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Query: UMLS:C0276640 (TEM)
 20,729 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The Gomori aldehyde-fuchsin (AF) method for selective staining of neurosecretory substance (NSS) has been adapted to tissue previously prepared for both scanning and transmission electron microscopy (SEM/TEM). The procedure results in precise correlation of light microscopic (LM) histochemistry with SEM/TEM of the same tissue.
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PMID:Demonstration of neurosecretory substance in previously scanned specimens: adaptation of the Gomori aldehyde-fuchsin method for correlative SEM/TEM/LM histochemistry. 8 9

Cells cultured on transparent conductive substrates (glass coated with indium oxide) were fixed with aldehyde and osmium tetroxide and then treated with tannic acid, uranyl acetate and lead citrate. The same cell culture preparation could be sequentially studied by light microscopy (in water immersed condition), SEM (after dehydration and critical point drying) and TEM (after embedding in an epoxy resin). This method ensures the preservation of intact cell morphology, cell surface topography and intracellular structures. The treatments used render the cells conductivity and permit to carry out successfully SEM of uncoated cells cultured on conductive substrates. This method also provides a higher contrast of TEM images.
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PMID:[A method for the chemical treatment of cells for the purpose of the subsequent study of the same cell culture preparation by light, scanning and transmission electron microscopy]. 170 38

Collagen shields have been studied in the enhancement of the initial healing of epithelial defects, as an adjunct in the treatment of dry eye, and as a reservoir and delivery system for topical ocular medications. The authors used collagen shields to collect information on the numbers and types of free cells populating the normal and postoperative ocular surface. In addition, correlative microscopic techniques were used to study details of the mechanisms responsible for the dissolution of the shields when applied to the human eye. Collagen shields were applied as a bandage lens on the eyes of patients who underwent extracapsular cataract extraction (n = 10) or penetrating keratoplasty (n = 10) and on normal volunteers (n = 10). The shields were collected at the 1-day postoperative examination and fixed in aldehyde mixtures. Specimens then were processed for correlative light (LM), transmission (TEM), and scanning (SEM) microscopy. Cell accumulation was shown by SEM on both anterior and posterior shield surfaces. Cell adherence occurred primarily on the posterior shield periphery for approximately 2 mm, with the central zone relatively clean. Both LM and TEM evaluation revealed cell counts ranging from 0.066 cells/10(4) microns2 (standard deviation, +/- 0.256) in healthy eyes compared with shields placed on postoperative eyes (194.25 +/- 7.32 cells/10(4) microns2). Various correlative microscopy techniques revealed that most cells were polymorphonuclear leukocytes with a low number of other hematogenous (lymphocytes and monocytes) and exfoliated epithelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Collagen shields as a vehicle for collecting and studying migratory cells on human corneas. 174 Mar 59

Long-standing release of locally cytotoxic aldehyde concentrations is responsible for lack of spontaneous endothelialization and increased calcification of glutaraldehyde fixed bovine pericardium. Postfixation treatment with amino acids made in vitro endothelialization of bioprosthetic heart valves possible. Such treated pericardium calcified significantly less (13 +/- 4 micrograms/mg dry weight) than did conventionally processed pericardium (114 +/- 25 micrograms/mg) after 63 days of subcutaneous implantation in rats. To test the ability for spontaneous in vivo endothelialization, 5 sheep had 6 mm grafts made from postfixation treated pericardium (PTP) implanted into the carotid artery, compared to PTFE grafts on the contralateral side, which spontaneously endothelialize in animal models. In a pregnant animal, both grafts occluded. All remaining pericardial grafts remained patent, but one additional PTFE graft occluded and another one was stenosed. The area covered with red thrombus was significantly smaller in the PTP grafts (3.05 +/- 3.9%) than in the PTFE grafts 42 +/- 14% (p = 0.0036); TEM and SEM showed endothelial cells growing directly on the PTP, but only on myofibroblasts in PTFE grafts. Postfixation treatment of glutaraldehyde fixed pericardium aids spontaneous endothelialization and decreases tissue calcification.
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PMID:Improved biocompatibility by postfixation treatment of aldehyde fixed bovine pericardium. 225 83

We recently reported structural changes in nodes of Ranvier of frog dorsal roots that are associated with activation at 20 or 50 Hz for 15 min. The structural effects co-occur with alterations in the latency, amplitude, and waveform of the compound action potential. The alterations in nodal morphology include formation of large extracellular vacuoles separating myelin terminal lammelae of the paranodal apparatus. Vacuolization was not apparent when the nerve was allowed to recover for 30 min prior to fixation and processing for transmission electron microscopy, indicating that it is a transient phenomenon. We first observed this effect in axons from nerve roots fixed with aldehydes and processed conventionally for TEM. The possibility that the changes noted actually occur later during tissue processing, reflecting differences in sensitivity to fixation or dehydration, remained. To address this concern we examined activated frog nodes processed by three different methods: (i) rapid freezing/freeze-substitution, (ii) direct osmication, and (iii) primary fixation in a mixture of aldehydes and osmium. Under these conditions paranodal vacuolization is still quite evident, but there are subtle differences, primarily in the appearance of the vacuoles. Also the nonstimulated control nodes fixed by direct osmication and by mixed aldehyde-osmium tended to have more vacuolized paranodes than did either the conventionally fixed or freeze-substituted preparations. Regardless, the changes are similar to those seen previously and very likely reflect changes occurring in situ. We interpret these results in the context of a functional model involving accumulation of potassium in the interstices of the compact myelin during impulse propagation. During periods of extended activity this system may be overloaded promoting focal edema and mechanical disruption of some paranodal structures. These changes could then bias subsequent nodal functions such as activation threshold, firing patterns, and conduction velocity.
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PMID:Activity associated ultrastructural changes in peripheral nodes of Ranvier are independent of fixation. 326 May 60

Surface coat material (SCM) has been illustrated in association with the apical surfaces of numerous epithelia during morphogenesis. This study investigates the development of a SCM associated with the invaginating otic placode/vesicle in the chick. Glycoconjugate containing SCM was retained by the inclusion of cetylpyridinium chloride (CPC) in the fixative, histochemically visualized by using ruthenium red (RR) staining, and viewed by scanning (SEM) or transmission (TEM) electron microscopy. Initial characterization of the glycoconjugates present in this material was elucidated by using lectins conjugated to fluorescein isothiocyanate. Lectins utilized included concanavalin A (Con A), wheat germ agglutinin (WGA), and soybean agglutinin (SBA). Invagination of the otic placode was apparent as early as stage 12. By stage 15 the vesicle was beginning to separate from the surface ectoderm as evidenced by its aperture, which was altered in shape and reduced in size. All embryos fixed with glutaraldehyde containing either CPC or RR were shown to possess SCM associated with the surface ectoderm, particularly in the area of the otic placode/vesicle. Additional embryos were processed by cryofixation without prior aldehyde fixation; these also exhibited SCM. All lectins labelled the epithelium of the otic placode/vesicle. However, their binding patterns were not identical. The binding of Con A and WGA remained constant over the stages studied, while SBA increased as the otic vesicle developed. The data clearly indicate that otic placode morphogenesis is accompanied by the synthesis of SCM rich in glycoconjugates.
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PMID:Surface coat material associated with the developing otic placode/vesicle in the chick. 335 62

In aldehyde-fixed liver and renal cortex of rat and mouse several variations of postfixation with osmium tetroxide plus potassium ferrocyanide ( FeII ) were tried. Depending on the ferrocyanide concentration different staining patterns were observed in TEM. -Osmium-High Ferrocyanide [40 mM (approximately 1%) OsO4 + 36 mM (approximately 1.5%) FeII , pH 10.4], stains membranes and glycogen. Cytoplasmic ground substance, mitochondrial matrices and chromatin are partially extracted, cell surface coats remain unstained. Membrane contrast, but extraction too, are higher with solutions containing cacodylate- than phosphate-buffer. -Osmium-Low Ferrocyanide [40 mM (approximately 1%) OsO4 + 2 mM (approximately 0.08%) FeII , pH 7.4], stains cell surface coats and basal laminae, but not glycogen, except for special cases. The trilaminar structure of membranes is poorly delineated. Signs of cytoplasmic extraction are not visible. The surface coat staining is stronger and more widespread with solutions containing phosphate- instead of cacodylate-buffer; it is enhanced by section staining with lead citrate. The cell surface coat stain does not traverse tight junctions nor permeate membranes.
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PMID:Electron staining of the cell surface coat by osmium-low ferrocyanide. 620 62

The ependymal lining of the lateral ventricles of the brain of rats, rabbits, and man was investigated at several times after death. In contrast to control material that was fixed by the aldehyde perfusing method, the following post-mortem (p.m.) changes were found: (1) Cytoplasmic protrusions of ependymal cells appear 15 min p.m. They are present up to several hours after death. (2) The formation of these protrusions causes the tufts of cilia to clump together and later to become integrated within the ependymal cell. This may simulate an unciliated surface. (3) Small porelike holes, which are present 15 min p.m. in the ependymal cell membrane, enlarge and in later stages produce a meshwork of fibers instead of a closed ependymal lining. (4) TEM observation shows that ependymal cells are separated from each other very soon after death by intercellular gaps. Cell junctions between ependymal cells resist separation over a longer p.m. period. In animal or human material that is fixed at any time after death, such modifications have to be considered very critically. In human p.m. autopsy material they are mostly the expression of a p.m. alteration.
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PMID:Post-mortem modifications of the ependyma of the lateral ventricular wall. 743 37

This study was designed to investigate patterns of fibrils organization in histochemically stained otoconia. Transmission electron microscope and video imaging were used. These data indicate that otoconia of the chick (Gallus domesticus) inner ear may have central cores in vivo. The data also show that the ultrastructural organization of fibrils fixed with aldehydes and histochemical stains follows trajectories that conform to the hexagonal shape of otoconia. These changes in direction may contribute to the formation of a central core. The existence of central cores is important for the in vivo buoyancy of otoconia. Packing of fibrils is tighter after phosphotungstic acid (PTA) stained otoconia than with other histochemical stains, which usually produce looser packing of fibrils and seemingly larger central core. TEM of tilted and untilted material showed that turning of fibrils occurs at the points where the face angles of otoconia form and where central cores exist. Video image processing of the images allowed reconstructing a template which, if assumed to repeat and change trajectories, would fit the pattern of fibrils seen in fixed otoconia. Since it is highly unlikely that aldehyde primary fixation or PTA stain caused such drastic change in the direction of fibrils, the template derived from these results may closely approximate patterns of otoconia fibrils packing in vivo. However, if the above is correct, the perfect crystallographic diffraction pattern of unfixed otoconia do not correspond to patterns of fixed fibrils.
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PMID:High resolution and image processing of otoconia matrix. 835 80

Clavulanic acid, the therapeutically important inhibitor of beta-lactamases containing a nucleophilic serine residue at their active sites, inhibits Escherichia coli TEM-2 beta-lactamase via a complex mechanism. Electrospray ionization mass spectrometry (ESIMS) studies revealed that a minimum of four different modified proteins are formed upon incubation of clavulanate with the TEM-2 enzyme. These exhibit mass increments relative to the unmodified TEM-2 beta-lactamase of 52, 70, 88, and 155 Da. Time course studies implied that no long-lived forms of clavulanate-inhibited TEM-2 beta-lactamase retain the carbons of the oxazolidine ring of clavulanate. The absence of a 199 Da increment to unmodified TEM-2 suggests rapid decarboxylation of clavulanate upon binding to the enzyme. Proteolytic digestions of purified forms of clavulanate inhibited TEM-2 beta-lactamase followed by analyses using high-performance liquid chromatography coupled to ESIMS (HPLC-ESIMS) and chemical sequencing were used to provide positional information on the modifications to the enzyme. Increments of 70 and 80 Da increments were shown to be located in a peptide containing Ser-70. A further 70 Da mass increment, assigned as a beta-linked acrylate, was localized to a peptide containing Ser-130. A mechanistic scheme for the reaction of clavulanate with TEM-2 beta-lactamase is proposed in which acylation at Ser-70 and subsequent decarboxylation is followed either by cross-linking with Ser-130 to form a vinyl ether or by reformation of unmodified enzyme via a Ser-70 linked (hydrated) aldehyde. Purified cross-linked vinyl ether was observed to slowly convert under acidic conditions to a Ser-70 linked (hydrated) aldehyde with concomitant conversion of Ser-130 to a dehydroalanyl residue.
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PMID:Inhibition of TEM-2 beta-lactamase from Escherichia coli by clavulanic acid: observation of intermediates by electrospray ionization mass spectrometry. 882 77


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