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At least 145 records · Page 8Linked to original sources
PubMed1999

Comparative Study of Field Emission-Scanning Electron Microscopy and Atomic Force Microscopy to Assess Self-assembled Monolayer Coverage on Any Type of Substrate.

: In this work, we show how field emission-scanning electron microscopy (FE-SEM) can be a useful tool for the study of self-assembled monolayer systems. We have carried out a comparative study using FE-SEM and atomic force microscopy (AFM) to assess the morphology and coverage of self-assembled monolayers (SAM) on different substrates. The results show that FE-SEM images present the same qualitative information obtained by AFM images when the SAM is deposited on a smooth substrate (e.g., mica). Further experiments with rough substrates (e.g., Al grains on glass) show that FE-SEM is capable of unambiguously identifying SAMs on any type of substrate, whereas AFM has significant difficulties in identifying SAMs on rough surfaces.

Journal Article↗
PubMed2000

Atomic force microscopy with carbon nanotube probe resolves the subunit organization of protein complexes.

Among many scanning probe microscopies, atomic force microscopy (AFM) is a useful technique to analyse the structure of biological materials because of its applicability to non-conductors in physiological conditions with high resolution. However, the resolution has been limited to an inherent property of the technique; tip effect associated with a large radius of the scanning probe. To overcome this problem, we developed a carbon nanotube probe by attaching a carbon nanotube to a conventional scanning probe under a well-controlled process. Because of the constant and small radius of the tip (2.5-10 nm) and the high aspect ratio (1:100) of the carbon nanotube, the lateral resolution has been much improved judging from the apparent widths of DNA and nucleosomes. The carbon nanotube probes also possessed a higher durability than the conventional probes. We further evaluated the quality of carbon nanotube probes by three parameters to find out the best condition for AFM imaging: the angle to the tip axis; the length; and the tight fixation to the conventional tip. These carbon nanotube probes, with high vertical resolution, enabled us to clearly visualize the subunit organization of multi-subunit proteins and to propose structural models for proliferating cell nuclear antigen and replication factor C. This success in the application of carbon nanotube probes provides the current AFM technology with an additional power for the analyses of the detailed structure of biological materials and the relationship between the structure and function of proteins.

Carbon↗
PubMed1992

Imaging and nanodissection of individual supercoiled plasmids by atomic force microscopy.

The atomic force microscope (AFM) was used to image supercoiled plasmid DNA deposited on a mica surface in either a hydrated or desiccated state. Hydrated plasmid was precisely cut by the scanning tip at a location determined by the instrument operator. Small pieces of DNA (100-150 nm in length) were excised and deposited adjacent to the dissected plasmid, demonstrating that it is possible to remove and manipulate genomic DNA fragments, unresolvable by light microscopy, from defined chromosomal locations by AFM.

DNA, Superhelical↗
PubMed1992

High-resolution imaging of chromosome-related structures by atomic force microscopy.

An atomic force microscope (AFM) was combined with a conventional optical microscope. The optical microscope proved to be very convenient for locating objects of interest. In addition, the high-resolution AFM image can be compared directly with the traditional optical image. The instrument was used to study chromosome structures. High-resolution chromosome images revealed details of the 30-nm chromatide structure, confirming earlier electron microscopic observations. Chromosomes treated with trypsin revealed a banding pattern in height which is very similar to the optical image observed after staining with Giemsa. Furthermore, it is shown that the AFM can be used to locate DNA probes on in situ hybridized chromosomes. Images of the synaptonemal complex isolated from rat spermatocytes revealed details that improve the understanding of the three-dimensional structure of this protein.

Animals↗
PubMed1992

Molecular-resolution images of aspirin crystals with atomic force microscopy.

The atomic force microscope was developed in order to image arrays of molecules on the (001), (100) and (011) faces of aspirin crystals in water. Lattice spacings and symmetries of methyl groups and the part of the phenyl groups on the (001) face in water are consistent with X-ray diffraction data. Surfaces of the (100) face that show the best-developed cleavage, in general, are difficult to image. Although the lattice constant derived from (100) image is consistent with the X-ray diffraction data, the c-glide plane symmetry is lost and diffused spots are observed that are 1.8-3.1 times more elongated than (001) spots. It is not possible to image the (011) surface. This initial success in imaging the molecular crystal surfaces of drug compounds has clarified the different submolecular structures of the three different crystal faces, and the close relationship between the imaging difficulties and the differences in the dissolution process in water.

Aspirin↗
PubMed2000

Subatomic Features on the Silicon (111)-(7x7) Surface Observed by Atomic Force Microscopy.

The atomic force microscope images surfaces by sensing the forces between a sharp tip and a sample. If the tip-sample interaction is dominated by short-range forces due to the formation of covalent bonds, the image of an individual atom should reflect the angular symmetry of the interaction. Here, we report on a distinct substructure in the images of individual adatoms on silicon (111)-(7x7), two crescents with a spherical envelope. The crescents are interpreted as images of two atomic orbitals of the front atom of the tip. Key for the observation of these subatomic features is a force-detection scheme with superior noise performance and enhanced sensitivity to short-range forces.

Journal Article↗
PubMed2005

Surface heterogeneity of polystyrene latex particles determined by dynamic force microscopy.

Atomic force microscopy (AFM) was employed to characterize the surface chemistry distribution on individual polystyrene latex particles. The particles were obtained by surfactant-free emulsion polymerization and contained hydrophilic quaternary ammonium chloride, sodium sulfonate, or hydroxyethyl groups. The phase shift in dynamic force mode AFM is sensitive to charge/chemical interactions between an oscillating atomic force microscope tip and a sample surface. In this work, the phase imaging technique distinguished phase domains of 50-100 nm on the surfaces of dried latex particles in ambient air. The domains are attributed to the separation of ion-rich and ion-poor components of the polymer on the particle surface.

Journal Article↗
PubMed1988

Scanning tunneling microscopy and atomic force microscopy: application to biology and technology.

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.

Animals↗
PubMedDate not supplied

[Functionalization of aminomodified probes for atomic force microscopy].

Probes for atomic force microscopy functionalized by bovine serum albumin were obtained, which may be used for molecular recognition studies. The procedure of the modification and functionalization of probes includes three stages. First, amino probes are obtained by modification in vapors of amino silane derivative. Then a homobifunctional amino reactive cross-linker is covalently linked to surface amino groups of the amino probe. And finally, the probe with the covalently attached cross-linker is functionalized by bovine serum albumin molecules. The probes obtained were characterized at different stages of the modification by atomic force microscopy: the adhesion force and the work of adhesion force were determined from histograms. The modification of probe surface was confirmed by visualization of bovine serum albumin and supercoiled pGEMEX DNA molecules immobilized on the amino mica and amino mica modified by cross-linker.

Amines↗
PubMed2006

Cytoskeletal response of microvessel endothelial cells to an applied stress force at the submicrometer scale studied by atomic force microscopy.

Cytoskeleton fibers form an intricate three-dimensional network to provide structure and function to microvessel endothelial cells. During accommodation to blood flowing, stress fiber bundles become more prominent and align with the direction of blood flow. This network either mechanically resists the applied shear stress (lateral force) or, if deformed, is dynamically remodeled back to a preferred architecture. However, the detailed response of these stress fiber bundles to applied lateral force at submicrometer scales are as yet poorly understood. In our in vitro study, the tip, topography probe in lateral force microscopy of atomic force microscopy, acted as a tool for exerting quantitative vertical and lateral force on the filaments of the cytoskeleton. Moreover, the authors developed a formula to calculate the value of lateral force exerted on every point of the filaments. The results show that cytoskeleton fibers of healthy tight junctions in rat cerebral microvessel endothelial cells formed a cross-type network, and were reinforced and elongated in the direction of scanning under lateral force of 15-42 nN. Under peroxidation (H(2)O(2) of 300 micromol/L), the cytoskeleton remodeled at intercellular junctions, and changed over the meshwork structures into a dense bundle, that redistributed the stress. Once mechanical forces were exerted on an area, the cells shrank and lost morphologic tight junctions. It would be useful in our understanding of certain pathological processes, such as cerebral ischemia/reperfusion injury, which maybe caused by biomechanical forces and which are overlooked in current disease models.

Animals↗
PubMed2004

Characterization of ionizing radiation-induced ring chromosomes by atomic force microscopy.

We applied atomic force microscopy (AFM) to the structural analysis of radiation-induced ring chromosomes. Constrictions observed on the metaphase ring chromosome were found to correspond to the centromere regions of the ring chromosome in comparison with the AFM image of the centromere of rod chromosomes and with the fluorescence in situ hybridization (FISH) technique. Section analysis by AFM revealed that some ring-like chromosome fragments and ring-like chromatid fragments were thicker than standard chromosomes or chromatids, suggesting that they were ring chromosomes viewed edge on. Topographic analysis by AFM makes it possible to distinguish a ring viewed edge on that is difficult to recognize as a ring by light microscopy and to discriminate between a centric ring chromosome and an acentric ring chromosome using the same slides prepared for light microscopy.

Centromere↗
PubMed1994

Viscoelasticity of living cells allows high resolution imaging by tapping mode atomic force microscopy.

Application of atomic force microscopy (AFM) to biological objects and processes under physiological conditions has been hampered so far by the deformation and destruction of the soft biological materials invoked. Here we describe a new mode of operation in which the standard V-shaped silicon nitride cantilever is oscillated under liquid and damped by the interaction between AFM tip and sample surface. Because of the viscoelastic behavior of the cellular surface, cells effectively "harden" under such a tapping motion at high frequencies and become less susceptible to deformation. Images obtained in this way primarily reveal the surface structure of the cell. It is now possible to study physiological processes, such as cell growth, with a minimal level of perturbation and high spatial resolution (approximately 20 nm).

Animals↗
PubMed2003

A general model for amyloid fibril assembly based on morphological studies using atomic force microscopy.

Based on atomic force microscopy analysis of the morphology of fibrillar species formed during fibrillation of alpha-synuclein, insulin, and the B1 domain of protein G, a previously described model for the assembly of amyloid fibrils of immunoglobulin light-chain variable domains is proposed as a general model for the assembly of protein fibrils. For all of the proteins studied, we observed two or three fibrillar species that vary in diameter. The smallest, protofilaments, have a uniform height, whereas the larger species, protofibrils and fibrils, have morphologies that are indicative of multiple protofilaments intertwining. In all cases, protofilaments intertwine to form protofibrils, and protofibrils intertwine to form fibrils. We propose that the hierarchical assembly model describes a general mechanism of assembly for all amyloid fibrils.

Amyloid↗
PubMed2005

Hierarchy of DNA immobilization and hybridization on poly-L-lysine using an atomic force microscopy study.

The atomic force microscopy has been used to analyze the immobilization of single stranded DNA on poly-L-lysine-coated glass and subsequent hybridization with complimentary DNA with the Z-threshold parameter and fractal analysis methods. The poly-L-lysine layer, which has a thickness of approximately 7 nm, presents nano-defects that could be critical for DNA immobilization by acting as a nucleation sites for ssDNA and subsequently for dsDNA aggregates. The Z-threshold for the dsDNA aggregates is much larger than for ssDNA, but the statistical fractal dimension is very similar, suggesting a conformal increase of the dimensions of the dsDNA aggregates mainly in the Z-direction, due to an effective ssDNA-ccDNA molecular recognition. This study demonstrates the use of fractal analysis in conjunction with the distribution of heights to evaluate the efficiency of DNA-DNA molecular recognition on surfaces and the impact of nanodefects.

Adsorption↗
PubMed2005

Antigen binding forces of single antilysozyme Fv fragments explored by atomic force microscopy.

We used atomic force microscopy (AFM) to explore the antigen binding forces of individual Fv fragments of antilysozyme antibodies (Fv). To detect single molecular recognition events, genetically engineered histidine-tagged Fv fragments were coupled onto AFM tips modified with mixed self-assembled monolayers (SAMs) of nitrilotriacetic acid- and tri(ethylene glycol)-terminated alkanethiols while lysozyme (Lyso) was covalently immobilized onto mixed SAMs of carboxyl- and hydroxyl-terminated alkanethiols. The quality of the functionalization procedure was validated using X-ray photoelectron spectroscopy (surface chemical composition), AFM imaging (surface morphology in aqueous solution), and surface plasmon resonance (SPR, specific binding in aqueous solution). AFM force-distance curves recorded at a loading rate of 5000 pN/s between Fv- and Lyso-modified surfaces yielded a distribution of unbinding forces composed of integer multiples of an elementary force quantum of approximately 50 pN that we attribute to the rupture of a single antibody-antigen pair. Injection of a solution containing free Lyso caused a dramatic reduction of adhesion probability, indicating that the measured 50 pN unbinding forces are due to the specific antibody-antigen interaction. To investigate the dynamics of the interaction, force-distance curves were recorded at various loading rates. Plots of unbinding force vs log(loading rate) revealed two distinct linear regimes with ascending slopes, indicating multiple barriers were present in the energy landscape. The kinetic off-rate constant of dissociation (k(off) approximately = 1 x 10(-3) s(-1)) obtained by extrapolating the data of the low-strength regime to zero force was in the range of the k(off) estimated by SPR.

Antigen-Antibody Complex↗
PubMed2005

Imaging proteins with atomic force microscopy: an overview.

Atomic force microscopy (AFM) has become a common tool for biophysical studies of proteins; mainly due its property to perform characterizations near physiological conditions. The tertiary and quaternary structures, forces driving folding-unfolding processes, and secondary structure elements can be studied in their native environments allowing high resolution level associated with small distortions. This review outlines the operational principles and applications of AFM for protein biophysics.

Crystallography, X-Ray↗
PubMed1999

Surface processes in the crystallization of turnip yellow mosaic virus visualized by atomic force microscopy.

In situ atomic force microscopy (AFM) was used to investigate surface evolution during the growth of single crystals of turnip yellow mosaic virus (TYMV). Growth of the (101) face of TYMV crystals proceeded by two-dimensional nucleation. The molecular structure of the step edges and adsorption of individual virus particles and their aggregates on the crystalline surface were recorded. The surfaces of individual virions within crystals were visualized and seen to be quite distinctive with the hexameric and pentameric capsomers of the T = 3 capsids being clearly resolved. This, so far as we are aware, is the first direct visualization of the capsomere structure of a virus by AFM. In the course of recording the in situ development of the crystals, a profound restructuring of the surface arrangement was observed. This transformation was highly cooperative in nature, but the transitions were unambiguous and readily explicable in terms of an organized loss of classes of virus particles from specific lattice positions. In some cases areas of a single crystal surface were recorded in which were captured successive phases of the transition. We believe this provides the first visual record of a cooperative restructuring of the surface of a supramolecular crystal.

Capsid↗
PubMed2006

Dynamic response of glucagon/anti-glucagon pairs to pulling velocity and pH studied by atomic force microscopy.

We used atomic force microscopy (AFM) to measure the unbinding force between antigen coupled to an AFM tip and antibody coated on the substrate surface. Dynamic responses of glucagon/anti-glucagon pairs with multiple pull-off steps to pH and pulling velocity were studied by AFM. Force-distance curves of a specific glucagon-anti-glucagon interaction system with mono-, di-, and multi-unbinding events were recorded, which may be attributed to a single, sequential or multiple breaking of interacting bond(s) between glucagon and anti-glucagon. We studied the dynamic response of glucagon-anti-glucagon pairs to various pulling velocities (16.7-166.7 nm/s). It was found that the mean value of the unbinding force was shifted toward higher values with increasing pulling velocity at each pH. This indicates that the friction force between glucagon and anti-glucagon may contribute to the unbinding force. Moreover, the dynamic response of glucagon-anti-glucagon pairs to pH (4-10) with different pulling velocities was studied. Within the acid range, the bond strength between the glucagon/anti-glucagon complex showed a rapid increase from pH 4 to 7 and reached a maximum (256.4+/-48.9 pN at 166.7 nm/s) at neutrality, followed by a sharp decrease with increasing pH (pH 7-10). This could be attributed to the conformational change that occurred in glucagon when the pH value in solution was varied from the reference level at neutrality. This study demonstrated that the pH dependence of multiple antigen-antibody bond-rupture forces could be measured by a force-based AFM biosensor. Unraveling the relationship between inter-molecular force and intra-molecular conformational change in acid, neutral, and alkaline environments may provide new directions for future application of force measurements by AFM in proteomics or in the development of a clinical cantilever-based mechanical biosensor.

Antibodies↗