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PubMed1982

Scanning electron microscopy of retinal ganglion cells by applying D,L-2-aminoadipic acid.

Retinal ganglion cells of the cat were clearly demonstrated in scanning electron microscopy. For identification of the cells, D,L-2-aminoadipic acid, a gliotoxic compound, was applied. Müller cells as well as astrocytes were destroyed by intravitreal injection of the aminoadipic acid prior to the enucleation. Balloon-shaped retinal ganglion cells with an axonal process were visualized in the normal cat retina. Flattened and sometimes indented ganglion cells were observed in the pathologic retina which was made 2 weeks after optic nerve transection. Pretreatment with the aminoadipic acid was a very effective method for the identification of the retinal ganglion cells by scanning electron microscopy.

2-Aminoadipic Acid↗
PubMed1976

Vascular changes in the urinary bladder wall after irradiation (an experimental study in rabbits with angiography and scanning electron microscopy of vascular casts).

Seven adult rabbits were used to study radiation-induced vascular changes in the urinary bladder by angiography and scanning electron microscopy of vascular casts. The angiographic studies, in which a magnification techniques was used, revealed a state of hypervascularization in the irradiated area of the bladder. With the angiographic method, however, it was not possible to visualize specific radiation-induced vascular changes within the irradiated area of the urinary bladder. Scanning electron microscopy of vascular casts made possible a detailed study of finer vessels in which typical radiation changes, teleangiectases, could be demonstrated.

Angiography↗
PubMed1994

Meshwork structures in bovine glomerular and tubular basement membrane as revealed by ultra-high-resolution scanning electron microscopy.

We examined the ultrastructure of the bovine glomerular basement membrane (GBM) and tubular basement membrane (TBM) using ultra-high-resolution scanning electron microscopy after conductive staining without metal coating. Purified basement membranes (BMs) were obtained by sonication and acellular BMs by detergent treatments. Purified GBMs (PGBMs) and acellular GBMs (AGBMs) showed similar meshwork structures composed of regular round or oval pores and branching strands. Pore diameters were 10.2 +/- 2.4 nm (mean +/- SD) in PGBMs and 9.8 +/- 3.1 nm in AGBMs. Purified TBMs (PTBMs) and acellular TBMs (ATBMs) exhibited heterogeneous meshwork structures in which compact meshes in the cristae were combined with coarse meshes in the invaginations on the epithelial surfaces. Pore diameters were 12.6 +/- 5.2 nm in PTBMs and 11.4 +/- 4.0 nm in ATBMs which were significantly larger than those in GBMs. The width of the strands ranged from 3 to 15 nm in all BMs. A 4 M guanidine hydrochloride extraction of the acellular BMs revealed large polygonal networks in the invaginations of the TBM and twisted strands which were considered to be type IV collagen fibrils. Ultra-high-resolution scanning electron microscopy and conductive staining were useful for the study of three-dimensional architectures of BMs and revealed heterogeneous meshwork structures in the GBM and TBM which were probably caused by a different ratio of the major components.

Animals↗
PubMed1984

Scanning electron microscopy of vascular corrosion casts--technique and applications.

The present paper states very briefly the main steps leading to the technique of scanning electron microscopy (SEM) of vascular corrosion casts. From the terms presently used (injection method, microcorrosion cast, injection replica, vascular corrosion cast, vascular cast) the use of "vascular corrosion cast" for lymphatic and blood vessels is recommended. Specification and pretreatment (kind, volume, dosage of anticoagulants, vasoactive substances and spasmolytica used) of the animals examined are referenced as they are available from the literature. The recommendation is given to pay more attention to these parameters than done so far. The steps necessary for producing reasonable and suitable vascular corrosion casts are critically described. Special attention is paid to the physical and chemical properties of the casting media and their significance for polymerizations, shrinkage, casting quality, corrosion resistance, and thermal and spatial stability. Emphasis is also focused on the advantages of cutting the vascular corrosion casts embedded in an ice block by a band saw and a self constructed multi-blade cutting device offer. From the drying methods presently used freeze-drying is stressed because of minimal specimen damage. To render casts conductive in most cases sputtercoating is sufficient. It is recommended to run the SEM with 5-10 kV since the resolution received still reveals all details the casting media presently can replicate. Lastly the application of scanning electron microscopy of vascular corrosion casts in fully differentiated normal tissue, in pathologic tissue as well as in developing tissues and organs is stated.

Animals↗
PubMed2002

Scanning electron microscopy of vascular corrosion casts and histologic examination of pulmonary microvasculature in dogs with dirofilariosis.

OBJECTIVE: To characterize structural changes in pulmonary vessels of dogs with dirofilariosis. ANIMALS: 8 dogs with dirofilariosis and 2 unaffected control dogs. PROCEDURE: Pulmonary artery pressure was measured in affected dogs, and dogs then were euthanatized. Scanning electron microscopy was used to examine vascular corrosion casts of pulmonary vasculature. Tissue sections of pulmonary vasculature were evaluated by use of histologic examination. RESULTS: Pulmonary artery pressure was higher in dogs with severely affected pulmonary vessels. In tissue sections, dilatation, as well as lesions in the tunica intima and proliferative lesions resulting in constriction or obstruction, were frequently observed in branches of the pulmonary artery. Numerous dilated bronchial arteries were observed around affected pulmonary arteries. Hyperplastic venous sphincters were observed in small pulmonary veins and venules. In corrosion casts, affected pulmonary lobar arteries had dilatation, pruning, abnormal tapering, constriction, and obstruction. In small arteries and arterioles, surface structures representing aneurisms and edema were seen. Bronchial arteries were well developed and extremely dilated, and they formed numerous anastomoses with pulmonary arteries at all levels, from the pulmonary trunk to peripheral vessels. Capillaries in the lungs were dilated with little structural change. Small pulmonary veins and venules had irregular annular constrictions that were caused by hyperplastic smooth muscle cells of venous sphincters. CONCLUSIONS AND CLINICAL RELEVANCE: Scanning electron microscopy of microvascular casts delineated links between the bronchial and pulmonary circulations in dogs with dirofilariosis. Results of scanning electron microscopy provided a structural explanation for the development of pulmonary circulatory disturbances and pulmonary hypertension in dogs affected by dirofilariosis.

Animals↗
PubMed1989

Scanning electron microscopy of dysplastic Barrett's epithelium.

Twenty-six patients with Barrett's esophagus (BE) were followed prospectively by endoscopic examination and biopsy. Two biopsies were taken from each of 4 areas of BE. One was processed for light microscopy (LM) and one for scanning electron microscopy (SEM). Those in whom dysplastic BE was demonstrated by LM were reexamined at 6-mo intervals, and the others at yearly intervals. One patient had low grade dysplasia (LGD) by LM on entry, and in 2 others, LGD was recognized on the second examination. These changes have persisted in semiannual examinations over 3, 2, and 2 yr, respectively. SEM prints were examined without knowledge of LM findings, and features that might correlate with LGD by LM were sought. SEM findings were similar to those of Zwas et al. (Gastroenterology 90:1932, 1986) in that most glandular cells had surface features unlike either gastric or intestinal cells but unique to BE. In the patient with LGD on entry, there was an aggregate of very large cells covered by short microvilli with bald patches. In the other patients with LGD, there was more variation in size and shape of cells than in nondysplastic cases.

Adult↗
PubMed2005

Fine needle aspiration cytology diagnosis of gastrointestinal stromal tumors utilizing scanning electron microscopy.

BACKGROUND: Digestive stromal neoplasms are the most frequent undifferentiated mesenchymal tumors. The outcome of these malignancies is difficult to predict and the histogenesis is still controversial. However, the frequent and specific expression of CD117 (c-kit) by these tumors could imply an origin from interstitial cells of Cajal. Our objective was to analyze the role of fine needle aspiration cytology, cell block preparation, and immunocytochemistry in the interpretation of gastrointestinal stromal tumors, and to establish scanning electron microscopy as a useful research aid for pathologic changes of the surface cells of gastrointestinal stromal tumors, not totally appreciated by light microscopy. MATERIAL AND METHODS: Twelve cases of gastrointestinal stromal tumors were included in this study, in which fine needle aspiration cytology was performed. RESULTS: On aspirated material, the tumor cells formed closely packed cohesive tissue fragments with high cellular density often in bloody background, or fascicles with parallel side-by-side arrangements of the nuclei. On cell block biopsy material, gastrointestinal stromal tumors were highly cellular spindle or epithelioid tumors with basophilic appearance. Immunocytochemically, they were CD117 positive in all twelve cases, CD34 positive in nine, weakly smooth muscle actin-positive in five, and S-100 and GFAP-negative in all cases. The scanning electron microscopy study showed a strong correlation with the cytomorphological profile. CONCLUSIONS: Gastrointestinal stromal tumors show a broad morphologic variety, but nuclear pleomorphism by cytology alone, rarely correlates with malignant potential. In the appropriate clinical and radiological setting, a confident diagnosis of gastrointestinal stromal tumors can be documented by fine needle aspiration cytology, cell block, immunocytochemical, and scanning electron microscopy results.

Biopsy, Needle↗
PubMed1987

In vitro excystation of Giardia from humans: a scanning electron microscopy study.

The in vitro excystation of Giardia lamblia on cysts isolated from human feces was studied. After purification by sucrose gradient, cysts were incubated in a pepsin-acid solution, then placed in a modified HSP3 medium where excystation occurred within a few minutes. The excystation procedure was studied by continuous observations by light microscopy and sequential observations by scanning electron microscopy (SEM). The in vitro excystation was stopped at timed intervals during incubation by addition of a large amount of 1% glutaraldehyde. The excystation process began by the cyst wall opening at one pole. Flagella protruded rapidly, the parasite emerged progressively from the cyst envelope, posterior end first, the empty cyst collapsed and shrank. Although flagella emerging from the organism were distinguishable, the cell body had not yet shown all the morphological features of the G. lamblia trophozoite. A radical rearrangement of the organism occurred gradually: initially oval in shape, the parasite became round, then elongated, flattened, and underwent cytokinesis. The daughter trophozoites acquired their typical morphological features: the shape, the adhesive disc with the C-shaped structure distinctly visible on the ventral surface, and the definite placement of the flagella. These observations obtained on G. lamblia by SEM were comparable to those obtained with G. muris.

Giardia↗
PubMed1992

Scanning electron microscopy of the small intestine during gluten-challenge in celiac disease.

The celiac disease syndrome is characterized by structural and ultrastructural alterations of the small intestine mucosa. According to criteria by European Society of Paediatric Gastroenterology and Nutrition, the conclusive diagnosis of celiac disease in children depends on the demonstration of histological relapse of the mucosa after reintroduction of gluten in the diet, as this syndrome is a permanent condition of gluten intolerance. Under these diseased conditions, the structure of the intestinal villi has been studied by light microscopy; morphological alterations were revealed only when the gluten challenge induced a clinical relapse. Scanning electron microscopy analyses of the intestinal mucosa in celiac diseased patients showed a strikingly uniform destruction of the villi with changes in their dimensions and arrangement. At high magnification the enterocytes were irregular in size and shape with a decrease and disruption of the glycocalyx. Reductions in length and density of microvilli were also clearly identified. Although these scanning electron microscopy findings could not demonstrate a relationship between the degrees of mucosal atrophy and the duration of the gluten challenge, they nevertheless revealed early stages of fine villous alterations that cannot be detected by the presently employed low resolution light microscopic techniques.

Autopsy↗
PubMed1986

[Scanning electron microscopy and light microscopy studies on the retina of chick embryos in cell culture].

Histological and scanning electron microscopy studies (SEM) suggest a classification of neural elements into five main types: receptor cells, bipolar cells, horizontal, amacrine, and ganglion cells in dissociated cell cultures of the retina from 7 to 10 days old chick embryos. By use of SEM the development of the receptor cell inner segments was observed. At the same time many protrusions were noticed at the receptor cell surface. Specific synaptic contacts between the axons of the receptor cells and the dendrites of the bipolar neurons, as well as unspecific contacts between the pericarya of the receptor cells and other dendrites were demonstrated. The bipolar neurons showed smooth cell surfaces, however, the horizontal cells appeared rough on the cell surfaces. Müller cells were always found adjacent to the photoreceptor cells.

Animals↗
PubMed1987

[Scanning electron microscopy studies of the structure of tissue in the cochlear opening of the cochlear aqueduct].

The structure of the internal and external tissue of the cochlear opening of the cochlear aquaeduct was examined by light microscopy on semithin sections and by scanning electron microscopy. The whole area is filled with a net of mesenchymal cells. The cell axes are randomly orientated inside the aquaeduct. On the outside of the cochlear aquaeduct fibrocytic tissue fills a space which is triangular in cross-section between the basal part of the cochlea wall of the tympanic scala and the middle portion of the round window membrane. In this area the direction of the net is uniform and it gives the impression of anchorage of the round window membrane on the perilymphatic side. The rim bordering the perilymphatic space is a dense net but not fully closed. The scanning electron microscopic pictures taken perpendicular to this border structure show clearly a texture of mesenchymal cells with open spaces. No closed "membrana limitans" was found. The possible function of the fixation of the round window membrane to the perilymphatic space giving rise to an asymmetric perilymph movement is discussed with regard to the physiology of sound transmission.

Animals↗
PubMed1999

Evaluation of initial attachment of human gingival fibroblast cells to biodegradable membranes in vitro by light and scanning electron microscopy.

Guided tissue regeneration procedures using resorbable membranes have become accepted therapy for treating periodontal defects. Resorbable collagen and synthetic polylactide and polyglycolide copolymer membranes have been found to support regeneration and preclude the need for surgical removal. This study was undertaken to assess and compare the initial attachment of human gingival fibroblast cells to four collagen-based membranes (fascia lata, fascia temporalis, dura mater, and Type I bovine collagen) and a synthetic polylactic acid-based membrane (resolut). Human gingival fibroblasts were grown from explants of normal tissue obtained during surgical reduction of retromolar tissues. Membrane specimens were placed in separate culture wells and incubated with fibroblasts for one hour. The number of adherent cells was evaluated by light microscopy using an ocular grid system and detailed examination was performed by scanning electron microscopy. The results of evaluation by light microscopy indicated that initial cell attachment was significantly less in the polylactic acid-based membrane group than in the collagen-based membrane groups (P < 0.01). However, no significant differences were found among the collagen membrane groups in terms of fibroblast attachment (P > 0.01). Scanning electron microscopy examination of fibroblasts cultured directly on barrier membranes indicated that the collagen-based membranes appeared to facilitate cell attachment, whereas the polylactic acid-based membrane exhibited a morphology that was not conducive to attachment of human gingival fibroblasts. Based on these limited in vitro results, it appears that collagen-based membranes offer greater potential than polylactic acid-based membranes for guided tissue regeneration at surgical sites.

Absorbable Implants↗
PubMed2002

Use of light and scanning electron microscopy to examine colonisation of barley rhizosphere by the nematophagous fungus Verticillium chlamydosporium.

Barley roots were readily colonised by the nematophagous fungus Verticillium chlamydosporium. Light microscopy (LM) but also low temperature scanning electron microscopy (LTSEM) revealed details of the colonisation process. Hyphae were found on the rhizoplane often with dictyochlamydospores. Hyphae of V. chlamydosporium penetrated epidermal cells, often by means of appressoria. A hyphal network was formed in epidermal and cortical cells. Likewise, hyphal coils were found within root cells next to transverse cell walls. Cortical cells were the limits of fungal colonisation, since no hyphae were seen in the vascular cylinder. Modifications of root cell contents (phenolic droplets and callose appositions) were common three weeks after inoculation with V. chlamydosporium. These features may indicate induction of plant defence reactions in late stages of root colonisation by the fungus. Both LTSEM and LM have proved extremely useful to describe root colonisation by the fungus. The results found may have implications in the mode action of nematophagous fungi against plant parasitic nematodes.

Animals↗
PubMed2005

A vision-based, 3D reconstruction technique for scanning electron microscopy: direct comparison with atomic force microscopy.

High-resolution, detailed 3D reconstructions of biological specimens obtained from scanning electron microscopy stereo-micrographs and proprietary software were compared with Tapping-Mode AFM datasets of the same fields. The reconstruction software implements several original solutions including a neural adaptive point-matching technique, the ability to build an irregular triangulated mesh rather than a regular orthogonal grid, and the ability to re-map one of the original images exactly onto the reconstructed surface. The technique was applied to human nerve tissue to obtain 1,424 x 968-pixel, texture-mapped datasets, which were subsequently compared against 512 x 512-pixel AFM datasets from the same viewfields. Accounting for the inherent differences of the two techniques, direct comparison revealed an excellent visual match. The correspondence was also quantified by calculating the cross-correlation coefficient between corresponding altimetric profiles in SEM and AFM data, which consistently exceeded a figure of 0.9, with a rate of point mismatch in the order of 0.01%. Research is still underway to improve the robustness of the technique when applied to arbitrary images

Humans↗