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PubMed1986

Scanning electron microscopy of Barrett's epithelium and its correlation with light microscopy and mucin stains.

The surface epithelial cells of Barrett's esophagus were characterized using quantitative scanning electron microscopy and light microscopy with mucin histochemical stains. Fifty-one biopsy specimens of Barrett's esophagus from 15 patients and 31 control specimens of the stomach and intestines from 9 patients were examined. Three distinct surface cell types, in addition to the goblet cell, were recognized in Barrett's epithelium: the gastric-like cell in 31% of specimens, which was similar to the normal gastric surface cell by quantitative scanning electron microscopy; the intestinal-like cell in 41%, which was most similar to the normal small intestinal surface cell; and the variant cell in 80%, which had a range of surface features. By light microscopy, all specimens with variant and intestinal-like cells were classified as specialized columnar epithelium. The surface mucous cells in Barrett's epithelium displayed a variety of mucin staining patterns with acid nonsulfated (small intestinal-like) mucin present in 90% of specimens and acid sulfated (colonic-like) mucin in 43% of specimens. Quantitative scanning electron microscopy and mucin histochemical stains reveal a striking cellular heterogeneity not apparent by routine light microscopy.

Barrett Esophagus↗

PubMed1975

New immunolatex spheres: visual markers of antigens on lymphocytes for scanning electron microscopy.

New immunochemical reagents consisting of antibodies bound to small latex spheres were used as visual markers for the detection and localization of cell surface antigens by scanning electron microscopy. Cross-linked latex spheres of various sizes from 300 to 3,4000 A in diameter were synthesized by aqueous emulsion copolymerization of methacrylate derivatives containing hydroxyl and carboxyl functional groups. Proteins and other molecules containing primary amino groups were covalently bonded to the acrylic spheres under a variety of mild conditions by the aqueous carbodiimide, cyanogen bromide, and glutaraldehyde methods. For use in the indirect immunochemical-labeling technique, goat antibodies directed against rabbit immunoglobulins were bonded to the spheres. These immunolatex reagents were shown to bind only to cells (red blood and lymphocytes) which had previously been sensitized with rabbit antibodies against cell surface antigens. Mouse spleen lymphocytes with exposed immunoglobulins on their surface (B cells) were labeled with these spheres and distinguished from unlabeled or T lymphocytes by scanning electron microscopy. The distribution of Ig receptors on lymphocytes was also studied using the spheres as visual markers. When lymphocytes were fixed with glutaraldehyde and subsequently labeled with the immunolatex reagents, a random distribution was observed by scanning electron microscopy; a patchy distribution was observed when unfixed lymphocytes were used. These results are consistent with studies using ferritin-labeled antibodies (S. De Petris and M. Raff. 1973. Nature [Lond.]. 241:257.) and support the view that Ig receptors on lymphocytes undergo translational diffusion. In addition to serving as visual markers for scanning electron microscopy, these latex spheres tagged with fluorescent or radioactive molecules have applications as highly sensitive markers for fluorescent microscopy and as reagents for quantitative studies of cell surface antigens and other receptors.

Animals↗

PubMed1982

Scanning electron microscopy of Giemsa-stained chromosomes and surface-spread chromosomes.

Giemsa-stained chromosomes as prepared for light microscopy, and including G-banded, C-banded, and FPG-stained chromosomes, were examined by scanning electron microscopy. Although suitable for light microscopy, these chromosomes were too flat for a close examination of their fine structure by scanning electron microscopy. The surface of Giemsa-positive regions was rough and bright, whereas that of unstained or poorly stained regions was smoother and less bright. Giemsa-staining, therefore, seems to produce the bulkiness of the chromosomes. On topographical examination by scanning electron microscopy, the transparent chromosomes as observed with the light microscope proved to be "footprints". Stereographical examinations of surface-spread chromosomes showed that minimally stretched chromosomes were composed of a mass of nodular and twisted looping fibers with an average diameter of about 300 A. The substructure of these chromosome fibers was not determined. The kinetochore region was discernible as a constriction in the mass of the chromosome fibers, and was distinguishable from gaps by the presence of several chromosome fibers parallel to the axis of the chromatid. The organization of the chromosome fibers, however, was disordered rather than regular.

Azure Stains↗

PubMed1990

A new method of bacterial identification using gold immunolabelling and scanning electron microscopy.

This study sought to develop an immunolabelling technique to identify specific bacteria by scanning electron microscopy. Bacterial suspensions were prepared of Actinobacillus actinomycetemcomitans, Porphyromonas (Bacteroides) gingivalis and mixtures of these species with other genera of common oral microorganisms. This method was also used to examine subgingival plaques from 6 subjects with chronic adult periodontitis. Sample preparation consisted of prefixation of the bacterial suspensions with 0.2% glutaraldehyde, incubation with species-specific rabbit antisera and goat anti-rabbit IgG conjugated with colloidal gold particles, postfixation in 2% glutaraldehyde and dehydration in ethanol. Finally, the samples were dried, coated with evaporated carbon and examined by scanning electron microscopy. Pure cultures and artificial mixtures of A. actinomycetemcomitans and Bact. gingivalis were specifically labelled by gold probes as demonstrated by both secondary and back-scattered imaging. These species were also evident in samples of subgingival plaque. The findings indicate that this new technique can be used to identify specific microorganisms both in plaque samples and on the root surface of extracted teeth.

Actinobacillus↗

PubMed1995

Scanning electron microscopy of the human esophagus: application to Barrett's esophagus, a precancerous lesion.

In Barrett's esophagus, metaplastic columnar epithelium replaces the normal squamous epithelium. The importance of this lesion lies in the increased incidence of adenocarcinoma of the esophagus occurring in patients with Barrett's esophagus. We characterized the surface epithelial cells of Barrett's esophagus using quantitative scanning electron microscopy. Three distinct surface cell types, in addition to the globlet cell, were recognized in Barrett's epithelium: the gastric-like cell and the intestinal-like cell, both of which were similar to normal gastric and small intestinal surface cells, respectively, by quantitative scanning electron microscopy, and the variant cell which had a range of surface features. In four biopsy specimens from the squamo-Barrett's junction in three patients, we found the distinctive cell that had features intermediate between those of squamous and columnar epithelium. On the distinctive cell's surface there are two disparate structures not normally present on the same cell in the gastrointestinal tract: microvilli (a scanning electron microscopy feature of glandular epithelium) and intercellular ridges (a scanning electron microscopy feature of squamous epithelium). The surface characteristics of this cell were almost identical to those of cells found in the transformation zone of the uterine cervix, an area in which squamous epithelium physiologically replaces columnar epithelium. Scanning electron microscopy of Barrett's esophagus has increased our understanding of this precancerous lesion by showing striking cellular heterogeneity. It has also identified the distinctive cell which may represent an intermediate step in the development of Barrett's epithelium during which the surface characteristics of two different cell types, columnar and squamous, coexist in the same cell.

Barrett Esophagus↗

PubMed1996

Observations of scanning electron microscopy detected abnormalities of non-lubricated latex condoms.

To describe surface abnormalities found by scanning electron microscopy of non-lubricated latex condoms, thirty samples of 15 condoms were photographed at low (26-94x), medium (240-940x) and high (> 1000x) magnification. Each sample was initially scanned at low magnification to determine the presence and location of surface abnormalities. Photographs taken at medium and high magnification were of the area most representative of the abnormality. Nine samples (30%) were completely normal at all magnifications. Of the total viewed at low power, 18 (60%) were normal, 11 (37%) revealed ridging and 1 (3%) revealed melting. Of the total viewed at medium power, 14 (47%) were normal, 3 (10%) showed cracking, 2 (7%) showed melting or a combination of ridging and cracking, and 11 (37%) showed ridging. At high power, 10 (34%) were normal, 11 (38%) revealed either melting or a combination of two defects, 5 (17%) showed ridging, and 3 (10%) revealed cracking. Fifteen (50%) of the samples revealed a surface abnormality (cracking or melting) other than ridging. The majority of condoms viewed under scanning electron microscopy revealed surface abnormalities. The implication of the detected abnormalities needs further evaluations. While ridging may represent a minor abnormality associated with condom handling, multiple abnormalities or melting may represent a more significant atypicality.

Condoms↗

PubMed1985

Critical-point drying versus freeze drying for scanning electron microscopy: a quantitative and qualitative study on isolated hepatocytes.

Critical-point drying and freeze drying were compared both quantitatively and qualitatively as preparative procedures for scanning electron microscopy. Isolated hepatocytes were used as model cells. Nomarski differential interference contrast microscopy was used for light microscopic measurements of the hepatocytes in the unfixed, the glutaraldehyde fixed, the glutaraldehyde + OsO4 fixed, the critical-point dried and the freeze dried states. Critical-point dried hepatocytes were found to shrink to 38% of glutaraldehyde + OsO4 fixed volume, whereas optimal freeze dried hepatocytes (frozen in water saturated with chloroform and freeze dried at 183 K for 84 h) were found to shrink to 51% of glutaraldehyde + OsO4 fixed volume. Transmission and scanning electron micrographs of the critical-point dried cells showed well-preserved ultrastructure and surface structure. Micrographs of the freeze dried cells showed ultrastructure destroyed by internal ice crystals and surface structure destroyed by external ice crystals. Double-fixed isolated hepatocytes were shown to swell during storage in buffer and to shrink during storage after critical-point drying. For low magnification scanning electron microscopy (up to about 3000 times) both critical-point drying and freeze drying can be used. However, for high magnification scanning electron microscopy, critical-point drying is superior to freeze drying.

Animals↗

PubMed1993

High-resolution scanning electron microscopy of rat incisor ameloblasts.

The internal three-dimensional organization of secretory and maturation stage ameloblasts was examined using field emission scanning electron microscopy. Particular attention was given to the structure of the Golgi apparatus, the distribution of smooth membrane tubulo-vesicular elements and their relationship with endosomal/lysosomal components. Rat incisors were fixed by intracardiac perfusion with glutaraldehyde, decalcified and divided into segments. The tissues were cryoprotected with glycerol and freeze-fractured. They were then macerated in osmium, and after conductive staining with osmium/tannic acid, the samples were critical-point dried and sputtered with gold. High-resolution scanning electron microscopy showed that ameloblasts contain a well-developed Golgi apparatus, even in the maturation stage where cells are generally believed not to be actively involved in protein secretion. Smooth tubulo-vesicular structures formed a complex network which extended throughout the cell. In secretory stage ameloblasts, this network reached into Tomes' process and consisted of small and large tubules, and distended portions. The smaller tubules radiated from a central core of organelles towards the plasma membrane. Numerous lysosomal/endosomal elements were observed in the Golgi region, and in some cases smooth tubular portions were seen at the surface of multivesicular bodies. These data show that high-resolution scanning electron microscopy can be applied to correlate three-dimensional structural detail with the secretory and resorptive functions of ameloblasts.

Ameloblasts↗

PubMed1975

Detection of particles in intravenous fluids using scanning electron microscopy.

A study was conducted to compare the particles in intravenous solutions found in glass and plastic containers, and to investigate the use of scanning electron microscopy in detecting and identifying such particulate contamination. The method used was to filter one liter of each fluid from glass and plastic containers through a membrane filter, attach the membrane to aluminum studs and coat the specimens with a thin layer of gold-palladium in a vacuum evaporator. Scanning electron microscopy was then used to examine the specimens, and photomicrographs were prepared. The results showed that the particles found in glass and in plastic containers have distinctive differences and that scanning electron microscopy is useful in detecting and characterizing these particles.

Drug Contamination↗

PubMed1995

Section directed cryosectioning of specimens for scanning electron microscopy: a new method to study cardiac development.

A new method to study the developing heart was developed. Using this "section directed" cryosectioning method, appropriate fixed embryos can be trimmed optimally to obtain sectional planes that, if necessary, can be matched with histologically treated sections. As a result, the morphological information obtained with the scanning electron microscope can be compared in detail with the information on the molecular phenotypes of the subpopulations of cells as deducted from staining patterns of the sections. This method allows combination of the specific advantages of sophisticated histological techniques, such as immunohistochemistry and in situ hybridisation, with those of the scanning electron microscope.

Animals↗

PubMed1984

The value of graphic reconstructions--comparison with scanning electron microscopy.

Because knowledge of spatial morphology of the human embryonic heart mainly derives from serial microscopic sections and from reconstructions based on such series, it might be questioned whether the results obtained are comparable with animal studies in which scanning electron microscopy is used. Therefore, pig embryo hearts of the same developmental stage were processed for routine histological sectioning as well as for scanning electron microscopy. A graphic reconstruction was made of a right ventricular view, to be compared with a similar scanning micrograph. The two images, produced by much different techniques, were strikingly similar. In addition, the graphic reconstruction technique is complementary to scanning electron microscopy, because histological details can be introduced.

Animals↗

PubMed1996

Evidence for aggregation in oxalate stone formation: atomic force and low voltage scanning electron microscopy.

PURPOSE: The aim of this investigation was to differentiate between aggregation and crystal growth by studying the structure of oxalate stones at high spatial resolution using recently developed microscopy techniques. MATERIALS AND METHODS: Sections from 6 complete human oxalate stones and 4 stone fragments were prepared by ultramicrotomy and examined by both low voltage scanning electron microscopy and atomic force microscopy. RESULTS: The scanning electron microscopy showed lamellar structures up to 10 microns. in size, consistent with previous results, and provided evidence that these structures were composed of smaller particles. The atomic force microscopy clearly showed arrays of the small particles, whose size varied between 500A and 2800A. CONCLUSION: Our images suggest that an ordered aggregation of small crystallites is responsible for oxalate stone formation.

Humans↗

PubMed1979

Scanning electron microscopy of rectal mucosa in Crohn's disease.

Rectal biopsies from healthy control subjects, and from patients with Crohn's disease whose rectal mucosa appeared normal by sigmoidoscopy and light microscopy, were examined by scanning electron microscopy. Appearances within both groups were quite variable, and no clear differences between the groups could be distinguished.

Adolescent↗

PubMed1993

High-resolution visualization by field emission scanning electron microscopy of Enterococcus faecalis surface proteins encoded by the pheromone-inducible conjugative plasmid pCF10.

Enterococcus faecalis can acquire antibiotic resistance and virulence genes by transfer of pheromone-inducible conjugative plasmids such as pCF10, which encodes tetracycline resistance. Two pCF10-encoded cell surface proteins, Sec10 and Asc10, have been previously shown to play an important role in the transfer of this plasmid. We used high-resolution, field emission scanning electron microscopy to visualize these proteins on the surfaces of a series of isogenic strains of E. faecalis. Immunogold labeling, using both 6- and 12-nm colloidal gold, unambiguously demonstrated the expression and distribution of Sec10 and Asc10 on the surface of the E. faecalis cells. On unlabeled E. faecalis cells which expressed either Sec10 or Asc10, the former appeared to be more readily detected. Immunogold labeling of E. faecalis cells expressing both Asc10 and Sec10 clearly demonstrated the abundance and intermixing of both proteins on the cell surface except at septal regions. Sec10 was observed to be distributed over the cell surface. At regions of cell-cell contact, fine strands representing Asc10 were observed directly attaching adjacent cells to one another.

Antibodies, Monoclonal↗

PubMed1980

Chemical and enzymatic changes in the cell walls of Candida albicans and Saccharomyces cerevisiae by scanning electron microscopy.

Candida albicans and Saccharomyces cerevisiae cells were examined by scanning scanning electron microscopy before and after extraction of the mannans of the cell wall. The surfaces of control cells were smooth; after mannan extraction they were rough and showed erosions which were particularly striking within the area of the scars. Helicase digested irregular holes through the cell wall within 20 min; these increased in size during an additional 40 min of digestion. These holes were not localized in or on the bud scars, which remained intact even after the long digestion period. The results were used to construct a model for yeast cell wall structure.

Candida albicans↗

PubMed1999

Quality assessment of atomic force microscopy probes by scanning electron microscopy: correlation of tip structure with rendered images.

While image quality from instruments such as electron microscopes, light microscopes, and confocal laser scanning microscopes is mostly influenced by the alignment of optical train components, the atomic force microscope differs in that image quality is highly dependent upon a consumable component, the scanning probe. Although many types of scanning probes are commercially available, specific configurations and styles are generally recommended for specific applications. For instance, in our area of interest, tapping mode imaging of biological constituents in fluid, double ended, oxide-sharpened pyramidal silicon nitride probes are most often employed. These cantilevers contain four differently sized probes; thick- and thin-legged 100 microm long and thick- and thin-legged 200 microm long, with only one probe used per cantilever. In a recent investigation [Taatjes et al. (1997) Cell Biol. Int. 21:715-726], we used the scanning electron microscope to modify the oxide-sharpened pyramidal probe by creating an electron beam deposited tip with a higher aspect ratio than unmodified tips. Placing the probes in the scanning electron microscope for modification prompted us to begin to examine the probes for defects both before and after use with the atomic force microscope. The most frequently encountered defect was a mis-centered probe, or a probe hanging off the end of the cantilever. If we had difficulty imaging with a probe, we would examine the probe in the scanning electron microscope to determine if any defects were present, or if the tip had become contaminated during scanning. Moreover, we observed that electron beam deposited tips were blunted by the act of scanning a hard specimen, such as colloidal gold with the atomic force microscope. We also present a mathematical geometric model for deducing the interaction between an electron beam deposited tip and either a spherical or elliptical specimen. Examination of probes in the scanning electron microscope may assist in interpreting images generated by the atomic force microscope.

DNA↗

PubMed1991

Polymorphic behavior of sprayed lipid micropellets and its evaluation by differential scanning calorimetry and scanning electron microscopy.

Considering the importance of polymorphism occurring in solid dosage forms causing instability, the polymorphic behavior of spray-dried and -congealed lipid micropellets was examined by differential scanning calorimetry and scanning electron microscopy. The results showed that both of the spraying processes exert an important effect on their polymorphic and crystallization properties. In spray-drying, due to the rapid solvent evaporation, the obtained lipid micropellets possess an unstable polymorphic form. This unstable form transforms gradually toward a stable form by storage at elevated temperatures. The same modifications were observed with spray-congealed lipid micropellets. The type of glyceride (composition, chain length), solvent and drugs (estradiol cypionate, medroxyprogesterone acetate) and, further, the presence of a stabilizing agent such as lecithin affect the polymorphic transition and its rate.

Calorimetry, Differential Scanning↗

PubMed1994

Particles found in lung and brain following subureteral injection of polytetrafluoroethylene paste are not teflon particles.

We injected 0.3 ml. of polytetrafluoroethylene paste (Polytef) behind the left submucosal ureter in 4 mini-pigs and 4 dogs, and 2 dogs and 2 mini-pigs underwent sham operation and acted as controls. Two mini-pigs were injected with 0.5 ml. polytetrafluoroethylene (Teflon) suspended in 50 ml. of normal saline into a peripheral vein and 2 dogs were injected with 0.5 ml. polytetrafluoroethylene into a bladder vein. In addition, 4 dogs were injected with 0.1 ml. polytetrafluoroethylene paste suspended in 20 ml. saline into the right carotid artery. The lungs and brain from half of the animals who had subureteral and intravascular injection of polytetrafluoroethylene paste as well as sham operated animals were dissolved in sodium hypochlorite solution. The resulting organ suspensions were then centrifuged and the smear preparations of the precipitate were examined by polarized light microscopy, scanning electron microscopy and x-ray microanalysis. Polytetrafluoroethylene paste suspended in saline acted as positive control for polytetrafluoroethylene particles. Lungs and brain from the remaining animals were fixed in formalin solution. The brain and lungs of animals who underwent subureteral injection with a minimal amount of polytetrafluoroethylene paste carefully placed in the submucosal plane showed no evidence of polytetrafluoroethylene on histological examination, polarized light microscopy, scanning electron microscopy and x-ray microanalysis.

Animals↗