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The scanning tunneling microscope is revolutionizing the study of surfaces. In ultra-high vacuum it is capable not only of imaging individual atoms but also of determining energy states on an atom-by-atom basis. It is now possible to operate this instrument in water. Aqueous optical microscopy is confined to a lateral resolution limit of about 2000 angstroms, and aqueous x-ray microscopy has yielded a lateral resolution of 75 angstroms. With a scanning tunneling microscope, an image of a graphite surface immersed in deionized water was obtained with features less than 3 angstroms apart clearly resolved. Further, an image measured in saline solution demonstrated that the instrument can be operated under conditions useful for many biological samples.
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Electron tunneling spectroscopy has been used to characterize the degradation of beta-D-fructose after electron bombardment in a scanning electron microscope. The decrease in intensity of various vibrational bands is correlated with structural changes in the molecule, thereby providing a detailed picture of the degradation process.
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The processes involved in the dissolution and growth of crystals are closely related. Atomic force microscopy (AFM) of faceted pits (called negative crystals) formed during quartz dissolution reveals subtle details of these underlying physical mechanisms for silicates. In imaging these surfaces, the AFM detected ledges <1 nanometer (nm) high that were spaced 10 to 90 nm apart. A dislocation pit, invisible to optical and scanning electron microscopy measurements and serving as a ledge source, was also imaged. These observations confirm the applicability of ledge-motion models to dissolution and growth of silicates; coupled with measurements of dissolution rate on facets, these methods provide a powerful tool for probing mineral surface kinetics.
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Metal-insulator-metal electron tunnel junctions can be doped with a solution of an organic compound by placing a drop of the solution on the insulator and spinning of the excess. Electrical measurement of the second derivative of voltage with respect to current, as a function of applied voltage, then gives a spectrum of vibrational modes equivalent to an infrared or Raman spectrum, but with the use of only micrograms of sample.
Assuntos
Espalhamento de Radiação , Análise Espectral , Animais , Bovinos , DNA , Escherichia coli , Lasers , Métodos , Fenilalanina , RNA Bacteriano , RNA de Transferência , Espectrofotometria Infravermelho , Timo , UracilaRESUMO
The height fluctuations on top of the protein lysozyme adsorbed on mica were measured locally with an atomic force microscope operated in tapping mode in liquid. Height fluctuations of an apparent size of 1 nanometer that lasted for about 50 milliseconds were observed over lysozyme molecules when a substrate (oligoglycoside) was present. In the presence of the inhibitor chitobiose, these height fluctuations decreased to the level without the oligoglycoside. The most straightforward interpretation of these results is that the height fluctuations correspond to the conformational changes of lysozyme during hydrolysis. It is also possible, however, that the height fluctuations are, at least in part, the result of a different height or elasticity of the transient complex of lysozyme plus the substrate.
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Microscopia/métodos , Muramidase/ultraestrutura , Conformação Proteica , Adsorção , Silicatos de Alumínio , Muramidase/química , Muramidase/metabolismoRESUMO
Atomically resolved images of pressed powder samples have been obtained with the atomic force microscope (AFM). The technique was successful in resolving the particle, domain, and atomic structure of pismo clam (Tivela stultorum) and sea urchin (Strongylocentrotus purpuratus) shells and of commercially available calcium carbonate (CaCO(3)) and strontium carbonate (SrCO(3)) powders. Grinding and subsequent pressing of the shells did not destroy the microstructure of these materials. The atomic-resolution imaging capabilities of AFM can be applied to polycrystalline samples by means of pressing powders with a grain size as small as 50 micrometers. These results illustrate that the AFM is a promising tool for material science and the study of biomineralization.
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We have measured and assigned the inelastic tunneling spectra of hexanoic acid chemisorbed onto an oxidized aluminum file. We present evidence for gauche-trans as well as-trans conformers in the monolayer and evidence in support of a recent theory of tunneling intensities.
Assuntos
Ácidos Graxos , Caprilatos , Luz , Ácidos Mirísticos , Espalhamento de Radiação , Espectrofotometria Infravermelho , Análise EspectralRESUMO
The atomic force microscope (AFM) was used to image an electrode surface at atomic resolution while the electrode was under potential control in a fluid electrolyte. A new level of subtlety was observed for each step of a complete electrochemical cycle that started with an Au(111) surface onto which bulk Cu was electrodeposited. The Cu was stripped down to an underpotential-deposited monolayer and finally returned to a bare Au(111) surface. The images revealed that the underpotential-deposited monolayer has different structures in different electrolytes. Specifically, for a perchloric acid electrolyte the Cu atoms are in a close-packed lattice with a spacing of 0.29 +/- 0.02 nanometer (nm). For a sulfate electrolyte they are in a more open lattice with a spacing of 0.49 +/- 0.02 nm. As the deposited Cu layer grew thicker, the Cu atoms converged to a (111)-oriented layer with a lattice spacing of 0.26 +/- 0.02 nm for both electrolytes. A terrace pattern was observed during dissolution of bulk Cu. Images were obtained of an atomically resolved Cu monolayer in one region and an atomically resolved Au substrate in another in which a 30 degrees rotation of the Cu monolayer lattice from the Au lattice is clearly visible.
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The high resolution of the scanning tunneling microscope (STM) makes it a potentially important tool for the study of biomaterials. Biological materials can be imaged with the STM by a procedure in which fluid, nonconductive biomaterials are replaced by rigid and highly conductive freeze-fracture replicas. The three-dimensional contours of the ripple phase of dimyristoylphosphatidylcholine bilayers were imaged with unprecedented resolution with commercial STMs and standard freeze-fracture techniques. Details of the ripple amplitude, asymmetry, and configuration unobtainable by electron microscopy or x-ray diffraction can be observed relatively easily with the STM.
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Dimiristoilfosfatidilcolina , Bicamadas Lipídicas , Membranas Artificiais , Microscopia/métodos , Técnica de Fratura por Congelamento , Membranas/ultraestrutura , Microscopia Eletrônica , Difração de Raios XRESUMO
A scanning ion-conductance microscope (SICM) has been developed that can image the topography of nonconducting surfaces that are covered with electrolytes. The probe of the SICM is an electrolyte-filled micropipette. The flow of ions through the opening of the pipette is blocked at short distances between the probe and the surface, thus, limiting the ion conductance. A feedback mechanism can be used to maintain a given conductance and in turn determine the distance to the surface. The SICM can also sample and image the local ion currents above the surfaces. To illustrate its potential for imaging ion currents through channels in membranes, a topographic image of a membrane filter with 0.80-micrometer pores and an image of the ion currents flowing through such pores are presented.
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Íons , Microscopia/instrumentação , Membrana Celular/ultraestrutura , Condutividade Elétrica , Eletrodos , Canais Iônicos/metabolismo , SoluçõesRESUMO
The scanning tunneling microscope (STM) and the atomic force microscope (AFM) are scanning probe microscopes capable of resolving surface detail down to the atomic level. The potential of these microscopes for revealing subtle details of structure is illustrated by atomic resolution images including graphite, an organic conductor, an insulating layered compound, and individual adsorbed oxygen atoms on a semiconductor. Application of the STM for imaging biological materials directly has been hampered by the poor electron conductivity of most biological samples. The use of thin conductive metal coatings and replicas has made it possible to image some biological samples, as indicated by recently obtained images of a recA-DNA complex, a phospholipid bilayer, and an enzyme crystal. The potential of the AFM, which does not require a conductive sample, is shown with molecular resolution images of a nonconducting organic monolayer and an amino acid crystal that reveals individual methyl groups on the ends of the amino acids. Applications of these new microscopes to technology are demonstrated with images of an optical disk stamper, a diffraction grating, a thin-film magnetic recording head, and a diamond cutting tool. The STM has even been used to improve the quality of diffraction gratings and magnetic recording heads.
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Microscopia Eletrônica de Varredura , Microscopia , Animais , Cristalização , DNA/metabolismo , DNA/ultraestrutura , Bicamadas Lipídicas , Microscopia/instrumentação , Microscopia/métodos , Microscopia Eletrônica de Varredura/instrumentação , Microscopia Eletrônica de Varredura/métodos , Recombinases Rec A/metabolismoRESUMO
Atomic force microscope images of polymerized monolayers of n-(2-aminoethyl)-10,12-tricosadiynamide revealed parallel rows of molecules with a side-by-side spacing of approximately equal to 0.5 nanometer. Forces used for imaging (10(-8) newton) had no observable effect on the polymer strands. These results demonstrate that atomic force microscope images can be obtained for an organic system.
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Amidas , Ácidos Graxos Insaturados , Microscopia/instrumentação , Polímeros , Microscopia Eletrônica , Alcamidas Poli-Insaturadas , Propriedades de SuperfícieRESUMO
Reproducible images of uncoated DNA in the atomic force microscope (AFM) have been obtained by imaging plasmid DNA on mica in n-propanol. Specially sharpened AFM tips give images with reproducible features several nanometers in size along the DNA. Plasmids can be dissected in propanol by increasing the force applied by the AFM tip at selected locations.
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DNA/ultraestrutura , Plasmídeos , DNA Viral/ultraestrutura , Microanálise por Sonda Eletrônica/métodos , Microscopia Eletrônica de Varredura/métodos , PropranololRESUMO
The adsorption of neutral molecules and ions on the surfaces of zeolites was observed in real time with an atomic force microscope (AFM). Direct imaging of the surface of the zeolite clinoptilolite was possible by using a diluted tert-butyl ammonium chloride solution as a medium. Images of the crystal in different liquids revealed that molecules could be bound to the surface in different ways; neutral molecules of tert-butanol formed an ordered array, whereas tert-butyl ammonium ions formed clusters. These absorbed molecules were not rearranged by the AFM tip when used in an imaging mode. However, when a sufficiently large force was applied, the tip of the AFM could rearrange the tert-butyl ammonium ions on the zeolite surface. This demonstration of molecular manipulation suggests new applications, including biosensors and lithography.
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The atomic force microscope (AFM) can be used to image the surface of both conductors and nonconductors even if they are covered with water or aqueous solutions. An AFM was used that combines microfabricated cantilevers with a previously described optical lever system to monitor deflection. Images of mica demonstrate that atomic resolution is possible on rigid materials, thus opening the possibility of atomic-scale corrosion experiments on nonconductors. Images of polyalanine, an amino acid polymer, show the potential of the AFM for revealing the structure of molecules important in biology and medicine. Finally, a series of ten images of the polymerization of fibrin, the basic component of blood clots, illustrate the potential of the AFM for revealing subtle details of biological processes as they occur in real time.
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Cristalografia , Microscopia/instrumentação , Polímeros , Água , Fibrina , Humanos , Técnicas In Vitro , Peptídeos , Trombina , Gravação em VídeoRESUMO
Reproducible, high-contrast, nanometer-resolution AFM images of uncoated plasmid DNA can be obtained with nanogram quantities of DNA with the help of two advances in sample preparation: (1) Heating a DNA solution at 35 degrees C for 10 to 20 minutes before deposition on mica helps separate and spread the DNA, and (2) Using 5 microliter drops of the heated DNA solution in the concentration range of 2 to 10 nanogram/microliter in contact with a specially prepared mica surface for 5 to 10 minutes gives optimal coverage with only nanograms of DNA.
Assuntos
DNA/química , Plasmídeos/química , Silicatos de Alumínio/química , Microscopia Eletrônica de VarreduraRESUMO
We have imaged DNA from the calf thymus nucleosome using a scanning tunneling microscope (STM) operated in water. The fragments are deposited onto the interface between a buffer solution and an epitaxially grown gold surface using an electrochemical tecnique. Most of the fragments are fairly straight, and when individual polymers can be identified, their length is consistent with the expected 146 basepairs (approximately 500 A). The resolution is often adequate to show signs of the 36 A helical pitch. Some images show a structure which appears to have abrupt kinks of the sort predicted by Crick and Klug (Nature 255, 530-533, 1975). In order to check that this shape is not a consequence of binding to underlying structure on the gold substrate, we have also made images of kinked structures using an atomic force microscope (AFM) with the DNA bound to glass.
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DNA/ultraestrutura , Microscopia Eletrônica de Varredura , Nucleossomos/ultraestrutura , Animais , Bovinos , Conformação MolecularRESUMO
An atomic force microscope (AFM) was used to investigate a specific recognition reaction: the binding of streptavidin to a biotinylated lipid bilayer. Prior to the recognition reaction, the phase coexistence of the lipid bilayer was clearly observed: fluid domains were lower than the crystalline domains. After introducing to the bilayer a very dilute solution of streptavidin to give a final concentration of approximately 0.5 microM, the recognition reaction was imaged in real time. Several hours later, we observed a contrast reversal, i.e., the previously lower fluid domains grew so much in height that they became higher than the crystalline domains. We found that the streptavidin molecules bound almost exclusively to the biotin in the fluid domain (less than 0.25% coverage of the crystalline domains). The apparent structure of the few streptavidin molecules bound to the crystalline domain of the bilayer is shown to depend on the applied force. Finally, in a 2-dimensional quasi-crystal in which the streptavidin molecules were compressed at the air-water interface molecular resolution was achieved.
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Proteínas de Bactérias/ultraestrutura , Proteínas de Bactérias/metabolismo , Biotina , Bicamadas Lipídicas/metabolismo , Microscopia/métodos , Ligação Proteica , EstreptavidinaRESUMO
AFM was used to investigate dialysis membranes made of regenerated cellulose by the cuoxam process. The membranes were either Cuprophan or experimental samples, modified with different amounts of diethylaminoethylcellulose (DEAE). Atomic force microscopes with optical-lever detection systems were used to image the dry membranes in air as received from the manufacturer as well as wet membranes in a swollen state under water. Differences could be observed between modified and unmodified as well as between dry and wet membranes.