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Fallopian tubes from women of reproductive and postmenopausal ages were examined by scanning electron microscopy. The surface topography of these Fallopian tubes is described and illustrated.
Topographic features of eight struvite calculi were investigated with scanning electron microscopy (SEM) equipped with energy-dispersive X-ray microanalysis (EDX). Perpendicularly cracked fragments showed concentric laminations composed of compact and loosely packed strata alternately. Magnesium and phosphorus were detected in the compact strata with their characteristic dispersive X-ray spectra. There existed numerous spherular crystals with smooth or porous surfaces and scattered penta- or hexa-hedral coffin-lid shaped crystals in the loosely packed strata. The former crystals showed the dispersive X-ray spectra of calcium and phosphorus, and were estimated to be calcium phosphate (CaP). The latter ones were presumed to be magnesium ammonium phosphate (MAP) with EDX. The surfaces of the fragments were cracked like eggshells and displayed numerous CaP crystals and scattered MAP crystals in most cases, while in only 1 case some faces of pieces demonstrated wavy MAP phases, sundry areas which were rimmed with aggregated CaP spherulites. The mean molar ratios of Mg/P and Ca/P in each case were 0.88-1.03 and 1.25-1.52, respectively. Though EDX was inadequate to determine the accurate chemical formula of CaP and MAP crystals by detecting their molar ratios of Ca/P and Mg/P with EDX, SEM/EDX is useful to observe these urolith crystals and to surmise them to be CaP or MAP crystals by detecting their atomic elements with EDX.
Air-dried and ethanol-fixed buccal epithelial cell smears from five subjects were observed by scanning electron microscopy. The mucous pellicle was precipitated as a smooth haze covering the cells, and outlines of bacteria were found embedded within it. Rinsing the preparations under running water gradually diminished the mucous pellicle but not the cell-adherent bacteria. A more complete dissolution of the pellicle was accomplished by washing the buccal epithelial cells before smearing. After a chlorhexidine mouthrinse the buccal cells appeared distorted, with only a few adherent bacteria. Three days after the rinsing, the denatured appearance still persisted on many cells, however, simultaneously with the emergence of undenatured epithelial cells with adherent bacteria. The method introduced in this study is useful to investigate the bacteria-mucus-epithelial cell interactions. A possible mode of antibacterial activity of chlorhexidine in vivo may be that it destroys bacterial adhesins. The substantivity of chlorhexidine in the oral cavity may be linked to the turnover rate of the oral epithelial cells.
Scanning electron microscopy was used to examine cryofracture surfaces of ventricular myocardium from glutaraldehyde fixed rat and rabbit hearts subjected to intravascular injection of polymerizing acrylic resin. This allowed simultaneous observation of morphological features of cardiac muscle cells and the functional state of their associated small blood vessels. Because the resin injected to identify capillaries accessible to flow might be soluble in commonly used tissue dehydrating agents, alternative preparation methods using the cryoprotectants dimethylsulfoxide (DMSO) and glycerol were investigated. Provided a high performance backscattered electron detector and simple environmental cell were used to abolish specimen charging and circumvent potential instrument contamination, immersion in 2.82 M DMSO for 12 hr prior to cryofracture and freeze-drying gave the best results. The SEM appearance of specimens dehydrated in this way differed little from that of specimens prepared by ethanol dehydration and freeze-drying or by acetone dehydration and critical-point drying. Tissue shrinkage was 26.5 +/- 9.4%, comparable to that found after standard methods using solvent dehydration and critical-point drying.
The shape and arrangement of the developing nephrons were studied three-dimensionally by scanning electron microscopy (SEM) of the neonatal mouse kidney. The specimens were treated with the KOH digestion method in order to remove extracellular connective tissue components, thus enabling the direct observation of the developing nephrons at various stages. At the subcapsular region of the renal cortex, the ureteric ducts were observed as branched tubules with terminal swellings or ampullae. Newly formed blood vessels were often associated with terminals of these ureteric ducts. The cup-shaped renal corpuscles had aggregations of mesangial cells with blood vessels in the groove. At the vascular pole of mature nephrons, extraglomerular mesangial cells were observed as a cellular sheet, which was continuous with the smooth muscle layer of afferent and efferent blood vessels. The present study also demonstrated the shape of the immature podocytes in relation to the endothelial morphology of glomerular capillaries.
Intracellular structures of the odontoblasts were studied by scanning electron microscopy (SEM) using a modified AODO (aldehyde prefixed-osmium-DMSO-osmium) method. Well-developed flattened and layered rER (rough endoplasmic reticulum), paved with its associated ribosomes on its outer surface, were clearly observed in the odontoblast. Branched tubular mitochondria with nodules and swollen endings, interposing between and passing through the fenestrated layered rER, were demonstrated in the functional cells. Oblique and cross-sections of both the rER system and tubular mitochondria showed orthodox configurations similar to those usually described in transmission electron microscopy (TEM) studies. Many finger-like projections constructing the cristae directing towards the inner mitochondrial chamber were observed, and external chamber extending into the tubular cristae was also demonstrated.
Microwave irradiation (MWI) has been applied to the development of rapid methods to process biological samples for scanning electron microscopy (SEM). In this paper we propose two simple and quick techniques for processing bacteria (Proteus mirabilis and Vibrio mimicus) for SEM using MWI. In the simplest methodology, the bacteria were placed on a cover-glass, air-dried, and submitted to conductivity stain. The reagent used for the conductivity stain was the mordant of a light microscopy staining method (10 ml of 5% carbolic acid solution, 2 g of tannic acid, and 10 ml of saturated aluminum sulfate 12-H2O). In the second method the samples were double fixed (glutaraldehyde and then osmium), submitted to conductivity stain, dehydrated through a series of ethanol solutions of increasing concentration, treated with hexamethyldisilazine (HMDS), and dried at 35 degrees C for 5 minutes. In both methods the steps from fixation to treatment with HMDS were done under MWI for 2 minutes in an ice-water bath, in order to dissipate the heat generated by the MWI. Although both techniques preserve bacterial morphology adequately, the latter, technique showed the best preservation, including the appearance of flagella, and that process was completed in less than 2 hours at temperatures of MWI between 4 to 5 degrees C.
We observed DNA double helix and nucleosomes in the chromatin of chicken erythrocytes by ultrahigh-resolution scanning electron microscopy. Specimens were prepared according to a modified microspreading technique in combination with the carbon plate method and observed without metal coating. A part of the DNA fibers without nucleosomes showed "left-handed" double-strands with twisting appearances and regular periodicities of the helix. Linker DNA between the nucleosomes showed "right-handed" appearances. Most of the nucleosome particles appeared as prolate ellipsoidal shapes of various sizes. DNA appeared to enter and exit the nucleosome particles on opposite sides winding around the histone core.
Conjunctival and corneal epithelial surfaces of normal rabbit eyes with their associated mucus were studied by scanning electron microscopy before and after treatment with the mucolytic agent N-acetylcysteine (AC). Four groups received topically one 50-microliters drop of either (Group A) 0.1 MAC, (Group B) 0.1 M AC every 5 min for 1 hr, (Group C) 0.1 M AC every 5 min for 2 hr, or (Group D) three drops of 20% AC over 15 min. The effects of the instillation of AC on mucus removal and cellular lesions increased in the order (A) less than (B) less than (C) less than (D). Treatment A had no effect on cornea and conjunctiva. Treatment B cleaned away mucosal debris without alteration of either conjunctival or corneal epithelium. Treatment C had a similar effect on the mucus but was associated with focal necrosis, and treatment D produced widespread necrosis, desquamation of epithelial cells, and inflammation.
The formation of dental calculus in dogs is a major problem. Scanning electron microscopy of tartar specimens from dogs revealed on the outer surface of the plaque polymorphic configurations, more or less arranged as free filaments, corn-cobs or swab-like structures. Uninhabited bacterial recesses were found on the inner surface of the calculus. Calcification may occur between or within the bacteria. Elucidating the mechanisms of calculus formation should help in the development of prophylactic measures.
Human carotid atheroma has been examined by scanning electron microscopy (SEM) utilizing both secondary and backscattered electron detectors and transmission electron microscopy (TEM) both at conventional and high voltage. Different cytochemical techniques have been used to map elastic fibers, proteoglycans, calcium, non esterified cholesterol. By immunocytochemistry the distribution of factor VIII related antigen and actin has been studied. With SEM it was possible to detect aspects of carotid plaques not appreciated when using other conventional techniques. With TEM some modifications of the structural and/or functional features of connective tissue macromolecules have been observed. The occurrence of anomalous collagen has been shown. The fine investigation of endothelial cells, smooth muscle cells and the intercellular matrix components has supplied information of particular morpho-functional significance, thus allowing the development of new ideas on the complex pathogenetic mechanisms of atherosclerosis and suggesting some important anatomo-clinical correlations.
Scanning electron microscopy investigations of the surface of rat peritoneum were performed after application of isotonic saline solution and peritoneal dialysis solutions with osmolalities of 375, 528, and 680 mosmol x kg-1 and intraabdominal stay periods of 15, 30, and 45 minutes. Severe morphological changes of the peritoneal surface were shown as clotting and atrophy of microvilli, detachment of surface cells from the basement layer, destruction of the basal lamina and appearance of rough-fibrous submembranous connective tissue, which was observed even after application of isotonic saline solution into the abdominal cavity. In conclusion, sclerosing peritonitis is caused by a fruitless inflammation reaction of the organism against the unphysiological function of the peritoneum as dialysis membrane forced upon peritoneal dialysis.
This paper reviews the contribution of scanning electron microscopy (SEM) to our understanding of platelet physiology and pathology. Observations of platelet shape changes which accompany activation and the ability to visualize and analyze platelet aggregation and adhesion in three dimensions make this experimental medium an important tool in the evaluation of healthy and diseased platelets. While SEM adds a valuable third dimension to the study of morphology and ultrastructure, its greatest contribution is realized when studies are correlated directly with light and/or transmission electron microscopic observations and with studies of functional capacity.
Consecutive stages of renal glomerulogenesis were studied in white rats using scanning electron microscopy of microvessel corrosion casts and specimens dried through critical point. The splitting of ingrowing capillaries was shown to be the leading mechanism of glomerulogenesis. The differentiation of endothelial and epithelial components of glomerular filter involves two main processes: progressive spreading and arborization of cells. A suggestion is put forward that the growth and spatial reorganization of cytoskeletal elements in the endothelial and epithelial cells are the basic mechanisms of their differentiation during glomerulogenesis.
The labial organization of adult Baylisascaris procyonis was studied by scanning electron microscopy, and found to be similar for males and females. The apical part of each lip was smooth, and the basal part reticulated. The dorsal lip possessed two dorsolateral double papillae and two internal labial papillae; the two subventral lips each had one ventrolateral double and one externolateral papilla, two internal labial papillae, and an amphid. The small papilla of each double set was dome-shaped and smooth, whereas the large papilla was broad and had a prominent central pore. The externolateral papillae had raised, highly sculptured surfaces with numerous slits and creases present. Amphids resembled those of previously studied nematodes. The internal labial papillae consisted of pits. Denticles arose as a single row from the apical edge of the inner labial surface, were usually evenly spaced, and pyramidal or conoidal in shape. They were typically unicuspid, but bicuspid denticles were occasionally seen. Denticle shape and size varied between specimens and on each specimen. A pit was seen in the cuticle in the central region of the denticular row of all lips. Several of these findings represent new information concerning ascaridoid nematodes.
Scanning electron microscopy disclosed differences in various areas of the surface of the adult tapeworm, and in larval and adult scoleces of M. endothoracicus. Microtriches with apically crooked spikes were found in the circlet of hooks of the adult tapeworm, and in the area between the outer and inner sucker margins, i.e., in sites important for intestinal attachment. Dimorphic microtriches were present on the inner margin of the suckers and around the genital pore, while the only area to contain dimorphic microtriches in the larva was the surface of the rostellum. An illustration is given of the egg and its fixation to uterine branches.
Colloid resorption from thyroid follicles in control and in calcitonin-treated rabbits was studied with the scanning electron microscope. Thus, an intense cell surface activity with several vesicles (exocytotic vesicles) and large pseudopods, sometimes cylindroids, is predominant on the thyroid follicular cells of calcitonin-treated rabbits. The pseudopods are irregular, mostly elongated, cup or dome-shaped and operate by phagocytosis or engulfment of colloid droplets (endocytosis). Only microvilli are visible on the cell surface of thyroid follicles in control rabbits. Comparative ultrastructural studies also revealed the occurrence of small exocytotic vesicles and large pseudopods in thyroid glands stimulated by calcitonin. The present findings demonstrate that the scanning electron microscopy better visualizes the colloid resorption and provides more details for the role of follicular cell surface in thyroid physiology.
Spermiation and epididymal maturation of spermatozoa were studied in the bonnet monkey (Macaca radiata) by light and scanning electron microscopy. The morphology, location, and release of residual body were observed during spermiation. Along the epididymal duct, the shape of spermatozoa changed, the constriction at the anulus disappeared, the marginal thickening diminished in length, and the cytoplasmic droplet regressed and moved toward the posterior end of the middle piece. Mature spermatozoa were very similar to those of other Cercopithecidae, and they showed a forward motility and a drop in eosin stainability.