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At least 163 records · Page 9Linked to original sources

Improved transfer of two-dimensional crystals from the air/water interface to specimen support grids for high-resolution analysis by electron microscopy.

Electron crystallographic analysis of two-dimensional crystals grown on lipid layers at the air/water interface has been limited by loss or damage during transfer of the crystals to an electron microscope support grid. Two methods of transfer are described which are applicable on a small scale (10 microliters of protein solution) and which give greatly improved results for streptavidin crystals on biotinylated lipid layers. In the first method, a hydrophobic grid surface was produced by coating a carbon support film with a thin layer of SiO2, followed by alkylation with dimethyloctadecylchlorosilane. The transfer efficiency of protein crystals approached 50% coverage of the alkylated grid surface. The degree of order of crystals transferred to the alkylated grid surface and preserved in negative stain was significantly improved over that of crystals transferred directly to a carbon support film. In the second method, crystals at the air/water interface were transferred to a holey carbon support film. The efficiency of transfer across the holes was virtually 100% as nearly every hole was completely covered with crystals. After preservation of the crystals in 1% glucose and cooling to liquid nitrogen temperature, electron diffraction was obtained that extended to 1/2.8 A-1 resolution. This demonstrates that two-dimensional crystals grown on lipid layers at the air/water interface can be sufficiently well-ordered, even after transfer to a support grid, to yield high-resolution structural information.

Bacterial Proteins↗

The surface coat of chylomicrons: electron microscopy.

Electron microscope studies were performed on thoracic duct lymph and on washed chylomicrons from dogs fed corn oil. High-resolution electron micrographs showed the presence of a surface coat that differed from the core material and did not resemble a plasma membrane. This was true for both chylomicrons in whole lymph and those that had been subjected to repeated washing. Apparently, the chylomicrons, while passing from the intracellular to the extracellular space, do not acquire their surface coat from pinched off cellular membrane.

Animals↗

Morphological forms and viability of Campylobacter species studied by electron microscopy.

Electron microscopic studies of Campylobacter revealed that different morphological forms predominate at different parts of a colony. At the periphery, cells were almost all spirals, while in the center of the colony cells were mainly coccus shaped. Unusual ring-shaped cells, "donuts", were observed in the raised, peripheral region of the colony. Donut or ring forms have not previously been reported for Campylobacter organisms. Our data indicate that young or actively growing cells are mainly spiral shaped. Older cells undergo a degenerative change to coccoid forms. The donut shape appears to be an intermediate stage between spirals and cocci. Comparisons of plate counts of actively growing and inactive cells confirmed that coccoid cells are probably nonviable.

Campylobacter↗

Annular structures in erythrocyte membranes of various animal species as revealed by electron microscopy.

Electron microscopic examination of negatively stained erythrocyte membranes revealed the presence of annular structures having an overall diameter of 33 nm and a ring-thickness of 9 nm. Each annular structure appeared to be composed of eleven globular subunits. The structures were most conspicuous in human erythrocyte membranes which showed, on the average, 160 per membrane. Appreciably smaller numbers of what appeared to be the same structures were seen in erythrocyte membranes derived from rhesus monkey, cat, guinea pig, rabbit, horse, dog and chicken. None at all could be found in erythrocyte membranes from sheep, deer, goat and ox. The size and shape of these structures readily distinguishes them from ring-like entities whose formation is induced by treatment of membranes with a variety of haemolytic agents.

Animals↗

[Mucous membrane changes in the anterior ethmoid bone in chronic sinusitis and mycoses of the paranasal sinuses. An endoscopy, light and electron microscopy study. 2. Electron microscopy findings].

In the second part of this paper the findings of electron microscopical studies on ultrastructural changes of the cilia of the ethmoidal mucous membrane in patients with mycotic chronic sinusitis are presented. Many pathological variations of the normal 9 + 2 pattern can be demonstrated, along with compound cilia and central axis deviation beyond 25 degrees. This together with the endoscopical and lightmicroscopical findings results in conditions which are different from those in acute sinusitis. It may as well explain, why hardly a conservative therapy--aiming mostly for vasoconstriction and decrease of mucosal edema--is successful in chronic sinusitis. Endoscopical surgery of the diseased anterior ethmoid in most of the cases brings along cure of the dependent frontal and maxillary sinuses as well.

Chronic Disease↗

Characterization of the annealed (0001) surface of sapphire (alpha-Al2O3) and interaction with silver by reflection electron microscopy and scanning reflection electron microscopy.

Annealed (0001) surfaces of single-crystal sapphire (alpha-Al2O3) rod have been studied in the electron microscope using reflection electron microscopy (REM), scanning reflection electron microscopy (SREM), and reflection high energy electron diffraction (RHEED). Annealed surfaces of (0001) sapphire are vicinal and characterized by close-packed (0001)-oriented terraces separated by faceted multiple-height steps, with edges parallel to energetically preferred low-index directions (less than 1010 greater than and less than 1120 greater than). These structural features are not seen on cleaved surfaces or polished surfaces treated at temperatures less than 1,250 degrees C. Oxygen-annealing produces clean surfaces which prove useful for investigating the interaction of deposited metals with the (0001) sapphire. Both REM and SREM (with microdiffraction spots) techniques have been used to observe fine structure of flat Ag islands on the scale of 1-100 nm on the (0001)-oriented terraces as well as aggregations at the steps. A preliminary result on interaction with Cu is also included.

Aluminum Oxide↗

Choroidal vasculature changes in spontaneously hypertensive rats - transmission electron microscopy and scanning electron microscopy with casts.

PURPOSE: The purpose of the present study was to elucidate the specific morphological changes of the choroidal vasculature in long-term hypertension. METHODS: The choroidal vessels of spontaneously hypertensive (SHR) rats were examined with corrosion casts/scanning electron microscope (SEM) and the transmission electron microscope (TEM). RESULTS: In 18-month-old SHRs, corrosion casts and SEM showed tortuosity, caliber irregularity and generalized narrowing of the choroidal arteries. The draining venules were prominently decreased in number. The ampullae of the vortex veins were narrow, but arteriovenous anastomoses, crossing defects and obstruction were not seen. The choriocapillaris was elongated and engorged. TEM of the choroidal vessels showed hypertrophy of smooth muscle cells and irregular thickening of the basement membrane and narrow lumen of the choriocapillaris. The RPE and Bruch's membrane were intact. CONCLUSION: The choroidal vascular bed showed extensive morphological changes. This study revealed that hypertensive choroidopathy has specific features such as tortuosity, caliber irregularity, generalized narrowing of arteries, decreased number of draining venules and elongated and engorged choriocapillaris. These findings indicate that hypertensive choroidopathy is as important as retinopathy.

Animals↗

Correlative Instrumental Neutron Activation Analysis, Light Microscopy, Transmission Electron Microscopy, and X-ray Microanalysis for Qualitative and Quantitative Detection of Colloidal Gold Spheres in Biological Specimens.

: Colloidal gold, conjugated to ligands or antibodies, is routinely used as a label for the detection of cell structures by light (LM) and electron microscopy (EM). To date, several methods to count the number of colloidal gold labels have been employed with limited success. Instrumental neutron activation analysis (INAA), a physical method for the analysis of the elemental composition of materials, can be used to provide a quantitative index of gold accumulation in bulk specimens. Given that gold is not naturally found in biological specimens in any substantial amount and that colloidal gold and ligand conjugates can be prepared to yield uniform bead sizes, the amount of label can be calculated in bulk biological samples by INAA. Here we describe the use of INAA, LM, transmission EM, and X-ray microanalysis (EDX) in a model to determine both distribution (localization) and amount of colloidal gold at the organ, tissue, cellular, and ultrastructural levels in whole animal systems following administration. In addition, the sensitivity for gold in biological specimens by INAA is compared with that of inductively coupled plasma-mass spectrometry (ICP-MS). The correlative use of INAA, LM, TEM, and EDX can be useful, for example, in the quantitative and qualitative tracking of various labeled molecular species following administration in vivo.

Journal Article↗

Optimizing parameters for correlative immunogold localization by video-enhanced light microscopy, high-voltage transmission electron microscopy, and field emission scanning electron microscopy.

Correlative video-enhanced light microscopy, high-voltage transmission electron microscopy, and low-voltage high resolution scanning electron microscopy were used to examine the binding of colloidal gold-labeled fibrinogen to platelet surfaces. Optimal conditions for the detection of large (18 nm) and small (3 nm) gold particles are described.

Blood Platelets↗

The fine structure of fenestrated adrenocortical capillaries revealed by in-lens field-emission scanning electron microscopy and scanning transmission electron microscopy.

Cell biologists probing the physiologic movement of macromolecules and solutes across the fenestrated microvascular endothelial cell have used electron microscopy to locate the postulated pore within the fenestrae. Prior to the advent of in-lens field-emission high-resolution scanning electron microscopy (HRSEM) and ultrathin metal coating technology, quick-freeze, platinum-carbon replica and grazing thin-section transmission electron microscopy (TEM) methods provided two-dimensional or indirect imaging methods. Wedge-shaped octagonal channels composed of fibrils interwoven in a central mesh were depicted as the filtering structures of fenestral diaphragms in images of platinum replicas enhanced by photographic augmentation. However, image accuracy was limited to replication of the cell surface. Subsequent to this, HRSEM technology was developed and provided a high-fidelity, three-dimensional topographic image of the fenestral surface directly from a fixed and dried bulk adrenal specimen coated with a 1 nm chromium film. First described from TEM replicas, the "flower-like" structure comprising the fenestral pores was readily visualized by HRSEM. High-resolution images contained particulate ectodomains on the lumenal surface of the endothelial cell membrane. Particles arranged in a rough octagonal shape formed the fenestral rim. Digital acquisition of analog photographic recordings revealed a filamentous meshwork in the diaphragm, thus confirming and extending observations from replica and grazing section TEM preparations. Endothelial cell pockets, first described in murine renal peritubular capillaries, were observed in rhesus and rabbit adrenocortical capillaries. This report features recent observations of fenestral diaphragms and endothelial pockets fitted with multiple diaphragms utilizing a Schottky field-emission electron microscope. In-lens staging of bulk and thin section specimens allowed tandem imaging in HRSEM and scanning TEM modes at 25 kV.

Adrenal Cortex↗

Low-energy electron microscopy (LEEM) and mirror electron microscopy (MEM) of biological specimens: preliminary results with a novel beam separating system.

Low-energy electron microscopy (LEEM) and mirror electron microscopy (MEM) utilize a parallel beam of slow-moving electrons backscattered from the specimen surface to form an image. If the electrons strike the surface an LEEM image is produced and if they are turned back just before reaching the surface an MEM image results. The applications thus far have been in surface physics. In the present study, applications of LEEM and MEM in the biological sciences are discussed. The preliminary results demonstrate the feasibility of forming images of uncoated cultured cells and cellular components using electrons in the threshold region (i.e. 0-10 V). The results also constitute a successful test of a novel beam-separating system for LEEM and MEM.

Animals↗

Analytical transmission electron microscopy and electron diffraction for characterization of multiphase Ni-P-Ti surface layer on Ti-6Al-4V alloy.

The microstructure, chemical and phase composition of the hard Ni-P-Ti layer formed on the Ti-6Al-4V alloy after duplex surface treatment were investigated by light microscopy, X-ray diffraction, scanning electron microscopy and analytical/high-resolution transmission electron microscopy. These investigations showed that the improved mechanical and tribological properties of the surface-treated alloy were related to the presence of a multilayered microstructure containing several phases from the Ni-Ti-P-Al system.

Journal Article↗