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PubMed2005

The attachment and entry of Leishmania (Leishmania) major into macrophages: observation by scanning electron microscope.

Leishmaniasis, a zoonotic protozoan disease, starts with the inoculation of the Leishmania promastigotes into the skin at the time of blood ingestion by a female sandfly. The infection of leishmaniasis is established when the Leishmania organisms start their own intracellular multiplication after having been phagocytized by the host's macrophages. In the earliest stage of the infection, therefore, the attachment of the promastigates to the macrophages is essential. We incubated a mixed culture of macrophages (JM774-1A) and Leishmania (Leishmania) major for 6 hours in vitro and observed the process of the attachment between the parasite and host cell by scanning electron microscope. We found for the first time that the attachment between the two occurred at the site of the parasite body, in addition to the previously reported sites of the flagellar tip, flagellar base, and aflagellar tip (posterior pole).

Animals↗

PubMed2003

Scanning electron microscopic study of cold-stored small bowel: comparison of Euro-Collins and Lactated Ringer's solutions.

The increasing use of the small bowel for transplantation necessitates the development of optimum preservation techniques. The aim of our study was to investigate time-related morphological changes of the rat small bowel during preservation of the organ in hypothermic Euro-Collins and Lactated Ringer's solutions. It is the only study comparing these widely used preservation solutions with scanning electron microscopy, which is especially suitable to show the intestinal mucosa, one of the tissues of the intestine most susceptible to ischemia. Small bowels were perfused with either Euro-Collins, Lactated Ringer's or physiological saline solution and placed in respective preservation solutions at 4 degrees C for 0, 3, 6 and 12 hours. Biopsy samples were prepared for examination at the light and scanning electron microscopic levels. According to the results of this study we conclude that, even Lactated Ringer's solution showed better tissue preservation than Euro-Collins solution, both preservation solutions are suitable for short term preservation of the small bowel and that further ameliorations of the preservation solutions and/or techniques are required for long term preservation.

Animals↗

PubMed2001

Beam characteristics for various sizes of annular aperture on scanning electron microscope.

Using an analogy between light optics and electron optics, we have calculated beam characteristics such as the beam profile and the optical transfer function for several sizes of annular and circular apertures on a scanning electron microscope (SEM). It has been found that an annular aperture improves the image quality with regard to several kinds of image resolution and the depth of focus at the price of good low-frequency (nu) contrast. In contrast with conventional circular-aperture SEM images, a combination of a low-nu-pass filtered, circular-aperture SEM image with a high-nu-pass filtered, annular-aperture SEM image has the potential to enhance the image quality in terms of both the image resolution and the depth of focus.

Journal Article↗

PubMed1999

Scanning electron microscopic analysis of diseased and healthy dental hard tissues after Er:YAG laser irradiation: in vitro study.

The aim of this study was to treat carious lesions, sound dentin, and enamel either with conventional methods or with an Er:YAG laser and to compare the results. Thirty freshly extracted carious human teeth were divided into two groups. In the first group, lesions were treated with burs in a dental handpiece; in the second group, we used the Er:YAG laser with parameters recommended by the manufacturer (Kavo). After treatment, the teeth were prepared for scanning electron microscopic observation. On laser-treated teeth, scaly, flaky, rough surfaces were seen: surfaces were clean with several morphological reliefs that may enhance bonding resin restoration. The Er:YAG laser beam can ablate carious dentin with an energy level of 250 mJ at 2 Hz. Sound dentin can be cut at 300 mJ and 2 Hz; for enamel, 350 mJ and 3 Hz are required. The Er:YAG laser seems to be effective in the treatment of carious lesions and in cavity preparation in vitro.

Dental Caries↗

PubMed1985

Morphologic changes in the guinea pig cochlea following cochleostomy--a preliminary scanning electron microscope study.

Fistulization of the membranous labyrinth via a surgically created endolymphatic-perilymphatic shunt (cochleosacculotomy) is currently being advocated as a treatment for endolymphatic hydrops. As a preliminary study to the investigation of the effects of labyrinthine fistulization on the hydropic condition, we sought first to reproduce the cochleosacculotomy procedure in a series of normal animals. Fistulization of the cochlear duct was performed in 15 normal animals using a tungsten wire directed via the round window through the cochlear partition in the region of the organ of Corti. The animals were sacrificed at regular intervals and the cochleas examined for histopathologic changes with the scanning electron microscope. Observations suggest that discrete permanent fistulization of the cochlear duct in the normal guinea pig results in progressive sensory cell degeneration. The implication of these observations and the apparent difference between these results and the experience thus far reported in humans are discussed.

Animals↗

PubMed2000

Angioarchitectural form, functional distributive pattern and classification of the fungiform papillae on the crossbred Japanese cat tongue anterodorsal surface in scanning electron microscopic specimens.

The purpose of this study was to undertake a three-dimensional comparative observation of the angioarchitectural form, functional distributive pattern, and to classify the fungiform papillae (FuP) found sporadically and geometrically on the oblique lines of the filiform papillae (FiP) on the entire dorsal surface of the crossbred Japanese cat foretongue in microvascular cast specimens (MVCS). By means of the corrosive resin casting technique, the MVCS of the FuP of the cat tongue were prepared and examined in detail under the scanning electron microscope (SEM). With regard to the arrangement of FiPs and FuPs on the frontal half of the antero-dorsal surface of the cat foretongue, FiP types I-V and FuP types I-IV are arranged a A form with the point in the direction of the apex and in an oblique line running from the antero-central portion to both postero-peripheral portions. In the rear half of the antero-dorsal surface of the foretongue, FiP types I-V and types FuP I-IV are arranged in a V form with the point in the direction of the radix and in an oblique line running from both antero-peripheral portions to the postero-median portion or towards the pharynx on the anterior centro-dorsal surface of the cat foretongue. With the FuP arranged in the medial zone (M) and peripheral zone (P) running from the central to the apical parts, both the peripheral parts and the pharyngeal can be classified into four types (FuP types I-IV) according to the shape and size of the main process (MP), the number of the accessory processes (AP) and the regional position of the lozenge arrangement. FuP Types I-III consisted of a MP containing a large half oval, network-like process, and the AP contained a bundle of spine-like processes arranged radially in a circle at the anterior basal margin of the MP. The FuP Type IV was made up of only one MP. In the central zone (C) of a lozenge part arranged in the A and V forms, there was no FuP as a taste organ. It was assumed that in the lozenge arrangement of the A and V forms, except for the C zone, FuPs could be classified into four types (FuP types I-IV) from the frontal portion of the antero-dorsal surface toward the pharynx on the foretongue, and would play an assistant functional role in receiving the sense of taste of foods and liquids.

Animals↗

PubMed2000

Scanning electron microscopic observations on advanced third-stage larva of Gnathostoma spinigerum after in vitro exposure to albendazole sulphoxide.

Gnathostomiasis is the parasitic disease caused by the migration of an advanced third-stage larva of Gnathostoma spinigerum. To date, albendazole is claimed to be the effective drug in preventing the reoccurrence of migratory swelling in patients. After being exposed to 1 and 2 micrograms/ml albendazole sulphoxide (AlbSO) in vitro, the parasites moved deteriorately, however, no dead larva was found even exposed to these concentrations for 21 consecutive days. The topographical alterations after 21 days of albendazole sulphoxide exposure are described using a scanning electron microscope. The marked changes in surface morphology were observed in both neck and body regions. The tegumental surface on the neck region was swollen and covered with fuzzy materials, whereas, the spines on the posterior region of the body were dislodged. These changes would probably lead to reduction of intermittent cutaneous migratory swelling in human gnathostomiasis patients.

Albendazole↗

PubMed1999

Effects of Nd:YAG lasers on root canal walls: a light and scanning electron microscopic study.

OBJECTIVE: This study evaluated amounts of debris, smear layer, and recrystallized dentin on root canal walls treated with an Nd:YAG laser. The presence of carbonized material was also assessed. METHOD AND MATERIALS: The root canals of 36 extracted incisors were prepared by stepdown and stepback techniques and divided into 4 groups. One group was not treated further, while 3 others were exposed to the laser beam at an energy density of 159, 239, or 318 J/cm2. The roots were split longitudinally, photographed, and examined using a scanning electron microscope. The amounts of debris, smear layer, and recrystallized dentin were recorded from horizontal bands around the root, 2, 6, and 10 mm from the apices. RESULTS: Mean smear layer scores were similar for all groups at the 3 levels examined, ranging from 2.157 to 2.851 on a scale of 0.0 to 3.0. Mean debris scores were similar for all groups at the 3 levels examined, ranging from 0.774 to 1.408 on a scale of 0.0 to 3.0. Recrystallized dentin was recorded in all 3 irradiated groups at the 2-mm level, but only in the group irradiated with 318 J/cm2 at the 6- and 10-mm levels. Severe carbonization was recorded at the 2-mm level. CONCLUSION: Use of the Nd:YAG laser did not reduce the mean amounts of debris and smear layer compared to a nonirradiated group. Carbonization was recorded in the apical part of the canal, but this is not obvious when only scanning electron photomicrographs are examined.

Dental Pulp Cavity↗

PubMed2002

Organization and distribution of intramuscular connective tissue in normal and immobilized skeletal muscles. An immunohistochemical, polarization and scanning electron microscopic study.

Collagen fiber network is a major contributor to the coherence and tensile strength of normal skeletal muscle. Despite the well-recognized importance of the intramuscular connective tissue to the normal integrity and function of the skeletal muscle, the specific architecture including the location and three-dimensional orientation of the intramuscular connective tissue within the muscle tissue is poorly described. The structure of the intramuscular connective tissue was studied by immunohistochemistry, polarization microscopy (the crimp length and angle of the collagen fibers) and scanning electron microscope (SEM) in rat skeletal muscles (gastrocnemius, soleus and tibialis anterior) in normal situation and after 3 weeks of disuse (immobilization). Three separate networks of collagen fibers were distinguished by SEM in the normal endomysium; fibers running longitudinally on the surface of the muscle fibers (the main collagen orientation), fibers running perpendicularly to the long axis of the muscle fibers and having contacts with adjacent muscle fibers, and fibers attached to the intramuscular nerves and arteries. Similarly, the SEM analysis also disclosed three distinct collagen fiber networks running in different directions in the perimysium, but, contrary to the endomysium, the main fiber orientation could not be established. Immobilization resulted in a marked increase in the endo- and perimysial connective tissue, the majority of the increased endomysial collagen being deposited directly on the sarcolemma of the muscle cells. Immobilization also resulted in substantial increase in the number of perpendicularly oriented collagen fibers with contacts to two adjacent muscle fibers in the endomysium. Further, immobilization clearly disturbed the normal structure of the endomysium making it impossible to distinguish the various networks of fibers from each other. In the perimysium, immobilization-induced changes were similar, the number of longitudinally oriented collagen fibers was increased, the connective tissue was very dense, the number of irregularly oriented collagen fibers was markedly increased, and consequently, the different networks of collagen fibers could not be distinguished from each other. Of the three studied intact muscles, the crimp angle of the collagen fibers was lowest in the soleus and highest in the gastrocnemius muscle, and the crimp angle decreased over 10% in all muscles after the immobilization-period. Altogether, the above described quantitative and qualitative changes in the intramuscular connective tissue are likely to contribute to the deteriorated function and biomechanical properties of the immobilized skeletal muscle.

Animals↗

PubMed2002

A scanning electron microscopic study of phenotypic plasticity and surface structural changes of aortal smooth muscle cells in primary culture.

Phenotypic modulation of smooth muscle cells (SMCs) from a contractile to a synthetic state characterised by active proliferation appears to be an early event in the pathogenesis of atherosclerosis. A similar transition occurs when SMCs are established in culture. In this study the phenotypic plasticity and surface structural changes of aortal smooth muscle cells during the transition from the contractile to the synthetic state and during maturation have been structurally assessed by scanning electron microscope (SEM). The experiments were performed on SMCs obtained from aorta of neonatal rats after enzymatic digestion and then cultured on glass coverslips. SEM observations revealed a three-dimensional appearance characteristic for different stages of SMCs. Intensively proliferating cells from monolayer region were large, polygonal in shape with lamellipodia and well spread. Long, uniform in diameter, finger-like microvilli were densely arranged on the surface of these cells. In the thickened region of culture, the cells were rather small, generally spindle-shaped, not well spread, with low density of short, bubble-like microvilli on the surface. Numerous plasma membrane structural alterations in apoptotic cells were observed by SEM: loss of cellular adhesion, smoothing, shrinkage and outpouching of membrane segments have been recognised as markers associated with the cell injury and death. It was concluded that scanning microscopy observations would allow a more complete understanding of SMCs and their changes in culture and atherosclerotic disease.

Animals↗

PubMedDate not supplied

Measurement technique for the incident electron current in secondary electron detectors and its application in scanning electron microscopes.

A measurement technique for incident electron current in secondary electron (SE) detectors, especially the Everhart-Thornley (ET) detector, based on signal-to-noise ratio (SNR), which uses the histogram of a digital scanning electron microscope (SEM) image, is described. In this technique, primary electrons are directly incident on the ET detector. This technique for measuring the correlation between incident electron current and SNR is applicable to the other SE detectors. This correlation was applied to estimate the efficiency of the ET detector itself, to evaluate SEM image quality, and to measure the geometric SE collection efficiency and the SE yield. It was found that the geometric SE collection efficiency at each of the upper and lower detectors of a Hitachi S-4500 SEM was greater than 0.78 at all working distances.

Journal Article↗

PubMed1993

Studies on Echinococcus granulosus using the scanning electron microscope. I. Preparations of the parasite for infection of the final host.

The development of the protoscoleces of Echinococcus granulosus was studied after chemical stimulation, which mimics that offered by the final host. This stimulation resulted in some morphological changes in the parasite and to its evagination. These changes prepare the parasite to infect the intestine of its final host. The external morphology of the protoscolex body and of the germinal layer of the cyst and brood capsule were studied using the scanning electron microscope (SEM). Findings were related to other studies using the transmission electron microscope (TEM). The morphology of the flame cell is described and four cases of teratogenesis are reported.

Animals↗

PubMed1990

A light and scanning electron microscopic study of enamel decalcification in children living in a water-fluoridated area.

Eleven children, each having one or two pairs of premolars to be extracted for orthodontic purposes, participated in the study. The model involved placement of a special orthodontic band that allowed the accumulation of plaque in a defined area between the band and the buccal enamel. Examination of enamel changes was carried out in experimental teeth that had been exposed to local plaque accumulation for one, two, four, eight or 14 days. The specimens were examined under the light (LM) and the scanning electron microscope (SEM). All teeth had signs of very mild dental fluorosis. No indications of demineralization were noted after one day. SEM examination showed signs of crystal dissolution in some of the two-day specimens. Six of eight four-day specimens exhibited surface dissolution. All eight- and 14-day specimens showed signs of surface demineralization in the LM as well as in the SEM. These observations documented that undisturbed bacterial deposits are capable of initiating enamel demineralization within short time periods, even in children living in a water-fluoridated area.

Adolescent↗

PubMedDate not supplied

Plaster of Paris and hair cell morphology. A scanning electron microscopic study of an alternative implant material for ear surgery.

Many techniques have been proposed for reconstruction of the posterior canal wall and/or obliteration of the mastoid bowl after radical mastoidectomy. The variety of materials that have been used, biological as well as foreign materials of different kinds, indicates that the ideal solution has yet to be found. Plaster of Paris, a biocompatible, degradable ceramic material prepared from CaSO4, may have an osteogenic property and become an alternative implant material for ear surgery. However, its possible ototoxicity has not been studied previously. Plaster was implanted in the tympanic cavity of guinea pigs. On investigation in the scanning electron microscope after 4.5 to 8 months, no toxic damage to the hair cell morphology was observed. Thus, from an ototoxic point of view, plaster of Paris would seem suitable for use as an implant material in ear surgery. Studies concerning the possible osteogenic property of plaster of Paris and its clinical application in human ear surgery are in progress.

Animals↗

PubMed1997

[Morphological development of the intrinsic laryngeal muscles in rat--a scanning electron microscopic and histochemical study].

The postnatal development of the subneural apparatus (SNA) and differentiation of muscle fibers of the intrinsic laryngeal muscles (ILMs) in the rat were investigated on the 1st, 5th, 15th, 30th and 90th days after birth. The cricothyroid (CT), thyroarytenoid (TA) and posterior cricoarytenoid (PCA) muscles in the ILMs were examined. Furthermore, we compared the development of these muscles with that of the skeletal muscles of the hindlimb, the extensor digitorum longus (EDL) and soleus (SOL) muscles. The SNA was observed with a scanning electron microscope. The SNA was quantitatively analyzed by use of two parameters: the ratio of the length of the secondary synaptic cleft to its maximal width (L/W ratio) and the ratio of the area of the secondary synaptic clefts to that of the primary synaptic cleft(s) (ASC/APC ratio). The muscle fiber types were classified according to the density of each muscle fiber by the ATPase stain. On the 15th postnatal day, the SNAs of the CT and TA muscles had a tendency to show more advanced development than those of the PCA muscle. The SNAs of the ILMs appeared to develop earlier than those of the EDL and SOL muscles. The L/W and ASC/APC ratios of the ILMs inclined to be higher than those of the hindlimb muscles. On the 30th day, the SNAs of the PCA muscle were still immature, while those of the CT and TA muscles had become nearly mature. The EDL muscle showed completely mature development and outstripped the ILMs. In contrast, the SNA of the SOL muscle was the least mature of all muscles. The L/W ratios of the ILMs were not significantly different from each other, while the ASC/APC ratio of the PCA muscle was significantly lower than those of both the CT and TA muscles. The L/W and ASC/APC ratios of the EDL muscle were the highest of all muscles examined and those of the SOL muscle were the lowest. The muscle fiber types were not significantly different in the ILMs throughout the course. On the 30th day, the EDL muscle had reached the mature adult proportion of the muscle fiber types, while the ILMs and SOL muscle needed further development. Phylogenetically, the TA muscle appeared as a sphincter of the lower airway and then the PCA muscle, as a dilator, appeared. The CT muscle arose after separation of the thyroid and cricoid cartilages. These findings infer that the lag in development of the PCA to the TA muscle in early postnatal life was due to the importance of the protective function of the lower airway and the phylogenetical difference. Moreover, these results suggest that the ILMs, which are indispensable for sucking, develop early in life, followed by rapid development of the EDL muscle to start quadruped walking after the 15th day. Thus, different muscles need to develop as an animal grows because distinct functions of muscles are necessary for normal living. In conclusion, the ILMs may have a specific mode of postnatal development in contrast with those of the hindlimb muscles.

Animals↗

PubMed1982

The scanning electron microscope in craniofacial remodeling research: application of the topographic principle.

Growth and development studies utilizing craniofacial skeletal material are best done by documenting evidence of the bone growth mechanisms responsible for morphogenesis. One primary mechanism is remodeling which involves coordinated bone resorption and deposition in localized areas. Some histological studies have been performed on human facial remodeling, but these studies have been limited in extent because of the destructive nature of the technique. For obvious reasons, large skeletal research collections cannot be utilized in histological work. A new technique is presented which does not damage specimens. This involves making high resolution replicas of subadult craniofacial bone which are then examined with the scanning electron microscope (SEM) for evidence of the characteristic microscopic surface topography of remodeling bone. The Topographic ("T") principle is introduced as a precedent for discriminating remodeling bone activity states with the SEM. These activity states in vivo specify characteristic microscopic surface topographies. The three distinctive surfaces are resorptive, depository, and resting. These surfaces can be mapped on a coordinate representation of the bone replica to obtain results which are similar to those of histological studies. Application of the "T" principle and SEM/replica methodological approach offers great promise to the study of craniofacial morphogenesis.

Animals↗

PubMed1993

Two-dimensional crystallization of streptavidin by nonspecific binding to a surface film: study with a scanning electron microscope.

A two-dimensional (2D) crystal of streptavidin has been obtained by a nonspecific binding method. The protein molecules were bound and formed a dense packing on the film of poly(1-benzyl-L-histidine) spread at the surface of protein solution. The surface film was moderately heated to stimulate crystallization of bound streptavidin. A potential of this method for obtaining 2D crystals of soluble proteins is demonstrated. The present 2D crystal structure of streptavidin resembles that previously obtained by specific binding to biotinylated lipid. We show in addition that the 2D array of protein with usual size approximately 50 A can be imaged using a high resolution scanning electron microscope (HR-SEM) and subject to structural analysis at low resolution. Various limitations in HR-SEM degrade considerably the image quality. However, the usability of a bulk plate as specimen support would make HR-SEM a convenient tool, when such a substrate must be considered in application of protein arrays, and if an intrinsic low resolution is acceptable.

Bacterial Proteins↗

PubMed1992

[Effects of uro-angiographic contrast media on blood coagulation--a scanning electron microscopic study on time dependent changes of blood clot and a clinical study].

Anticoagulant activity of ionic and nonionic contrast media (CM) was investigated in vitro and in vivo. Based on the time course of FPA and TAT generations and gross examinations of the blood clots on the catheters placed in CM-blood mixtures [an 2 to 8 ratio (20% v/v)]. It was demonstrated that blood coagulation was activated during the period of 20 to 30 minutes when nonionic CM (iopamidol, iohexol) was employed, but no activation of blood coagulation was noted with ionic CM (diatrizoate, ioxaglate). Scanning electron microscopic examinations of the clots on the catheters revealed that fine fibrin meshwork fibers, in which many red blood cells were trapped in bound, were observed with nonionic CM. In contrast, no fibrin mesh was formed with ionic CM after 30 minutes. In vivo, antithrombin III and fibrinogen significantly decreased in the patients who underwent infusion of nonionic CM. Our studies confirmed that nonionic CM show weaker anticoagulant activity than do ionic CM. And these findings account for previous reported thromboembolic complications with the use of nonionic CM. Extreme caution should be therefore exercised when nonionic CM are employed during prolonged angiographic and interventional procedures.

Blood Coagulation↗