the fourth ventricular floors of all nine rats showed tears. Four rats that exhibited immediate, permanent apnea had tears in the caudal fourth ventricle floor near the obex, whereas five rats with no or only transient apnea had tears located more anteriorly, near the aqueduct or laterally. Changes in cerebrospinal fluid flow or pressure dynamics may have caused these tears. Light microscopy, focused near the area postrema, revealed a shearing defect through the ependyma of the fourth ventricular floor into the subjacent neuropil with a disruption of axonal pathways. CONCLUSIONS: Respiratory neuronal network components lying within 2 mm of the area postrema may well have been disrupted by the caudal tears producing permanent apnea. A similar phenomenon could account for the transient or permanent postinjury apnea seen in humans with severe head injury.

Animals↗

Scanning electron microscopy of glomerular and non glomerular red blood cells.

Phase contrast microscopic examination of the urine has been recently shown to be of value in predicting whether hematuria is due to glomerulonephritis or lesions of the lower urinary tract. Glomerular red cells show variations in size and shape and have distorted surfaces. Non glomerular red cells are uniform in size and shape and have smooth surfaces. Scanning electron microscopy was performed on urine sediment containing either glomerular or non glomerular red cells to better define their surface characteristics. Glomerular red cells exhibited a variety of forms, most cells having lumpy projections from the surface, some showing fragmentation of the membrane and others showing gross distortion. In contrast non glomerular red cells show smooth surfaces and usually maintain the normal biconcave disc shape of peripheral red blood cells. Scanning electron microscopy can better define surface structural abnormalities of urinary glomerular and non glomerular red blood cells.

Analgesics↗

Use of scanning electron microscopy to confirm the identity of lice infesting communally grazed goat herds.

Lice have been described on goats in commercial farming systems in South Africa but not from flocks on communal grazing. During a longitudinal survey on the causes of goat kid mortality, conducted in Jericho district, North West Province, lice were collected from communally grazed indigenous goats. These lice were prepared for and viewed by scanning electron microscopy, and micro-morphological taxonomic details are described. Three species of lice were found in the study area and identified as Bovicola caprae, Bovicola limbatus and Linognathus africanus. Sucking and biting lice were found in ten of the 12 herds of goats examined. Lice were found on both mature goats and kids. Bovicola caprae and L. africanus were the most common biting and sucking lice respectively in all herds examined. Scanning electron microscopy revealed additional features which aided in the identification of the louse species. Photomicrographs were more accurate aids to identification than the line drawings in the literature and facilitated identification using dissecting microscope.

Animals↗

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↗

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↗

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↗

Detection of calcifications in breast biopsies by scanning electron microscopy.

The identification of microcalcifications in breast biopsy specimens is a common and important task for the surgical pathologist that may sometimes be problematic. Although visualized by preoperative mammographic and operative specimen radiography, some cases may fail to reveal calcifications by histopathologic examination or occasion laborious and time-consuming study to confirm sparse calcifications. The present study demonstrates how conventional bright-field and polariscopic light microscopy did require considerable effort to confirm rare calcification. On the other hand, scanning electron microscopy was employed directly on a routine slide and provided definitive visual and physical proof, e.g., by means of energy dispersive x-ray microanalysis of a rare calcification on a slide previously judged to be negative for calcium and in a short period of time. Recent instrument advances in high voltage beam stabilization and in the ability to vary the pressure of the scanning electron microscope in the vicinity of the specimen now allow complete insertion of a diagnostic glass slide with only trivial preparation. The report, although brief, raises important questions regarding the extent and prevalence of breast calcifications.

Adult↗

[Scanning electronic microscopy of the small intestine in persistent diarrhea].

Persistent diarrhea very often leads children to malnutrition. It has become the major cause of death resulting from acute diarrhea episodes in developing countries. In order to determine the ultrastructural alterations of the small bowel that occur in the syndrome, 16 infants with severe persistent diarrhea were studied, utilizing light microscopy and the scanning electron microscope. Stool and jejunal fluid samples were collected for culture, rotavirus, ova and parasite search. Enteropathogenic agents were isolated in stools from 11 (68.7%) patients and bacterial proliferation in the small bowel was detected in 11 (68.7%) patients. EPEC strains were the most frequent enteropathogenic agent isolated both from stool and jejunal fluid cultures. The stool cultures revealed the presence of the following enteropathogenic microorganisms: EPEC 0111 in four, EPEC 0119 in one, EAggEC in five, Shigella flexneri in two, and Shigella sonnei in one; mixed infections due to EAggEC associated with EPEC 0111 were seen in two patients. The light microscopic analysis revealed that 56.2% of the patients suffered moderate villous atrophy most frequently associated with effacement of the microvilli, intracytoplasmatic vacuolization, increased number of multivesicular bodied and increased lymphocytic and eosinophylic infiltration in the lamina propria. The scanning electron microscopic analysis revealed in all cases shortening of the villi and enterocyte derangements; very often there was a total lack and/or effacement of the microvilli; in half of the patients there was a mucoid material covering the enterocytes tightly adhered to the apical epithelium surface. The scanning ultrastructural alterations observed in these patients are probably due to an association of factors brought about by the presence of enteropathogenic microorganisms and the resulting food intolerance that is responsible for perpetuation of diarrhea.

Diarrhea, Infantile↗