Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Microscopy, Scanning Probe”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,459 records · Page 81Linked to original sources
PubMed1998

Undifferentiated HL-60 cells internalize an antitumor alkyl ether phospholipid more rapidly than resistant K562 cells.

In this study, we confirmed a previous finding that 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (methyl-PAF) expresses higher antineoplastic activity against the promyelocytic leukemia cell line HL-60, than against the erythroleukemic cell line K562, and intended to clarify the reason for this. Using an albumin back-exchange method, we measured the rates of binding and internalization of [3H]methyl-PAF by HL-60 and K562 cells. We found that methyl-PAF associated very rapidly and to similar extents with the two types of cells at low concentrations of extracellular bovine serum albumin, but that when bound to the cell surface, it was internalized into HL-60 cells faster than into K562 cells. The internalization of methyl-PAF by HL-60 cells was concentration-independent, intracellular ATP-independent and susceptible to thiol group-modifying reagents and cytochalasin B. Thus the inward transbilayer movement of methyl-PAF seems to occur by cytochalasin B-sensitive protein-mediated mechanism based on passive diffusion not requiring energy, in which SH-groups of protein play a critical role. We also found that the internalization of 1-hexadecanoyl-2-(4,4-difluoro-5,7- dimethyl-4-bora-3a, 4a-diaza-s-indacene-3-pentanoyl)-sn-glycero-3-phosphocholine (Bodipy-C5-PC), whose structure resembles that of methyl-PAF, into HL-60 cells was faster than that into K562 cells. Using a combination of an albumin back-exchange method and observation by confocal laser scanning microscopy, we next examined the intracellular distribution of this fluorescent phospholipid probe after its internalization. Intracellular membranes, especially those peripheral to nuclei, were fluorescence-labeled in both HL-60 and K562 cells, but fluorescence of the nuclear membranes was weak, suggesting that this probe seems mainly to accumulate in intracellular granules, and may interact directly with several key enzymes for phospholipid metabolism, leading to cell injury. Because the difference between the internalization rates of methyl-PAF in HL-60 and K562 cells was correlated with their different susceptibilities to the cytotoxic effect of methyl-PAF, we suggest that the capacities for uptake of methyl-PAF and its accumulation in intracellular membranes are critical factor for its induction of apoptosis. (c) 1998 Elsevier Science B.V.

Adenosine Triphosphate↗
PubMed2001

Membrane-initiated Ca(2+) signals are reshaped during propagation to subcellular regions.

An important aspect of Ca(2+) signaling is the ability of cells to generate intracellular Ca(2+) waves. In this study we have analyzed the cellular and subcellular kinetics of Ca(2+) waves in a neuroendocrine transducer cell, the melanotrope of Xenopus laevis, using the ratiometric Ca(2+) probe indo-1 and video-rate UV confocal laser-scanning microscopy. The purpose of the present study was to investigate how local Ca(2+) changes contribute to a global Ca(2+) signal; subsequently we quantified how a Ca(2+) wave is kinetically reshaped as it is propagated through the cell. The combined kinetics of all subcellular Ca(2+) signals determined the shape of the total cellular Ca(2+) signal, but each subcellular contribution to the cellular signal was not constant in time. Near the plasma membrane, [Ca(2+)](i) increased and decreased rapidly, processes that can be described by a linear and exponential function, respectively. In more central parts of the cell slower kinetics were observed that were best described by a Hill equation. This reshaping of the Ca(2+) wave was modeled with an equation derived from a low-pass RC filter. We propose that the differences in spatial kinetics of the Ca(2+) signal serves as a mechanism by which the same cellular Ca(2+) signal carries different regulatory information to different subcellular regions of the cell, thus evoking differential cellular responses.

Animals↗
PubMed1999

Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation.

Multiphoton excitation (MPE) of fluorescent probes has become an attractive alternative in biological applications of laser scanning microscopy because many problems encountered in spectroscopic measurements of living tissue such as light scattering, autofluorescence, and photodamage can be reduced. The present study investigates the characteristics of two-photon excitation (2PE) in comparison with confocal one-photon excitation (1PE) for intracellular applications of fluorescence correlation spectroscopy (FCS). FCS is an attractive method of measuring molecular concentrations, mobility parameters, chemical kinetics, and fluorescence photophysics. Several FCS applications in mammalian and plant cells are outlined, to illustrate the capabilities of both 1PE and 2PE. Photophysical properties of fluorophores required for quantitative FCS in tissues are analyzed. Measurements in live cells and on cell membranes are feasible with reasonable signal-to-noise ratios, even with fluorophore concentrations as low as the single-molecule level in the sampling volume. Molecular mobilities can be measured over a wide range of characteristic time constants from approximately 10(-3) to 10(3) ms. While both excitation alternatives work well for intracellular FCS in thin preparations, 2PE can substantially improve signal quality in turbid preparations like plant cells and deep cell layers in tissue. At comparable signal levels, 2PE minimizes photobleaching in spatially restrictive cellular compartments, thereby preserving long-term signal acquisition.

Animals↗
PubMed1996

Intraneuronal aluminum potentiates iron-induced oxidative stress in cultured rat hippocampal neurons.

Aluminum can facilitate Fe-mediated oxidative injury, which may contribute to Al neurotoxicity. It has been reported that Al potentiates Fe-induced oxidative stress in cultured granule cells, suggesting a mechanism for Al facilitation of Fe-mediated oxidative injury. However, the relationship of intracellular Al concentration to Fe-induced oxidative stress has not been reported. In the present study, neuronal oxidative stress and survival were investigated. Embryo rat hippocampal neuron cultures were treated with Al2(SO4)3 and/or FeSO4. An ionophore, A23187, was utilized to facilitate cellular Al uptake. Intraneuronal Al concentration was ascertained by laser microprobe mass spectrometry (LMMS). Neuronal oxidative stress was measured by confocal laser scanning microscopy, using 2,7-dichlorofluorescin diacetate (DCFH-DA) as a probe. The study showed that neuronal Al uptake was facilitated by the ionophore and that an increase of intraneuronal Al concentration potentiated Fe-induced oxidative stress and neuronal death. The results indicate that Al potentiation of Fe-induced oxidative stress might contribute to Al facilitation of oxidative injury, and thus to Al neurotoxicity.

Aluminum↗
PubMed1998

Changes in three-dimensional architecture of microfilaments in cultured vascular smooth muscle cells during phenotypic modulation.

To investigate changes in the three-dimensional microfilament architecture of vascular smooth muscle cells (SMC) during the process of phenotypic modulation, rabbit aortic SMCs cultured under different conditions and at different time points were either labelled with fluorescein-conjugated probes to cytoskeletal and contractile proteins for observation by confocal laser scanning microscopy, or extracted with Triton X-100 for scanning electron microscopy. Densely seeded SMCs in primary culture, which maintain a contractile phenotype, display prominent linear myofilament bundles (stress fibres) that are present throughout the cytoplasm with alpha-actin filaments predominant in the central part and beta-actin filaments in the periphery of the cell. Intermediate filaments form a meshed network interconnecting the stress fibres and linking directly to the nucleus. Moderately and sparsely seeded SMCs, which modulate toward the synthetic phenotype during the first 5 days of culture, undergo a gradual redistribution of intermediate filaments from the perinuclear region toward the peripheral cytoplasm and a partial disassembly of stress fibres in the central part of the upper cortex of the cytoplasm, with an obvious decrease in alpha-actin and myosin staining. These changes are reversed in moderately seeded SMCs by day 8 of culture when they have reached confluence. The results reveal two changes in microfilament architecture in SMCs as they undergo a change in phenotype: the redistribution of intermediate filaments probably due to an increase in synthetic organelles in the perinuclear area, and the partial disassembly of stress fibres which may reflect a degradation of contractile components.

Actin Cytoskeleton↗
PubMed2006

Side-by-side characterization of electron tunneling through monolayers of isomeric molecules: a combined experimental and theoretical study.

We report a new experimental method for measuring relatively small differences in electron tunneling through two distinct monolayers. We place them side by side using scanning probe nanolithography and compare the tunneling currents by conductive probe atomic force microscopy under identical force, voltage, and tip contamination conditions. We demonstrate the validity of our approach by applying it to two isomeric molecules with similar length and functional groups, with only the position of two functional groups, one aromatic and the other aliphatic, being inverted with respect to each other. The relative values of the two tunneling currents, calculated using density functional theory and the Tersoff-Hamann approach, compare very well with the experimental data, providing us with an example of theory vs experiment agreement that is rather uncommon in this field.

Letter↗
PubMedDate not supplied

[Wear of the polyethylene components of endoprostheses following clinical application].

13 various hip joint and knee joint polyethylene components, previously removed from patients after an implantation period of 1.5 to 4 years, were tested and showed an obvious damage of the sliding surface. Macroscopic and microscopic investigations performed by light and scanning electron microscopy revealed an extreme wear of the polyethylene surface respectively an unregular wear at the component's border up to the mm-range. Additional specific X-ray and infrared spectrometric analyses proved in the clear majority of all cases the contribution of bone cement to increased abrasion and its influence upon the intensity of wear. Surplus bone cement which had not been completely removed from the prosthesis component or loosening of prosthesis components are considered as reason for inclusion of bone cement particles between the sliding surfaces.

Bone Cements↗
PubMed2002

Temperature control of electronic channels through a single atom.

The elastic electronic channels through a single hydrogen atom adsorbed on a Ge(111)-c(2 x 8) surface have been investigated by scanning tunneling microscopy and I(V) spectroscopy, whereas inelastic channels have been probed by the vertical and horizontal manipulation of individual hydrogen atoms. The substrate temperature, over the range 30-300 K, has proven to be a powerful parameter to freeze specific electronic channels, offering the possible control of elastic and inelastic channels through a single atom. This opens up very interesting perspectives for controlling the operation of nanodevices.

Journal Article↗
PubMed2000

Intra- and extracellular measurement of reactive oxygen species produced during heat stress in diaphragm muscle.

Skeletal muscles are exposed to increased temperatures during intense exercise, particularly in high environmental temperatures. We hypothesized that heat may directly stimulate the reactive oxygen species (ROS) formation in diaphragm (one kind of skeletal muscle) and thus potentially play a role in contractile and metabolic activity. Laser scan confocal microscopy was used to study the conversion of hydroethidine (a probe for intracellular ROS) to ethidium (ET) in mouse diaphragm. During a 30-min period, heat (42 degrees C) increased ET fluorescence by 24 +/- 4%, whereas in control (37 degrees C), fluorescence decreased by 8 +/- 1% compared with baseline (P < 0.001). The superoxide scavenger Tiron (10 mM) abolished the rise in intracellular fluorescence, whereas extracellular superoxide dismutase (SOD; 5,000 U/ml) had no significant effect. Reduction of oxidized cytochrome c was used to detect extracellular ROS in rat diaphragm. After 45 min, 53 +/- 7 nmol cytochrome c. g dry wt(-1). ml(-1) were reduced in heat compared with 22 +/- 13 nmol. g(-1). ml(-1) in controls (P < 0.001). SOD decreased cytochrome c reduction in heat to control levels. The results suggest that heat stress stimulates intracellular and extracellular superoxide production, which may contribute to the physiological responses to severe exercise or the pathology of heat shock.

1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium ↗
PubMed1994

The spatial localization of 18 S rRNA genes, in relation to the descent of the cells, in the root cortex of Petunia hybrida.

The 3-D localization of transcription inactive 18 S rRNA genes was studied in interphase nuclei of Petunia hybrida root tip cells. To enable a cell type (i.e. cortex)-specific study in which also the orientation and descent of the cells could be taken into account, a method was developed to preserve the spatial organization of the root meristem. The ribosomal genes were detected by fluorescence in situ hybridization using a biotinylated cDNA probe. 3-D images of 81 nuclei, obtained by confocal scanning laser microscopy, were processed with newly developed computer software. 3-D nucleolar and nuclear dimensions, and the localization of the FISH-spots, were recorded interactively. We compared the absolute and relative position of the genes within and between files of cells of the cortex region of several roots, taking into account the genealogical relationship of the cells. Statistical analysis showed that both the relative and absolute positions of the inactive genes were random, also in more closely related cells within a file of cells. A 'relict telophase orientation' of the genes (i.e. the position of the genes in the daughter cells are mirror images of each other) could only be observed in the G0/1 phase of 'true' daughter cells; the orientation was not preserved throughout the next cell cycle.

Cell Nucleolus↗
PubMed2000

[Regenerative potential in the organ of Corti after otic intoxication].

The auditory sensory cells are sensitive to a variety of influences such as noise, ototoxic drugs and aging. In the cochlea of mammals, the destroyed sensory cells are not replaced by new sensory cells. That leads to cochlear deafness, a frequent disease in human. Unfortunately, such auditory impairment is out of reach of treatment. The development of new therapeutic strategies in this field requires a precise knowledge of the mechanisms involved in auditory sensory cells disappearance and in organ of Corti's degeneration. The aim of our study was to characterize cellular and molecular changes in the cochlea of rats which had been intoxicated with the ototoxic antibiotic amikacin. The animals were sacrificed at different survival times during and after the antibiotic treatment and their cochleas were investigated using transmission and scanning electron microscopy and using confocal microscopy after tissue labellings with different fluorescent probes. The results revealed the existence of three periods. The first one corresponds to the disappearance of the sensory cells which die by apoptosis. During the second period, the organ of Corti undergoes a scarring process; concomitantly, a contingent of nonsensory supporting cells attempts to transdifferentiate directly into sensory cells. This process however fails, and the supporting cells never reach the status of hair cells. A general process of dedifferentiation of all the epithelial cells of the organ of Corti followed by a massive apoptosis of numerous epithelial cells and of most ganglion cells occurs during the third period. After that, the organ of Corti is definitely reduced to a simple monolayered epithelium. On the basis of these data, experimental strategies aimed i) to protect the sensory cells against apoptosis and ii) to promote sensory cell regeneration are now under study. They might have important implications in human therapy.

Amikacin↗
PubMed2002

[Structural and metabolic status and functional properties of erythrocytes in schizophrenic patients].

Structural metabolic properties and functional state of blood erythrocytes have been studied in 118 patients with paranoid schizophrenia and 22 patients with residual schizophrenia. Control group comprised 103 normals. Thin layer chromatography, fluorescent probing Na+, K(+)-ATP-ase activity evaluation, scanning and transmission electron microscopy, laser difractometry and selectometry were used. A distinct increase of cholesterol and lysophosphatidylcholin, a decrease of phosphatidyl ethanolamine, higher organisation of both integral lipid biolayer and annular lipid fraction, structural modification of outer membrane layers, Na+, K(+)-ATP-ase activity decrease, disorganization of surface architectonics and ultrastructure of erythrocytes, decrease of their deformatability and aggretablity enhancement were found in erythrocyte membranes of the patients with paranoid schizophrenia who had not received psychopharmacocorrective therapy for 6 months before examination. The expression of the indices studied did not correlate with either the disease exacerbation or remission. A similar pattern of erythrocyte disturbances was detected in patients with paranoid and residual schizophrenia on long-term treatment with neuroleptics.

Adult↗
PubMed2003

[Principles of changes of structural organization of cell membranes and functional properties of erythrocytes in neurotic disorders].

Investigation into structural, metabolic, and functional conditions of red blood cells was performed in 24 patients with a neurosis (neurasthenia, disturbance of asaptation) with the aid of electrophoretic division of proteins of the erythrocyte membrane, thin-layer chromatography, fluorescent probing of membranes, evaluation of peroxidative oxidation process, scanning and transmission electron microscopy, laser diphractometry, photometry. The patients with neurotic disorders at the early period after the influence of psychogenic factors (up to 3 months) revealed disorganization of lipid and protein composition of the red cell membrane, increase in microviscosity of its lipid phase, impairment of surface architectonics and ultrastructure of red cells, decrease of a deformation ability and increase of aggregate properties of erythrocytes. The authors treat stability of erythrocytes' homeostasis under the long-term influence of psychogenic factors from a viewpoint of adaptive changes in organism under the influence of neurogenic factors.

Adult↗
PubMed1989

[Endocytosis and cytotoxin effect induced by the attachment of the glycolipoprotein of leptospires to vascular endothelial cell in culture].

The Glycolipoprotein (GLP) of various virulent leptospires was extracted, purified and identified. The GLP was combined with fluorescence rhodamine B and the colloidal gold as the probe. Meanwhile, the bovine aortic endothelial cell (EN) was isolated and cultured in the medium 199 "Nissui". The GLP-rhodamine B and GLP-colloidal gold probe were added to cultured EN and incubated at 4 degrees C for 2 h then at 37 degrees C for 2 h. The interaction between GLP-rhodamine B, GLP-CG probe and EN were observed and photography by fluorescence microscopy (Leitz) and scanning electronmicroscopy. The releasing rate of lactic acid dehydrogenase (LDH), acid phosphatase (AcP) and protein from the EN were detected. The results indicated that the GLP of Leptospira interrogans serovar Lai, strain 017 attached to EN and a endocytosis was happened subsequently. The LDH and AcP releasing rate were significantly higher than control and the cell protein was much less than control. The results showed that the GLP of L. interrogans strain 017 had the cytotoxin effect through a specific attachment, endocytosis in EN. The endocytosis probably is a receptor-mediated endocytosis.

Animals↗
PubMed2000

Effect of 10% carbamide peroxide bleaching gel on enamel microhardness, microstructure, and mineral content.

The effect of 10% carbamide peroxide on human enamel subsurface layers was examined. Microhardness, microstructure, and mineral content were studied in such a manner that control and test side were located on the same tooth. Longitudinally sectioned samples were obtained from six teeth and Vickers microhardness of the enamel was measured. Enamel microstructure was analyzed using a scanning electron microscope and subjected to electron probe microanalysis for chemical analysis of calcium (Ca) and phosphorus (P). The concentration of Ca in the bleaching gel was measured spectrophotometrically and the concentration of P was measured photometrically. Results showed that a bleaching gel of 10% carbamide peroxide did not significantly affect the microhardness of the enamel. Scanning electron microscopic analysis showed local changes in enamel microstructure similar to those of initial caries. Electron probe microanalysis showed lowered concentrations of Ca and P; in addition the Ca:P ratio was lowered. There was some Ca and P in the bleaching gel after use. It is concluded that 10% carbamide peroxide causes local microstructural and chemical changes in enamel that are likely not clinically significant.

Adolescent↗
PubMed1980

Alzheimer's disease: X-ray spectrometric evidence of aluminum accumulation in neurofibrillary tangle-bearing neurons.

The elemental content of neurons of the hippocampus was studied by a combination of scanning electron microscopy and x-ray spectrometry in autopsy-derived brain tissue from three cases of senile dementia (Alzheimer type) and three nondemented elderly controls. Foci of aluminum were detected within the nuclear region of a high percentage of neurons containing neurofibrillary tangles from the cases of senile dementia as well as the elderly controls. The adjacent normal-appearing neurons from both groups of patients were virtually free of detectable aluminum. These findings suggest that the association of aluminum to Alzheimer's disease extends to the neuronal level.

Aluminum↗
PubMed2005

Cementum on Smilodon sabers.

The maxillary canines of Smilodon californicus Bovard, 1907 have a deeply curved cementoenamel junction. The gingiva of modern cats is attached to the tooth at the cementoenamel junction and provides tactile and other dental information to the animal. The presence of cementum at the cervix of the maxillary canines, also called sabers, would indicate that the gingiva in Smilodon was attached in this region. Such an attachment would be advantageous, providing stability and sensory input for the large tooth. Also, gingiva at the cervix would impact the manner in which the teeth were used. Previous study using scanning electron microscopy of dental casts was indirect. The purpose of this study was to confirm by direct methods the presence of cementum at the cervix of Smilodon californicus sabers. Parts of three Smilodon californicus sabers were sectioned and examined with light and scanning electron microscopy (EDS). In addition, percent weight of calcium and phosphorus was measured in enamel, dentin, and cementum using electron dispersive spectroscopy. Cementum was identified in the cervical region of each saber. Spectroscopy confirmed that the tissue is calcified and the mineral is hydroxyapatite. Percent calcium and percent phosphorus of individual tissues were highly variable between specimens. However, the ratios of calcium to phosphorus were not significantly different from the hydroxyapatite standard. In the future, bite models will have to take the presence of soft tissues into account.

Animals↗
PubMed2000

Alumina powder/Bis-GMA composite: effect of filler content on mechanical properties and osteoconductivity.

Three composites consisting of alumina powder dispersed in a bisphenol-a-glycidyl methacrylate (Bis-GMA) matrix were prepared and evaluated to assess the effect of alumina powder content on the mechanical properties and osteoconductivity of the composite. The alumina powder composites (APC) consisted of alumina powder (AL-P) as the inorganic filler dispersed in a Bis-GMA matrix that was solidified by a radical polymerization process. Prior to polymerization the AL-P was mixed with the monomers in proportions of 50%, 70%, and 80% by weight (APC50, APC70, and APC80). A fused silica-glass-filled composite containing 70% glass by weight (SGC70) was used as a control. The compressive and bending strengths, the elastic modulus in bending, and the bending strain of the composites increased as the AL-P content increased. We also evaluated the composites in vivo by implanting them into the medullary canals of rat tibiae. To compare the osteoconductivity of the composites, an affinity index was calculated for each composite; the affinity index equals the length of a bone in direct apposition to the composite and is expressed as a percentage of the total length of the composite surface. Microradiographic examination for periods of up to 26 weeks after implantation revealed that APC50, APC70, and APC80 all exhibited excellent osteoconductivity and made direct contact with the bone with no interposed soft tissues. However, the higher the AL-P content of the composite, the higher the osteoconductivity, especially at 4 weeks after the operation. Moreover, the amount of bone directly apposed to the composite surface increased with time. In contrast, little bone formation was seen on the surface of SGC70, even after 26 weeks. Observation by scanning electron microscope-energy dispersive X-ray microanalysis demonstrated that bone made direct contact with the APC surface through a layer containing calcium, phosphorus, and alumina powder. These results suggest that APC shows promise as a basis for developing mechanically strong and highly osteoconductive composites.

Aluminum Oxide↗