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Results for “Atomic force microscopy (AFM)”

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PubMed2004

Adsorption and desorption of polymer/surfactant mixtures at solid-liquid interfaces: substitution experiments.

The adsorption of mixtures of aqueous solutions of cationic hydroxyethylcellulose polymer JR400 and anionic surfactant, sodium dodecyl sulfate, using atomic force microscopy (AFM) has been studied. Samples with various compositions from different regions of the ternary phase diagram presented in our previous work were imaged by atomic force microscopy on freshly cleaved mica, and hydrophobically modified mica and silica in soft-contact mode. A series of "washing" (subsequent injection of compositions with gradually decreasing polymer/surfactant ratio) and "scratching" (mechanical agitation of the surface material with an AFM tip) experiments were performed. It was revealed that the morphology of the adsorbed layer altered in a manner following the changes in morphology in the bulk solution. These changes were evidenced in cluster formation in the layer. The results suggest that the influence of the surface was limited to the formation of the adsorbed layer where the local concentrations of polymer and surfactant were higher than those in the bulk. All further modifications were driven by changes in the mixture composition in bulk. Force measurements upon retraction reveal the formation of network structures within the surface aggregates that will greatly slow structural reequilibration.

Adsorption↗

PubMed2006

Structure of the rhodopsin dimer: a working model for G-protein-coupled receptors.

G-protein-coupled receptors (GPCRs) participate in virtually all physiological processes. They constitute the largest and most structurally conserved family of signaling molecules. Several class C GPCRs have been shown to exist as dimers in their active form and growing evidence indicates that many, if not all, class A receptors also form dimers and/or higher-order oligomers. High-resolution crystal structures are available only for the detergent-solubilized light receptor rhodopsin (Rho), the archetypal class A GPCR. In addition, Rho is the only GPCR for which the presumed higher-order oligomeric state has been demonstrated, by imaging native disk membranes using atomic force microscopy (AFM). Based on these data and the X-ray structure, an atomic model of Rho dimers has been proposed, a model that is currently scrutinized in various ways. AFM has also been used to measure the forces required to unfold single Rho molecules, thereby revealing which residues are responsible for Rho's stability. Recent functional analyses of fractions from solubilized disk membranes revealed that higher-order Rho oligomers are the most active species. These and other results have enhanced our understanding of GPCR structure and function.

Animals↗

PubMed2003

AFM characterization of tilt and intrinsic flexibility of Rhodobacter sphaeroides light harvesting complex 2 (LH2).

Atomic force microscopy (AFM) has developed into a powerful tool to investigate membrane protein surfaces in a close-to-native environment. Here we report on the surface topography of Rhodobacter sphaeroides light harvesting complex 2 (LH2) reconstituted into two-dimensional crystals. These photosynthetic trans-membrane proteins formed cylindrical oligomeric complexes, which inserted tilted into the lipid membrane. This peculiar packing of an integral membrane protein allowed us to determine oligomerization and tilt of the LH2 complexes, but also protrusion height and intrinsic flexibility of their individual subunits. Furthermore the surface contouring reliability and limits of the atomic force microscopy could be studied. The two-dimensional crystals examined had sizes of up to 5 microm and, as revealed by a 10 A cryo electron microscopy projection map, p22(1)2(1) crystal symmetry. The unit cell had dimensions of a = b = 150 A and gamma = 90 degrees, and housed four nonameric complexes, two pointing up and two pointing down. AFM topographs of these 2D crystals had a lateral resolution of 10 A. Further, the high vertical resolution of approximately 1 A, allowed the protrusion height of the cylindrical LH2 complexes over the membrane to be determined. This was maximally 13.1 A on one side and 3.8 A on the other. Interestingly, the protrusion height varied across the LH2 complexes, showing the complexes to be inserted with a 6.2 degree tilt with respect to the membrane plane. A detailed analysis of the individual subunits showed the intrinsic flexibility of the membrane protruding peptide stretches to be equal and independent of their protrusion height. Furthermore, our analysis of membrane proteins within this peculiar packing confirmed the high vertical resolution of the atomic force microscopy on biological samples, and led us to conclude that the image acquisition function was equally accurate for contouring protrusions with heights up to approximately 15 A.

Amino Acid Sequence↗

PubMed1999

Corrosion and biocompatibility of orthodontic wires.

With the increasing number of orthodontic treatments using devices containing nickel and the growing prevalence of nickel allergy in the average population, biocompatibility studies of these devices have become a topic of major interest. The corrosion behavior of orthodontic wires is a decisive factor determining their biocompatibility. Therefore four nickel-titanium guiding arches, a titanium-molybdenum and a stainless steel wire were analyzed for corrosion behavior under realistic conditions. Pure potentiostatic, pure mechanical and combined potentiostatic and mechanical stresses were applied to the specimens. Subsequently, the surfaces of the wires were investigated employing atomic force microscopy (AFM) and nickel loss was measured with an atomic absorption spectrophotometer. The results yield information about the relative corrosion tendency of the wires under in vitro conditions. The wires examined can be classified into two groups, one with a high and a second group with a low tendency towards corrosion, that is American Orthodontics Memory wire as well as GAC Neo Sentalloy and Ormco Ni-Ti as well as Unitek Nitinol respectively. Although corrosion behavior under clinical conditions can not be directly derived from these results, analyses of wires after clinical usage indicate that changes of wire surfaces might show the same characteristics under in vitro conditions.

Journal Article↗

PubMed2003

Tablet surface characterisation by various imaging techniques.

The aim of this study was to characterise tablet surfaces using different imaging and roughness analytical techniques including optical microscopy, scanning electron microscopy (SEM), laser profilometry and atomic force microscopy (AFM). The test materials compressed were potassium chloride (KCl) and sodium chloride (NaCl). It was found that all methods used suggested that the KCl tablets were smoother than the NaCl tablets and higher compression pressure made the tablets smoother. Imaging methods like optical microscopy and SEM can give useful information about the roughness of the sample surface, but they do not provide quantitative information about surface roughness. Laser profilometry and AFM on the other hand provide quantitative roughness data from two different scales, laser profilometer from 1 mm and atomic force microscope from 90 microm scale. AFM is a powerful technique but other imaging and roughness measuring methods like SEM, optical microscopy and laser profilometry give valuable additional information.

Compressive Strength↗

PubMed2006

Quantification of the number of EP3 receptors on a living CHO cell surface by the AFM.

The distribution of EP3 receptors on a living cell surface was quantitatively studied by atomic force microscopy (AFM). Green fluorescent protein (GFP) was introduced to the extracellular region of the EP3 receptor on a CHO cell. A microbead was used as a probe to ensure certain contact area, whose surface was coated with anti-GFP antibody. The interactions between the antibodies and GFP molecules on the cell surface were recorded to observe the distribution of the receptors. The result indicated that EP3 receptors were distributed on the CHO cell surface not uniformly but in small patches coincident with immunohistochemical observation. Repeated measurements on the same area of cell surface gave confirmation that it was unlikely that the receptors were extracted from the cell membrane during the experiments. The measurement of single molecular interaction between GFP and the anti-GFP antibody was succeeded on the cell surface using compression-free force spectroscopy. The value of separation work required to break a single molecular pair was estimated to be about 1.5 x 10(-18)J. The number of EP3 receptor on the CHO cell surface was estimated using this value to be about 1 x 10(4) under the assumption that the area of the cell surface was about 5,000 microm(2). These results indicated that the number of receptors on a living cell surface could be quantified through the force measurement by the AFM.

Animals↗

PubMed1994

Atomic force microscopy studies of the interaction of a dentin adhesive with tooth hard tissue.

The usefulness of atomic force microscopy (AFM) in the study of dental biomaterials was investigated. Two basic points were addressed: the first is the use of an atomic force microscope for the imaging of tooth hard tissue; the second is a study by AFM of the interaction of a liquid adhesive (Gluma) with dentin. Results show that dental tissues are readily imaged by AFM and that high-resolution images of the dentin ultrastructure can be obtained. As to the dentin adhesive, it was possible to elucidate the mechanism of action of Gluma. In particular, polymerization of the hydroxyethylmethacrylate present in the Gluma mixture was observed to occur in < 30 seconds.

Dentin↗

PubMed2005

Indentation and adhesive probing of a cell membrane with AFM: theoretical model and experiments.

In probing adhesion and cell mechanics by atomic force microscopy (AFM), the mechanical properties of the membrane have an important if neglected role. Here we theoretically model the contact of an AFM tip with a cell membrane, where direct motivation and data are derived from a prototypical ligand-receptor adhesion experiment. An AFM tip is functionalized with a prototypical ligand, SIRPalpha, and then used to probe its native receptor on red cells, CD47. The interactions prove specific and typical in force, and also show in detachment, a sawtooth-shaped disruption process that can extend over hundreds of nm. The theoretical model here that accounts for both membrane indentation as well as membrane extension in tip retraction incorporates membrane tension and elasticity as well as AFM tip geometry and stochastic disruption. Importantly, indentation depth proves initially proportional to membrane tension and does not follow the standard Hertz model. Computations of detachment confirm nonperiodic disruption with membrane extensions of hundreds of nm set by membrane tension. Membrane mechanical properties thus clearly influence AFM probing of cells, including single molecule adhesion experiments.

Animals↗

PubMedDate not supplied

AFM and STM study of beta-amyloid aggregation on graphite.

Atomic force microscopy (AFM) and scanning tunneling microscopy (STM) have been employed in situ and ex situ to directly study the aggregation of beta-amyloid(1-42) (Abeta42) peptide on hydrophobic graphite. From in situ AFM images, Abeta42 peptides were seen to aggregate into the sheets that preferred to three orientations with characteristic 3-fold symmetry (Proc. Natl. Acad. Sci. USA 96 (1999) 3688). The sheets were formed by parallel narrow lines with a height of 0.8-1.0nm and a width of 12-14nm. The narrow lines looked like beaded chains and have a right-handed axial periodicity. The high-resolution ex situ AFM and STM images showed that some fibrils of beta-amyloid had a characteristic domain texture, indicating they were formed through the association of protofibrils and monomers. The fibril containing lateral associated filaments that exhibited right-handed twist was clearly observed in the STM image. These results provide important clues to study the detailed structure of beta-amyloid aggregates and the mechanism of the Abeta fibrils formation on hydrophobic surface.

Alzheimer Disease↗

PubMed1998

Magnetic Effects on Thickness of Adsorbed Layer in Aqueous Solutions Evaluated Directly by Atomic Force Microscope.

The short-range interaction force between a mica surface and an AFM probe tip in electrolyte solutions was measured by atomic force microscopy (AFM) to directly evaluate the thickness of the adsorbed layer on the surface. The magnetic effects were estimated by comparing the thickness in the magnetized solution with that in nonmagnetized solution. It was found that (i) the magnetic exposure thickens the adsorbed layer on the surface in electrolyte solutions, (ii) the magnetic effects are easily destroyed by the external disturbance, (iii) there exists a memory in the magnetic effects, which remains for at least a day, (iv) the magnetic effects appear mostly in the solutions of structure-disordering cations, such as Cs+, Rb+, and K+, and (v) the results are consistent with those obtained in preceding macroscopic experiments. It is hypothesized from these results that the water molecules weekly bound around the structure-disordering ions are quasistabilized and structured by the magnetic exposure and that the magnetic thickening of the adsorbed layer is caused by the adsorption of those structured ions on the surface. Copyright 1998 Academic Press.

Journal Article↗

PubMed2005

Dehydration damage of domain-exhibiting supported bilayers: an AFM study on the protective effects of disaccharides and other stabilizing substances.

Atomic force microscopy (AFM) has been applied to characterize hydrated sphingomyelin/dioleoylphosphatidylcholine/cholesterol supported bilayers, after dehydration either in the absence or in the presence of several stabilizing substances. Such a study provides information about the effect of extreme environmental conditions on biological membranes and, in particular, on lipidic microdomains. Dehydration stress, indeed, is thought to cause both macroscopical damage and alterations of microdomains in biomembranes, leading to deleterious effects. These phenomena can be avoided if disaccharides are added during dehydration. In this work, we apply AFM imaging to directly visualize damage caused to supported lipid bilayers by water removal. We compare the efficiency of sucrose, trehalose, dextran, dimethyl sulfoxide, and glucose in preserving the structural integrity of domain-exhibiting model membranes. Finally, in addition to confirming previous findings, our results provide further insight into damage and alteration of microdomains in membranes as a consequence of stressful drying conditions.

Disaccharides↗

PubMed2000

Elasticity mapping of living fibroblasts by AFM and immunofluorescence observation of the cytoskeleton.

Using the force mapping mode of atomic force microscopy (AFM), we measured spatial distribution of elastic moduli of living mouse fibroblasts (NIH3T3) in a physiological condition. The nuclear portion of the cellular surface is about 10 times softer than the surroundings. Stiffer fibers are confirmed in the elastic images. In order to investigate origin of the softer nuclear portion and the stiffer fibers, we fixed the identical cells imaged by the AFM, and carried out immunofluorescence observation for three types of cytoskeletal filaments--actin filaments, microtubules, and intermediate filaments, using confocal laser scanning microscopy (CLSM). A comparison between the AFM and the CLSM images revealed that the elasticity of the cells was concerned not only with the distribution of actin network, but also with intermediate filaments, whereas microtubules had no large effect on the measured elasticity.

3T3 Cells↗

PubMed2006

AFM images of short oligonucleotides on a surface of supported lipid films.

Atomic force microscopy (AFM) was used to study the topography of lipid films on a gold support with immobilized 19 mer single stranded DNA (ssDNA) chemically modified by oleylamine and after hybridization with complementary DNA. The topography of various surfaces was analyzed, including alkanethiol layer chemisorbed on a gold support, lipid films formed on alkanethiol layer without and with immobilized single or double stranded DNA (dsDNA). The value of root means square roughness (RMS) for each surface was determined. RMS value for sBLM with immobilized ssDNA was 2.98 nm, while slightly higher value of 3.37 nm was typical for dsDNA. The analysis of AFM images revealed that both ssDNA and dsDNA form clusters. The clusters formed by ssDNA are not uniform, but that formed by dsDNA are almost of circular shape with diameter of 13.6+/-0.5 nm. Formation of the clusters could be consequence of lower hydration of lipids and DNA at an air. The water deficit and hence increased ion concentration probably facilitate the attraction between oligonucleotides.

Adsorption↗

PubMed1998

Immobilization strategies for biological scanning probe microscopy.

Biological atomic force microscopy (AFM) is now established as a method for studying the structure and function of biomolecular objects at the solid-liquid interface. Major progress in this field is linked to new developments in instrumentation, a better understanding of tip-sample interactions, and improved sample preparation techniques. In this review, the most common strategies for biomolecular immobilization with respect to biological AFM applications are summarized.

Microscopy, Atomic Force↗

PubMed2001

A tapping mode AFM study of collapse and denaturation in dentinal collagen.

OBJECTIVES: Tapping mode atomic force microscopy (AFM) was used to investigate the surface changes of collagen exposed to phosphoric acid treatment. We focus on denaturation and collapse following demineralization and exposure to air. METHODS: Unpolished dentin disks, obtained from freshly extracted human molars, were etched in 37% phosphoric acid for 15s, rinsed ultrasonically and gently blotted with soft paper; the specimens were then continuously observed using tapping mode AFM. RESULTS: Immediately after the removal of bulk water, the surface consisted of a porous network of banded collagen fibrils, having periodicities of 67nm. After approximately 8min of subsequent air-drying, the spacing between fibrils was lost, and the surface was observed to consist of a dense array of closely spaced fibrils. The banding periodicity was still observable. SIGNIFICANCE: The air drying of etched dentin results in the collapse of the collagen network, but not in the denaturation of the collagen fibrils. This study indicates that collapse and denaturation are separate phenomena. It further shows that water loss occurs rapidly, and disrupts the native conformation of the collagen network. This would have adverse effects on adhesion.

Acid Etching, Dental↗

PubMed2003

Nanoscale tripodal 1,3,5,7-tetrasubstituted adamantanes for AFM applications.

The synthesis of four novel nanoscale 1,3,5,7-tetrasubstituted adamantanes 22 and 25-27 designed for atomic force microscopy (AFM) applications is described. Each tetrahedrally shaped molecule incorporates a broad tripodal base made up of three identical legs that terminate with a sulfur-containing moiety, which is either a 4-acetylsulfanylmethylphenyl unit or else a (1,2,5-dithiazepan-1-yl)phenyl unit. The sulfur atoms are intended for eventual binding of the molecule multivalently to the apex of a gold-coated commercial AFM tip through formation of multiple S-Au bonds. In each molecule, the fourth terminus is a para-substituted benzoic acid methyl ester that is designed to scan the sample. We demonstrate that 27 is sufficiently large and rigid to be imaged by a conventional AFM tip. Adamantanes 22 and 25-27 may also find application as chemically well-defined nanoscale objects for calibration of AFM tips.

Adamantane↗

PubMed2006

Physical structures of lipid layers on pyrite.

The physical structures of lipid layers on pyrite (FeS2), a ubiquitous sulfide mineral, were studied in air and in water by atomic force microscopy (AFM). Egg PC, a phospholipid that forms bilayer structures on atomically flat substrates, was investigated, and our experimental observations show that this lipid formed bilayers on an atomically rough pyrite surface, as inferred by a measured layer thickness of 5.0 +/- 0.2 nm. The surface coverage of the lipid coating increased from approximately 15% to 80% when the concentration of the lipid suspension was increased from 0.014 to 0.15 mM. Although further increases up to 1.5 mM resulted in an incremental increase in surface coverage of only 5%, multilayer structures of 20- to 40-nm height formed on top of the first bilayer. The findings provide a structural explanation for the results of earlier kinetic studies showing that the presence of the lipid decreases the rate of pyrite oxidation in air and in water. Lipid coatings applied to iron-sulfide bearing minerals are a possible approach to preventing oxidation and acidification and thereby mitigating environmental damage that can result from acid mine drainage.

Iron↗

PubMed2004

AFM studies of the crystallization and habit modification of an excipient material, adipic acid.

Atomic force microscopy (AFM) has been used to investigate the (1 0 0) face of crystalline adipic acid, both in air and liquid environments. In air, surface reorganization occurred during scanning of the AFM probe, which has been investigated using single point force-distance analysis under a controlled relative humidity (RH) environment. We suggest such reorganization can be attributed to the influence of a network of water molecules bound to the hydrophilic (1 0 0) surface permitting local AFM tip-enhanced dissolution and reorganization of the solute. In situ imaging was also carried out on the crystals, revealing etch-pit formation during dissolution, and rapid growth at higher levels of supersaturation (sigma), both of which are direct consequences of the hydrophilic nature of the (1 0 0) face. Also presented here are nanoscale observations of the effect of octanoic acid, a structurally-related habit modifier, on crystalline adipic acid. Using AFM, we have been able to show that the presence of octanoic acid at low concentration has little observable affect on the development of the (1 0 0) face; however, as this concentration is increased, there are clear changes in step morphology and growth mode on the (1 0 0) face of the crystal. At a concentration of 1.26 mmol dm(-3) (a concentration corresponding to a molar ratio of approximately 1:175 octanoic acid:adipic acid), growth on the (1 0 0) face is inhibited, with in situ AFM imaging indicating this is a direct consequence of octanoic acid binding to the surface, and pinning the monomolecular growth steps.

Adipates↗