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PubMedDate not supplied

Imaging of anisotropic cellulose suspensions using environmental scanning electron microscopy.

The effect of concentration on anisotropic phase behavior of acid-hydrolyzed cellulose suspensions has been examined using conventional polarizing microscopy and the novel technique of environmental scanning electron microscopy (ESEM). Microcrystalline cellulose dispersed in water formed biphasic suspensions in a narrow concentration range, 4-12 wt % for a suspension pH of 4, where the upper and lower phases were isotropic and anisotropic (chiral nematic), respectively. It is known from previous work that within the biphasic regime total suspension concentration affects only the volume fractions of the two phases, not phase concentration or interfacial packing. As the total suspension concentration surpassed the upper critical limit (c), however, a single anisotropic phase of increasing concentration was observed. It was evident from polarizing microscopy that the chiral nematic pitch of the anisotropic phase decreased with increasing concentration, which has been attributed to a reduction in the electrostatic double layer thickness of the individual rods, thus increasing intermolecular interactions. Chiral nematic textures were also visible using ESEM. This technique has the advantage of studying individual rod orientation within the liquid crystalline phase as it permits the high resolution of electron microscopy to be applied to hydrated samples in their natural state. To our knowledge this is the first time such lyotropic systems have been observed using electron microscopy.

Anisotropy↗

PubMed2003

Vascular system of intramural leiomyomata revealed by corrosion casting and scanning electron microscopy.

BACKGROUND: The vascular system of leiomyomata, the most common benign tumours in women, is an important factor controlling development and growth of the tumour. It has not been, however, investigated morphologically using the best currently available technique, corrosion casting combined with scanning electron microscopy. METHODS: Myomatous uteri collected upon autopsy were perfused via afferent vessels with fixative followed by Mercox resin and corroded after polymerization of the resin. The obtained vascular casts visualizing all vessels including capillaries were examined using scanning electron microscopy. RESULTS: The smallest (1-3 mm) fibroids were avascular, in larger ones (<1 cm) a few small vessels invaded the lesion from the periphery. The largest tumours (>1 cm) contained irregular networks of blood vessels with density similar to or lower than that of normal myometrium. Such tumours were surrounded by an extremely dense vascular layer ('vascular capsule') which was the source of larger vessels supplying and draining the tumour. CONCLUSIONS: During development of leiomyoma, the pre-existing blood vessels undergo regression and new vessels invade the tumour from the periphery, where intense angiogenesis, probably promoted by growth factors secreted by the tumour, leads to the formation of a 'vascular capsule' responsible for supply of blood to the growing tumour.

Adult↗

PubMed1990

Identification of Actinobacillus actinomycetemcomitans by gold immunolabeling and scanning electron microscopy.

Recent evidence suggests that dental plaque is not a homogeneous bacterial mass but, on the contrary, specific bacterial morphotypes and species may be preferentially located within certain microenvironments. The aim of the present study was to develop combined gold immunolabeling and scanning electron microscopic (SEM) techniques for the identification of periodontal pathogens in subgingival dental plaque and on the root surfaces of extracted teeth. Suspensions of pure A. actinomycetemcomitans cultures or suspensions of A. actinomycetemcomitans mixed together with other oral bacteria were prepared, labeled with goat anti-rabbit IgG conjugated with 5 nm or 40 nm colloidal gold particles, and observed by SEM using both secondary and back-scattered imaging. Scanning electron microscopy demonstrated that the A. actinomycetemcomitans bacterial cell surface was specifically labelled. There was no cross-reaction with any of the other bacterial morphotypes. Only the labeled A. actinomycetemcomitans were visible in mixtures examined by back-scattered imaging scanning electron microscopy. The combined techniques of gold immunolabeling and SEM may, therefore, be useful in identifying A. actinomycetemcomitans in subgingival plaque samples and on the root surfaces of extracted teeth as well as in studies of bacterial ecology in dental plaque, in general.

Actinobacillus↗

PubMed1986

Scanning electron microscopy of conjunctival surfaces in patients with ocular cicatricial pemphigoid.

The conjunctival surfaces of ten patients with active, ocular cicatricial pemphigoid, three patients with drug-controlled ocular cicatricial pemphigoid, and six patients with normal conjunctivas were studied using scanning electron microscopy. A homogeneous granular sheet of amorphous mucin-like material was observed covering extensive areas of the conjunctiva in eight of ten patients with active ocular cicatricial pemphigoid. This sheet of amorphous material was absent on drug-controlled ocular cicatricial pemphigoid and normal conjunctival specimens. Our study demonstrates that patients with active ocular cicatricial pemphigoid possess ocular surface mucus that appears thicker and more continuous than normal ocular mucus when observed with scanning electron microscopy. This observation is in agreement with clinical observations of thick mucus strands in the inferior fornix of patients with active ocular cicatricial pemphigoid.

Conjunctiva↗

PubMed1997

[Study on the organization of chromatin and chromosome in mouse spermatogenic cells by scanning electron microscopy].

It remains unclear about the intermediate construction of chromosome due to its highly compact nature and the limitation in methods. The present study was designed to investigate the construction of chromatin and mitotic chromosome in situ with scanning electron microscopy. Mouse testes were selected as the material, because of in which the spermatogenic cells divide actively and successively to form the sperm. Such a feature would be able to study the structure of mammalian chromatin and chromosomes along with the change of nuclear cycle. The animal were perfused with 200 ml of 0.075 mol/L KCl hypotonic solution to remove blood and placed for 15-20 min on ice followed by 0.5% glutaraldehyde and 0.5% formaldehyde for fixing. Through treated by the routine process of fractured and freeze dried with t-butyl alcohol, the specimens were then coated with a 3 nm thick platinum and observed with Hitachi S-430 scanning electron microscopy. It was found that the hypotonic treatment with 0.075 mol/L KCl solution was suit for demonstrating the nuclear structure, when the organelles were well preserved. The chromatin fibers of 10-30 nm and 80-125 nm in diameter could be recognized in the interphase nuclei, which were arranged losely at the region of euchromatin, and folded with each other into chromatin masses at the region of heterochromatin, while the chromatin fibers with the diameter of 80-125 nm often could be viewed on the mitotic chromosomes. Since its presence in interphase nuclei and mitotic chromosomes, it was considered that the chromatin fibers with 80-125 nm in diameter might play a role in the condensation of chromosome, serve as a type of the intermediate structure.

Animals↗

PubMed1996

Dynamics of salivary secretion studied by confocal laser and scanning electron microscopy.

Plasma membrane events during secretion of the parotid and submandibular glands of humans and rats were observed in living cells by confocal microscopy and from the cytoplasmic side by scanning electron microscopy after removal of the cell organelles by the OsO4 maceration method. These new microscopic techniques revealed in acinar cells two distinct exocytosis-endocytosis coupling mechanisms elicited in response to different secretory stimuli. Beta-adrenergic stimulation with isoproterenol and its second messenger analog dibutyryl cyclic AMP evoked exocytosis after which fused granule membranes were individually removed from the luminal plasma membrane within several minutes. On the fused membrane area, small endocytotic vesicles about 100-150 nm in diameter were abundant. Muscarinic stimulation with carbachol also induced exocytosis, but in this case the fused granule membranes coalesced to form enlarged invaginations which stayed on the luminal membrane for more than 30 min. These invaginations, almost devoid of small endocytotic vesicles, were then pinched off from the luminal membrane and dispersed into the cytoplasm in the form of light microscopically detectable vesicles. After treatment with isoproterenol and carbachol, acinar cells exhibited different distributional changes of F-actin around the fused membrane area, suggesting the involvement of microfilaments in regulating the membrane events following exocytosis.

Actin Cytoskeleton↗

PubMed1985

Relationship of Mallory bodies to intermediate filaments in hepatocytes. A scanning electron microscopy study.

Livers from 12 mice fed griseofulvin for 4 to 6 months were perfused in situ with a detergent solution to extract lipid membranes leaving the cytoskeleton intact. Seven control mice were similarly studied. After 30 to 120 minutes perfusion, liver samples were examined by scanning electron microscopy and transmission electron microscopy. By light microscopy, Mallory bodies (MBs) were observed in pericentral hepatocytes. These were confirmed by transmission electron microscopy. Intermediate filaments (IFs) were observed in close apposition to MBs. Numerous IFs were seen throughout the cytoplasm. The 3-dimensional organization of the cytoskeleton and MBs were clearly visualized by scanning electron microscopy. The IFs were disorganized in the hepatocytes and formed small MBs in griseofulvin-treated mice. In the case of hepatocytes containing large MBs, there was an apparent decrease in the concentration of IFs. Transition forms of dense networks of IFs between the normal cytoskeleton and the MBs were noted in the cytoplasm between small MBs and the nucleus and also at the cell border. The IFs connected to the nucleus and invaded the MBs. The MBs appeared to form as a result of condensation or collapse of the IFs.

Animals↗

PubMed1999

On the rugose area and caudal papillae distribution in Dirofilaria immitis (Leidy, 1856) Railliet & Henry, 1911, by scanning electron microscopy.

The objective of this study was to obtain a better characterization of Dirofilaria immitis (Leidy, 1856) Railliet & Henry, 1911, in Brazil, ratifying previous accounts and adding new data on the rugose area on the ventral surface of the spiralled posterior portion of males observed by scanning electron microscopy, which presents a different arrangement compared to other species of the genus, and differs also from the genera Litomosoides Chandler, 1931, and Wuchereria Silva Araújo, 1877, adding another taxonomic character to Dirofilariinae and Onchocercinae. Scanning electron microscopy studies also showed some aspects of the distribution of caudal papillae in D. immitis that have not been described before.

Animals↗

PubMed1993

Delaying effects of dietary eicosapentaenoic-docosahexaenoic acids on development of "fatty streaks" in hypercholesterolaemic rabbits. A morphological study by scanning electron microscopy.

Besides causing functional and clinical damage, hypercholesterolaemia also causes morphological alterations of vascular endothelium. Rabbits fed a diet with 1% cholesterol for 4, 6, or 8 weeks are experimental models for hypercholesterolaemia, with pathological structural changes in vascular luminal surface. Morphological investigation by scanning electron microscopy was performed to reveal the tridimensional growth of these lesions and the differences in this growth induced by concomitant dietary assimilation of fish-oil (2 g/day). Macroscopic reduction in fatty-streak production was clearly seen in rabbits fed fish-oil. Scanning electron microscopy confirmed that the area of intimal lesions was only 21 +/- 6% in this group, while in the group fed cholesterol without fish oil, the lesioned area attained 76 +/- 5%. Endothelial swelling was less marked, probably due to reduced intracellular lipid accumulation into the foam cells. Adherent macrophages were also fewer. The differences might be correlated with protection against the lipoproteins' atherogenic effects and to hemorheological benefits produced by the Omega-3 fatty acid (85%) present in fish-oil.

Animals↗

PubMed2000

Hydrophilic bile salts protect bile duct epithelium during cold preservation: a scanning electron microscopy study.

Prolonged graft preservation is associated with postoperative bile duct strictures after liver transplantation. We previously showed that hydrophilic bile salts mitigate bile duct preservation injury in a pig model. Because this injury occurs at the epithelial level, scanning electron microscopy was performed to further characterize this effect in vitro. Swine livers were harvested after the intravenous infusion of 1 of 3 solutions: saline (n = 7), tauroursodeoxycholate ([TUDC] hydrophilic; n = 4), or taurodeoxycholate ([TDC] hydrophobic; n = 4). Livers were perfused with University of Wisconsin solution. The bile ducts were flushed retrograde, and the liver was stored at 0 degrees C to 1 degrees C for 20 hours. Bile duct samples were obtained at the time of harvest and 8, 12, 16, and 20 hours thereafter. In saline-infused controls at time 0, the epithelium was intact and composed of uniform cuboidal cells covered with fine regular microvilli. There were no spaces between individual cells. After 8 to 12 hours of preservation, cells were more irregular in shape, with loss of cell-cell contact. The cell surfaces showed fewer microvilli. Surface erosions suggested loss of cell-wall integrity. TUDC was protective, evidenced by normal-appearing cells with uniform microvilli after 16 hours. In contrast, TDC accelerated the injury process, causing cell-surface erosions, blebs, and loss of microvilli as early as time 0. Scanning electron microscopy is an excellent tool to study injury to bile duct epithelium. This study supports the hypothesis that hydrophilic bile salts protect bile ducts during preservation. To determine whether treatment with hydrophilic bile salts can prevent postoperative stricture, in vivo transplantation studies are needed.

Animals↗

PubMed2006

Scanning electron microscopy of the three different types of cementum in the molar teeth of the guinea pig.

The aim of this study was to observe the three-dimensional distribution and structural characteristics of the three different types of cementum in the molar teeth of guinea pig by means of scanning electron microscopy. Twenty-five 4-week-old male guinea pigs were used in this study. Using decalcified and undecalcified specimens with or without NaOH maceration, we examined the mandibles, maxillae and extracted molars by scanning electron microscopy. Guinea pig molars consist of two longitudinal, deeply folded lamina cores covered by enamel on all surfaces, except the buccal surface of the upper molars and the lingual surface of the lower molars. In the regions without enamel, we observed continuous thin belt-like layers of conventional acellular cementum on the dentin surface. On the enamel-covered surfaces, two different types of coronal cementum were found: small circular islands of coronal cementum called cementum pearls, which were distributed widely at almost regular intervals on the peripheral enamel surface from the apical fifth to the occlusal surface; and cartilage-like cementum, which occupied almost all of the occlusal half of the two longitudinally folded grooves. The present study demonstrated the unique distribution pattern of the three different types of cementum in guinea pig molars. These cementum types may contribute to the requirements for many different functions such as mastication, anchorage and continuous tooth eruption.

Animals↗

PubMed1991

Alcoholic cirrhosis and osteoporosis in men: a light and scanning electron microscopy study.

Subjects with chronic alcoholism are associated with a higher prevalence of bone fractures, compared with age-matched controls. However, the pathogenesis of alcoholic osteopathy remains poorly understood. In this study, the bone cells activities and the bone matrix were studied using different techniques such as bone morphometry, scanning electron microscopy and computer reconstruction. Male patients (N = 20), aged 59.1 +/- 10.1 years, presenting a chronic decompensated alcoholic cirrhosis, were admitted into this study. A histomorphometric analysis of a transiliac bone biopsy was done after a double tetracycline labeling of the bone. A scanning electron microscopy (SEM) study was performed on eight out of the 20 patients on an additional biopsy. The bone mass was significantly decreased in cirrhotic patients. A marked defect in the osteoblastic function was observed with reduced osteoid parameters, lower mean wall thickness, and slower bone formation rate leading to a thinning of bone trabeculae. Conversely, trabecular resorption surfaces were markedly increased. SEM examination of bone biopsies was also consistent with delayed and impaired osteoblastic activity leading to extended and scalloped resorption surfaces covered by unusually thin layers of calcified collagen fibers. The reduced osteoblastic activity associated with normal osteoclastic function appears to play a major role in the pathogenesis of alcoholic osteoporosis leading to decreased bone mass with thinner trabeculae.

Aged↗

PubMed1995

Bone resorption in human cholesteatoma: morphological study with scanning electron microscopy.

The detailed mechanism of bone resorption in cholesteatoma was investigated by means of eroded ossicles obtained during middle ear surgery for cholesteatoma. In the light microscopic study, multinucleate osteoclasts with ruffled borders were found in contact with the eroded bone, which appeared to be osteoclastic lacunae. Scanning electron microscopy revealed the lacunae to be many absorption bays, 30 to 100 microns in diameter, clustered on the surface of eroded areas. Numerous cone-shaped stubs of digested collagen fiber bundles, consisting of scores of collagen fibrils and degenerated extracellular organic substances, were visible at higher magnification on the bottom of the absorption bay. The pattern of fusion and twining of the disarranged collagen fibrils at the top of the partly digested fiber bundles was clearly rendered by the alkali-water maceration method for scanning electron microscopy. We infer from the morphological evidence that osteoclastic resorption may be one of the major mechanisms of bone destruction in cholesteatoma, and that demineralization and degeneration of extracellular organic substances precede disruption of collagen fibrils at the front of bone resorption.

Bone Resorption↗

PubMed2005

Three-dimensional ultrasound biomicroscopy, environmental and conventional scanning electron microscopy investigations of the human zonula ciliaris for numerical modelling of accommodation.

PURPOSE: Biomechanical modelling of the accommodation process is a useful tool for studying the mechanism of accommodation and presbyopia and can aid in the development of accommodative lens-replacing materials. Existing biomechanical models, however, use a very simplified zonula structure. The aim of this study was to use three-dimensional ultrasonic imaging and scanning electron microscopy to provide a more detailed, three-dimensional description of the structure of the human zonula to improve the modelling of accommodation. METHODS: Five human eyes were examined without invasive manipulation using a custom-made three-dimensional ultrasonic imaging technique that allows scanning of features with a spatial resolution of 30 microm. Environmental and conventional scanning electron microscopy (SEM) provided information to complement the ultrasonic images for use in development of a more anatomically correct finite-element model of the zonula structures. These data along with the material properties of the ocular tissue structures were used to construct an advanced geometric model for finite-element simulation of the accommodation process. RESULTS: Images were obtained through three-dimensional ultrabiomicroscopy (3D-UBM) of anatomical features heretofore not directly imagable in their native state. Ciliary processes and zonula structures were clearly separated by both the 3D-UBM and the SEM methods. It was found that fibres inserting on the anterior and posterior lens capsule emerge anteriorly at the ciliary body. Fibres emerging near the pars plana insert on the lens and the ciliary body. No X-shaped crossing fibres were found. Modelling of the accommodation process with both the simple and the more complex geometric models produced refractive power changes comparable with in vivo findings. CONCLUSIONS: The 3D-UBM allowed examination of zonula structures in their native state with minimized preparation artefacts. While these data were incorporated into a complex and more anatomically correct finite-element simulation of intraocular features including lens, zonular system and ciliary body it was found that a simplified zonular model is sufficient for the numerical simulation of the accommodation process.

Accommodation, Ocular↗

PubMed2000

Different patterns of collagen-proteoglycan interaction: a scanning electron microscopy and atomic force microscopy study.

The extracellular matrix of unfixed, unstained rat corneal stroma, visualized with high-resolution scanning electron microscopy and atomic force microscopy after minimal preliminary treatment, appears composed of straight, parallel, uniform collagen fibrils regularly spaced by a three-dimensional, irregular network of thin, delicate proteoglycan filaments. Rat tail tendon, observed under identical conditions, appears instead made of heterogeneous, closely packed fibrils interwoven with orthogonal proteoglycan filaments. Pre-treatment with cupromeronic blue just thickens the filaments without affecting their spatial layout. Digestion with chondroitinase ABC rids the tendon matrix of all its interconnecting filaments while the corneal stroma architecture remains virtually unaffected, its fibrils always being separated by an evident interfibrillar spacing which is never observed in tendon. Our observations indicate that matrix proteoglycans are responsible for both the highly regular interfibrillar spacing which is distinctive of corneal stroma, and the strong interfibrillar binding observed in tendon. These opposite interaction patterns appear to be distinctive of different proteoglycan species. The molecular details of proteoglycan interactions are still incompletely understood and are the subject of ongoing research.

Animals↗

PubMed1996

Morphometry, scanning electron microscopy and X-ray spectral microanalysis of protostylid pits on human lower third molars.

Protostylid pits on 50 lower third molars were studied. The depth and width of the pits, the thickness of the enamel at the pit bottom and the concavity of the dentino-enamel junction under the pit were measured by light microscopy. The pit content was examined by scanning electron microscopy and X-ray spectral microanalysis. The results show that the morphometric variables of protostylid pits are similar to those of occlusal fissures and pits. The protostylid pits were seen to contain globular formations, belonging mainly to dental calculus; organic rod-shaped bodies, which could represent remnants of the enamel organ--or more probably--foreign bodies, were frequently observed. The study suggests that the development of the most common form of protostylid is basically similar to that of an occlusal cusp, with variations attributable to the low expression of the protostylid and particularities of cusp development in the lateral tooth wall.

Calcium↗

PubMed1989

Comparison of backscattered scanning electron microscopy and microradiography of secondary caries.

Carious lesions are usually studied using light microscopy and/or microradiography which require preparation of thin sections. Backscattered scanning electron microscopy (BSEM) has received little attention although it provides information similar to that obtained with microradiographs, with the potential for higher resolution. Recently, microscopes have been introduced that can be used to study wet or nonconducting specimens, offering techniques for studying specimens without desiccation or preparation of thin sections. This investigation sought to determine if secondary carious lesions have the same characteristics when studied by microradiography as when using the "wet" BSEM mode. Microradiographs were made of thin sections from restored teeth with secondary caries induced in an artificial caries system. The thin sections were also studied by BSEM with a partial pressure in the specimen chamber to prevent specimen charging. Comparisons of the lesion size and shape were made using the two methods. Lesion depth measurements in enamel were the same; lesions that penetrated into dentin appeared to be of similar size and shape, but lesion depths measured by BSEM were slightly greater (paired t-test, p less than .05). This was a result of cracks at the carious enamel-dentin interface that probably developed during storage of the samples. Variations in the surface enamel rod structure and the development of subsurface lesions were apparent. Several zones were also apparent in the carious dentin, demonstrating loss of dentinal tubule detail in the depth of the lesion, collapse of tubules, and hypermineralized regions near the advancing front of the lesion. Several additional samples of natural carious teeth were examined. They demonstrated the characteristic structural features of the carious process.(ABSTRACT TRUNCATED AT 250 WORDS)

Dental Caries↗

PubMed1997

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↗